Patent Publication Number: US-2016231341-A1

Title: Multidrug analysis in urine by liquid chromatography-tandem mass spectrometry

Description:
TECHNICAL FIELD 
     The subject matter disclosed herein relates to a method of detection and/or quantification of multiple drugs and/or metabolites from a sample of body fluid by liquid chromatography-tandem mass spectrometry in a single run. 
     BACKGROUND 
     Promoting mental health and preventing mental and/or substance abuse disorders are fundamental to SAMHSA&#39;s mission to reduce the impact of behavioral health conditions in America&#39;s communities. 
     Mental and substance abuse disorders can have a powerful effect on the health of individuals, their families, and their communities. In 2012, an estimated 9.6 million adults aged 18 and older in the United States had a serious mental illness, and 2.2 million youth aged 12 to 17 had a major depressive episode during the year 2011. In 2012, an estimated 23.1 million Americans aged 12 and older needed treatment for substance abuse. These disorders are among the top conditions that cause disability and carry a high burden of disease in the United States, resulting in significant costs to families, employers, and publicly funded health systems. By 2020, mental and substance abuse disorders will surpass all physical diseases as a major cause of disability worldwide. 
     In addition, drug and alcohol abuse can lead to other chronic diseases such as diabetes and heart disease. Addressing the impact of substance use alone is estimated to cost Americans more than $600 billion each year. 
     Preventing mental and/or substance abuse disorders and related problems in children, adolescents, and young adults are critical to Americans&#39; behavioral and physical health. Behaviors and symptoms that signal the development of a behavioral disorder often manifest two to four years before a disorder is present. In addition, people with a mental health issue are more likely to use alcohol or drugs than those not affected by a mental illness. If communities and families can intervene early, behavioral health disorders might be prevented, or symptoms can be mitigated. 
     Data have shown that early intervention following the first episode of a serious mental illness can make an impact. Coordinated, specialized services offered during or shortly after the first episode of psychosis are effective for improving clinical and functional outcomes. 
     In addition, the Institute of Medicine and National Research Council&#39;s Preventing Mental, Emotional, and Behavioral Disorders Among Young People report—2009 notes that cost-benefit ratios for early treatment and prevention programs for addictions and mental illness programs range from 1:2 to 1:10. This means a $1 investment yields $2 to $10 savings in health costs, criminal and juvenile justice costs, educational costs, and lost productivity. 
     In different areas of human activities people are confronted with the illegal use of doping in order to enhance the output of the doped species. In sports this can be the improvement of the endurance of the athlete (like the swimmer, cyclist, triathlist, etc.), or (also) of the animal used in the sport (like the horse). Breeders of animals are sometimes known to illegally use doping to enhance the breeding process or the breeding product (like a faster growth of the animal). In order to combat such activities, authorities need fast and reliable equipment in order to detect such abuse. Generally the determination of the use of drugs in doping control is performed by analyzing the used drug or its metabolites in the body fluid of the treated species. 
     Confirmation of identity of forensically relevant compounds, such as drugs of abuse, is a necessary step in medico-legal event controls of people involved in crimes, workplace accidents and driving under the influence of drugs (DUID). Plasma is a useful medium in determining the short-term use of illicit drugs and its analysis is mandatory in the case of DUID in many countries. Urine has been the sample of choice for monitoring drug abuses in workplaces and is subjective to strict regulations. 
     The guidelines from the US Substance Abuse and Mental Health Services Administration (SAMHSA), effective October 2010, require LC/MS/MS methods for confirmation of initial drug tests. 
     Several methods are available in literature for drug-detection and quantification in body fluids. 
     WO 2007/134711 discloses a comprehensive multi-dimensional gas chromatography mass spectroscopic method for determining drug-metabolite in a body fluid by prior treatment of body fluid with alkyl haloformate. 
     CN 100381812 discloses liquid chromatography-tandem mass spectrometry (LC-MS/MS) used for drug detection from urine, which involves sample pre-treating and solid phase extraction. The method is reliable in the quantative detection of 19 drugs. 
     Journal article ‘Neurology (1972), 22(5), 540-50, discloses determination of multiple anticonvulsant drug levels in human serum by gas-liquid chromatography. 
     Journal article ‘Forensic Science International (2005), 150(2-3), 227-238’, discloses analysis of multiple illicit basic drugs in preserved oral fluid by solid-phase extraction and liquid chromatography-tandem mass spectrometry. 
     Journal article ‘Talanta (2009), 78(2), 377-387’ discloses use of ultra high-pressure liquid chromatography with a single quadrupole mass spectrometer for investigation of several cytochromes P 450 (CYP450) substrates and respiratory metabolites. 
     Journal article ‘Chromatographia (2012), 75(1-2), 55-63′ discloses determination of illicit drugs in urine and plasma by micro-SPE followed by HPLC-MS/MS. 
     Journal article ‘Archives of Pharmacal Research (2014), 37(6), 760-772’ discloses a method of screening multiple drugs of abuse and metabolites in urine using LC/MS/MS with polarity switching electrospray Ionization. This method involves simultaneous analysis of 35 drugs of abuse and relevant metabolites. The drugs and metabolites in urine were extracted by using mixed mode strong cation exchange polymeric solid phase extraction cartridges after enzymic hydrolysis and were then injected into the LC/MS/MS system. 
     Journal article reference ‘Journal of Analytical Toxicology (2007), 31(7), 359-368’ discloses determination of multiple drugs of abuse and relevant metabolites in urine by LC-MS-MS. The method is developed for the quantitative analysis of 30 drugs from classes such as opiates, barbiturates, amphetamines, cocaine, cannabinoids, phencyclidine, methadone, and benzodiazepines. This method uses solid-phase extraction (SPE) on an Oasis HLB column followed by liquid chromatography-tandem mass spectrometry. 
     Journal article reference ‘Rapid Communications in Mass Spectrometry (2014), 28(19), 2043-2053, discloses identification of multiple drugs of abuse and relative metabolites in urine samples using liquid chromatography/triple quadrupole mass spectrometry coupled with a library search with two multiple reaction monitoring (MRM) transitions per compound. The quantification and identification performance for 13 drugs of abuse and their metabolites were evaluated. 
     Journal article reference ‘Chromatographia (2001), 54(5/6), 345-349’ discloses method for simultaneous determination of multiple antiepileptic drugs in human serum. 
     Journal article reference ‘Journal of Mass Spectrometry (2008), 43(7), 980-992’ discloses multicomponent screening method for diuretics, masking agents, central nervous system (CNS) stimulants and opiates in human urine by UPLC-MS/MS. 
     Thus, the prior art methods of determining multiple drugs of abuse and their metabolites requires the special apparatus for extraction for sample preparation. In some prior art it is necessary to perform the derivatization of analyte, which subsequently will add to the cost and time for analysis. Though prior art methods discloses multiple drug analysis and their detection, the number of drugs and metabolites analyzed is limited to the class of drugs and number of drugs. 
     As a result, there is a need to provide a simple, cost effective, improved, highly efficient, and reliable method for quantitative analysis of a large number of drug or metabolites belonging to different chemical and toxicological classes in a biological sample. 
     The disclosed methods provide a rapid and cost effective method of drug-metabolite detection belonging to different chemical and toxicological classes form the body fluids such as urine. The disclosed methods meet SAMHSA guidelines to demonstrate linearity, limit of detection (LOD), accuracy and precision, as well as measurement of matrix effect, extraction recovery and overall process efficiency. Methods disclosed herein are suitable for all classes of SAMHSA-regulated drug. 
     SUMMARY 
     In view of the problems of the related art discussed above, disclosed herein is an improved method of quantitative analysis of a drug or a metabolite in a biological sample. Moreover, an improved method of analysis is provided that is relatively salt free, which is important for mass spectrometry analysis. 
     The disclosed method facilitates the detection of 63 different drugs belonging to different chemical and toxicological classes in urine by liquid chromatography-tandem mass spectrometry method. 
     The disclosed method pertains to a sample preparation method that may be used for the quantitative analysis of a drug and/or metabolite in a biological sample. Moreover, also disclosed herein is a method for preparing a sample, without use of derivatization and/or extraction procedure, for quantitative analysis of a drug and/or metabolite in a biological sample. 
     The disclosed method can be a fast and reliable confirmatory method for the determination of multiple drugs of abuse belonging to different chemical and toxicological classes: opiates/opioids (28)-selected from 6-Monoacetylmorphine (6-MAM), Codeine, Dihydrocodeine, Hydrocodone, Hydromorphone, Morphine, Oxycodone, Oxymorphone, Buprenophrine, Carisoprodol, Desmethyl Tapentadol, Desmethyl Tramadol, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), Meperidine, Meprobamate, Methadone, Norbuprenophrine, Normeperidine, Tapentadol, Tramadol, Fentanyl, Norfentanyl, Norpropoxyphene, Propoxyphene, Dextromethophan, Dextrophan, Desomorphine, Nalaxone; benzodiazepines (12)-7-AminoClonazepam, Diazepam, Flunitrazepam, 4-HydroxyAlprazolam, Nordiazepam, Oxazepam, Temazepam, Chloradiazepoxide, OH-et-flunizepam, Lorazepam, Triazolam, Midazolam; barbiturates (2)-Butalbital, Phenobarbital; amphetamines (4)-Amphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-methamphetamine (MDMA), Methamphetamine; tricyclic antidepressants (8)-Desipramine, Imipramine, Nortriptyline, Ritalinic Acid, Sertraline, Cyclobenzaprine, Amitriptyline, Methyl phenidate; illicit drugs (3) Tetrahydrocannabinolic acid (THCA), Benzoylecgonine, Phencyclidine (PCP); Z drugs (4)-Zolpidem, Zaleplon, Zopiclone, Zolpidem-COOH and Antiepileptics (2)-Pregabilan, Gabapentin. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is an optimized multi-step gradient elution method for LC-MS/MS analysis.  FIG. 2  is a scheduled MRM transitions in positive and negative mode.  FIG. 3 , panels labeled A, is a 7-points calibration curve for 6-Monoacetylmorphine (6-MAM) (left panel) and Buprenorphine (right panel) in positive mode.  FIG. 3 , panels labeled B, is a 7-points calibration curve for THCA (left panel) and Butalbital (right panel) in negative mode. 
     
    
    
     DETAILED DESCRIPTION 
     Advantages of the present invention will become more apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. The invention will be described in more detail hereinbelow by making reference to its particularly preferred embodiments. 
     Disclosed herein is highly selective and specific method for detection or determination of multiple drugs of abuse potential and their metabolite species belonging to different chemical and toxicological classes from a sample of body fluid comprising the steps: a) mixing the sample with internal standard, b) hydrolyzing the drug metabolite in the sample by β-Glucuronidase enzyme, c) centrifugation of the mixture of step (b) and diluting the clear supernatant liquid with deionized water and (d) analyzing said sample using liquid chromatography tandem mass spectrometer (LC-MS-MS) to determine the concentration of different drug metabolites; wherein, the method is devoid of derivatization and/or solid/liquid phase extraction of samples. 
     The term “accuracy” is art-recognized and describes the degree of conformity of a measure, i.e., the quantity, to a standard or a true value. For example, an increase in the accuracy of analyte quantification refers to an improvement in obtaining a measured value that is closer to the actual or true value. This improvement may be identified/described by reference to a percent increase in accuracy with respect to the accuracy obtainable using existing methods of measurement. 
     The term “analyte” as used herein, refers to any chemical or biological compound or substance that is subject to the analysis of the disclosed methods. Analytes of the disclosed methods include, but are not limited to, small organic compounds, amino acids, peptides, polypeptides, proteins, nucleic acids, polynucleotides, biomarkers, synthetic or natural polymers, or any combination or mixture thereof. 
     The term “analyte derivative,” as used herein, describes an analyte that is functionalized with another moiety in order to convert the analyte into a derivative thereof. It is the analyte derivative that is detected for use in determining the unknown quantity of an analyte in a sample, using a response factor calculation. 
     The term “analyzing” or “analysis” is used herein to describe the method by which the quantity of each of the individual analytes described herein is detected. Such analysis may be made using any technique that distinguishes between the analyte (or analyte derivative) and the analyte standard (or analyte derivative standard). In one embodiment of the disclosed methods, the analysis or act of analyzing includes liquid chromatography-tandem mass spectrometry (LC-MS-MS). 
     The term “chromatographic separation” is art-recognized, and describes the process in which a chemical mixture carried by a liquid or gas is separated into components as a result of differential distribution of the solutes as they flow around or over a stationary liquid or solid phase. For example, chromatographic separations suitable for use in the disclosed methods include, but are not limited to liquid chromatographic (including HPLC) methods such as normal-phase HPLC, RP-HPLC, HILIC, and size-exclusion chromatography (SEC), including gel permeation chromatography (GPC). Other suitable methods include additional HPLC methods and related liquid chromatographic techniques, including, e.g., ultra-performance liquid chromatography (HPLC), fast performance liquid chromatography (FPLC) and the like. 
     The term “internal standard,” as used herein, describes a collection of one or more functionalized chemical or biological compounds or substances, e.g., one or more analytes functionalized with another moiety in order to convert such compounds or substances into a derivative thereof. Internal standards of the disclosed methods are present in known concentrations and added to the sample to form a sample mixture. The addition of the internal standard allows for the detection of and comparison between the known concentrations of one or more known analytes, with the unknown concentrations of analytes in the original sample. As such, the internal standards of the disclosed methods provide a novel way to measure the absolute quantity of an analyte in sample using a response factor calculation. 
     The term “liquid chromatography” is art-recognized and includes chromatographic methods in which compounds are partitioned between a liquid mobile phase and a solid stationary phase. Liquid chromatographic methods are used for analysis or purification of compounds. The liquid mobile phase can have a constant composition throughout the procedure (an isocratic method), or the composition of the mobile phase can be changed during elution (e.g., a gradual change in mobile phase composition such as a gradient elution method). 
     The term “mass spectrometry” and “mass spectroscopy” are art-recognized and used herein, interchangeably to describe an instrumental method for identifying the chemical constitution of a substance by means of the separation of gaseous ions according to their differing mass and charge. A variety of mass spectrometry systems can be employed to analyze the analyte molecules of a sample subjected to the methods disclosed herein. For example, mass analyzers with high mass accuracy, high sensitivity and high resolution may be used and include, but are not limited to, atmospheric chemical ionization (APCI), chemical ionization (CI), electron impact (EI), fast atom bombardment (FAB), field desorption/field ionization (FD/FI), electrospray ionization (ESI), thermospray ionization (TSP), matrix-assisted laser desorption (MALDI), matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometers, ESI-TOF mass spectrometers, and Fourier transform ion cyclotron mass analyzers (FT-ICR-MS). In addition, it should be understood that any combination of MS methods could be used in the methods described herein to analyze an analyte in a sample. In certain embodiments, the MS technique used for analysis of the analyte described herein is one that is applicable to most polar compounds, including amino acids, e.g., ESI. 
     The term “mobile phase” is art-recognized, and describes a liquid solvent system used to carry a compound of interest into contact with a solid phase (e.g., a solid phase in a solid phase extraction (SPE) cartridge or HPLC column) and to elute a compound of interest from the solid phase. 
     The term “precision” is art-recognized and describes the reproducibility of a result. It is measured by comparison of successive values obtained for a measurement to the prior values, where more precise measurements (or those with greater precision) will be demonstrated by successive measurements that are more consistently closer to the prior measurements. 
     The terms “quantitative” and “quantitatively” are art-recognized and used herein to describe measurements of quantity or amount. For example, the term “quantification” describes the act of measuring the quantity or amount of a particular object, e.g., an analyte. However, in the embodiments of the disclosed methods, the quantitative analysis is a measurement of an absolute amount, as opposed to relative amount, i.e., the total amount of analyte may be quantified absolutely in order to determine the actual amount of the analyte. 
     The term “sample” is used herein to describe a representative portion of a larger whole or group of components that are capable of being separated and detected by the methods disclosed herein. Exemplary samples include chemically or biologically derived substances, e.g., analytes of the disclosed methods. In particular embodiments, the components of the sample include, but are not limited to small organic compounds, amino acids, peptides, polypeptides, proteins, nucleic acids, polynucleotides, biomarkers, synthetic or natural polymers, or any combination or mixture thereof. 
     The term “sample mixture,” as used herein, describes the resultant product when a sample is mixed or combined with one or more analyte derivative standards, e.g., of a known concentration. 
     The term “Standard” as used herein, describes a reference material possessing one or more properties that are sufficiently well established that it can be used to prepare calibrators. 
     The term “Calibrator” as used herein, describes a solution, either prepared from the reference material or purchased, used to calibrate the assay. Where possible, calibrators should be prepared in a matrix similar to that of the specimens. 
     The term “Control” as used herein, describes a solution either prepared from the reference material (separately from the calibrators; that is, weighed or measured separately), purchased, or obtained from a pool of previously analyzed samples. Controls from any of these sources are used to determine the validity of the calibration; that is, the stability of a quantitative determination over time. Where possible, controls should be matrix-matched to specimens and calibrators, as indicated above. 
     The term “β-Glucuronidases” as used herein, describes a routinely used for the enzymatic hydrolysis of glucuronides from urine, plasma, and other fluids prior to analysis by enzyme immunoassay, mass spectrometry, gas chromatography, high performance liquid chromatography, or other means. Typically, between 1 and 20 units of glucuronidase is used per μl of plasma, urine, or bile for the enzymatic hydrolysis of glucuronides present in these samples. The exact amount needed will depend on the specific conditions used. 
     According to the methods disclosed herein the sample is a bodily fluid selected from the group consisting of oral fluids (saliva), sweat, urine, blood, serum, plasma, spinal fluid, and combination thereof. 
     According to the methods disclosed herein liquid chromatography tandem mass spectrometer (LC-MS-MS) comprises matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) MS analysis or electrospray ionization (ESI) MS. 
     In one another embodiment the methods disclosed herein involve a method for determination of drug/metabolite in a urine sample including: 
     (a) the sample were treated with enzyme and internal standard and mixture is allowed to react at room temperature 
     (b) calibrators were prepared in synthetic urine, 
     (c) detecting or determining the drug-metabolite by LC-MS-MS. 
     In some embodiments, the samples are independently selected from a saliva sample, a blood sample, a serum sample, a plasma sample, or a urine sample. There can be numerous advantages of the disclosed methods, such as
     1. Rapid analysis process—8 minute/run   2. High number of drugs analyzed concurrently—63 drugs   3. Polarity switching for basic and acidic compounds   4. Sample volume required—100 μL   5. Results show high accuracy and precision for all the analytes   6. No extraction or derivatization steps required   7. Significantly reduced cost by using less consumables and reagents   8. Reduced man hours necessary for sample preparation   9. Reduced risk for errors in sample mix-ups and cross contamination   

