**SW-846 Update V 3620C - 1 Revision 4**

**October 2012 METHOD 3620C** 

**FLORISIL CLEANUP**

**SW-846 is not intended to be an analytical training manual. Therefore, method procedures** 

**are written based on the assumption that they will be performed by analysts who are formally** 

**trained in at least the basic principles of chemica l analysis and in the use of the subject** 

**technology.**

**In addition, SW-846 methods, with the exception of required method use for the analysis of** 

**method-defined parameters, are intended to be guida nce methods which contain general** 

**information on how to perform an analytical procedu re or technique which a laboratory can use as** 

**a basic starting point for generating its own detai led Standard Operating Procedure (SOP), either** 

**for its own general use or for a specific project a pplication. The performance data included in this** 

**method are for guidance purposes only, and are not intended to be and must not be used as** 

**absolute QC acceptance criteria for purposes of lab oratory accreditation.** 

**1.0 SCOPE AND APPLICATION**

**1.1 Florisil®, a registered trade name of U. S. Sil ica Co., is a magnesium silicate with** 

**basic properties. It is used to separate analytes from interfering compounds prior to sample** 

**analysis by a chromatographic method.**

**1.2 Florisil® has been used for the cleanup of pest icide residues and other chlorinated** 

**hydrocarbons; the separation of nitrogen compounds from hydrocarbons; the separation of** 

**aromatic compounds from aliphatic-aromatic mixtures ; and similar applications for use with fats,** 

**oils, and waxes. Additionally, Florisil® is consid ered good for separations with steroids, esters,** 

**ketones, glycerides, alkaloids, and some carbohydra tes.** 

**1.3 Florisil® cleanup may be accomplished by either using a glass chromatographic** 

**column packed with Florisil® or using solid-phase e xtraction cartridges containing Florisil®.** 

**1.4 This method includes procedures for cleanup of sample extracts containing the** 

**following analyte groups:**

**Phthalate esters Chlorinated hydrocarbons** 

**Nitrosamines Organochlorine pesticides** 

**Nitroaromatics Organophosphates** 

**Haloethers Organophosphorus pesticides** 

**Aniline and aniline derivatives PCBs** 

**Other analytes may potentially be cleaned up using this method provided that adequate** 

**performance is demonstrated.**

**1.5 Prior to employing this method, analysts are ad vised to consult the base method for** 

**each type of procedure that may be employed in the overall analysis (e.g., Methods 3500, 3600,** 

**5000, and 8000) for additional information on quali ty control procedures, development of QC** 

**acceptance criteria, calculations, and general guid ance. Analysts also should consult the** 

**disclaimer statement at the front of the manual and the information in Chapter Two for guidance** 

**on the intended flexibility in the choice of method s, apparatus, materials, reagents, and supplies,** **SW-846 Update V 3620C - 2 Revision 4**

**October 2012 and on the responsibilities of the analyst for demo**  **nstrating that the techniques employed are**

**appropriate for the analytes of interest, in the ma trix of interest, and at the levels of concern.** 

**In addition, analysts and data users are advised th at, except where explicitly specified in a** 

**regulation, the use of SW-846 methods is** not **mandatory in response to Federal testing**

**requirements. The information contained in this me thod is provided by EPA as guidance to be** 

**used by the analyst and the regulated community in making judgments necessary to generate** 

**results that meet the data quality objectives for t he intended application.** 

**1.6 This method is restricted to use by, or under t he supervision of, appropriately** 

**experienced and trained analysts. Each analyst mus t demonstrate the ability to generate** 

**acceptable results with this method.**

**2.0 SUMMARY OF METHOD**

**2.1 This method describes procedures for Florisil® cleanup of solvent extracts of** 

**environmental samples. It provides the option of u sing either traditional column chromatography** 

**techniques or solid-phase extraction cartridges. G enerally, the traditional column** 

**chromatography technique uses larger amounts of ads orbent and, therefore, has a greater** 

**cleanup capacity.**

**2.2 In the column cleanup protocol, the column is p acked with the appropriate amount of** 

**adsorbent, topped with a water adsorbent, and then loaded with the sample extract. Elution of** 

**the analytes is effected with a suitable solvent(s) , leaving the interfering compounds on the** 

**column. The eluate may be further concentrated pri or to gas chromatographic analysis.** 

**2.3 The cartridge cleanup protocol uses solid-phase extraction cartridges containing 40**  **μm particles of Florisil® (60 Å pores). Each cartr idge is washed with solvent immediately prior to** 