     The advantages of this fast polarity switching, robust, sensitive, and rapid method yield a positive impact on a laboratory&#39;s goals in providing an accurate and lean process for the analysis of multiple drugs in a single run. 
     EXAMPLES 
     Multidrug Analysis from Urine 
     Standards, deuteriated internal standards (IS) were purchased from Cerilliant, Inc. and standard stock solution is prepared with all the analytes of interest at appropriate concentrations in methanol and stored in the freezer. Five levels of working standards are prepared form the stock solution and working standards in aliquots are stored as per manufactures recommendations, and are stable until manufactures listed expiration date. 
     Deuteriated internal standards (IS) are purchased from Cerilliant, Inc and are diluted to appropriate concentrations from which a stock IS solution is prepared. The stock contains 45 spiked with different concentrations. Aliquots of stock IS mix are kept at −4° C. Working IS prepared to acetate buffer and was stable for up to 30 days. 
     Negative control was purchased from Utak Laboratories Inc. Positive Threshold control (in house control prepared from Cerilliant standards with different lot numbers from the Calibrators). Pain Management control (PM 100) was purchased from Utak Laboratories and reconstituted as per manufacturer&#39;s instructions. Benzodiazepines (100 ng/ml) were obtained from Utak Laboratories. Bio-Rad Urine Toxicology Controls are employed for selected assays. A range of acceptable concentrations (±30%) was calculated from a series of assays and recorded. 
     All other chemicals and solvents were of the highest purity available from commercial sources and used without further purification. 
     Method: 
     Urine samples were treated with P-Glucuronidase followed by minimum 90-minute incubation before centrifuging. Calibration standards and controls contain opiates/opioids (30); benzodiazepines (12); barbiturates (2) amphetamines (4); tricyclic antidepressants (8); illicit drugs (3); and Z drugs (4). Deuterated standards were used as the internal standards in the procedure. High performance liquid chromatography (Schimadzu LC-20) separation utilized gradient elution with a total run time of 8 minutes including a post run equilibration. Using positive and negative mode of ionization, an ABSciex 4500 triple-quadruple mass spectrometer was used to monitor the precursor and major product ions for each drug. Sequential MRM mode allowed for monitoring of multiple transitions based on an analyte specific retention time window. When a polarity switching experiment was utilized to obtain the maximum amount of information from a single injection, overall data quality was comparable to dedicated positive or negative experiments when intensity, signal-to-noise and reproducibility were compared. The number of data points measured across a chromatographic peak proved to be significantly high enough to achieve good resolution, precision and accuracy. Mobile phase comprised of water, methanol and acetonitrile with 0.1% Formic acid and 0.1% ammonium formate. Data analysis was performed on the Multiquant software version 3.0. 
     Sample Preparation 
     The calibrators, controls and the sample prepared in the same method. All the samples and the working solutions (calibrators, controls, and internal standards) should be allowed to get into room temperature.
         a) Qualitatively aliquot 100 μL of the calibrators, controls, and specimens into the vial and label the tubes.   b) If a dilution is necessary, record the dilution factor, volume of negative urine, and the volume of the specimen used to make the dilution on the work list.   c) Add 100 μL of the working internal standard (made of Acetate buffer pH 4.1) to the vial.   d) Add 25 VL of β-Glucuronidase enzyme to the vial   e) Call all the tubes and vortex   f) Incubate all tubes in 55° C. in the oven for 1.5 hrs.   g) Remove all the tubes from the oven to allow cooling to room temperature   h) Centrifuge the tubes at 14,000 RPM for 15 minutes   i) Transfer 100 μL to a clean vial and add 500 μL of DI H2O   j) Vortex and analyze by LC/MS/MS       

     Instruments and Equipment 
     LC Conditions: 
     The chromatographic separation and detection were performed using Kinetex 2.6u XB-C18 100A (50×3.0 mm) column (Phenomenox, USA). A mutli-step gradient elution method with an aqueous DI H2O containing 0.1% formic acid &amp; ammonium formate (Mobile phase A) and 50% Acetonitrile &amp; 50% Methanol containing 0.1% formic acid (Mobilephase B) at a flow rate of 0.50 mL/min to separate all the 63 compounds in positive and negative ionization mode. With a low sample injection volume of 10 μL and no sample preconcentraion, the presented method demonstrated excellent signal-to-noise (S/N) ratios due to the enhanced sensitivity of the Absicex 4500 Triple Quadrupole LC/MS/MS. 
     Column temperature set at 45° C. during whole run and the injection volume as 10 μL. The gradient elution method for chromatographic separation is provided in Table 1. The graphical view of optimized multi-step gradient elution method for LC/MS/MS is shown in  FIG. 1 . 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Gradient elution method for the separation 
               
            
           
           
               
               
               
            
               
                 Time 
                   
                   
               
               
                 (mins) 
                 Pump A 
                 Pump B 
               
               
                   
               
            
           
           
               
               
               
            
               
                 0.5 
                 95% 
                  5% 
               
               
                 2.00 
                 75% 
                 25% 
               
               
                 4.50 
                 15% 
                 85% 
               
               
                 4.51 
                  2% 
                 98% 
               
               
                 5.50 
                  2% 
                 98% 
               
               
                 5.51 
                 95% 
                  5% 
               
               
                 6.50 
                 95% 
                  5% 
               
               
                   
               
            
           
         
       
     
     Mass Spectrometer Conditions 
     ABsciex source provides a range of capabilities for testing samples. The precursor and product ions, along with optimized fragment and collision energy and other optimal voltages for each of the analytes are described in the method program. The ABSciex-Triple Quadrupole System consists of an ion source, enhanced desolvation technology followed by ion optics that transfer the ions to the first quadrupole (the Precursor Quad filter Q1), to the collision cell (Q2), and then to the third quadrupole (product Quad Filter Q3). The non-reactive inert gas nitrogen is used as collision gas. The precursor ion is selected using the first quadrupole and is sent to the collision cell for fragmentation. Fragment ions are derived from the precursor and therefore represent the structure of the precursor molecule. A specific precursor ion and specific product ions are thus selected and monitored. This type of analysis is termed Selected Reaction Monitoring (SRM). A triple quadrupole MS instrument running multiple SRMs for the same precursor ions is called Multiple Reaction Monitoring (MRM). 
     The Analyst software, a quantitation Wizard Program, is used to quantitate the analytes. A quantitation method is created with an algorithm (Intelliquan) which will generate quantitation tables. Integration of the peaks, regression and linearity of the calibration curve and accuracy of the standards and controls must be reviewed and verified. 
     The MS parameters for each analyte are obtained by infusing 10 ng/mL and optimized the parameters based on the sensitivity. The optimized parameters for the mass spectrometry were given in the following Table 2. 
     The specific parameters such as Declustering Potential (DP) decluster ions, Entrance Potential (EP), Collision Cell Entrance Potential (CE) and Collision Cell exit potential (CXP), parent ion, daughter ion and the retention time are mentioned in the Table 3 with 60 analytes in positive mode and 3 analytes in negative mode. 
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Optimized parameter for the mass spectrometry. 
               
            
           
           
               
               
               
            
               
                   
                 Parameters 
                 Ranges 
               
               
                   
                   
               
            
           
           
               
               
               
            
               
                   
                 Curtain gas (CUR) 
                 35 
               
               
                   
                 Collision Gas (CAD) 
                 7 
               
               
                   
                 Ionspray Voltage (IS) 
                 2500 
               
               
                   
                 Temperature (TEM) 
                 550 
               
               
                   
                 Ion source Gas 1 (GS1) 
                 60 
               
               
                   
                 Ion Source Gas 2 (GS 2) 
                 60 
               
               
                   
                   
               
            
           
         
       
     
     Determination: the sample were analyzed by liquid chromatography-tandem quadrupole mass spectrometer measured which enables 63 kinds of drugs belonging to different chemical and toxicological class, where barbiturates were analyzed in negative ion mode of analysis and all other compounds were analyzed using positive ion mode of analysis. 
     MS conditions and parameters for positive ion mode and negative ion mode can be used as are known in the art without affecting the overall concept of the methods disclosed herein. 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 List of Internal Standards (IS) with Q1, Q3 and their 
               
               
                 parameters for both positive and negative mode 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 Internal Standard 
                 Q1 
                 Q3 
                 RT 
                   
                   
                   
                   
               
               
                 (IS) 
                 (m/z) 
                 (m/z) 
                 (min) 
                 DP 
                 EP 
                 CE 
                 CXP 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 6-MAM-D6 
                 334.3 
                 165.2 
                 2.8 
                 61.0 
                 10.0 
                 49.0 
                 14.0 
               
               
                 Hydrocodone-D6 
                 306.2 
                 202.2 
                 2.7 
                 81.0 
                 10.0 
                 39.0 
                 4.0 
               
               
                 Hydromorphone-D3 
                 289.2 
                 128.1 
                 1.5 
                 81.0 
                 10.0 
                 73.0 
                 4.0 
               
               
                 Morphine-D3 
                 289.2 
                 152.1 
                 1.0 
                 66.0 
                 10.0 
                 77.0 
                 4.0 
               
               
                 Oxycodone-D3 
                 319.1 
                 244.1 
                 2.6 
                 81.0 
                 10.0 
                 39.0 
                 7.0 
               
               
                 Oxymorphone-D3 
                 305.2 
                 230.0 
                 1.2 
                 70.0 
                 10.0 
                 35.0 
                 16.0 
               
               
                 Carisoprodol-D7 
                 268.1 
                 183.1 
                 4.5 
                 56.0 
                 10.0 
                 13.0 
                 15.0 
               
               
                 Tapentadol-D3 
                 225.2 
                 121.0 
                 3.4 
                 51.0 
                 10.0 
                 27.0 
                 4.0 
               
               
                 EDDP-D3 
                 281.3 
                 234.2 
                 4.1 
                 61.0 
                 10.0 
                 39.0 
                 4.0 
               
               
                 Meperidine-D4 
                 252.2 
                 224.2 
                 3.6 
                 46.0 
                 10.0 
                 29.0 
                 4.0 
               
               
                 Meprobamate-D7 
                 226.2 
                 165.3 
                 3.8 
                 41.0 
                 10.0 
                 11.0 
                 4.0 
               
               
                 Methadone-D3 
                 313.2 
                 105.0 
                 4.4 
                 31.0 
                 10.0 
                 35.0 
                 4.0 
               
               
                 Normeperidine-D4 
                 238.2 
                 164.2 
                 3.6 
                 36.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                 Tramadol-D3 
                 267.2 
                 58.0 
                 3.4 
                 60.0 
                 10.0 
                 75.0 
                 8.0 
               
               
                 Fentanyl-D5 
                 342.2 
                 105.1 
                 3.9 
                 46.0 
                 11.5 
                 53.0 
                 4.0 
               
               
                 Norfentanyl-D5 
                 238.2 
                 84.0 
                 3.3 
                 75.0 
                 10.0 
                 24.0 
                 20.0 
               
               
                 Norpropoxyphene-D5 
                 313.3 
                 105.0 
                 4.4 
                 31.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                 Propoxyphene-D5 
                 345.2 
                 271.3 
                 4.4 
                 16.0 
                 10.0 
                 15.0 
                 4.0 
               
               
                 7-AminoClonazepam-D4 
                 290.2 
                 121.1 
                 3.3 
                 71.0 
                 10.0 
                 41.0 
                 8.0 
               
               
                 Diazepam-D5 
                 290.4 
                 198.0 
                 4.9 
                 76.0 
                 9.0 
                 41.0 
                 4.0 
               
               
                 4-HydroxyAlprazolam-D5 
                 330.2 
                 302.1 
                 4.5 
                 86.0 
                 10.0 
                 27.0 
                 4.0 
               
               
                 Nordiazepam-D5 
                 276.1 
                 165.1 
                 4.7 
                 71.0 
                 10.0 
                 39.0 
                 10.0 
               
               
                 Oxazepam-D5 
                 292.1 
                 246.0 
                 4.5 
                 76.0 
                 10.0 
                 23.0 
                 10.0 
               
               
                 OH-et-Flurazepam-D4 
                 337.1 
                 113.1 
                 4.6 
                 61.0 
                 10.0 
                 43.0 
                 4.0 
               
               
                 Lorazepam-D4 
                 325.2 
                 279.0 
                 4.6 
                 76.0 
                 6.5 
                 25.0 
                 4.0 
               
               
                 Zolpidem-D7 
                 315.0 
                 242.0 
                 3.6 
                 121.0 
                 10.0 
                 45.0 
                 12.0 
               
               
                 Amphetamine-D6 
                 142.2 
                 93.1 
                 2.6 
                 41.0 
                 10.5 
                 21.0 
                 4.0 
               
               
                 MDA-D5 
                 185.2 
                 168.0 
                 2.7 
                 41.0 
                 10.0 
                 29.0 
                 4.0 
               
               
                 MDMA-D5 
                 199.2 
                 165.2 
                 2.8 
                 41.0 
                 10.5 
                 15.0 
                 4.0 
               
               
                 Methamphetamine-D5 
                 155.2 
                 92.0 
                 2.8 
                 26.0 
                 10.0 
                 27.0 
                 4.0 
               
               
                 Methylphenidate-D9 
                 243.2 
                 93.0 
                 3.4 
                 46.0 
                 10.0 
                 31.0 
                 4.0 
               
               
                 Desipramine-D3 
                 270.2 
                 193.2 
                 4.3 
                 46.0 
                 10.5 
                 45.0 
                 4.0 
               
               
                 Imipramine-D3 
                 284.2 
                 89.1 
                 4.3 
                 46.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                 Nortriptyline-D3 
                 267.2 
                 91.0 
                 4.4 
                 46.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                 Sertraline-D3 
                 309.2 
                 275.1 
                 4.5 
                 66.0 
                 10.0 
                 15.0 
                 4.0 
               
               
                 Pregabalin-D6 
                 166.0 
                 148.1 
                 2.4 
                 40.0 
                 10.0 
                 13.0 
                 6.0 
               
               
                 Gabapentin-D10 
                 182.1 
                 164.1 
                 2.4 
                 21.0 
                 10.0 
                 21.0 
                 14.0 
               
               
                 Butalbital-D5 
                 228.0 
                 42.0 
                 4.1 
                 −45.0 
                 −12.0 
                 −36.0 
                 −11.0 
               
               
                 Phenobarbital-D5 
                 236.0 
                 42.0 
                 3.9 
                 −50.0 
                 −8.0 
                 −36.0 
                 −9.0 
               
               
                 THC-COOH D3 
                 346.2 
                 302.3 
                 5.7 
                 −100.0 
                 −9.0 
                 −30.0 
                 −9.0 
               
               
                 Benzoylecgonine-D3 
                 293.0 
                 171.2 
                 3.3 
                 70.0 
                 10.0 
                 26.0 
                 8.0 
               
               
                 PCP-D5 
                 249.3 
                 96.0 
                 3.8 
                 30.0 
                 10.0 
                 41.0 
                 16.0 
               
               
                   
               
               
                 Q1—Quadrapole one, 
               
               
                 Q3—Quadrapole three, 
               
               
                 RT: Retention time, 
               
               
                 DP—declustering potentials, 
               
               
                 EP—entrance potentials, 
               
               
                 CE—collision energies, 
               
               
                 CXP—collision cell exit potentials, 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 List of analytes with Q1, Q3 and their parameters for both positive and negative mode 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Drug-Metabolite 
                 Drug -metabolite 
                 Q1 
                 Q3 
                 RT 
                   
                   
                   
                   
               
               
                 category 
                 Name 
                 (m/z) 
                 (m/z) 
                 (min) 
                 DP 
                 EP 
                 CE 
                 CXP 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Opiates/opoids 
                 6MAM 
                 328.1 
                 165.2 
                 2.8 
                 85.0 
                 10.0 
                 49.0 
                 12.0 
               
               
                   
                   
                 328.1 
                 211.2 
                 2.8 
                 85.0 
                 10.0 
                 35.0 
                 16.0 
               
               
                   
                 Codeine 
                 300.1 
                 152.1 
                 2.4 
                 106.0 
                 10.0 
                 73.0 
                 10.0 
               
               
                   
                   
                 300.1 
                 115.0 
                 2.4 
                 106.0 
                 10.0 
                 91.0 
                 8.0 
               
               
                   
                 Dihydrocodeine 
                 302.3 
                 128.2 
                 2.4 
                 66.0 
                 10.0 
                 81.0 
                 4.0 
               
               
                   
                   
                 302.3 
                 199.1 
                 2.4 
                 66.0 
                 10.0 
                 41.0 
                 4.0 
               
               
                   
                 Hydrocodone 
                 300.1 
                 199.1 
                 2.7 
                 56.0 
                 10.0 
                 39.0 
                 4.0 
               
               
                   
                   
                 300.1 
                 128.1 
                 2.7 
                 56.0 
                 10.0 
                 69.0 
                 4.0 
               
               
                   
                 Hydromorphone 
                 286.2 
                 185.0 
                 1.5 
                 56.0 
                 10.0 
                 39.0 
                 4.0 
               
               
                   
                   
                 286.2 
                 128.0 
                 1.5 
                 56.0 
                 10.0 
                 73.0 
                 4.0 
               
               
                   
                 Morphine 
                 286.2 
                 152.0 
                 1.0 
                 56.0 
                 10.0 
                 73.0 
                 4.0 
               
               
                   
                   
                 286.2 
                 165.0 
                 1.0 
                 56.0 
                 10.0 
                 47.0 
                 4.0 
               
               
                   
                 Oxycodone 
                 316.1 
                 241.0 
                 2.6 
                 50.0 
                 10.0 
                 43.0 
                 15.0 
               
               
                   
                   
                 316.1 
                 256.0 
                 2.6 
                 50.0 
                 10.0 
                 38.0 
                 15.0 
               
               
                   
                 Oxymorphone 
                 302.1 
                 227.0 
                 1.2 
                 75.0 
                 10.0 
                 37.0 
                 8.0 
               
               
                   
                   
                 302.1 
                 198.1 
                 1.2 
                 75.0 
                 10.0 
                 55.0 
                 8.0 
               
               
                   
                 Buprenophrine 
                 468.3 
                 396.1 
                 4.1 
                 55.0 
                 10.0 
                 53.0 
                 14.0 
               
               
                   
                   
                 468.3 
                 414.2 
                 4.1 
                 55.0 
                 10.0 
                 43.0 
                 6.0 
               
               
                   
                 Carisoprodol 
                 261.2 
                 176.2 
                 4.5 
                 26.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                   
                   
                 261.2 
                 97.2 
                 4.5 
                 26.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                 Desmethyl 
                 208.2 
                 107.2 
                 3.4 
                 51.0 
                 10.0 
                 31.0 
                 4.0 
               
               
                   
                 Tapentadol 
                 208.2 
                 121.2 
                 3.4 
                 51.0 
                 10.0 
                 25.0 
                 4.0 
               
               
                   
                 Desmethyl 
                 250.3 
                 58.0 
                 2.9 
                 26.0 
                 10.0 
                 13.0 
                 4.0 
               
               
                   
                 Tramadol 
                 250.3 
                 91.0 
                 2.9 
                 26.0 
                 10.0 
                 55.0 
                 4.0 
               
               
                   
                 EDDP 
                 278.2 
                 234.2 
                 4.1 
                 100.0 
                 10.0 
                 67.0 
                 4.0 
               
               
                   
                   
                 278.2 
                 249.2 
                 4.1 
                 100.0 
                 10.0 
                 43.0 
                 4.0 
               
               
                   
                 Meperidine 
                 248.2 
                 220.0 
                 3.5 
                 95.0 
                 10.0 
                 43.0 
                 8.0 
               
               
                   
                   
                 248.2 
                 174.1 
                 3.5 
                 95.0 
                 10.0 
                 47.0 
                 20.0 
               
               
                   
                 Meprobamate 
                 219.1 
                 157.9 
                 3.8 
                 46.0 
                 10.0 
                 11.0 
                 4.0 
               
               
                   
                   
                 219.1 
                 97.0 
                 3.8 
                 46.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                   
                 Methadone 
                 310.2 
                 265.2 
                 4.4 
                 44.0 
                 10.0 
                 33.0 
                 15.0 
               
               
                   
                   
                 310.2 
                 105.0 
                 4.4 
                 44.0 
                 10.0 
                 54.0 
                 17.0 
               
               
                   
                 Norbuprenophrine 
                 414.1 
                 211.0 
                 3.8 
                 95.0 
                 10.0 
                 50.0 
                 8.0 
               
               
                   
                   
                 414.1 
                 187.0 
                 3.8 
                 95.0 
                 10.0 
                 45.0 
                 8.0 
               
               
                   
                 Normeperidine 
                 234.1 
                 160.2 
                 3.6 
                 36.0 
                 10.0 
                 21.0 
                 4.0 
               
               
                   
                   
                 234.1 
                 188.1 
                 3.6 
                 36.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                   
                 Tapentadol 
                 222.1 
                 107.2 
                 3.4 
                 116.0 
                 10.0 
                 39.0 
                 18.0 
               
               
                   
                   
                 222.1 
                 121.2 
                 3.4 
                 116.0 
                 10.0 
                 27.0 
                 12.0 
               
               
                   
                 Tramadol 
                 264.2 
                 58.1 
                 3.3 
                 82.0 
                 10.0 
                 55.0 
                 30.0 
               
               
                   
                   
                 264.2 
                 42.1 
                 3.3 
                 60.0 
                 10.0 
                 107.0 
                 20.0 
               
               
                   
                 Fentanyl 
                 337.2 
                 105.1 
                 3.9 
                 51.0 
                 9.0 
                 53.0 
                 4.0 
               
               
                   
                   
                 337.2 
                 188.1 
                 3.9 
                 51.0 
                 9.0 
                 29.0 
                 4.0 
               
               
                   
                 Norfentanyl 
                 233.2 
                 84.1 
                 3.3 
                 41.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                   
                 233.2 
                 150.0 
                 3.3 
                 41.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                 Norpropoxyphene 
                 308.2 
                 100.1 
                 4.3 
                 43.0 
                 10.0 
                 30.0 
                 8.0 
               
               
                   
                   
                 308.2 
                 143.2 
                 4.3 
                 31.0 
                 10.0 
                 29.0 
                 10.0 
               
               
                   
                 Propoxyphene 
                 340.2 
                 266.3 
                 4.3 
                 57.0 
                 10.0 
                 19.0 
                 23.0 
               
               
                   
                   
                 340.2 
                 91.1 
                 4.3 
                 16.0 
                 10.0 
                 67.0 
                 14.0 
               
               
                   
                 Dextromethophan 
                 272.2 
                 171.0 
                 4.0 
                 28.0 
                 10.0 
                 55.0 
                 26.0 
               
               
                   
                   
                 272.2 
                 147.0 
                 4.0 
                 28.0 
                 10.0 
                 55.0 
                 26.0 
               
               
                   
                 Dextrophan 
                 257.9 
                 157.5 
                 3.3 
                 51.0 
                 10.0 
                 49.0 
                 13.0 
               
               
                   
                   
                 257.9 
                 133.0 
                 3.3 
                 51.0 
                 10.0 
                 35.0 
                 13.0 
               
               
                   
                 Desomorphine 
                 273.0 
                 216.0 
                 2.6 
                 101.0 
                 10.5 
                 35.0 
                 6.0 
               
               
                   
                   
                 273.0 
                 196.0 
                 2.6 
                 101.0 
                 10.5 
                 41.0 
                 8.0 
               
               
                   
                 Nalaxone 
                 328.3 
                 310.2 
                 2.4 
                 46.0 
                 10.0 
                 21.0 
                 4.0 
               
               
                   
                   
                 328.3 
                 212.1 
                 2.4 
                 46.0 
                 10.0 
                 57.0 
                 4.0 
               
               
                 Benzodiazepines 
                 7-AminoClonazepam 
                 286.0 
                 121.2 
                 3.3 
                 68.0 
                 10.0 
                 36.0 
                 16.0 
               
               
                   
                   
                 286.0 
                 222.1 
                 3.3 
                 68.0 
                 10.0 
                 34.0 
                 14.0 
               
               
                   
                 Diazepam 
                 285.1 
                 154.0 
                 4.9 
                 101.0 
                 10.0 
                 39.0 
                 26.0 
               
               
                   
                   
                 285.1 
                 193.1 
                 4.9 
                 101.0 
                 10.0 
                 45.0 
                 10.0 
               
               
                   
                 Flunitrazepam 
                 314.1 
                 268.1 
                 4.6 
                 91.0 
                 10.0 
                 50.0 
                 22.0 
               
               
                   
                   
                 314.1 
                 239.1 
                 4.6 
                 91.0 
                 10.0 
                 62.0 
                 14.0 
               
               
                   
                 4-HydroxyAlprazolam 
                 325.1 
                 297.2 
                 4.5 
                 71.0 
                 10.0 
                 27.0 
                 4.0 
               
               
                   
                   
                 325.1 
                 216.1 
                 4.5 
                 71.0 
                 10.0 
                 53.0 
                 4.0 
               
               
                   
                 Nordiazepam 
                 271.0 
                 140.0 
                 4.7 
                 71.0 
                 10.0 
                 37.0 
                 10.0 
               
               
                   
                   
                 271.0 
                 165.1 
                 4.7 
                 71.0 
                 10.0 
                 37.0 
                 14.0 
               
               
                   
                 Oxazepam 
                 287.0 
                 241.1 
                 4.5 
                 76.0 
                 10.0 
                 33.0 
                 11.0 
               
               
                   
                   
                 287.0 
                 269.1 
                 4.5 
                 76.0 
                 10.0 
                 23.0 
                 10.0 
               
               
                   
                 Temazepam 
                 301.1 
                 255.1 
                 4.7 
                 70.0 
                 10.0 
                 31.0 
                 20.0 
               
               
                   
                   
                 301.1 
                 283.0 
                 4.7 
                 70.0 
                 10.0 
                 21.0 
                 14.0 
               
               
                   
                 Chloradiazepoxide 
                 300.0 
                 283.1 
                 4.1 
                 70.0 
                 5.0 
                 25.0 
                 4.0 
               
               
                   
                   
                 300.0 
                 227.2 
                 4.1 
                 70.0 
                 5.0 
                 33.0 
                 3.0 
               
               
                   
                 OH-et Flurazepam 
                 333.1 
                 109.1 
                 4.6 
                 61.0 
                 10.0 
                 43.0 
                 4.0 
               
               
                   
                   
                 333.1 
                 211.3 
                 4.6 
                 61.0 
                 10.0 
                 43.0 
                 4.0 
               
               
                   
                 Lorazepam 
                 321.0 
                 275.0 
                 4.6 
                 75.0 
                 10.0 
                 31.0 
                 14.0 
               
               
                   
                   
                 321.0 
                 229.0 
                 4.6 
                 75.0 
                 10.0 
                 43.0 
                 16.6 
               
               
                   
                 Triazolam 
                 344.9 
                 317.0 
                 4.6 
                 111.0 
                 10.0 
                 37.0 
                 14.0 
               
               
                   
                   
                 344.9 
                 310.0 
                 4.6 
                 111.0 
                 10.0 
                 35.0 
                 14.0 
               
               
                   
                 Midazolam 
                 326.1 
                 291.1 
                 4.1 
                 101.0 
                 10.0 
                 37.0 
                 22.0 
               
               
                   
                   
                 326.1 
                 249.1 
                 4.1 
                 101.0 
                 10.0 
                 47.0 
                 18.0 
               
               
                 Z drugs 
                 Zolpidem 
                 308.0 
                 235.1 
                 3.6 
                 85.0 
                 10.0 
                 65.0 
                 45.0 
               
               
                   
                   
                 308.0 
                 219.0 
                 3.6 
                 51.0 
                 10.0 
                 77.0 
                 6.0 
               
               
                   
                 Zaleplon 
                 306.1 
                 264.0 
                 4.3 
                 66.0 
                 10.0 
                 21.0 
                 4.0 
               
               
                   
                   
                 306.1 
                 236.0 
                 4.3 
                 66.0 
                 10.0 
                 45.0 
                 4.0 
               
               
                   
                 Zopiclone 
                 389.2 
                 244.9 
                 3.3 
                 15.0 
                 10.0 
                 27.0 
                 10.0 
               
               
                   
                   
                 389.2 
                 217.0 
                 3.3 
                 15.0 
                 10.0 
                 45.0 
                 8.0 
               
               
                   
                 Zolpidem-COOH 
                 337.8 
                 293.1 
                 3.1 
                 126.0 
                 10.0 
                 37.0 
                 12.0 
               
               
                   
                   
                 337.8 
                 265.1 
                 3.1 
                 126.0 
                 10.0 
                 47.0 
                 13.0 
               
               
                 Amphetamines 
                 Amphetamine 
                 136.1 
                 91.0 
                 2.6 
                 64.0 
                 10.0 
                 33.0 
                 12.0 
               
               
                   
                   
                 136.1 
                 119.0 
                 2.6 
                 60.0 
                 10.0 
                 17.0 
                 8.0 
               
               
                   
                 MDA 
                 180.1 
                 163.0 
                 2.7 
                 16.0 
                 10.0 
                 29.0 
                 4.0 
               
               
                   
                   
                 180.1 
                 133.0 
                 2.7 
                 16.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                 MDMA 
                 194.1 
                 163.2 
                 2.9 
                 65.0 
                 10.0 
                 31.0 
                 14.0 
               
               
                   
                   
                 194.1 
                 105.2 
                 2.9 
                 65.0 
                 10.0 
                 40.0 
                 18.0 
               
               
                   
                 Methamphetamine 
                 150.1 
                 91.2 
                 2.8 
                 53.0 
                 10.0 
                 42.0 
                 38.0 
               
               
                   
                   
                 150.1 
                 119.2 
                 2.8 
                 50.0 
                 10.0 
                 22.0 
                 19.0 
               
               
                 Tricyclic 
                 Methyl phenidate 
                 234.3 
                 84.1 
                 3.4 
                 60.0 
                 10.0 
                 38.0 
                 20.0 
               
               
                 antidepressants 
                   
                 234.3 
                 56.1 
                 3.4 
                 46.0 
                 10.0 
                 63.0 
                 14.0 
               
               
                   
                 Desipramine 
                 267.3 
                 72.1 
                 4.3 
                 46.0 
                 10.0 
                 33.0 
                 20.0 
               
               
                   
                   
                 267.3 
                 193.1 
                 4.3 
                 46.0 
                 10.0 
                 45.0 
                 4.0 
               
               
                   
                 Imipramine 
                 281.3 
                 86.1 
                 4.3 
                 50.0 
                 10.0 
                 30.0 
                 15.0 
               
               
                   
                   
                 281.3 
                 58.1 
                 4.3 
                 50.0 
                 10.0 
                 55.0 
                 15.0 
               
               
                   
                 Nortriptyline 
                 264.2 
                 117.2 
                 4.4 
                 56.0 
                 10.0 
                 25.0 
                 4.0 
               
               
                   
                   
                 264.2 
                 233.2 
                 4.4 
                 56.0 
                 10.0 
                 17.0 
                 4.0 
               
               
                   
                 Ritalinic Acid 
                 220.2 
                 84.1 
                 3.2 
                 52.0 
                 10.0 
                 34.0 
                 15.0 
               
               
                   
                   
                 220.2 
                 56.1 
                 3.2 
                 52.0 
                 10.0 
                 60.0 
                 14.0 
               
               
                   
                 Sertraline 
                 306.1 
                 159.1 
                 4.5 
                 30.0 
                 11.0 
                 33.0 
                 14.0 
               
               
                   
                   
                 306.1 
                 275.1 
                 4.5 
                 30.0 
                 11.0 
                 17.0 
                 17.0 
               
               
                   
                 Cyclobenzaprine 
                 276.2 
                 215.0 
                 4.3 
                 90.0 
                 10.0 
                 50.0 
                 8.0 
               
               
                   
                   
                 276.2 
                 216.2 
                 4.3 
                 90.0 
                 10.0 
                 35.0 
                 8.0 
               
               
                   
                 Amitriptyline 
                 278.4 
                 91.0 
                 4.4 
                 56.0 
                 10.0 
                 33.0 
                 8.0 
               
               
                   
                   
                 278.4 
                 117.1 
                 4.4 
                 56.0 
                 10.0 
                 31.0 
                 8.0 
               
               
                 Antiepileptics 
                 Pregabalin 
                 160.1 
                 142.0 
                 2.4 
                 40.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                   
                 160.1 
                 97.0 
                 2.4 
                 40.0 
                 10.0 
                 23.0 
                 4.0 
               
               
                   
                 Gabapentin 
                 172.1 
                 95.1 
                 2.4 
                 64.0 
                 10.0 
                 30.0 
                 15.0 
               
               
                   
                   
                 172.1 
                 137.2 
                 2.4 
                 64.0 
                 10.0 
                 21.0 
                 13.0 
               
               
                 Barbiturates 
                 Butalbital 
                 223.1 
                 42.0 
                 4.1 
                 −55.0 
                 −12.0 
                 −40.0 
                 −11.0 
               
               
                   
                   
                 223.1 
                 180.0 
                 4.1 
                 −55.0 
                 −12.0 
                 −16.0 
                 −5.0 
               
               
                   
                 Phenobarbital 
                 231.1 
                 42.0 
                 3.9 
                 −55.0 
                 −8.0 
                 −42.0 
                 −5.0 
               
               
                   
                   
                 231.1 
                 188.0 
                 3.9 
                 −55.0 
                 −8.0 
                 −14.0 
                 −9.0 
               
               
                 Illicit drugs 
                 THCA 
                 343.1 
                 299.1 
                 5.7 
                 −115.0 
                 −9.0 
                 −30.0 
                 −11.0 
               
               
                   
                   
                 343.1 
                 245.1 
                 5.7 
                 −115.0 
                 −9.0 
                 −40.0 
                 −13.0 
               
               
                   
                 Benzoylecgonine 
                 290.1 
                 168.1 
                 3.3 
                 61.0 
                 10.0 
                 38.0 
                 30.0 
               
               
                   
                   
                 290.1 
                 105.0 
                 3.3 
                 61.0 
                 10.0 
                 45.0 
                 16.0 
               
               
                   
                 PCP 
                 244.2 
                 91.0 
                 3.8 
                 21.0 
                 10.0 
                 41.0 
                 4.0 
               
               
                   
                   
                 244.2 
                 86.0 
                 3.8 
                 21.0 
                 10.0 
                 41.0 
                 4.0 
               
               
                   
               
               
                 Q1—Quadrapole one, 
               
               
                 Q3—Quadrapole three, 
               
               
                 RT: Retention time, 
               
               
                 DP—declustering potentials, 
               
               
                 EP—entrance potentials, 
               
               
                 CE—collision energies, 
               
               
                 CXP—collision cell exit potentials, 
               
            
           
         
       
     
     The limit of quantification varies from 2 to 100 ng/mL depends on the analytes. The list of analytes shown in the Table 5 based on the classification. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Analytes based on classification 
               
            
           
           
               
               
               
            
               
                   
                 Drug Name 
                 LOQ 
               
               
                   
                   
               
            
           
           
               
               
               
            
               
                   
                 6MAM 
                 2 
               
               
                   
                 Codeine 
                 50 
               
               
                   
                 Dihydrocodeine 
                 50 
               
               
                   
                 Hydrocodone 
                 50 
               
               
                   
                 Hydromorphone 
                 50 
               
               
                   
                 Morphine 
                 50 
               
               
                   
                 Oxycodone 
                 50 
               
               
                   
                 Oxymorphone 
                 50 
               
               
                   
                 Buprenophrine 
                 5 
               
               
                   
                 Carisoprodol 
                 50 
               
               
                   
                 Desmethyl Tapentadol 
                 50 
               
               
                   
                 DesmethylTramadol 
                 50 
               
               
                   
                 EDDP 
                 50 
               
               
                   
                 Meperidine 
                 50 
               
               
                   
                 Meprobamate 
                 50 
               
               
                   
                 Methadone 
                 50 
               
               
                   
                 Norbuprenophrine 
                 10 
               
               
                   
                 Normeperidine 
                 50 
               
               
                   
                 Tapentadol 
                 50 
               
               
                   
                 Tramadol 
                 50 
               
               
                   