**use. The sample extract is loaded onto the cartrid ge which is then eluted with suitable** 

**solvent(s). A vacuum manifold is necessary to obta in reproducible results. The eluate may be** 

**further concentrated prior to gas chromatographic a nalysis.** 

**3.0 DEFINITIONS**

**Refer to Chapter One and the manufacturer's instruc tions for definitions that may be** 

**relevant to this method.**

**4.0 INTERFERENCES**

**4.1 Solvents, reagents, glassware, and other sample processing hardware may yield** 

**artifacts and/or interferences to sample analysis. All of these materials must be demonstrated to** 

**be free from interferences under the conditions of the analysis by analyzing method blanks.** 

**Specific selection of reagents and purification of solvents by distillation in all glass systems may** 

**be necessary. Refer to each method to be used for specific guidance on quality control** 

**procedures and to Chapter Four for general guidance on the cleaning of glassware.** 

**4.2 A reagent blank should be prepared and analyzed for the compounds of interest** 

**prior to the use of this method. The level of inte rferences must meet the QC acceptance criteria** 

**for blanks that are specified in a project planning document, e.g. QAPP, SAP, or in a laboratory** 

**SOP. SW-846 Update V 3620C - 3 Revision 4** 

**October 2012 4.3 The procedures for reagent purification outline**  **d here should be considered to be the**

**minimum needed for successful use of this method. More extensive procedures may be** 

**necessary to achieve acceptable levels of interfere nces for some analytes. However, during the** 

**evaluation of the cartridge cleanup procedure, phth alate esters were detected in the Florisil®** 

**cartridge method blanks at concentrations of up to 400 ng per cartridge. Therefore, complete** 

**removal of the phthalate esters from Florisil® cart ridges may not be possible. Phthalate ester** 

**contamination may be a problem with certain cartrid ges. The more inert the column and/or** 

**cartridge material (i.e., glass or polytetrafluoroe thylene (PTFE)), the fewer problems with** 

**phthalates contamination will occur. Phthalates cr eate interference problems for all method** 

**analytes, not just the phthalate esters themselves.**

**5.0 SAFETY**

**This method does not address all safety issues asso ciated with its use. The laboratory is** 

**responsible for maintaining a safe work environment and a current awareness file of OSHA** 

**regulations regarding the safe handling of the chem icals listed in this method. A reference file of** 

**material safety data sheets (MSDSs) should be avail able to all personnel involved in these** 

**analyses.**

**6.0 EQUIPMENT AND SUPPLIES**

**The mention of trade names or commercial products i n this manual is for illustrative** 

**purposes only, and does not constitute an EPA endor sement or exclusive recommendation for** 

**use. The products and instrument settings cited in SW-846 methods represent those products** 

**and settings used during method development or subs equently evaluated by the Agency.** 

**Glassware, reagents, supplies, equipment, and setti ngs other than those listed in this manual** 

**may be employed provided that method performance ap propriate for the intended application has** 

**been demonstrated and documented.**

**This section does not list common laboratory glassw are (e.g., beakers and flasks).** 

**6.1 Chromatography column -- 300 mm x 10 mm ID, fit ted with a polytetrafluoroethylene** 

**(PTFE) stopcock.**

**NOTE : Columns equipped with fritted glass discs are dif**  **ficult to clean once the column has been**

**used to process highly contaminated extracts. Colu mns without frits may be** 

**purchased, and a small pad of glass wool may be use d to retain the adsorbent.** 

**Prewash the glass wool pad with 50 mL of acetone fo llowed by 50 mL of elution solvent** 

**prior to packing the column with adsorbent.**

**6.2 Muffle furnace -- Capable of maintaining 400** ° **C.**

**6.3 Vials -- Glass, 10-mL and 25-mL capacity, fitte d with polytetrafluoroethylene** 

**(PTFE)-lined screw caps or crimp tops.**

**6.4 Vacuum manifold -- VacElute Manifold SPS-24 (An alytichem International), Visiprep** 

**(Supelco, Inc.), or equivalent, consisting of glass vacuum basin, collection rack and funnel,** 

**collection vials, replaceable stainless steel deliv ery tips, built-in vacuum bleed valve and gauge.** 

**The system is connected to a vacuum pump or water a spirator through a vacuum trap made from** 

**a 500-mL sidearm flask fitted with a one-hole stopp er and glass tubing. The manifold is** 

**necessary for use of the cartridge cleanup protocol**

**. SW-846 Update V 3620C - 4 Revision 4** 

**October 2012 6.5 Top loading balance -- Capable of weighing to 0**  **.01 g.**

**7.0 REAGENTS AND STANDARDS.**

**7.1 Reagent grade chemicals must be used in all tes ts. Unless otherwise indicated, it is** 

**intended that all reagents conform to the specifica tions of the Committee on Analytical Reagents** 

**of the American Chemical Society, where such specif ications are available. Other grades may** 

**be used, provided it is first ascertained that the reagent is of sufficiently high purity to permit it**  **s**

**use without lessening the accuracy of the determina tion. Reagents should be stored in glass to** 

**prevent the leaching of contaminants from plastic c ontainers.