                 Fentanyl 
                 2 
               
               
                   
                 Norfentanyl 
                 2 
               
               
                   
                 Norpropoxyphene 
                 50 
               
               
                   
                 Propoxyphene 
                 50 
               
               
                   
                 Dextromethophan 
                 50 
               
               
                   
                 Dextrophan 
                 50 
               
               
                   
                 Desomorphine 
                 50 
               
               
                   
                 Nalaxone 
                 10 
               
               
                   
                 7-AminoClonazepam 
                 10 
               
               
                   
                 Diazepam 
                 10 
               
               
                   
                 Flunitrazepam 
                 10 
               
               
                   
                 HydroxyAlprazolam 
                 10 
               
               
                   
                 Nordiazepam 
                 10 
               
               
                   
                 Oxazepam 
                 10 
               
               
                   
                 Temazepam 
                 10 
               
               
                   
                 Chloradiazepoxide 
                 10 
               
               
                   
                 OH-et-Flurazepam 
                 10 
               
               
                   
                 Lorazepam 
                 10 
               
               
                   
                 Triazolam 
                 10 
               
               
                   
                 Midazolam 
                 10 
               
               
                   
                 Z-DRUGS 
               
               
                   
                 Zolpidem 
                 10 
               
               
                   
                 Zaleplon 
                 10 
               
               
                   
                 Zopiclone 
                 10 
               
               
                   
                 Zolpidem-COOH 
                 10 
               
               
                   
                 TRICYCLIC 
               
               
                   
                 ANTIDEPRESSANTS 
               
               
                   
                 Desipramine 
                 20 
               
               
                   
                 Imipramine 
                 20 
               
               
                   
                 Methyl Phenidate 
                 50 
               
               
                   
                 Nortriptyline 
                 20 
               
               
                   
                 Ritalinic Acid 
                 100 
               
               
                   
                 Sertraline 
                 20 
               
               
                   
                 Cyclobenzaprine 
                 20 
               
               
                   
                 Amitriptyline 
                 20 
               
               
                   
                 ANTIEPILEPTICS 
               
               
                   
                 Pregabilan 
                 50 
               
               
                   
                 Gabapentin 
                 100 
               
               
                   
                 AMPHETAMINES 
               
               
                   
                 Amphetamine 
                 50 
               
               
                   
                 MDA 
                 50 
               
               
                   
                 MDMA 
                 50 
               
               
                   
                 Methamphetamine 
                 50 
               
               
                   
                 BARBITURATES 
               
               
                   
                 Butalbital 
                 50 
               
               
                   
                 Phenobarbital 
                 50 
               
               
                   
                 ILLICITS 
               
               
                   
                 THCA 
                 10 
               
               
                   
                 Benzoylecgonine 
                 50 
               
               
                   
                 PCP 
                 4 
               
               
                   
                   
               
            
           
         
       
     
     Results: 
     All drugs were analyzed simultaneously using the same procedure steps. Target concentrations varied between 2 ng/ml to 20,000 ng/ml among the analytes depending on their therapeutic and toxic levels. Approximately 173 transitions were monitored per run by sequential MRM. Assays with low level cutoffs (such as 2 ng/mL for certain opioids such as fentanyl and 20 ng/ml for benzodiazepines) were reproducible and met acceptable chromatographic criteria. An accuracy of 95-105% and a CV of 2.0-10.2 were achieved for each calibration point in a method validation study. Further statistical data indicated an accuracy of 96-108% for quality control samples. The limit of detection and the upper linearity limit results have been quantified and established for each component. 
     MRM Parameter 
     Each chemical compound supposes two female ions-sub-ion pair, is divided into 3 MRM channel collections. 
     The methods disclosed herein can achieve the lowest detection limit 2-200 ng/mL. Within 2-100 ng/mL concentration range, 63 kinds of drugs good linear correlation coefficient of 0.9771-0.9995. In urine, the recovery of most drugs between 70-130% and the recovery of a few drugs between 60-70%, RSD is less than 15.0%, to meet the needs of daily quantitative analysis. 
     SAMHSA guidelines require the use of one quantifier and at least one quantifier for both target compound and internal standard (Table 3) The detection methods disclosed herein to establish a liquid chromatography-tandem mass spectrometry detection in urine 63 kinds of common drugs. Can be used for Entry-Exit Inspection and Quarantine, Centers for Disease Control, the public security departments rapid detection and confirmation of positive results. 
     LC-MS Set Up: 
     ABSciex-Triple Quadrupole System 4500 consists of an ion source, enhanced desolvation technology with Electron Spray Ionization in positive mode. 
     For all MS-MS experiments, mass calibration and resolution adjustments (at 0.7 amu full width at half height) on both the resolving quadrupoles were automatically optimized using a poly(propylene)glycol 1 3 1024 mol/L solution introduced via the built-in infusion pump. In the ABSciex API3200 Triple Quad adjustable voltage, Declustering Potential (DP) declusters ions, Entrance Potential (EP) focuses ions, Collision Cell Entrance Potential (CEP) focuses ions into Q2, Collision Energy fragments ions and Collision Cell exit potential (CXP) assists ions going into Q3. All these voltages are optimized for each analyte in the assay by compound optimization and the values are incorporated into the acquisition method. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Chromatography parameters 
               
            
           
           
               
               
               
            
               
                   
                 Parameters 
                 Ranges 
               
               
                   
                   
               
               
                   
                 Chromatography mode 
                 Reverse phase 
               
               
                   
                 Isocratic/gradient method 
                 Gradient 
               
               
                   
                 Aqueous phase 
                 DI H2O with 0.1% Formic Acid 
               
               
                   
                   
                 and 0.1% Ammonium Formate 
               
               
                   
                 Organic phase 
                 50.0% Acetonitrile and 50.0% 
               
               
                   
                   
                 Methanol with 0.1% Formic Acid 
               
               
                   
                 Needle Wash 
                 40.0% Isopropanol + 40.0% 
               
               
                   
                   
                 Acetonitrile + 20.0% Acetone 
               
            
           
           
               
               
               
               
            
               
                   
                 Flow rate 
                 0.5 
                 mL/min 
               
               
                   
                 Run time 
                 6.5 
                 minutes 
               
               
                   
                 Sample injection volume 
                 10 
                 μL 
               
               
                   
                 Column temperature 
                 45° 
                 C. 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Ion source parameters 
               
            
           
           
               
               
               
            
               
                   
                 Parameters 
                 Ranges 
               
               
                   
                   
               
               
                   
                 Interface 
                 Electrospray ionization 
               
               
                   
                 Ionization mode 
                 Positive and negative 
               
            
           
           
               
               
               
               
            
               
                   
                 Source/Gas temperature 
                 550° 
                 C. 
               
               
                   
                 Ion source gas 1 
                 60 
                 PSI 
               
               
                   
                 Ion source gas 2 
                 60 
                 PSI 
               
               
                   
                 Curtain Gas 
                 35 
                 PSI 
               
               
                   
                 Collision gas 
                 7 
                 PSI 
               
            
           
           
               
               
               
            
               
                   
                 Ion Spray voltages 
                 2500 
               
               
                   
                   
               
            
           
         
       
     
     Method Validation 
     Method was validated for linearity, accuracy, precision, recovery, LOD and LOQ. 
     Calibration curves for drug/metabolite were made by serial dilution from a stock solution and were created in duplicates. Peak heights vs nominal concentrations were used to construe calibration curves. Curves were evaluated using least squares fitting and by linear regression analysis. 
     The calibration for concentration from 2 ng/ml to 10000 ng/ml and its linearity is shown in Table 8. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Linearity 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 No. 
                 Drug Name 
                 LOQ 
                 Cal 1 
                 Cal 2 
                 Cal 3 
                 Cal 4 
                 Cal 5 
                 ULOL 
               
               
                   
               
            
           
           
               
            
               
                 OPIATES 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 1 
                 6MAM 
                 2 
                 4 
                 20 
                 40 
                 100 
                 200 
                 200 
               
               
                 2 
                 Codeine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 3 
                 Dihydrocodeine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 4 
                 Hydrocodone 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 5 
                 Hydromorphone 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 6 
                 Morphine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 7 
                 Oxycodone 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 8 
                 Oxymorphone 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 9 
                 Buprenophrine 
                 5 
                 10 
                 50 
                 100 
                 250 
                 500 
                 2000 
               
               
                 10 
                 Carisoprodol 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 11 
                 Desmethyl Tapentadol 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 12 
                 DesmethylTramadol 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 13 
                 EDDP 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 14 
                 Meperidine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 7500 
               
               
                 15 
                 Meprobamate 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 16 
                 Methadone 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 17 
                 Norbuprenophrine 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 2000 
               
               
                 18 
                 Normeperidine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 19 
                 Tapentadol 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 20 
                 Tramadol 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 21 
                 Fentanyl 
                 2 
                 4 
                 20 
                 40 
                 100 
                 200 
                 800 
               
               
                 22 
                 Norfentanyl 
                 2 
                 4 
                 20 
                 40 
                 100 
                 200 
                 800 
               
               
                 23 
                 Norpropoxyphene 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 24 
                 Propoxyphene 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 25 
                 Dextromethophan 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 26 
                 Dextrophan 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 27 
                 Desomorphine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 28 
                 Nalaxone 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
            
           
           
               
            
               
                 BENZODIAZEPINES 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 29 
                 7-AminoClonazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 30 
                 Diazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 31 
                 Flunitrazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 32 
                 HydroxyAlprazolam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 33 
                 Nordiazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 34 
                 Oxazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 35 
                 Temazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 36 
                 Chloradiazepoxide 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 37 
                 OH-et-flunizepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 38 
                 Lorazepam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 39 
                 Triazolam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 40 
                 Midazolam 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
            
           
           
               
            
               
                 Z-DRUGS 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 41 
                 Zolpidem 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 42 
                 Zaleplon 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 43 
                 Zopiclone 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
               
                 44 
                 Zolpidem-COOH 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 5000 
               
            
           
           
               
            
               
                 TRICYCLIC ANTIDEPRESSANTS 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 45 
                 Desipramine 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
               
                 46 
                 Imipramine 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
               
                 47 
                 MethylPhenydate 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 5000 
               
               
                 48 
                 Nortriptyline 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
               
                 49 
                 Ritalinic Acid 
                 100 
                 200 
                 1000 
                 2000 
                 5000 
                 10000 
                 20000 
               
               
                 50 
                 Sertraline 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
               
                 51 
                 Cyclobenzaprine 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
               
                 52 
                 Amitriptyline 
                 20 
                 40 
                 200 
                 400 
                 1000 
                 2000 
                 5000 
               
            
           
           
               
            
               
                 ANTIEPILEPTICS 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 53 
                 Pregabilan 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 54 
                 Gabapentin 
                 100 
                 200 
                 1000 
                 2000 
                 5000 
                 10000 
                 10000 
               
            
           
           
               
            
               
                 AMPHETAMINES 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 55 
                 Amphetamine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 56 
                 MDA 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 57 
                 MDMA 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 58 
                 Methamphetamine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
            
           
           
               
            
               
                 BARBITURATES 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 59 
                 Butalbital 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 60 
                 Phenobarbital 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
            
           
           
               
            
               
                 ILLICITS 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 61 
                 THCA 
                 10 
                 20 
                 100 
                 200 
                 500 
                 1000 
                 1000 
               
               
                 62 
                 Benzoylecgonine 
                 50 
                 100 
                 500 
                 1000 
                 2500 
                 5000 
                 10000 
               
               
                 63 
                 PCP 
                 4 
                 8 
                 40 
                 80 
                 200 
                 400 
                 2000 
               
               
                   
               
            
           
         
       
     
     Precision 
       
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Intraday accuracy and precision studied 10 runs 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Actual 
                 Mean 
                 CV 
                   
               
               
                 Component Name 
                 (ng/mL) 
                 n = 10 
                 (%) 
                 Accuracy 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 6-MAM 
                 2 
                 1.85 
                 10.85 
                 92.28 
               
               
                   
                 4 
                 3.65 
                 9.01 
                 91.37 
               
               
                   
                 20 
                 18.21 
                 11.23 
                 91.07 
               
               
                   
                 40 
                 40.26 
                 7.96 
                 100.66 
               
               
                   
                 100 
                 91.66 
                 8.94 
                 91.66 
               
               
                   
                 200 
                 210.37 
                 10.27 
                 105.18 
               
               
                 OH-et-Flurazepam 
                 10 
                 9 
                 6.57 
                 90 
               
               
                   
                 20 
                 19 
                 9.65 
                 95.01 
               
               
                   
                 100 
                 93.86 
                 3.34 
                 93.86 
               
               
                   
                 200 
                 192.06 
                 4.03 
                 96.03 
               
               
                   
                 500 
                 450.87 
                 3.32 
                 90.17 
               
               
                   
                 1000 
                 1065.2 
                 5.14 
                 106.52 
               
               
                 4-Hydroxyalprazolam 
                 10 
                 10.74 
                 8.03 
                 107.43 
               
               
                   
                 20 
                 20.64 
                 6.22 
                 103.21 
               
               
                   
                 100 
                 98.42 
                 7.45 
                 98.42 
               
               
                   
                 200 
                 193.36 
                 7.21 
                 96.68 
               
               
                   
                 500 
                 462.43 
                 4.93 
                 92.49 
               
               
                   
                 1000 
                 1044.4 
                 8.52 
                 104.44 
               
               
                 7-Aminoclonazepam 
                 10 
                 10.31 
                 7.85 
                 103.13 
               
               
                   
                 20 
                 19.77 
                 8.26 
                 98.87 
               
               
                   
                 100 
                 94.07 
                 4.87 
                 94.07 
               
               
                   
                 200 
                 203.54 
                 8.48 
                 101.77 
               
               
                   
                 500 
                 461.98 
                 6.15 
                 92.4 
               
               
                   
                 1000 
                 1040.33 
                 6.09 
                 104.03 
               
               
                 Amitriptyline 
                 20 
                 21.82 
                 7.21 
                 109.08 
               
               
                   
                 40 
                 42.95 
                 6.67 
                 107.38 
               
               
                   
                 200 
                 193.08 
                 4.32 
                 96.54 
               
               
                   
                 400 
                 417.8 
                 5.29 
                 104.45 
               
               
                   
                 1000 
                 972.07 
                 4.95 
                 97.21 
               
               
                   
                 2000 
                 2012.28 
                 2.28 
                 100.61 
               
               
                 Amphetamine 
                 50 
                 57.62 
                 5.52 
                 115.24 
               
               
                   
                 100 
                 121.11 
                 7.26 
                 121.11 
               
               
                   
                 500 
                 633.3 
                 6.15 
                 126.66 
               
               
                   
                 1000 
                 1226.36 
                 3.48 
                 122.64 
               
               
                   
                 2500 
                 2419.79 
                 5.37 
                 96.79 
               
               
                   
                 5000 
                 4599.11 
                 6.75 
                 91.98 
               
               
                 Benzoylecgonine 
                 50 
                 59.19 
                 5.61 
                 118.37 
               
               
                   
                 100 
                 118.85 
                 4.33 
                 118.85 
               
               
                   
                 500 
                 576.98 
                 5.38 
                 115.4 
               
               
                   
                 1000 
                 1128.86 
                 4.12 
                 112.89 
               
               
                   
                 2500 
                 2510.59 
                 4.23 
                 100.42 
               
               
                   
                 5000 
                 4755.53 
                 3.24 
                 95.11 
               
               
                 Buprenorphine 
                 5 
                 4.45 
                 9.45 
                 88.97 
               
               
                   
                 10 
                 9.85 
                 12.31 
                 98.45 
               
               
                   
                 50 
                 46.85 
                 4.78 
                 93.7 
               
               
                   
                 100 
                 94.57 
                 6.19 
                 94.57 
               
               
                   
                 250 
                 236.92 
                 6.83 
                 94.77 
               
               
                   
                 500 
                 522.37 
                 6.19 
                 104.47 
               
               
                 Carisoprodol 
                 50 
                 58.55 
                 7.65 
                 117.1 
               
               
                   
                 100 
                 129.8 
                 5.96 
                 129.8 
               
               
                   
                 500 
                 608.97 
                 9.30 
                 121.79 
               
               
                   
                 1000 
                 1172.67 
                 7.12 
                 117.27 
               
               
                   
                 2500 
                 2524.38 
                 7.13 
                 100.98 
               
               
                   
                 5000 
                 4648.64 
                 9.13 
                 92.97 
               
               
                 Chlordiazepoxide 
                 10 
                 8.01 
                 9.73 
                 80.13 
               
               
                   
                 20 
                 15.91 
                 5.14 
                 79.55 
               
               
                   
                 100 
                 77.2 
                 6.72 
                 77.2 
               
               
                   
                 200 
                 166.47 
                 8.55 
                 83.23 
               
               
                   
                 500 
                 448.7 
                 6.63 
                 89.74 
               
               
                   
                 1000 
                 1113.71 
                 5.40 
                 111.37 
               
               
                 Codeine 
                 50 
                 47.91 
                 11.12 
                 95.82 
               
               
                   
                 100 
                 96.41 
                 8.78 
                 96.41 
               
               
                   
                 500 
                 479.82 
                 9.72 
                 95.96 
               
               
                   
                 1000 
                 936.27 
                 7.70 
                 93.63 
               
               
                   
                 2500 
                 2313.89 
                 6.08 
                 92.56 
               
               
                   
                 5000 
                 5275.7 
                 8.47 
                 105.51 
               
               
                 Cyclobenzaprine 
                 20 
                 22.22 
                 6.81 
                 111.09 
               
               
                   
                 40 
                 46.78 
                 6.59 
                 116.95 
               
               
                   
                 200 
                 219.64 
                 5.67 
                 109.82 
               
               
                   
                 400 
                 453.2 
                 4.38 
                 113.3 
               
               
                   
                 1000 
                 1024.32 
                 6.21 
                 102.43 
               
               
                   
                 2000 
                 1893.84 
                 2.56 
                 94.69 
               
               
                 Desipramine 
                 20 
                 20.07 
                 3.45 
                 100.34 
               
               
                   
                 40 
                 41.77 
                 5.62 
                 104.44 
               
               
                   
                 200 
                 212.74 
                 9.55 
                 106.37 
               
               
                   
                 400 
                 437.84 
                 4.63 
                 109.46 
               
               
                   
                 1000 
                 957.96 
                 6.11 
                 95.8 
               
               
                   
                 2000 
                 1989.62 
                 7.72 
                 99.48 
               
               
                 Desmethyltramadol 
                 50 
                 37.56 
                 9.94 
                 75.12 
               
               
                   
                 100 
                 107.72 
                 4.86 
                 107.72 
               
               
                   
                 500 
                 621.11 
                 7.49 
                 124.22 
               
               
                   
                 1000 
                 1055.88 
                 8.46 
                 105.59 
               
               
                   
                 2500 
                 1943.94 
                 9.43 
                 77.76 
               
               
                   
                 5000 
                 2617.24 
                 3.26 
                 52.34 
               
               
                 Desmethyltramadol 
                 50 
                 43.85 
                 7.11 
                 87.71 
               
               
                   
                 100 
                 101.33 
                 5.15 
                 101.33 
               
               
                   
                 500 
                 551.35 
                 7.12 
                 110.27 
               
               
                   
                 1000 
                 1072.2 
                 6.47 
                 107.22 
               
               
                   
                 2500 
                 2293.45 
                 5.74 
                 91.74 
               
               
                   
                 5000 
                 5090.48 
                 5.04 
                 101.81 
               
               
                 Desomorphine 
                 50 
                 59.4 
                 6.45 
                 118.8 
               
               
                   
                 100 
                 120.71 
                 12.89 
                 120.71 
               
               
                   
                 500 
                 586.47 
                 8.15 
                 117.29 
               
               
                   
                 1000 
                 1148.7 
                 5.54 
                 114.87 
               
               
                   
                 2500 
                 2370.25 
                 10.74 
                 94.81 
               
               
                   
                 5000 
                 4795.48 
                 5.81 
                 95.91 
               
               
                 DesTapentadol 
                 50 
                 35.32 
                 14.29 
                 70.64 
               
               
                   
                 100 
                 112.82 
                 5.81 
                 112.82 
               
               
                   
                 500 
                 601.01 
                 5.59 
                 120.2 
               
               
                   
                 1000 
                 1083.9 
                 5.14 
                 108.39 
               
               
                   
                 2500 
                 2057.18 
                 6.41 
                 82.29 
               
               
                   
                 5000 
                 4946.59 
                 2.43 
                 98.93 
               
               
                 DesTapentadol 
                 50 
                 41.3 
                 13.90 
                 70.64 
               
               
                   
                 100 
                 128.94 
                 5.70 
                 112.82 
               
               
                   
                 500 
                 653.56 
                 5.28 
                 120.2 
               
               
                   
                 1000 
                 1126.01 
                 4.59 
                 108.39 
               
               
                   
                 2500 
                 1937.11 
                 5.04 
                 82.29 
               
               
                   
                 5000 
                 3382.97 
                 3.65 
                 98.93 
               
               
                 Dextromethorphan 
                 50 
                 61.12 
                 5.61 
                 122.24 
               
               
                   
                 100 
                 126.92 
                 5.34 
                 126.92 
               
               
                   
                 500 
                 625.19 
                 8.29 
                 123.04 
               
               
                   
                 1000 
                 1248.79 
                 2.62 
                 124.88 
               
               
                   
                 2500 
                 2591.04 
                 3.53 
                 103.64 
               
               
                   
                 5000 
                 4425.94 
                 5.88 
                 88.52 
               
               
                 Dextrorphan 
                 50 
                 58.26 
                 9.27 
                 116.52 
               
               
                   
                 100 
                 110.63 
                 5.94 
                 110.63 
               
               
                   
                 500 
                 546.6 
                 6.57 
                 109.32 
               
               
                   
                 1000 
                 1015.62 
                 6.46 
                 101.56 
               
               
                   
                 2500 
                 2408.01 
                 3.79 
                 96.32 
               
               
                   
                 5000 
                 5010.87 
                 6.20 
                 100.22 
               
               
                 Diazepam 
                 10 
                 10.02 
                 2.31 
                 100.17 
               
               
                   
                 20 
                 20.45 
                 1.43 
                 102.24 
               
               
                   
                 100 
                 99.91 
                 2.10 
                 99.91 
               
               
                   
                 200 
                 203.53 
                 1.46 
                 101.76 
               
               
                   
                 500 
                 471.52 
                 1.58 
                 94.3 
               
               
                   
                 1000 
                 1024.58 
                 2.25 
                 102.46 
               
               
                 Dihydrocodeine 
                 50 
                 49.17 
                 5.85 
                 98.35 
               
               
                   
                 100 
                 93.29 
                 11.17 
                 93.29 
               
               
                   
                 500 
                 484.06 
                 10.80 
                 96.81 
               
               
                   
                 1000 
                 928.81 
                 8.43 
                 92.88 
               
               
                   
                 2500 
                 2424.79 
                 6.71 
                 96.99 
               
               
                   
                 5000 
                 5169.87 
                 7.31 
                 103.4 
               
               
                 EDDP 
                 50 
                 51.05 
                 2.70 
                 102.1 
               
               
                   
                 100 
                 98.72 
                 7.24 
                 98.72 
               
               
                   
                 500 
                 498.15 
                 4.64 
                 99.63 
               
               
                   
                 1000 
                 1008.52 
                 4.67 
                 100.85 
               
               
                   
                 2500 
                 2317.71 
                 1.87 
                 92.71 
               
               
                   
                 5000 
                 5175.86 
                 4.24 
                 103.52 
               
               
                 Fentanyl 
                 2 
                 1.93 
                 8.18 
                 96.59 
               
               
                   
                 4 
                 3.95 
                 5.60 
                 98.75 
               
               
                   
                 20 
                 20 
                 5.68 
                 100.02 
               
               
                   
                 40 
                 40.16 
                 4.36 
                 100.41 
               
               
                   
                 100 
                 94.55 
                 5.23 
                 94.55 
               
               
                   
                 200 
                 205.4 
                 4.16 
                 102.7 
               
               
                 Flunitrazepam 
                 10 
                 9.56 
                 12.16 
                 95.63 
               
               
                   
                 20 
                 19.37 
                 9.53 
                 96.87 
               
               
                   
                 100 
                 90.35 
                 8.30 
                 90.35 
               
               
                   
                 200 
                 188.97 
                 7.02 
                 94.48 
               
               
                   
                 500 
                 458.9 
                 4.13 
                 91.78 
               
               
                   
                 1000 
                 1062.85 
                 6.59 
                 106.28 
               
               
                 Gabapentin 
                 100 
                 100.97 
                 10.40 
                 100.97 
               
               
                   
                 200 
                 186.78 
                 5.25 
                 93.39 
               
               
                   
                 1000 
                 865.03 
                 4.93 
                 86.5 
               
               
                   
                 2000 
                 1722.23 
                 8.55 
                 86.11 
               
               
                   
                 5000 
                 4386.95 
                 7.11 
                 87.74 
               
               
                   
                 10000 
                 11038.04 
                 7.49 
                 110.38 
               
               
                 Hydrocodone 
                 50 
                 53.97 
                 11.54 
                 107.95 
               
               
                   
                 100 
                 107.92 
                 9.15 
                 107.92 
               
               
                   
                 500 
                 537.89 
                 5.65 
                 107.58 
               
               
                   
                 1000 
                 1066.67 
                 8.79 
                 106.67 
               
               
                   
                 2500 
                 2439.19 
                 6.23 
                 97.57 
               
               
                   
                 5000 
                 4944.35 
                 7.44 
                 98.89 
               
               
                 Hydromorphone 
                 50 
                 51.86 
                 2.04 
                 103.72 
               
               
                   
                 100 
                 99.52 
                 2.83 
                 99.52 
               
               
                   
                 500 
                 489.68 
                 5.05 
                 97.94 
               
               
                   
                 1000 
                 1007.19 
                 3.02 
                 100.72 
               
               
                   
                 2500 
                 2366.96 
                 4.67 
                 94.68 
               
               
                   
                 5000 
                 5134.79 
                 4.53 
                 102.7 
               
               
                 Imipramine 
                 20 
                 25.18 
                 7.20 
                 125.9 
               
               
                   
                 40 
                 51.14 
                 5.28 
                 127.85 
               
               
                   
                 200 
                 247.68 
                 5.48 
                 123.84 
               
               
                   
                 400 
                 476.31 
                 3.29 
                 119.08 
               
               
                   
                 1000 
                 983.45 
                 4.61 
                 98.35 
               
               
                   
                 2000 
                 1876.24 
                 5.30 
                 93.81 
               
               
                 Lorazepam 
                 10 
                 11.6 
                 5.96 
                 115.95 
               
               
                   
                 20 
                 24.9 
                 5.96 
                 124.51 
               
               
                   
                 100 
                 109.85 
                 6.87 
                 109.85 
               
               
                   
                 200 
                 211.98 
                 7.68 
                 105.99 
               
               
                   
                 500 
                 505.18 
                 4.46 
                 101.04 
               
               
                   
                 1000 
                 966.5 
                 4.17 
                 96.65 
               
               
                 MDA 
                 50 
                 58.97 
                 11.25 
                 117.95 
               
               
                   
                 100 
                 113.77 
                 12.69 
                 113.77 
               
               
                   
                 500 
                 517.22 
                 10.24 
                 103.44 
               
               
                   
                 1000 
                 1071.89 
                 12.18 
                 107.19 
               
               
                   
                 2500 
                 2341.24 
                 9.74 
                 93.65 
               
               
                   
                 5000 
                 5046.91 
                 17.14 
                 100.94 
               
               
                 MDMA 
                 50 
                 53.76 
                 5.72 
                 107.52 
               
               
                   
                 100 
                 102.58 
                 4.91 
                 102.58 
               
               
                   
                 500 
                 503.81 
                 4.51 
                 100.76 
               
               
                   
                 1000 
                 997.64 
                 5.81 
                 99.76 
               
               
                   
                 2500 
                 2312.37 
                 7.09 
                 92.49 
               
               
                   
                 5000 
                 5179.83 
                 6.16 
                 103.6 
               
               
                 Meperidine 
                 50 
                 52.23 
                 5.13 
                 104.45 
               
               
                   
                 100 
                 106.63 
                 2.34 
                 106.63 
               
               
                   
                 500 
                 519.44 
                 2.94 
                 103.89 
               
               
                   
                 1000 
                 1028.08 
                 3.55 
                 102.81 
               
               
                   
                 2500 
                 2377.1 
                 5.96 
                 95.08 
               
               
                   
                 5000 
                 5066.51 
                 5.20 
                 101.33 
               
               
                 Meprobamate 
                 50 
                 62.32 
                 4.42 
                 124.64 
               
               
                   
                 100 
                 118.12 
                 8.97 
                 118.12 
               
               
                   
                 500 
                 621.68 
                 7.31 
                 124.34 
               
               
                   
                 1000 
                 1249.61 
                 7.24 
                 124.96 
               
               
                   
                 2500 
                 2460.62 
                 7.27 
                 98.42 
               
               
                   
                 5000 
                 4637.66 
                 7.85 
                 92.75 
               
               
                 Methadone 
                 50 
                 50.22 
                 4.30 
                 100.43 
               
               
                   
                 100 
                 104.15 
                 2.24 
                 104.15 
               
               
                   
                 500 
                 520.93 
                 3.14 
                 104.19 
               
               
                   
                 1000 
                 1068.53 
                 4.35 
                 106.85 
               
               
                   
                 2500 
                 2347.98 
                 4.24 
                 93.92 
               
               
                   
                 5000 
                 5058.21 
                 4.40 
                 101.16 
               
               
                 Methamphetamine 
                 50 
                 53.77 
                 6.90 
                 107.55 
               
               
                   
                 100 
                 109.99 
                 10.11 
                 109.99 
               
               
                   
                 500 
                 543.03 
                 7.94 
                 108.61 
               
               
                   
                 1000 
                 1079.8 
                 9.14 
                 107.98 
               
               
                   
                 2500 
                 2428.15 
                 8.16 
                 97.13 
               
               
                   
                 5000 
                 4935.25 
                 8.83 
                 98.71 
               
               
                 Methylphenidate 
                 50 
                 40.65 
                 8.36 
                 81.3 
               
               
                   
                 100 
                 107.03 
                 5.18 
                 107.03 
               
               
                   
                 500 
                 577.36 
                 3.23 
                 115.47 
               
               
                   
                 1000 
                 1036.55 
                 3.53 
                 103.66 
               
               
                   
                 2500 
                 2214.58 
                 8.68 
                 88.58 
               
               
                   
                 5000 
                 5042.33 
                 4.89 
                 100.85 
               
               
                 Midazolam 
                 10 
                 7.76 
                 10.12 
                 77.57 
               
               
                   
                 20 
                 15.35 
                 2.68 
                 76.75 
               
               
                   
                 100 
                 75.27 
                 6.96 
                 75.27 
               
               
                   
                 200 
                 168.36 
                 6.37 
                 84.18 
               
               
                   
                 500 
                 448.16 
                 3.93 
                 89.63 
               
               
                   
                 1000 
                 1115.1 
                 6.18 
                 111.51 
               
               
                 Morphine 
                 50 
                 52.04 
                 4.79 
                 104.08 
               
               
                   
                 100 
                 103.13 
                 3.00 
                 103.13 
               
               
                   
                 500 
                 475.86 
                 4.15 
                 95.17 
               
               
                   
                 1000 
                 971.74 
                 3.47 
                 97.17 
               
               
                   
                 2500 
                 2314.21 
                 2.14 
                 92.57 
               
               
                   
                 5000 
                 5233.01 
                 2.22 
                 104.66 
               
               
                 Naloxone 
                 10 
                 11.01 
                 7.79 
                 110.06 
               
               
                   
                 20 
                 22.14 
                 8.03 
                 110.69 
               
               
                   
                 100 
                 105.01 
                 6.49 
                 105.01 
               
               
                   
                 200 
                 201.16 
                 8.25 
                 100.58 
               
               
                   
                 500 
                 494.8 
                 6.74 
                 98.96 
               
               
                   
                 1000 
                 995.88 
                 10.46 
                 99.59 
               
               
                 Norbuprenorphine 
                 10 
                 9.62 
                 11.12 
                 96.17 
               
               
                   
                 20 
                 21.54 
                 12.20 
                 107.71 
               
               
                   
                 100 
                 96.04 
                 5.48 
                 96.04 
               
               
                   
                 200 
                 192.75 
                 11.12 
                 96.37 
               
               
                   
                 500 
                 467.43 
                 7.99 
                 93.49 
               
               
                   
                 1000 
                 1042.63 
                 8.47 
                 104.26 
               
               
                 Nordiazepam 
                 10 
                 9.53 
                 3.44 
                 95.29 
               
               
                   
                 20 
                 18.75 
                 3.95 
                 93.74 
               
               
                   
                 100 
                 92.96 
                 3.57 
                 92.96 
               
               
                   
                 200 
                 190.63 
                 1.97 
                 95.31 
               
               
                   
                 500 
                 454.01 
                 3.03 
                 90.8 
               
               
                   
                 1000 
                 1064.13 
                 1.94 
                 106.41 
               
               
                 Norfentanyl 
                 2 
                 2.49 
                 6.82 
                 124.62 
               
               
                   
                 4 
                 4.78 
                 6.59 
                 119.42 
               
               
                   
                 20 
                 22.2 
                 4.35 
                 111.01 
               
               
                   
                 40 
                 43.69 
                 4.30 
                 109.23 
               
               
                   
                 100 
                 94.56 
                 4.76 
                 94.56 
               
               
                   
                 200 
                 198.28 
                 5.72 
                 99.14 
               
               
                 Normeperidine 
                 50 
                 40.77 
                 10.36 
                 81.53 
               
               
                   
                 100 
                 103.81 
                 8.14 
                 103.81 
               
               
                   
                 500 
                 584.21 
                 4.72 
                 116.84 
               
               
                   
                 1000 
                 1080.51 
                 8.67 
                 108.05 
               
               
                   
                 2500 
                 2132.03 
                 6.69 
                 85.28 
               
               
                   
                 5000 
                 4983.46 
                 11.42 
                 99.67 
               
               
                 Norpropoxyphene 
                 50 
                 51.15 
                 8.66 
                 102.29 
               
               
                   
                 100 
                 115.47 
                 6.79 
                 115.47 
               
               
                   
                 500 
                 529.06 
                 7.83 
                 105.81 
               
               
                   
                 1000 
                 1120.26 
                 6.56 
                 112.03 
               
               
                   
                 2500 
                 2482.96 
                 6.33 
                 99.32 
               
               
                   
                 5000 
                 4851.11 
                 5.91 
                 97.02 
               
               
                 Nortriptyline 
                 20 
                 20.79 
                 5.43 
                 103.96 
               
               
                   
                 40 
                 43.25 
                 9.28 
                 108.12 
               
               
                   
                 200 
                 203.92 
                 7.45 
                 101.96 
               
               
                   
                 400 
                 437.37 
                 4.05 
                 109.34 
               
               
                   
                 1000 
                 1002.25 
                 6.73 
                 100.22 
               
               
                   
                 2000 
                 1952.43 
                 3.75 
                 97.62 
               
               
                 Oxazepam 
                 10 
                 9.94 
                 7.25 
                 99.42 
               
               
                   
                 20 
                 18.96 
                 6.43 
                 94.8 
               
               
                   
                 100 
                 91.3 
                 6.77 
                 91.3 
               
               
                   
                 200 
                 195 
                 7.77 
                 97.5 
               
               
                   
                 500 
                 443.4 
                 5.30 
                 88.68 
               
               
                   
                 1000 
                 1071.41 
                 7.58 
                 107.14 
               
               
                 Oxycodone 
                 50 
                 59.12 
                 5.59 
                 118.24 
               
               
                   
                 100 
                 112.03 
                 5.90 
                 112.03 
               
               
                   
                 500 
                 521.75 
                 7.96 
                 104.35 
               
               
                   
                 1000 
                 1082.02 
                 6.71 
                 108.2 
               
               
                   
                 2500 
                 2438.31 
                 4.37 
                 97.53 
               
               
                   
                 5000 
                 4936.78 
                 4.39 
                 98.74 
               
               
                 Oxymorphone 
                 50 
                 49.56 
                 2.72 
                 99.11 
               
               
                   
                 100 
                 97.74 
                 2.61 
                 97.74 
               
               
                   
                 500 
                 487.94 
                 2.84 
                 97.59 
               
               
                   
                 1000 
                 1014 
                 2.59 
                 101.4 
               
               
                   