** 

**7.2 Organic free reagent water -- All references to water in this method refer to organic** 

**free reagent water, as defined in Chapter One.**

**7.3 Granular Florisil® -- For column cleanup proced ure. Florisil® is produced in four** 

**grades, two of which are appropriate for this proce dure. The differences between grades are** 

**primarily a function of the activation temperature, resulting in somewhat different chemical** 

**characteristics among the grades. Florisil® Grade PR is activated at 675**  ° **C and is most useful**

**for pesticide residue analyses. Florisil® Grade A is activated at 650**  ° **C and is generally used for**

**other analytes. Whichever grade is used, store Flo risil® in glass containers fitted with** 

**ground-glass stoppers or foil-liner screw caps.**

**7.4 Lauric acid -- Reagent grade. Used for the sta ndardization of the Florisil® activity.** 

**Weigh 10.00 g of lauric acid in a 500-mL volumetric flask. Add 50 mL of hexane to the flask to** 

**dissolve the lauric acid. Swirl the flask gently u ntil the lauric acid is dissolved, then dilute the** 

**solution in the flask to 500 mL with additional hex ane.** 

**7.5 Phenolphthalein indicator -- Dissolve 1 g of ph enolphthalein in ethanol and dilute to** 

**100 mL in a volumetric flask.**

**7.6 Sodium hydroxide, NaOH -- Weigh out 20 g of NaO H (pellets, reagent grade) in a** 

**500-mL volumetric flask. Dissolve in organic-free reagent water and dilute to 500 mL to make a** 

**1 N solution. Dilute 25 mL of the 1 N NaOH to 500 mL with water in a second 500-mL volumetric** 

**flask, yielding a 0.05N solution. The NaOH solutio n should be standardized against lauric acid,** 

**as follows.**

**7.6.1 Weigh 100 to 200 mg of lauric acid to the nea rest 1 mg in a 125-mL** 

**Erlenmeyer flask. Add 50 mL of ethanol to the flas k and swirl to dissolve the lauric acid.** 

**7.6.2 Add 3 drops of phenolphthalein indicator to t he flask, and titrate with the** 

**0.05 N NaOH solution to a permanent endpoint (i.e., the indicator color does not disappear** 

**when the solution is allowed to stand for 1 min).**

**7.6.3 Calculate the "strength" of the NaOH solution as the mg of lauric acid** 

**neutralized per mL of NaOH solution.**

**7.7 Deactivation/activation of Florisil®**

**7.7.1 Deactivation of Florisil® -- for cleanup of p hthalate esters. To prepare for** 

**use, place 100 ± 10 g of Florisil® into a 500-mL be aker and heat to 140**  ° **C for approximately**

**16 hrs. After heating, transfer to a 500-mL reagen t bottle. Tightly seal and cool to room** 

**temperature. When cool, add 3 ± 0.1 mL of organic free reagent water. Mix thoroughly by** 

**shaking or rolling for 10 min and let stand for at least 2 hrs. Keep the bottle sealed tightly.** **SW-846 Update V 3620C - 5 Revision 4**

**October 2012 7.7.2 Activation of Florisil® -- for all cleanups o**  **ther than phthalate esters. It is**

**advisable to treat both Florisil® Grade A and Grade PR prior to use to drive off any moisture** 

**adsorbed during storage and handling. Heat the Flo risil® in a glass container loosely** 

**covered with aluminum foil in an oven at 130** ° **C overnight. Cool the Florisil® in a**

**dessicator before use.**

**7.7.3 Florisil® from different batches or sources m ay vary in adsorptive capacity.** 

**To standardize the amount of Florisil® which is use d, use the lauric acid value, described** 

**below. The procedure determines the adsorption fro m a hexane solution of lauric acid (mg)** 

**per g of Florisil®.**

**7.7.3.1 Weigh 2.000 g of Florisil® in a 25-mL, glas s stoppered,** 

**Erlenmeyer flask. Cover loosely with aluminum foil and heat overnight at 130**  ° **C.**

**Stopper the flask and cool to room temperature.**

**7.7.3.2 Add 20.0 mL of the lauric acid solution to the flask, stopper, and** 

**shake occasionally for 15 min.**

**7.7.3.3 Let the Florisil® settle and using a volume tric pipet, transfer** 

**10.0 mL of supernatant liquid into a 125-mL Erlenme yer flask. Avoid including any** 

**Florisil®.**

**7.7.3.4 Add 60 mL of ethanol and 3 drops of the phe nolphthalein** 

**indicator solution to the flask.**

**7.7.3.5 Titrate the solution in the flask with the 0.05N NaOH solution** 

**until a permanent end point is reached (i.e., the i ndicator color does not disappear** 

**when the solution is allowed to stand for 1 min).**

**7.7.3.6 The lauric acid value is calculated as foll ows:** 

Lauric acid value  200 ‐ titration volume in mL o f NaOH strength of NaOH

**Where the strength of the NaOH is measured in Sec. 7.6.3 as the mg of lauric acid** 

**neutralized per mL of NaOH solution.**

**7.7.3.7 Use the following equation to obtain an equ ivalent quantity of** 

**any batch of Florisil®.**

110

lauric acid value  20 g  Required weight of Florisil

**NOTE : This equation was written incorrectly in the prev**  **ious version of this method.**

**7.8 Sodium sulfate (granular, anhydrous), Na 2**  **SO 4**  **-- Purify by heating at 400** ° **C for 4 hrs**

**in a shallow tray, or by precleaning the sodium sul fate with methylene chloride. A method blank** 

**should be analyzed in order to demonstrate that the re is no interference from the sodium sulfate.** 

**7.9 Florisil® cartridges -- 40 μm particles, 60 Å p ores. The cartridges from which this** 

**method were developed consist of 6-mL serological g rade polypropylene tubes, with the 1 g of** 

**Florisil® held between two polyethylene or stainles s steel frits with 20 μm pores. Cartridges** **SW-846 Update V 3620C - 6 Revision 4**