                 2500 
                 2298.61 
                 3.44 
                 91.94 
               
               
                   
                 5000 
                 5202.15 
                 3.48 
                 104.04 
               
               
                 PCP 
                 4 
                 4.47 
                 5.98 
                 111.71 
               
               
                   
                 8 
                 8.54 
                 6.85 
                 106.78 
               
               
                   
                 40 
                 40.16 
                 6.95 
                 100.39 
               
               
                   
                 80 
                 82.5 
                 6.07 
                 103.12 
               
               
                   
                 200 
                 189.19 
                 6.55 
                 94.59 
               
               
                   
                 400 
                 407.15 
                 7.34 
                 101.79 
               
               
                 Pregabalin 
                 50 
                 37.8 
                 16.92 
                 75.59 
               
               
                   
                 100 
                 109.4 
                 6.68 
                 109.4 
               
               
                   
                 500 
                 588.84 
                 6.08 
                 117.77 
               
               
                   
                 1000 
                 1076.22 
                 9.81 
                 107.62 
               
               
                   
                 2500 
                 2135.92 
                 9.65 
                 85.44 
               
               
                   
                 5000 
                 5134.68 
                 14.35 
                 102.69 
               
               
                 Pregabalin 
                 50 
                 51.89 
                 4.47 
                 103.78 
               
               
                   
                 100 
                 103.88 
                 8.33 
                 103.88 
               
               
                   
                 500 
                 500.98 
                 5.71 
                 100.2 
               
               
                   
                 1000 
                 959.36 
                 7.37 
                 95.94 
               
               
                   
                 2500 
                 2276.43 
                 6.31 
                 91.06 
               
               
                   
                 5000 
                 5257.47 
                 6.72 
                 105.15 
               
               
                 Propoxyphene 
                 50 
                 47.89 
                 11.61 
                 95.78 
               
               
                   
                 100 
                 102.21 
                 11.30 
                 102.21 
               
               
                   
                 500 
                 520.52 
                 7.66 
                 104.1 
               
               
                   
                 1000 
                 1011.32 
                 13.48 
                 101.13 
               
               
                   
                 2500 
                 2382.82 
                 13.76 
                 95.31 
               
               
                   
                 5000 
                 5085.24 
                 11.02 
                 101.7 
               
               
                 Ritalinic Acid 
                 100 
                 120.21 
                 7.20 
                 120.21 
               
               
                   
                 200 
                 246.88 
                 6.43 
                 123.44 
               
               
                   
                 1000 
                 1287.35 
                 4.06 
                 128.73 
               
               
                   
                 2000 
                 2080.99 
                 4.51 
                 104.05 
               
               
                   
                 5000 
                 4056.9 
                 7.55 
                 81.14 
               
               
                   
                 10000 
                 9839.57 
                 8.62 
                 98.4 
               
               
                 Sertraline 
                 20 
                 16.85 
                 4.59 
                 84.27 
               
               
                   
                 40 
                 33.39 
                 4.82 
                 83.47 
               
               
                   
                 200 
                 175.01 
                 5.65 
                 87.5 
               
               
                   
                 400 
                 393.38 
                 4.32 
                 98.34 
               
               
                   
                 1000 
                 986.91 
                 4.68 
                 98.69 
               
               
                   
                 2000 
                 2054.47 
                 4.69 
                 102.72 
               
               
                 Tapentadol 
                 50 
                 62.65 
                 5.69 
                 125.3 
               
               
                   
                 100 
                 128.26 
                 5.05 
                 128.26 
               
               
                   
                 500 
                 569.26 
                 4.01 
                 113.85 
               
               
                   
                 1000 
                 1236.2 
                 5.74 
                 123.62 
               
               
                   
                 2500 
                 2377.24 
                 3.49 
                 95.09 
               
               
                   
                 5000 
                 4644.8 
                 4.01 
                 92.9 
               
               
                 Temazepam 
                 10 
                 8.35 
                 7.78 
                 83.49 
               
               
                   
                 20 
                 16.84 
                 4.55 
                 84.22 
               
               
                   
                 100 
                 84.3 
                 7.19 
                 84.3 
               
               
                   
                 200 
                 180.92 
                 6.95 
                 90.46 
               
               
                   
                 500 
                 458.11 
                 3.71 
                 91.62 
               
               
                   
                 1000 
                 1081.48 
                 5.70 
                 108.15 
               
               
                 Tramadol 
                 50 
                 44.36 
                 9.82 
                 88.72 
               
               
                   
                 100 
                 103.96 
                 6.97 
                 103.96 
               
               
                   
                 500 
                 538.48 
                 9.18 
                 107.7 
               
               
                   
                 1000 
                 1059.56 
                 7.72 
                 105.96 
               
               
                   
                 2500 
                 2257.6 
                 9.31 
                 90.3 
               
               
                   
                 5000 
                 4634.21 
                 9.15 
                 92.68 
               
               
                 Triazolam 
                 10 
                 7.37 
                 6.93 
                 73.68 
               
               
                   
                 20 
                 14.3 
                 5.29 
                 71.51 
               
               
                   
                 100 
                 71.96 
                 8.60 
                 71.96 
               
               
                   
                 200 
                 160.84 
                 9.39 
                 80.42 
               
               
                   
                 500 
                 439.54 
                 2.26 
                 87.91 
               
               
                   
                 1000 
                 1135.99 
                 6.28 
                 113.6 
               
               
                 Zaleplon 
                 10 
                 12.21 
                 11.15 
                 122.1 
               
               
                   
                 20 
                 24.61 
                 7.91 
                 123.05 
               
               
                   
                 100 
                 123.78 
                 7.38 
                 123.78 
               
               
                   
                 200 
                 237.4 
                 8.23 
                 118.7 
               
               
                   
                 500 
                 497.49 
                 6.71 
                 99.5 
               
               
                   
                 1000 
                 929 
                 5.86 
                 92.9 
               
               
                 Zopiclone 
                 10 
                 7.92 
                 12.50 
                 79.2 
               
               
                   
                 20 
                 25.06 
                 7.91 
                 125.31 
               
               
                   
                 100 
                 116.69 
                 9.29 
                 116.69 
               
               
                   
                 200 
                 177.16 
                 7.16 
                 88.58 
               
               
                   
                 500 
                 435.26 
                 8.00 
                 87.05 
               
               
                   
                 1000 
                 945.84 
                 10.46 
                 94.58 
               
               
                 Zolpidem 
                 10 
                 10.28 
                 7.07 
                 102.8 
               
               
                   
                 20 
                 20.24 
                 5.94 
                 101.19 
               
               
                   
                 100 
                 100.19 
                 4.11 
                 100.19 
               
               
                   
                 200 
                 200.89 
                 4.50 
                 100.45 
               
               
                   
                 500 
                 468.89 
                 3.97 
                 93.78 
               
               
                   
                 1000 
                 1029.52 
                 2.76 
                 102.95 
               
               
                 Carboxy Zolpidem 
                 10 
                 12.27 
                 6.01 
                 122.7 
               
               
                   
                 20 
                 23.29 
                 3.49 
                 116.45 
               
               
                   
                 100 
                 123.21 
                 4.89 
                 123.21 
               
               
                   
                 200 
                 237.56 
                 5.47 
                 118.78 
               
               
                   
                 500 
                 481.45 
                 4.84 
                 96.29 
               
               
                   
                 1000 
                 946.15 
                 6.97 
                 94.62 
               
               
                 Butalbital 
                 50 
                 50.08 
                 7.48 
                 100.15 
               
               
                   
                 100 
                 100.86 
                 10.56 
                 100.86 
               
               
                   
                 500 
                 465.91 
                 5.93 
                 93.18 
               
               
                   
                 1000 
                 1000.05 
                 5.67 
                 100.01 
               
               
                   
                 2500 
                 2328.06 
                 7.40 
                 93.12 
               
               
                   
                 5000 
                 5205.04 
                 6.02 
                 104.1 
               
               
                 THC 
                 10 
                 10.78 
                 4.94 
                 107.77 
               
               
                   
                 20 
                 17.65 
                 4.14 
                 88.26 
               
               
                   
                 100 
                 81.09 
                 3.81 
                 81.09 
               
               
                   
                 200 
                 184.76 
                 2.33 
                 92.38 
               
               
                   
                 500 
                 533.47 
                 1.30 
                 106.69 
               
               
                   
                 1000 
                 1238.05 
                 1.70 
                 123.8 
               
               
                 Phenobarbital 
                 50 
                 52.37 
                 6.79 
                 104.73 
               
               
                   
                 100 
                 99.61 
                 5.91 
                 99.61 
               
               
                   
                 500 
                 490.34 
                 5.79 
                 98.07 
               
               
                   
                 1000 
                 971.15 
                 3.90 
                 97.12 
               
               
                   
                 2500 
                 2253.82 
                 4.60 
                 90.15 
               
               
                   
                 5000 
                 5282.71 
                 5.42 
                 105.65 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Inter-day accuracy and precision studied for 7 runs 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Actual 
                 Mean 
                 CV 
                   
               
               
                 Component Name 
                 (ng/mL) 
                 n = 7 
                 (%) 
                 Accuracy 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 6-MAM 
                 2 
                 1.93 
                 12.20 
                 96.5 
               
               
                   
                 4 
                 3.616 
                 8.61 
                 90.4 
               
               
                   
                 20 
                 19.019 
                 7.92 
                 95.095 
               
               
                   
                 40 
                 40.122 
                 6.88 
                 100.305 
               
               
                   
                 100 
                 95.868 
                 7.69 
                 95.868 
               
               
                   
                 200 
                 205.445 
                 7.63 
                 102.7225 
               
               
                 OH-et-Flurazepam 
                 10 
                 9.145 
                 12.89 
                 91.45 
               
               
                   
                 20 
                 18.462 
                 5.00 
                 92.31 
               
               
                   
                 100 
                 96.227 
                 1.89 
                 96.227 
               
               
                   
                 200 
                 197.936 
                 2.86 
                 98.968 
               
               
                   
                 500 
                 480.16 
                 4.42 
                 96.032 
               
               
                   
                 1000 
                 1028.071 
                 9.00 
                 102.8071 
               
               
                 4-Hydroxyalprazolam 
                 10 
                 10.293 
                 7.62 
                 102.93 
               
               
                   
                 20 
                 20.108 
                 4.27 
                 100.54 
               
               
                   
                 100 
                 100.205 
                 6.87 
                 100.205 
               
               
                   
                 200 
                 209.968 
                 6.08 
                 104.984 
               
               
                   
                 500 
                 482.232 
                 3.58 
                 96.4464 
               
               
                   
                 1000 
                 1007.194 
                 9.72 
                 100.7194 
               
               
                 7-Aminoclonazepam 
                 10 
                 9.83 
                 13.03 
                 98.3 
               
               
                   
                 20 
                 19.096 
                 8.23 
                 95.48 
               
               
                   
                 100 
                 93.704 
                 5.67 
                 93.704 
               
               
                   
                 200 
                 197.896 
                 9.11 
                 98.948 
               
               
                   
                 500 
                 485.943 
                 9.47 
                 97.1886 
               
               
                   
                 1000 
                 1023.531 
                 14.16 
                 102.3531 
               
               
                 Amitriptyline 
                 20 
                 21.327 
                 9.34 
                 106.635 
               
               
                   
                 40 
                 40.661 
                 3.10 
                 101.6525 
               
               
                   
                 200 
                 199.041 
                 7.35 
                 99.5205 
               
               
                   
                 400 
                 415.941 
                 3.17 
                 103.98525 
               
               
                   
                 1000 
                 998.277 
                 6.12 
                 99.8277 
               
               
                   
                 2000 
                 1984.752 
                 8.55 
                 99.2376 
               
               
                 Benzoylecgonine 
                 50 
                 59.513 
                 6.31 
                 119.026 
               
               
                   
                 100 
                 115.159 
                 6.75 
                 115.159 
               
               
                   
                 500 
                 579.174 
                 5.71 
                 115.8348 
               
               
                   
                 1000 
                 1124.089 
                 5.01 
                 112.4089 
               
               
                   
                 2500 
                 2558.48 
                 4.66 
                 102.3392 
               
               
                   
                 5000 
                 4713.587 
                 5.44 
                 94.27174 
               
               
                 Buprenorphine 
                 5 
                 5.143 
                 12.30 
                 102.86 
               
               
                   
                 10 
                 9.647 
                 12.18 
                 96.47 
               
               
                   
                 50 
                 47.809 
                 11.55 
                 95.618 
               
               
                   
                 100 
                 95.13 
                 5.13 
                 95.13 
               
               
                   
                 250 
                 252.313 
                 4.66 
                 100.9252 
               
               
                   
                 500 
                 504.958 
                 10.12 
                 100.9916 
               
               
                 Carisoprodol 
                 50 
                 59.774 
                 9.38 
                 119.548 
               
               
                   
                 100 
                 116.419 
                 10.01 
                 116.419 
               
               
                   
                 500 
                 574.089 
                 8.19 
                 114.8178 
               
               
                   
                 1000 
                 1172.398 
                 6.04 
                 117.2398 
               
               
                   
                 2500 
                 2513.025 
                 11.58 
                 100.521 
               
               
                   
                 5000 
                 4714.295 
                 13.34 
                 94.2859 
               
               
                 Chlordiazepoxide 
                 10 
                 9.379 
                 11.18 
                 93.79 
               
               
                   
                 20 
                 17.617 
                 11.81 
                 88.085 
               
               
                   
                 100 
                 90.256 
                 11.18 
                 90.256 
               
               
                   
                 200 
                 201.645 
                 11.43 
                 100.8225 
               
               
                   
                 500 
                 511.167 
                 13.46 
                 102.2334 
               
               
                   
                 1000 
                 999.889 
                 10.34 
                 99.9889 
               
               
                 Codeine 
                 50 
                 49.854 
                 8.63 
                 99.708 
               
               
                   
                 100 
                 95.894 
                 9.67 
                 95.894 
               
               
                   
                 500 
                 508.453 
                 8.69 
                 101.6906 
               
               
                   
                 1000 
                 1011.263 
                 9.92 
                 101.1263 
               
               
                   
                 2500 
                 2380.853 
                 10.16 
                 95.23412 
               
               
                   
                 5000 
                 5103.683 
                 9.48 
                 102.07366 
               
               
                 Cyclobenzaprine 
                 20 
                 23.485 
                 13.01 
                 117.425 
               
               
                   
                 40 
                 44.509 
                 5.12 
                 111.2725 
               
               
                   
                 200 
                 221.073 
                 7.97 
                 110.5365 
               
               
                   
                 400 
                 442.333 
                 4.72 
                 110.58325 
               
               
                   
                 1000 
                 998.628 
                 4.61 
                 99.8628 
               
               
                   
                 2000 
                 1929.973 
                 7.26 
                 96.49865 
               
               
                 Desipramine 
                 20 
                 21.342 
                 5.85 
                 106.71 
               
               
                   
                 40 
                 40.636 
                 4.64 
                 101.59 
               
               
                   
                 200 
                 223.338 
                 4.15 
                 111.669 
               
               
                   
                 400 
                 430.5 
                 6.55 
                 107.625 
               
               
                   
                 1000 
                 1013.344 
                 5.12 
                 101.3344 
               
               
                   
                 2000 
                 1930.84 
                 6.49 
                 96.542 
               
               
                 Desmethyltramadol 
                 50 
                 39.342 
                 7.94 
                 78.684 
               
               
                   
                 100 
                 105.554 
                 11.12 
                 105.554 
               
               
                   
                 500 
                 603.255 
                 9.63 
                 120.651 
               
               
                   
                 1000 
                 1073.545 
                 10.42 
                 107.3545 
               
               
                   
                 2500 
                 1985.786 
                 9.14 
                 79.43144 
               
               
                   
                 5000 
                 4714.284 
                 0.58 
                 94.28568 
               
               
                 Desomorphine 
                 50 
                 58.837 
                 12.17 
                 117.674 
               
               
                   
                 100 
                 127.902 
                 10.65 
                 127.902 
               
               
                   
                 500 
                 616.638 
                 8.90 
                 123.3276 
               
               
                   
                 1000 
                 1139.192 
                 6.40 
                 113.9192 
               
               
                   
                 2500 
                 2380.669 
                 10.36 
                 95.22676 
               
               
                   
                 5000 
                 4816.762 
                 9.79 
                 96.33524 
               
               
                 DesTapentadol 
                 50 
                 39.3 
                 9.19 
                 78.6 
               
               
                   
                 100 
                 106.631 
                 9.00 
                 106.631 
               
               
                   
                 500 
                 589.536 
                 5.96 
                 117.9072 
               
               
                   
                 1000 
                 1074.349 
                 9.62 
                 107.4349 
               
               
                   
                 2500 
                 2108.364 
                 9.17 
                 84.33456 
               
               
                   
                 5000 
                 4938.339 
                 2.33 
                 98.76678 
               
               
                 Dextromethorphan 
                 50 
                 57.101 
                 8.00 
                 114.202 
               
               
                   
                 100 
                 126.855 
                 5.70 
                 126.855 
               
               
                   
                 500 
                 567.615 
                 8.43 
                 113.523 
               
               
                   
                 1000 
                 1260.622 
                 7.32 
                 126.0622 
               
               
                   
                 2500 
                 2557.931 
                 8.40 
                 102.31724 
               
               
                   
                 5000 
                 4459.877 
                 8.10 
                 89.19754 
               
               
                 Dextrorphan 
                 50 
                 60.202 
                 13.50 
                 120.404 
               
               
                   
                 100 
                 119.585 
                 9.20 
                 119.585 
               
               
                   
                 500 
                 565.957 
                 6.99 
                 113.1914 
               
               
                   
                 1000 
                 1062.165 
                 14.11 
                 106.2165 
               
               
                   
                 2500 
                 2534.949 
                 11.67 
                 101.39796 
               
               
                   
                 5000 
                 4807.142 
                 8.92 
                 96.14284 
               
               
                 Diazepam 
                 10 
                 10.335 
                 6.02 
                 103.35 
               
               
                   
                 20 
                 20.196 
                 3.58 
                 100.98 
               
               
                   
                 100 
                 100.989 
                 4.50 
                 100.989 
               
               
                   
                 200 
                 205.221 
                 2.82 
                 102.6105 
               
               
                   
                 500 
                 496.036 
                 2.17 
                 99.2072 
               
               
                   