**October 2012 containing 0.5 g and 2.0 g of Florisil® are availab**  **le, however, the compound elution patterns**

**should be verified when cartridges containing other than 1 g of Florisil® are used.** 

**7.10 Elution solvents**

**The choice of solvent will depend on the analytes o f interest and no single solvent is** 

**universally applicable to all analyte groups . Whatever solvent system is employed**  including

**those specifically listed in this method, the analy st**  must **demonstrate adequate performance for**

**the analytes of interest, at the levels of interest . At a minimum, such a demonstration will** 

**encompass the initial demonstration of proficiency described in Method 3500, using a clean** 

**reference matrix. Method 8000 describes procedures that may be used to develop performance** 

**criteria for such demonstrations as well as for mat rix spike and laboratory control sample results.** 

**All solvents should be pesticide quality or equival ent. Solvents may be degassed prior to** 

**use.**

**7.10.1 Diethyl ether, C 2**  **H 5**  **OC 2**  **H 5**  **-- Must be free of peroxides as indicated by test**

**strips (EM Quant, or equivalent). Procedures for r emoval of peroxides are provided with** 

**the test strips. After cleanup, 20 mL of ethyl alc ohol preservative must be added to each** 

**liter of ether.**

**7.10.2 Pentane, CH 3**  **(CH 2**  **) 3**  **CH 3** 

**7.10.3 Hexane, C 6**  **H 14** 

**7.10.4 Methylene chloride, CH 2**  **Cl 2** 

**7.10.5 Acetone, CH 3**  **COCH 3** 

**7.10.6 Petroleum ether (boiling range of 30 - 60** ° **C)**

**7.10.7 Toluene, C 6**  **H 5**  **CH 3** 

**7.10.8 2-Propanol, (CH 3**  **) 2**  **CHOH**

**7.11 Florisil® cartridge phenol check solution (for the organochlorine pesticide technique)** 

**-- Prepare a solution of 2,4,5-trichlorophenol in a cetone at a concentration of 0.1 mg/L. See the** 

**note in Sec. 7.12.**

**7.12 Florisil® cartridge pesticide check solution - - Prepare a solution containing the** 

**following analytes in hexane:**

**α-BHC 5 μg/L**  **4,4'-DDD 10 μg/L** 

**Heptachlor 5 μg/L**  **4,4'-DDT 10 μg/L** 

**γ-BHC 5 μg/L**  **Methoxychlor 50 μg/L** 

**Endosulfan I 5 μg/L**  **Tetrachloro-** m **-xylene 20 μg/L** 

**Dieldrin 10 μg/L**  **Decachlorobiphenyl 20 μg/L** 

**Endrin 10 μg/L** 

**NOTE : The concentrations of these analytes were incorre**  **ctly listed in units of mg/L in the**

**previous version of this method. SW-846 Update V 3620C - 7 Revision 4** 

**October 2012 7.13 Chlorophenoxy acid herbicide check solution --**  **Prepare a solution containing**

**2,4,5-T methyl ester at 100 μg/L, pentachlorophenyl methyl ester at 50 μg/L, and picloram methyl** 

**ester at 200 μg/L. See the note in Sec. 7.12.**

**8.0 SAMPLE COLLECTION, PRESERVATION, AND STORAGE**

**See the introductory material to Chapter Four, "Org anic Analytes" and the specific** 

**determinative methods to be employed**

**9.0 QUALITY CONTROL**

**9.1 Refer to Chapter One for additional guidance on quality assurance (QA) and quality** 

**control (QC) protocols. When inconsistencies exist between QC guidelines, method-specific QC** 

**criteria take precedence over both technique-specif ic criteria and those criteria given in Chapter** 

**One, and technique-specific QC criteria take preced ence over the criteria in Chapter One. Any** 

**effort involving the collection of analytical data should include development of a structured and** 

**systematic planning document, such as a Quality Ass urance Project Plan (QAPP) or a Sampling** 

**and Analysis Plan (SAP), which translates project o bjectives and specifications into directions for** 

**those that will implement the project and assess th e results. Each laboratory should maintain a** 

**formal quality assurance program. The laboratory s hould also maintain records to document the** 

**quality of the data generated. All data sheets and quality control data should be maintained for** 

**reference or inspection.**

**9.2 Also refer to Method 3600 for cleanup QC proced ures, and Method 8000 and the** 

**specific determinative method to be used for inform ation on determinative QC procedures.** 

**9.3 Initially, before processing any samples, the a nalyst should demonstrate that all** 

**parts of the equipment in contact with the sample a nd reagents are interference-free. This is** 

**accomplished through the analysis of a method blank . As a continuing check, each time** 

**samples are extracted, cleaned up, and analyzed, an d when there is a change in reagents, a** 

**method blank should be prepared and analyzed for th e compounds of interest as a safeguard** 

**against chronic laboratory contamination.**

**9.4 The analyst must demonstrate that the compounds of interest are being** 

**quantitatively recovered by the cleanup technique b efore applying this method to actual samples.** 

**This test applies to both the column cleanup and ca rtridge cleanup procedures. A recovery** 

**check needs to be performed using standards of the target analytes at known concentration.** 

**9.4.1 This test needs to be conducted on each batch of Florisil® following its** 

**activation (Sec. 7.3).**

**9.4.2 The efficiency of each lot of the solid-phase extraction cartridges needs to** 

**be verified. Only lots of cartridges from which th e spiked analytes are quantitatively** 

**recovered may be used to process the samples. A ch eck should also be performed at least** 

**once on each individual lot of cartridges and at le ast once for every 300 cartridges of a** 

**particular lot, whichever frequency is greater.**

**9.4.3 Organochlorine pesticides --To check each new lot of Florisil® cartridges** 

**before use, perform the following in duplicate:**

**9.4.3.1 Combine 0.5 mL of the 2,4,5-trichlorophenol solution in Sec.** 

**7.11, 1.0 mL of the pesticide solution in Sec. 7.12 , and 0.5 mL of hexane in a vial.** **SW-846 Update V 3620C - 8 Revision 4**

**October 2012 9.4.3.2 Condition the cartridge as described in Sec**  **. 11.1 and then**

**perform the cartridge cleanup starting with Sec. 11 .7.** 

**9.4.3.3 Elute the cartridge with 9 mL of acetone/he xane (10/90, v/v)** 

**only. Reduce the volume to 1.0 mL and analyze by M ethod 8081.** 

**9.4.3.4 The lot of Florisil® cartridges is acceptab le if all pesticides are** 

**recovered at 80 to 110%, if the recovery of trichlo rophenol is less than 5%, and if no** 

**peaks interfering with the target analytes are dete cted.** 

**9.4.4 Chlorophenoxy acid herbicides -- To check eac h new lot of granular** 

**Florisil® perform the following:**

**9.4.4.1 Add 5 mL of the chlorophenoxy acid herbicid e check solution** 

**(Sec. 7.13) to a Florisil® column packed and washed as in Sec. 11.13.2.** 

**9.4.4.2 Elute Fractions 1 and 2 as described in Sec s. 11.13.3 and** 

**11.13.4, collecting each in a separate flask.**

**9.4.4.3 Elute the column with approximately 100 mL of diethyl ether** 

**and collect ten separate 10-mL fractions.**

**9.4.4.4 Concentrate Fraction 1 and Fraction 2 separ ately and** 

**concentrate each of the ten 10-mL diethyl ether fra ctions to 5 mL** 

**9.4.4.5 Analyze each of the 12 eluates by GC/ECD an d calculate the** 

**recovery of each analyte. Pentachlorophenyl methyl ether should be found in** 

**Fraction 1. 2,4,5-T methyl ester (and the methyl e sters of the other chlorophenoxy** 

**acids) should be found in Fraction 2. Determine th e volume of diethyl ether that is** 

**necessary to elute picloram methyl ester.**

**9.4.4.6 The lot of Florisil® is acceptable if the t arget analytes are** 

**quantitatively recovered and if the recovery of tri chlorophenol is less than 5%. No** 

**interferences should be detected in any of these el uates.** 

**9.5 Any method blanks, matrix spike samples, or rep licate samples should be subjected** 

**to the same analytical procedures (Sec. 11.0) as th ose used on actual samples. For sample** 

**extracts that are cleaned up using this method, the associated quality control samples should also** 

**be processed through this cleanup method.**

**10.0 CALIBRATION AND STANDARDIZATION**

**There are no calibration or standardization steps d irectly associated with this cleanup** 

**procedure. SW-846 Update V 3620C - 9 Revision 4** 

**October 2012 11.0 PROCEDURE** 

**Sec. 11.1 describes the procedures for assembling a nd conditioning the Florisil® cartridges.** 

**Sec. 11.2 describes general procedures for handling sample extracts prior to cleanup. Secs.** 

**11.3 through 11.13 describe the column and cartridg e procedures for phthalate esters (Secs. 11.3** 

**and 11.4); nitrosamines (Sec. 11.5); organochlorine pesticides, haloethers, and** 

**organophosphorus pesticides (Secs. 11.6 and 11.7); nitroaromatics and isophorone (Sec. 11.8);** 

**chlorinated hydrocarbons (Secs. 11.9 and 11.10); an iline and aniline derivatives (Sec. 11.11);** 

**organophosphates (Sec. 11.12); and derivatized chlo rophenoxy acid herbicides (Sec. 11.13).** 

**The column chromatography procedures employ a large r amount of Florisil® than the** 

**cartridge procedures and, therefore, have a greater cleanup capacity. Samples that exhibit** 

**greater degrees of interferences should be cleaned up using the column procedures. However,** 

**both techniques have limitations on the amount of i nterferences which can be removed.** 

**If the interference is caused by high boiling mater ials, then Method 3640 should be** 

**employed prior to Florisil® cleanup. If the interf erence is caused by relatively polar compounds** 

**in the same boiling range as the analytes of intere st, then multiple column or cartridge cleanups** 

**may be necessary. For additional cleanup of organo chlorine pesticides and PCBs, see Method** 

**3665. If crystals of sulfur are present in the ext ract, then Method 3660 should be employed prior** 

**to Florisil® cleanup.**

**Whenever Florisil® is used to fractionate groups of target compounds (rather than to simply** 

**remove potential interferants) it is critical that the specific fractionation scheme be validated usin**  **g**

**spiked solutions or spiked sample extracts that con tain most or all of the analytes of interest.** 

**This may be particularly important when the Florisi l® cartridge techniques are employed, because** 

**the differences between the various cartridge forma ts and manufacturers may affect the** 

**fractionation patterns. In addition, it may be use ful to archive any fractions not originally intende**  **d**

**for analysis, in the event that the fractionation s cheme chosen does not yield the intended results.** 

**Once the determinative analysis has been performed and demonstrates that the fractionation has** 

**been successful, such archived fractions may be dis posed of in an appropriate manner.** 

**However, if the fractionation did not perform as in tended, the analytes of interest may be** 

**contained in the archived fractions which may be an alyzed or combined with the other fraction(s)** 

**for reanalysis.**

**Following Florisil® cleanup, extracts may need furt her concentration and/or solvent** 

**exchange. Consult the appropriate determinative me thod and 3500 series extraction method for** 

**details.**

**11.1 Cartridge set-up and conditioning**

**11.1.1 Arrange the cartridges on the manifold in th e closed-valve position.** 

**11.1.2 Turn on the vacuum pump and set the vacuum t o 10 in (254 mm) of Hg.** 

**Do not exceed the manufacturer's recommendation for manifold vacuum. Flow rates may** 

**be controlled by opening and closing cartridge valv es.** 

**11.1.3 Condition the cartridges by adding 4 mL of h exane to each cartridge.** 

**Slowly open the cartridge valves to allow hexane to pass through the sorbent beds to the** 

**lower frits. Allow a few drops per cartridge to pa ss through the manifold to remove all air** 

**bubbles. Close the valves and allow the solvent to soak the entire sorbent bed for 5** 

**minutes. Do not turn off the vacuum. SW-846 Update V 3620C - 10 Revision 4** 

**October 2012 11.1.4 Slowly open cartridge valves to allow the he**  **xane to pass through the**

**cartridges. Close the cartridge valves when there is still at least 1 mm of solvent above the** 

**sorbent bed. Do not allow the cartridges to become dry. If cartridges go dry, repeat the** 

**conditioning step.