                 1000 
                 997.223 
                 5.70 
                 99.7223 
               
               
                 Dihydrocodeine 
                 50 
                 50.689 
                 6.86 
                 101.378 
               
               
                   
                 100 
                 96.589 
                 5.67 
                 96.589 
               
               
                   
                 500 
                 518.144 
                 9.90 
                 103.6288 
               
               
                   
                 1000 
                 1031.237 
                 9.39 
                 103.1237 
               
               
                   
                 2500 
                 2384.294 
                 9.75 
                 95.37176 
               
               
                   
                 5000 
                 5069.047 
                 10.55 
                 101.38094 
               
               
                 EDDP 
                 50 
                 52.373 
                 9.29 
                 104.746 
               
               
                   
                 100 
                 104.689 
                 5.74 
                 104.689 
               
               
                   
                 500 
                 513.104 
                 7.34 
                 102.6208 
               
               
                   
                 1000 
                 1034.748 
                 7.05 
                 103.4748 
               
               
                   
                 2500 
                 2502.021 
                 7.85 
                 100.08084 
               
               
                   
                 5000 
                 4943.064 
                 8.43 
                 98.86128 
               
               
                 Fentanyl 
                 2 
                 2.102 
                 6.68 
                 105.1 
               
               
                   
                 4 
                 3.959 
                 4.65 
                 98.975 
               
               
                   
                 20 
                 19.736 
                 6.07 
                 98.68 
               
               
                   
                 40 
                 40.809 
                 5.15 
                 102.0225 
               
               
                   
                 100 
                 99.57 
                 7.53 
                 99.57 
               
               
                   
                 200 
                 199.824 
                 5.01 
                 99.912 
               
               
                 Flunitrazepam 
                 10 
                 10.279 
                 12.47 
                 102.79 
               
               
                   
                 20 
                 20.037 
                 9.02 
                 100.185 
               
               
                   
                 100 
                 89.661 
                 14.99 
                 89.661 
               
               
                   
                 200 
                 191.934 
                 12.91 
                 95.967 
               
               
                   
                 500 
                 469.029 
                 12.40 
                 93.8058 
               
               
                   
                 1000 
                 1068.684 
                 7.77 
                 106.8684 
               
               
                 Gabapentin 
                 100 
                 113.582 
                 7.63 
                 113.582 
               
               
                   
                 200 
                 210.796 
                 11.85 
                 105.398 
               
               
                   
                 1000 
                 1001.313 
                 8.49 
                 100.1313 
               
               
                   
                 2000 
                 1938.305 
                 5.77 
                 96.91525 
               
               
                   
                 5000 
                 4793.645 
                 14.25 
                 95.8729 
               
               
                   
                 10000 
                 10242.359 
                 11.87 
                 102.42359 
               
               
                 Hydrocodone 
                 50 
                 56.75 
                 8.27 
                 113.5 
               
               
                   
                 100 
                 108.487 
                 6.51 
                 108.487 
               
               
                   
                 500 
                 535.285 
                 7.52 
                 107.057 
               
               
                   
                 1000 
                 1076.554 
                 4.47 
                 107.6554 
               
               
                   
                 2500 
                 2391.087 
                 6.39 
                 95.64348 
               
               
                   
                 5000 
                 4981.836 
                 7.59 
                 99.63672 
               
               
                 Hydromorphone 
                 50 
                 50.036 
                 5.29 
                 100.072 
               
               
                   
                 100 
                 101.853 
                 3.52 
                 101.853 
               
               
                   
                 500 
                 497.046 
                 3.97 
                 99.4092 
               
               
                   
                 1000 
                 1015.242 
                 3.44 
                 101.5242 
               
               
                   
                 2500 
                 2409.174 
                 5.04 
                 96.36696 
               
               
                   
                 5000 
                 5076.649 
                 7.21 
                 101.53298 
               
               
                 Imipramine 
                 20 
                 25.249 
                 6.92 
                 126.245 
               
               
                   
                 40 
                 49.328 
                 7.01 
                 123.32 
               
               
                   
                 200 
                 245.662 
                 3.32 
                 122.831 
               
               
                   
                 400 
                 481.357 
                 4.11 
                 120.33925 
               
               
                   
                 1000 
                 1014.236 
                 7.43 
                 101.4236 
               
               
                   
                 2000 
                 1844.167 
                 5.63 
                 92.20835 
               
               
                 Lorazepam 
                 10 
                 12.221 
                 2.68 
                 122.21 
               
               
                   
                 20 
                 23.327 
                 3.93 
                 116.635 
               
               
                   
                 100 
                 110.648 
                 7.02 
                 110.648 
               
               
                   
                 200 
                 223.622 
                 7.19 
                 111.811 
               
               
                   
                 500 
                 524.027 
                 5.76 
                 104.8054 
               
               
                   
                 1000 
                 936.155 
                 7.19 
                 93.6155 
               
               
                 MDA 
                 50 
                 56.424 
                 10.54 
                 112.848 
               
               
                   
                 100 
                 115.115 
                 14.11 
                 115.115 
               
               
                   
                 500 
                 577.012 
                 12.89 
                 115.4024 
               
               
                   
                 1000 
                 1027.623 
                 10.10 
                 102.7623 
               
               
                   
                 2500 
                 2415.519 
                 7.28 
                 96.62076 
               
               
                   
                 5000 
                 4958.307 
                 9.97 
                 99.16614 
               
               
                 MDMA 
                 50 
                 60.24 
                 17.77 
                 120.48 
               
               
                   
                 100 
                 112.654 
                 11.58 
                 112.654 
               
               
                   
                 500 
                 529.626 
                 14.07 
                 105.9252 
               
               
                   
                 1000 
                 1044.036 
                 11.34 
                 104.4036 
               
               
                   
                 2500 
                 2406.061 
                 8.12 
                 96.24244 
               
               
                   
                 5000 
                 4997.382 
                 10.02 
                 99.94764 
               
               
                 Meperidine 
                 50 
                 54.096 
                 5.96 
                 108.192 
               
               
                   
                 100 
                 106.027 
                 4.57 
                 106.027 
               
               
                   
                 500 
                 531.724 
                 5.46 
                 106.3448 
               
               
                   
                 1000 
                 1050.335 
                 4.88 
                 105.0335 
               
               
                   
                 2500 
                 2494.327 
                 7.30 
                 99.77308 
               
               
                   
                 5000 
                 4913.491 
                 7.53 
                 98.26982 
               
               
                 Meprobamate 
                 50 
                 58.591 
                 13.27 
                 117.182 
               
               
                   
                 100 
                 124.163 
                 12.01 
                 124.163 
               
               
                   
                 500 
                 620.061 
                 9.60 
                 124.0122 
               
               
                   
                 1000 
                 1198.158 
                 7.49 
                 119.8158 
               
               
                   
                 2500 
                 2510.349 
                 7.29 
                 100.41396 
               
               
                   
                 5000 
                 4628.679 
                 5.61 
                 92.57358 
               
               
                 Methadone 
                 50 
                 51.255 
                 3.85 
                 102.51 
               
               
                   
                 100 
                 102.709 
                 4.03 
                 102.709 
               
               
                   
                 500 
                 522.547 
                 4.66 
                 104.5094 
               
               
                   
                 1000 
                 1048.074 
                 4.51 
                 104.8074 
               
               
                   
                 2500 
                 2470.304 
                 2.87 
                 98.81216 
               
               
                   
                 5000 
                 4955.111 
                 2.91 
                 99.10222 
               
               
                 Methamphetamine 
                 50 
                 58.358 
                 10.68 
                 116.716 
               
               
                   
                 100 
                 110.745 
                 7.70 
                 110.745 
               
               
                   
                 500 
                 558.027 
                 11.54 
                 111.6054 
               
               
                   
                 1000 
                 1107.356 
                 6.84 
                 110.7356 
               
               
                   
                 2500 
                 2496.679 
                 6.06 
                 99.86716 
               
               
                   
                 5000 
                 4818.835 
                 7.39 
                 96.3767 
               
               
                 Methylphenidate 
                 50 
                 41.532 
                 7.54 
                 83.064 
               
               
                   
                 100 
                 78.541 
                 5.94 
                 78.541 
               
               
                   
                 500 
                 419.333 
                 4.81 
                 83.8666 
               
               
                   
                 1000 
                 1236.24 
                 7.47 
                 123.624 
               
               
                   
                 2500 
                 2786.336 
                 5.60 
                 111.45344 
               
               
                   
                 5000 
                 4629.931 
                 11.54 
                 92.59862 
               
               
                 Midazolam 
                 10 
                 8.281 
                 8.96 
                 82.81 
               
               
                   
                 20 
                 16.408 
                 5.54 
                 82.04 
               
               
                   
                 100 
                 84.021 
                 9.19 
                 84.021 
               
               
                   
                 200 
                 180.579 
                 9.69 
                 90.2895 
               
               
                   
                 500 
                 477.496 
                 7.69 
                 95.4992 
               
               
                   
                 1000 
                 1063.214 
                 13.88 
                 106.3214 
               
               
                 Morphine 
                 50 
                 51.924 
                 5.18 
                 103.848 
               
               
                   
                 100 
                 102.435 
                 5.23 
                 102.435 
               
               
                   
                 500 
                 501.113 
                 3.63 
                 100.2226 
               
               
                   
                 1000 
                 1008.631 
                 1.84 
                 100.8631 
               
               
                   
                 2500 
                 2437.596 
                 3.44 
                 97.50384 
               
               
                   
                 5000 
                 5048.301 
                 6.24 
                 100.96602 
               
               
                 Naloxone 
                 10 
                 11.592 
                 8.74 
                 115.92 
               
               
                   
                 20 
                 22.361 
                 9.36 
                 111.805 
               
               
                   
                 100 
                 109.608 
                 6.13 
                 109.608 
               
               
                   
                 200 
                 226.355 
                 5.99 
                 113.1775 
               
               
                   
                 500 
                 483.011 
                 4.40 
                 96.6022 
               
               
                   
                 1000 
                 977.073 
                 11.87 
                 97.7073 
               
               
                 Norbuprenorphine 
                 10 
                 9.619 
                 19.45 
                 96.19 
               
               
                   
                 20 
                 21.224 
                 12.12 
                 106.12 
               
               
                   
                 100 
                 107.124 
                 4.89 
                 107.124 
               
               
                   
                 200 
                 198.969 
                 7.87 
                 99.4845 
               
               
                   
                 500 
                 484.094 
                 8.81 
                 96.8188 
               
               
                   
                 1000 
                 1008.97 
                 6.72 
                 100.897 
               
               
                 Nordiazepam 
                 10 
                 9.635 
                 10.50 
                 96.35 
               
               
                   
                 20 
                 19.99 
                 6.79 
                 99.95 
               
               
                   
                 100 
                 98.472 
                 4.63 
                 98.472 
               
               
                   
                 200 
                 195.575 
                 4.81 
                 97.7875 
               
               
                   
                 500 
                 486.519 
                 5.25 
                 97.3038 
               
               
                   
                 1000 
                 1019.809 
                 5.67 
                 101.9809 
               
               
                 Norfentanyl 
                 2 
                 2.15 
                 4.98 
                 107.5 
               
               
                   
                 4 
                 4.177 
                 4.81 
                 104.425 
               
               
                   
                 20 
                 21.008 
                 4.66 
                 105.04 
               
               
                   
                 40 
                 41.422 
                 9.35 
                 103.555 
               
               
                   
                 100 
                 100.387 
                 14.25 
                 100.387 
               
               
                   
                 200 
                 196.856 
                 14.93 
                 98.428 
               
               
                 Normeperidine 
                 50 
                 62.319 
                 4.03 
                 124.638 
               
               
                   
                 100 
                 126.811 
                 9.47 
                 126.811 
               
               
                   
                 500 
                 642.542 
                 5.69 
                 128.5084 
               
               
                   
                 1000 
                 1200.314 
                 7.22 
                 120.0314 
               
               
                   
                 2500 
                 2724.204 
                 10.19 
                 108.96816 
               
               
                   
                 5000 
                 4651.934 
                 7.82 
                 93.03868 
               
               
                 Norpropoxyphene 
                 50 
                 57.505 
                 11.63 
                 115.01 
               
               
                   
                 100 
                 106.874 
                 8.33 
                 106.874 
               
               
                   
                 500 
                 569.853 
                 8.41 
                 113.9706 
               
               
                   
                 1000 
                 1075.36 
                 6.76 
                 107.536 
               
               
                   
                 2500 
                 2491.723 
                 7.10 
                 99.66892 
               
               
                   
                 5000 
                 4848.685 
                 10.49 
                 96.9737 
               
               
                 Nortriptyline 
                 20 
                 20.265 
                 7.54 
                 101.325 
               
               
                   
                 40 
                 41.048 
                 4.33 
                 102.62 
               
               
                   
                 200 
                 207.141 
                 4.25 
                 103.5705 
               
               
                   
                 400 
                 430.264 
                 3.23 
                 107.566 
               
               
                   
                 1000 
                 1001.463 
                 5.20 
                 100.1463 
               
               
                   
                 2000 
                 1959.819 
                 7.62 
                 97.99095 
               
               
                 Oxazepam 
                 10 
                 10.312 
                 12.48 
                 103.12 
               
               
                   
                 20 
                 19.77 
                 13.90 
                 98.85 
               
               
                   
                 100 
                 95.562 
                 13.26 
                 95.562 
               
               
                   
                 200 
                 206.874 
                 9.05 
                 103.437 
               
               
                   
                 500 
                 479.802 
                 6.71 
                 95.9604 
               
               
                   
                 1000 
                 1017.68 
                 8.78 
                 101.768 
               
               
                 Oxycodone 
                 50 
                 57.266 
                 7.12 
                 114.532 
               
               
                   
                 100 
                 114.272 
                 6.37 
                 114.272 
               
               
                   
                 500 
                 523.065 
                 7.15 
                 104.613 
               
               
                   
                 1000 
                 1086.639 
                 8.36 
                 108.6639 
               
               
                   
                 2500 
                 2466.262 
                 5.38 
                 98.65048 
               
               
                   
                 5000 
                 4902.496 
                 7.86 
                 98.04992 
               
               
                 Oxymorphone 
                 50 
                 51.45 
                 6.20 
                 102.9 
               
               
                   
                 100 
                 102.58 
                 3.63 
                 102.58 
               
               
                   
                 500 
                 525.212 
                 4.06 
                 105.0424 
               
               
                   
                 1000 
                 1023.084 
                 3.34 
                 102.3084 
               
               
                   
                 2500 
                 2483.991 
                 3.39 
                 99.35964 
               
               
                   
                 5000 
                 4963.683 
                 6.83 
                 99.27366 
               
               
                 PCP 
                 4 
                 4.326 
                 6.14 
                 108.15 
               
               
                   
                 8 
                 8.389 
                 10.13 
                 104.8625 
               
               
                   
                 40 
                 41.353 
                 7.14 
                 103.3825 
               
               
                   
                 80 
                 83.883 
                 7.82 
                 104.85375 
               
               
                   
                 200 
                 189.042 
                 6.93 
                 94.521 
               
               
                   
                 400 
                 405.007 
                 7.71 
                 101.25175 
               
               
                 Pregabalin 
                 50 
                 57.604 
                 13.24 
                 115.208 
               
               
                   
                 100 
                 123.443 
                 9.64 
                 123.443 
               
               
                   
                 500 
                 638.824 
                 5.52 
                 127.7648 
               
               
                   
                 1000 
                 1232.32 
                 4.73 
                 123.232 
               
               
                   
                 2500 
                 2485.475 
                 6.78 
                 99.419 
               
               
                   
                 5000 
                 4352.334 
                 9.06 
                 87.04668 
               
               
                 Propoxyphene 
                 50 
                 56.678 
                 10.60 
                 113.356 
               
               
                   
                 100 
                 104.987 
                 14.21 
                 104.987 
               
               
                   
                 500 
                 505.595 
                 14.65 
                 101.119 
               
               
                   
                 1000 
                 1064.326 
                 11.32 
                 106.4326 
               
               
                   
                 2500 
                 2450.388 
                 13.37 
                 98.01552 
               
               
                   
                 5000 
                 4968.026 
                 9.85 
                 99.36052 
               
               
                 Sertraline 
                 20 
                 14.54 
                 11.51 
                 72.7 
               
               
                   
                 40 
                 29.663 
                 14.13 
                 74.1575 
               
               
                   
                 200 
                 177.291 
                 9.21 
                 88.6455 
               
               
                   
                 400 
                 403.396 
                 6.10 
                 100.849 
               
               
                   
                 1000 
                 1023.69 
                 5.85 
                 102.369 
               
               
                   
                 2000 
                 2011.421 
                 6.22 
                 100.57105 
               
               
                 Triazolam 
                 10 
                 7.666 
                 11.27 
                 76.66 
               
               
                   
                 20 
                 14.838 
                 6.84 
                 74.19 
               
               
                   
                 100 
                 79.783 
                 9.45 
                 79.783 
               
               
                   
                 200 
                 181.93 
                 13.79 
                 90.965 
               
               
                   
                 500 
                 468.668 
                 11.18 
                 93.7336 
               
               
                   
                 1000 
                 1077.115 
                 14.49 
                 107.7115 
               
               
                 Zaleplon 
                 10 
                 11.648 
                 8.28 
                 116.48 
               
               
                   
                 20 
                 25.18 
                 6.51 
                 125.9 
               
               
                   
                 100 
                 120.886 
                 6.67 
                 120.886 
               
               
                   
                 200 
                 230.422 
                 5.85 
                 115.211 
               
               
                   
                 500 
                 522.275 
                 14.49 
                 104.455 
               
               
                   
                 1000 
                 917.59 
                 9.22 
                 91.759 
               
               
                 Zolpidem 
                 10 
                 10.411 
                 5.29 
                 104.11 
               
               
                   
                 20 
                 20.82 
                 4.36 
                 104.1 
               
               
                   
                 100 
                 103.137 
                 6.28 
                 103.137 
               
               
                   
                 200 
                 203.121 
                 8.46 
                 101.5605 
               
               
                   
                 500 
                 502.471 
                 4.36 
                 100.4942 
               
               
                   
                 1000 
                 990.041 
                 7.50 
                 99.0041 
               
               
                 Carboxy Zolpidem 
                 10 
                 12.582 
                 4.76 
                 125.82 
               
               
                   
                 20 
                 22.923 
                 3.01 
                 114.615 
               
               
                   