**

**11.2 Handling sample extracts**

**Most sample extracts have to be concentrated to a s maller volume prior to the use of** 

**Florisil® cleanup. The extract volume is a functio n of the analytical sensitivity necessary to meet** 

**the project objectives. The extract volume will al so affect the ability of the Florisil® to separate** 

**target analytes from potential interferences, parti cularly for the cartridge procedures, where** 

**applying large extract volumes to the cartridges ma y cause poor results. As noted in Sec. 11.0,** 

**consult the appropriate extraction and determinativ e methods for the details on final extract** 

**volumes, extract concentration techniques, and solv ent exchange procedures.** 

**11.2.1 Reduce the sample extract volume to 2 mL pri or to cleanup for:** 

**Phthalate esters Chlorinated hydrocarbons** 

**Nitrosamines Chlorophenoxy acid herbicides** 

**Nitroaromatics and isophorone Aniline and aniline derivatives** 

**The extract solvent should be hexane for the phthal ate esters, nitroaromatics,** 

**chlorinated hydrocarbons, and chlorophenoxy acid he rbicides, and methylene chloride for** 

**the nitrosamines and aniline and aniline derivative s.** 

**11.2.2 Reduce the sample extract volume to 10 mL pr ior to cleanup for:** 

**Organochlorine pesticides Organophosphates** 

**Haloethers PCBs** 

**Organophosphorus pesticides**

**The extract solvent should be hexane for these anal ytes. In most cases, given the** 

**sensitivity of the determinative methods, only 1 mL of the 10 mL extract needs to be** 

**subjected to the Florisil® cleanup procedure. The remaining 9 mL should be archived for** 

**later use, if needed.**

**11.2.3 If the extract was in cold storage, allow it to reach room temperature.** 

**Inspect the extract visually to ensure that there a re no particulates or phase separations and** 

**that no evaporative loss has taken place. If cryst als of sulfur are visible or if the presence of** 

**sulfur is suspected, proceed with Method 3660.**

**11.3 Column procedure for phthalate esters**

**11.3.1 Place approximately 10 g of deactivated Flor isil® (Sec. 7.7.1) into a 10 mm** 

**ID chromatographic column. Tap the column to settl e the Florisil® and add approximately** 

**1 cm of anhydrous sodium sulfate to the top.**

**11.3.2 Pre-elute the column with 40 mL of hexane. The rate for all elutions** 

**should be about 2 mL/min. Discard the eluate and, just prior to exposure of the sodium** 

**sulfate layer to the air; quantitatively transfer t he 2-mL sample extract onto the column using** 

**an additional 2 mL of hexane to complete the transf er.** **SW-846 Update V 3620C - 11 Revision 4**

**October 2012 11.3.3 Just prior to exposure of the sodium sulfate**  **layer to the air, add 40 mL of**

**hexane and continue the elution of the column. Dis card this hexane eluate.** 

**11.3.4 Elute the column with 100 mL of ethyl ether/ hexane (20/80, v/v) and collect** 

**this fraction in a flask (e.g., a 500-mL K-D flask equipped with a clean 10 mL concentrator** 

**tube). Concentrate the eluate to the volume listed in the determinative method, using the** 

**techniques described in the appropriate 3500 series method. No solvent exchange is** 

**necessary. Compounds that elute in this fraction a re:** 

**Bis(2-ethylhexyl) phthalate Diethyl phthalate** 

**Butyl benzyl phthalate Dimethyl phthalate** 

**Di-** n **-butyl phthalate Di-**  n **-octyl phthalate**

**11.4 Cartridge procedure for phthalate esters**

**11.4.1 Using 1-g Florisil® cartridges, condition th e cartridges with hexane as** 

**described in Sec. 11.1.**

**11.4.2 Transfer the extract (Sec. 11.2) to the cart ridge. Open the cartridge valve** 

**to allow the extract to pass through the cartridge bed at approximately 2 mL/min.** 

**11.4.3 When the entire extract has passed through t he cartridges, but before the** 

**cartridges become dry, rinse the sample vials with an additional 0.5 mL of solvent, and add** 

**the rinse to the cartridges to complete the quantit ative transfer.** 

**11.4.4 Close the cartridge valve and turn off the v acuum after the solvent has** 

**passed through, ensuring that the cartridge never g ets dry.** 

**11.4.5 Place a 5-mL vial or volumetric flask into t he sample rack corresponding to** 

**the cartridge position. Attach a solvent rinsed st ainless steel solvent guide to the manifold** 

**cover and align it with the collection vial.**

**11.4.6 If the sample is suspected to contain organo chlorine pesticides, elute the** 

**cartridge with methylene chloride/hexane (20/80, v/ v). Turn on the vacuum pump and** 

**adjust the pump pressure to 10 in (254 mm) of Hg. Allow the solvent to soak the sorbent** 

**bed for 1 min or less. Slowly open the cartridge v alve, and collect the eluate (this fraction** 

**contains the organochlorine pesticides, and should be discarded).** 

**11.4.7 Close the cartridge valve, replace collectio n vials, and add 10 mL of** 

**acetone/hexane (10/90, v/v) to the cartridge. Slow ly open the cartridge valve and collect** 

**the eluate into the collection vial. This fraction contains the phthalate esters, and should be** 

**retained for analysis.**

**11.4.8 Concentrate the eluate to the volume listed in the determinative method,** 

**using the techniques described in the appropriate 3 500 series method.** 

**11.5 Column procedure for nitrosamines**

**11.5.1 Add a weight of activated Florisil® (nominal ly 22 g) predetermined by** 

**calibration (Sec. 7.7.3.7) into a 20 mm ID chromato graphic column. Tap the column to** 

**settle the Florisil® and add about 5 mm of anhydrou s sodium sulfate to the top.** 

**11.5.2 Pre-elute the column with 40 mL of ethyl eth er/pentane (15/85, v/v).** 

**Discard the eluate and, just prior to exposure of t he sodium sulfate layer to the air,** **SW-846 Update V 3620C - 12 Revision 4**

**October 2012 quantitatively transfer the 2-mL sample extract (Se**  **c. 11.2) onto the column using an**

**additional 2 mL of pentane to complete the transfer**

**.**

**11.5.3 Just prior to the exposure of the sodium sul fate layer to the air, elute the** 

**column with 90 mL of ethyl ether/pentane (15/85, v/ v). Discard the eluate. This fraction** 

**will contain any diphenylamine present in the extra ct.** 

**11.5.4 Elute the column with 100 mL of acetone/ethy l ether (5/95, v/v), collecting** 

**the eluate in a flask (e.g., a 500 mL K-D flask equ ipped with a clean 10 mL concentrator** 

**tube). This fraction will contain all of the nitro samines listed in the scope of the method.** 

**11.5.5 Add 15 mL of methanol to the collected fract ion, and concentrate this** 

**fraction to the volume listed in the determinative method, using the techniques described in** 

**the appropriate 3500 series method.**

**11.6 Column procedure for organochlorine pesticides , haloethers, and** 

**organophosphorus pesticides (see Table 6 for fracti onation patterns of organophosphorus** 

**pesticides).**

**11.6.1 Add a weight of activated Florisil® (nominal ly 20 g), predetermined by** 

**calibration (Sec. 7.7.3.7), to a 20 mm ID chromatog raphic column. Settle the Florisil® by** 

**tapping the column. Add anhydrous sodium sulfate t o the top of the Florisil® to form a layer** 

**1 to 2 cm deep.**

**11.6.2 Pre-elute the column with 60 mL of hexane an d discard the eluate. Just** 

**prior to exposure of the sodium sulfate to air, qua ntitatively transfer the 10-mL sample** 

**extract (Sec. 11.2) onto the column, completing the transfer with two 1- to 2-mL rinses with** 

**hexane.**

**11.6.3 Place a flask (e.g., a 500-mL K-D flask equi pped with a clean concentrator** 

**tube) under the chromatographic column. Drain the column into the flask until the sodium** 

**sulfate layer is nearly exposed. Elute the column with 200 mL of ethyl ether/hexane (6/94,** 

**v/v) using a drip rate of about 5 mL/min. This is Fraction 1, and all of the haloethers are in** 

**this fraction. Remove the flask and set aside for later concentration.** 

**11.6.4 Elute the column again, using 200 mL of ethy l ether/hexane (15/85, v/v),** 

**into a second flask. This is Fraction 2.**

**11.6.5 Perform a third elution using 200 mL of diet hyl ether/hexane (50/50, v/v),** 

**collecting the eluate in a third flask. This is Fr action 3.** 

**11.6.6 Perform a final elution with 200 mL of 100% ethyl ether, collecting the** 

**eluate in a fourth flask. This is Fraction 4.**

**11.6.7 Concentrate the four eluates to the volume l isted in the determinative** 

**method, using the techniques described in the appro priate 3500 series method.** 

**11.7 Cartridge procedure for organochlorine pestici des and PCBs** 

**11.7.1 Using 1-g Florisil® cartridges, condition th e cartridges with hexane, as** 

**described in Sec. 11.1. SW-846 Update V 3620C - 13 Revision 4** 

**October 2012 11.7.2 Transfer the 1 mL (or other appropriate volu**  **me) of the extract (Sec. 11.2) to**

**the cartridge. Open the cartridge valve to allow t he extract to pass through the cartridge** 

**bed at approximately 2 mL/min.**

**11.7.3 When the entire extract has passed through t he cartridge, but before the** 

**cartridge becomes dry, rinse the sample vial with a n additional 0.5 mL of hexane, and add** 

**the rinse to the cartridge to complete the quantita tive transfer.** 

**11.7.4 Close the cartridge valve and turn off the v acuum after the solvent has** 

**passed through, ensuring that the cartridge never g oes dry.** 

**11.7.5 Place a 10-mL vial or volumetric flask into the sample rack corresponding** 

**to the cartridge position. Attach a solvent rinsed stainless steel solvent guide to the** 

**manifold cover and align with the collection vial.