                 100 
                 123.592 
                 7.91 
                 123.592 
               
               
                   
                 200 
                 231.672 
                 5.30 
                 115.836 
               
               
                   
                 500 
                 504.777 
                 11.98 
                 100.9554 
               
               
                   
                 1000 
                 927.454 
                 8.08 
                 92.7454 
               
               
                 Carisoprodol 1 
                 50 
                 59.281 
                 8.26 
                 118.562 
               
               
                   
                 100 
                 114.222 
                 9.91 
                 114.222 
               
               
                   
                 500 
                 576.377 
                 11.83 
                 115.2754 
               
               
                   
                 1000 
                 1169.596 
                 10.68 
                 116.9596 
               
               
                   
                 2500 
                 2580.468 
                 12.88 
                 103.21872 
               
               
                   
                 5000 
                 4650.057 
                 12.81 
                 93.00114 
               
               
                 Amphetamine 1 
                 50 
                 64.646 
                 10.52 
                 129.292 
               
               
                   
                 100 
                 125.28 
                 8.88 
                 125.28 
               
               
                   
                 500 
                 609.03 
                 11.52 
                 121.806 
               
               
                   
                 1000 
                 1136.826 
                 9.09 
                 113.6826 
               
               
                   
                 2500 
                 2402.718 
                 10.79 
                 96.10872 
               
               
                   
                 5000 
                 4791.499 
                 12.45 
                 95.82998 
               
               
                 Tapentadol 1 
                 10 
                 8.069 
                 9.81 
                 80.69 
               
               
                   
                 20 
                 15.677 
                 5.65 
                 78.385 
               
               
                   
                 100 
                 84.309 
                 9.47 
                 84.309 
               
               
                   
                 200 
                 179.573 
                 9.13 
                 89.7865 
               
               
                   
                 500 
                 478.155 
                 9.31 
                 95.631 
               
               
                   
                 1000 
                 1064.217 
                 12.45 
                 106.4217 
               
               
                 Tramadol 1 
                 50 
                 58.769 
                 13.72 
                 117.538 
               
               
                   
                 100 
                 110.987 
                 6.24 
                 110.987 
               
               
                   
                 500 
                 538.282 
                 13.75 
                 107.6564 
               
               
                   
                 1000 
                 1097.215 
                 12.08 
                 109.7215 
               
               
                   
                 2500 
                 2477.924 
                 9.88 
                 99.11696 
               
               
                   
                 5000 
                 4866.823 
                 9.38 
                 97.33646 
               
               
                 Temazepam 1 
                 10 
                 7.666 
                 10.67 
                 76.66 
               
               
                   
                 20 
                 14.838 
                 5.81 
                 74.19 
               
               
                   
                 100 
                 79.783 
                 9.41 
                 79.783 
               
               
                   
                 200 
                 181.93 
                 12.61 
                 90.965 
               
               
                   
                 500 
                 468.668 
                 10.79 
                 93.7336 
               
               
                   
                 1000 
                 1077.115 
                 14.37 
                 107.7115 
               
               
                 Zopiclone 1 
                 10 
                 10.411 
                 5.52 
                 104.11 
               
               
                   
                 20 
                 20.82 
                 7 
                 104.1 
               
               
                   
                 100 
                 103.137 
                 7.84 
                 103.137 
               
               
                   
                 200 
                 203.121 
                 8.2 
                 101.5605 
               
               
                   
                 500 
                 502.471 
                 7.26 
                 100.4942 
               
               
                   
                 1000 
                 990.041 
                 8.48 
                 99.0041 
               
               
                 Butalbital 1 
                 50 
                 56.133 
                 10.86 
                 112.266 
               
               
                   
                 100 
                 106.332 
                 8.65 
                 106.332 
               
               
                   
                 500 
                 547.55 
                 8.61 
                 109.51 
               
               
                   
                 1000 
                 1061.016 
                 5.75 
                 106.1016 
               
               
                   
                 2500 
                 2462.409 
                 6.38 
                 98.49636 
               
               
                   
                 5000 
                 4916.56 
                 10.12 
                 98.3312 
               
               
                 THCA 1 
                 10 
                 11.582 
                 4.67 
                 115.82 
               
               
                   
                 20 
                 22.923 
                 5.56 
                 114.615 
               
               
                   
                 100 
                 123.592 
                 7.19 
                 123.592 
               
               
                   
                 200 
                 231.672 
                 5.3 
                 115.836 
               
               
                   
                 500 
                 504.777 
                 10.89 
                 100.9554 
               
               
                   
                 1000 
                 927.454 
                 7.4 
                 92.7454 
               
               
                 Ritalinic acid 1 
                 50 
                 52.594 
                 5.18 
                 105.188 
               
               
                   
                 100 
                 100.343 
                 5.23 
                 100.343 
               
               
                   
                 500 
                 509.723 
                 3.63 
                 101.9446 
               
               
                   
                 1000 
                 1018.434 
                 1.84 
                 101.8434 
               
               
                   
                 2500 
                 2468.733 
                 3.44 
                 98.74932 
               
               
                   
                 5000 
                 5000.174 
                 6.24 
                 100.00348 
               
               
                 Phenobarbital 1 
                 50 
                 51.306 
                 3.61 
                 102.612 
               
               
                   
                 100 
                 98.075 
                 5.50 
                 98.075 
               
               
                   
                 500 
                 501.966 
                 2.96 
                 100.3932 
               
               
                   
                 1000 
                 988.116 
                 1.72 
                 98.8116 
               
               
                   
                 2500 
                 2468.619 
                 2.22 
                 98.74476 
               
               
                   
                 5000 
                 5041.918 
                 7.36 
                 100.83836 
               
               
                   
               
            
           
         
       
     
     Recovery 
     Recoveries were calculated by adding known concentrations of drug metabolites, to 3 different samples previously analyzed, and then the final concentrations were measured in duplicate. Results were measured as differences between the measured and the theoretical values and expressed as percentage of recovery. 
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Intra-day study of recovery with the quality 
               
               
                 controls and their yield (n = 10) 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Mean 
                 CV 
               
               
                   
                 Component Name 
                 QC Name 
                 n = 10 
                 (%) 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 6-MAM 
                 C3 
                 9.32 
                 8.59 
               
               
                   
                 OH-et-Flurazepam 
                 BZ 100 
                 79.38 
                 4.68 
               
               
                   
                 4-Hydroxyalprazolam 
                 BZ 100 
                 83.49 
                 4.98 
               
               
                   
                 7-Aminoclonazepam 
                 BZ 100 
                 112.24 
                 6.68 
               
               
                   
                 Amitriptyline 
                 POS 
                 138.38 
                 3.54 
               
               
                   
                 Amphetamine 
                 POS 
                 1567.18 
                 5.22 
               
               
                   
                 Benzoylecgonine 
                 C3 
                 196.79 
                 2.91 
               
               
                   
                 Buprenorphine 
                 PM 100 
                 86.11 
                 7.56 
               
               
                   
                 Carisoprodol 
                 POS 
                 329.32 
                 10.28 
               
               
                   
                 Chlordiazepoxide 
                 BZ 100 
                 60.23 
                 5.27 
               
               
                   
                 Codeine 
                 PM 100 
                 84.28 
                 6.31 
               
               
                   
                 Cyclobenzaprine 
                 POS 
                 152.42 
                 4.18 
               
               
                   
                 Desipramine 
                 POS 
                 188.98 
                 7.42 
               
               
                   
                 Desmethyltramadol 
                 POS 
                 330.79 
                 9.18 
               
               
                   
                 Desomorphine 
                 POS 
                 316.45 
                 8.60 
               
               
                   
                 DesTapentadol 
                 POS 
                 367.13 
                 3.69 
               
               
                   
                 Dextromethorphan 
                 POS 
                 390.96 
                 4.88 
               
               
                   
                 Dextrorphan 
                 POS 
                 302.44 
                 6.63 
               
               
                   
                 Diazepam 
                 BZ 100 
                 68.5 
                 1.29 
               
               
                   
                 Dihydrocodeine 
                 POS 
                 264.34 
                 9.32 
               
               
                   
                 EDDP 
                 PM 100 
                 97.64 
                 6.57 
               
               
                   
                 Fentanyl 
                 PM 100 
                 11.36 
                 7.49 
               
               
                   
                 Flunitrazepam 
                 BZ 100 
                 80.49 
                 7.86 
               
               
                   
                 Gabapentin 
                 POS 
                 1077.48 
                 8.11 
               
               
                   
                 Hydrocodone 
                 PM 100 
                 82.34 
                 7.76 
               
               
                   
                 Hydromorphone 
                 PM 100 
                 82.32 
                 3.58 
               
               
                   
                 Imipramine 
                 POS 
                 180.64 
                 5.29 
               
               
                   
                 Lorazepam 
                 BZ 100 
                 100.59 
                 4.64 
               
               
                   
                 MDA 
                 POS 
                 1392.91 
                 12.30 
               
               
                   
                 MDMA 
                 POS 
                 1413.43 
                 6.84 
               
               
                   
                 Meperidine 
                 POS 
                 275.67 
                 5.75 
               
               
                   
                 Meprobamate 
                 POS 
                 348.37 
                 6.07 
               
               
                   
                 Methadone 
                 C3 
                 332.6 
                 3.12 
               
               
                   
                 Methamphetamine 
                 POS 
                 1429.31 
                 8.88 
               
               
                   
                 Methylphenidate 
                 POS 
                 127.81 
                 5.11 
               
               
                   
                 Midazolam 
                 BZ 100 
                 68.1 
                 6.52 
               
               
                   
                 Morphine 
                 PM 100 
                 96.13 
                 4.18 
               
               
                   
                 Naloxone 
                 POS 
                 277.89 
                 8.12 
               
               
                   
                 Norbuprenorphine 
                 PM 100 
                 91.43 
                 12.49 
               
               
                   
                 Nordiazepam 
                 BZ 100 
                 83.01 
                 4.28 
               
               
                   
                 Norfentanyl 
                 PM 100 
                 13.25 
                 4.89 
               
               
                   
                 Normeperidine 
                 POS 
                 340.25 
                 8.69 
               
               
                   
                 Norpropoxyphene 
                 C3 
                 203.78 
                 9.01 
               
               
                   
                 Nortriptyline 
                 POS 
                 151.38 
                 4.22 
               
               
                   
                 Oxazepam 
                 BZ 100 
                 84.48 
                 5.13 
               
               
                   
                 Oxycodone 
                 PM 100 
                 84.11 
                 9.17 
               
               
                   
                 Oxymorphone 
                 PM 100 
                 83.94 
                 3.81 
               
               
                   
                 PCP 
                 C3 
                 27.26 
                 6.14 
               
               
                   
                 Pregabalin 
                 POS 
                 295.75 
                 9.69 
               
               
                   
                 Propoxyphene 
                 C3 
                 329.14 
                 9.77 
               
               
                   
                 Ritalinic Acid 
                 POS 
                 1360.81 
                 4.48 
               
               
                   
                 Sertraline 
                 POS 
                 152.82 
                 2.91 
               
               
                   
                 Tapentadol 
                 POS 
                 334.33 
                 5.01 
               
               
                   
                 Temazepam 
                 BZ 100 
                 76.93 
                 3.91 
               
               
                   
                 Tramadol 
                 POS 
                 259.3 
                 7.58 
               
               
                   
                 Triazolam 
                 BZ 100 
                 64.97 
                 4.14 
               
               
                   
                 Zaleplon 
                 POS 
                 355.86 
                 12.71 
               
               
                   
                 Zopiclone 
                 POS 
                 223.53 
                 10.72 
               
               
                   
                 Zolpidem 
                 POS 
                 263.9 
                 4.90 
               
               
                   
                 Carboxy Zolpidem 
                 POS 
                 334.25 
                 5.86 
               
               
                   
                 Butalbital 
                 C3 
                 210.27 
                 5.54 
               
               
                   
                 THC 
                 C3 
                 10.81 
                 6.80 
               
               
                   
                 Phenobarbital 
                 C3 
                 213.69 
                 7.88 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Inter-day study of recovery with the quality 
               
               
                 controls and their yield (n = 10) 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                   
                 Mean 
                 CV 
               
               
                   
                 Component Name 
                 QC Name 
                 n = 10 
                 (%) 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 6-MAM 
                 C3 
                 9.32 
                 8.59 
               
               
                   
                 OH-et-Flurazepam 
                 BZ 100 
                 79.38 
                 4.68 
               
               
                   
                 4-Hydroxyalprazolam 
                 BZ 100 
                 83.49 
                 4.98 
               
               
                   
                 7-Aminoclonazepam 
                 BZ 100 
                 112.24 
                 6.68 
               
               
                   
                 Amitriptyline 
                 POS 
                 138.38 
                 3.54 
               
               
                   
                 Amphetamine 
                 POS 
                 1558.05 
                 8.54 
               
               
                   
                 Benzoylecgonine 
                 C3 
                 196.79 
                 2.91 
               
               
                   
                 Buprenorphine 
                 C3 
                 0.27 
                 42.23 
               
               
                   
                 Carisoprodol 
                 POS 
                 329.32 
                 10.28 
               
               
                   
                 Chlordiazepoxide 
                 BZ 100 
                 60.23 
                 5.27 
               
               
                   
                 Codeine 
                 PM 100 
                 84.28 
                 6.31 
               
               
                   
                 Cyclobenzaprine 
                 POS 
                 152.42 
                 4.18 
               
               
                   
                 Desipramine 
                 POS 
                 188.98 
                 7.42 
               
               
                   
                 Desmethyltramadol 
                 POS 
                 330.79 
                 9.18 
               
               
                   
                 Desomorphine 
                 POS 
                 316.45 
                 8.60 
               
               
                   
                 DesTapentadol 
                 POS 
                 367.13 
                 3.69 
               
               
                   
                 Dextromethorphan 
                 POS 
                 390.96 
                 4.88 
               
               
                   
                 Dextrorphan 
                 POS 
                 302.44 
                 6.63 
               
               
                   
                 Diazepam 
                 BZ 100 
                 68.5 
                 1.29 
               
               
                   
                 Dihydrocodeine 
                 POS 
                 264.34 
                 9.32 
               
               
                   
                 EDDP 
                 PM 100 
                 97.64 
                 6.57 
               
               
                   
                 Fentanyl 
                 PM 100 
                 11.36 
                 7.49 
               
               
                   
                 Flunitrazepam 
                 BZ 100 
                 80.49 
                 7.86 
               
               
                   
                 Gabapentin 
                 POS 
                 1077.48 
                 8.11 
               
               
                   
                 Hydrocodone 
                 PM 100 
                 82.34 
                 7.76 
               
               
                   
                 Hydromorphone 
                 PM 100 
                 82.32 
                 3.58 
               
               
                   
                 Imipramine 
                 POS 
                 180.64 
                 5.29 
               
               
                   
                 Lorazepam 
                 BZ 100 
                 100.59 
                 4.64 
               
               
                   
                 MDA 
                 POS 
                 1338.53 
                 9.05 
               
               
                   
                 MDMA 
                 POS 
                 1413.43 
                 6.84 
               
               
                   
                 Meperidine 
                 POS 
                 275.67 
                 5.75 
               
               
                   
                 Meprobamate 
                 POS 
                 348.37 
                 6.07 
               
               
                   
                 Methadone 
                 C3 
                 332.6 
                 3.12 
               
               
                   
                 Methamphetamine 
                 POS 
                 1429.31 
                 8.88 
               
               
                   
                 Methylphenidate 
                 POS 
                 127.81 
                 5.11 
               
               
                   
                 Midazolam 
                 BZ 100 
                 60 
                 6.71 
               
               
                   
                 Morphine 
                 PM 100 
                 96.13 
                 4.18 
               
               
                   
                 Naloxone 
                 POS 
                 277.89 
                 8.12 
               
               
                   
                 Norbuprenorphine 
                 C3 
                 0.68 
                 64.20 
               
               
                   
                 Nordiazepam 
                 BZ 100 
                 83.01 
                 4.28 
               
               
                   
                 Norfentanyl 
                 PM 100 
                 13.25 
                 4.89 
               
               
                   
                 Normeperidine 
                 POS 
                 351.23 
                 6.38 
               
               
                   
                 Norpropoxyphene 
                 C3 
                 203.78 
                 9.01 
               
               
                   
                 Nortriptyline 
                 POS 
                 151.38 
                 4.22 
               
               
                   
                 Oxazepam 
                 BZ 100 
                 84.48 
                 5.13 
               
               
                   
                 Oxycodone 
                 PM 100 
                 84.11 
                 9.17 
               
               
                   
                 Oxymorphone 
                 PM 100 
                 83.94 
                 3.81 
               
               
                   
                 PCP 
                 C3 
                 27.26 
                 6.14 
               
               
                   
                 Pregabalin 
                 POS 
                 295.75 
                 9.69 
               
               
                   
                 Propoxyphene 
                 C3 
                 329.14 
                 9.77 
               
               
                   
                 Ritalinic Acid 
                 POS 
                 1360.81 
                 4.48 
               
               
                   
                 Sertraline 
                 POS 
                 152.82 
                 2.91 
               
               
                   
                 Tapentadol 
                 POS 
                 334.33 
                 5.01 
               
               
                   
                 Temazepam 
                 BZ 100 
                 76.93 
                 3.91 
               
               
                   
                 Tramadol 
                 POS 
                 259.3 
                 7.58 
               
               
                   
                 Triazolam 
                 BZ 100 
                 64.97 
                 4.14 
               
               
                   
                 Zaleplon 
                 POS 
                 355.86 
                 12.71 
               
               
                   
                 Zopiclone 
                 POS 
                 223.53 
                 10.72 
               
               
                   
                 Zolpidem 
                 POS 
                 263.9 
                 4.90 
               
               
                   
                 Carboxy Zolpidem 
                 POS 
                 334.25 
                 5.86 
               
               
                   
                 Butalbital 
                 C3 
                 210.27 
                 5.54 
               
               
                   
                 THC 
                 C3 
                 10.81 
                 6.80 
               
               
                   
                 Phenobarbital 
                 C3 
                 213.69 
                 7.88 
               
               
                   
                   
               
            
           
         
       
     
     An object of the presently disclosed subject matter having been stated hereinabove, and which is achieved in whole or in part by the presently disclosed subject matter, other objects and advantages will become evident to those of ordinary skill in the art after a study of the description of the presently disclosed subject matter, figures, and non-limiting examples.