**

**11.7.6 If there is no need to separate the organoch lorine pesticides from the** 

**PCBs, then add 9 mL of acetone/hexane (10/90, v/v) to the cartridge. Turn on the vacuum** 

**pump and adjust the pump pressure to 10 in (254 mm) of Hg. Allow the solvent to soak the** 

**sorbent bed for 1 min or less. Slowly open the car tridge valve and collect the eluate into the** 

**collection vial. Go directly to Sec. 11.7.8.**

**11.7.7 The following procedures are used to separat e the organochlorine** 

**pesticides from the PCBs.**

**11.7.7.1 Add 3 mL of hexane to the cartridge. Turn on the vacuum** 

**pump and adjust the pump pressure to 10 inches (254 mm) of Hg. Allow the solvent** 

**to soak the sorbent bed for 1 min or less. Slowly open the cartridge valve and** 

**collect the eluate into the collection vial. This is Fraction 1 and it will contain the** 

**PCBs and a few of the organochlorine pesticides (se e Table 5).** 

**11.7.7.2 Close the cartridge valve, replace the col lection vial, and add 5** 

**mL of methylene chloride/hexane (26/74, v/v) to the cartridge. Slowly open the** 

**cartridge valve and collect the eluate into the col lection vial. This is Fraction 2 and it** 

**will contain most of the pesticides.**

**11.7.7.3 Close the cartridge valve, replace collect ion vials, and add 5 mL** 

**of acetone/hexane (10/90, v/v) to the cartridge. S lowly open the cartridge valve and** 

**collect the eluate into the collection vial. This is Fraction 3 and it will contain the** 

**remaining pesticides.**

**11.7.8 As needed, perform a solvent exchange and ad just the final volume of the** 

**eluant to the volume listed in the determinative me thod, using the techniques described in** 

**the appropriate 3500 series method.**

**11.8 Column procedure for nitroaromatics and isopho rone** 

**11.8.1 Add a weight of activated Florisil® (nominal ly 10 g) predetermined by** 

**calibration (Sec. 7.7.3.7) into a 10 mm ID chromato graphic column. Tap the column to** 

**settle the Florisil® and add about 1 cm of anhydrou s sodium sulfate to the top.** 

**11.8.2 Pre-elute the column with methylene chloride /hexane (10/90, v/v) at about** 

**2 mL/min. Discard the eluate and, just prior to ex posure of the sodium sulfate layer to the** 

**air, quantitatively transfer the 2-mL sample extrac t (Sec. 11.2) onto the column using an** 

**additional 2 mL of hexane to complete the transfer. SW-846 Update V 3620C - 14 Revision 4** 

**October 2012 11.8.3 Just prior to exposure of the sodium sulfate**  **layer to the air, add 30 mL of**

**methylene chloride/hexane (10/90, v/v) and continue the elution of the column. Discard the** 

**eluate.**

**11.8.4 Elute the column with 90 mL of ethyl ether/p entane (15/85, v/v) and discard** 

**the eluate. This fraction will contain any dipheny lamine present in the extract.** 

**11.8.5 Elute the column with 100 mL of acetone/ethy l ether (5/95, v/v), and collect** 

**the eluate in a flask (e.g., a 500-mL K-D flask equ ipped with a 10-mL concentrator tube).** 

**This fraction will contain all of the nitrosamines listed in the scope of the method.** 

**11.8.6 Add 15 mL of methanol to the collected fract ion, and concentrate to the** 

**volume listed in the determinative method, using th e techniques described in the** 

**appropriate 3500 series method.**

**11.8.7 Elute the column with 30 mL of acetone/methy lene chloride (10/90, v/v),** 

**and collect the eluate in a flask (e.g., a 500-mL K -D flask equipped with a 10-mL** 

**concentrator tube). Concentrate the collected frac tion to the volume listed in the** 

**determinative method, using the techniques describe d in the appropriate 3500 series** 

**method, and exchanging the solvent to hexane. Comp ounds that elute in this fraction are:** 

**2,4-Dinitrotoluene Isophorone** 

**2,6-Dinitrotoluene Nitrobenzene** 

**11.9 Column procedure for chlorinated hydrocarbons**

**11.9.1 Add a weight of activated Florisil® (nominal ly 12 g) predetermined by** 

**calibration (Sec. 7.7.3.7) into a 10 mm ID chromato graphic column. Tap the column to** 

**settle the Florisil® and add about 1 to 2 cm of anh ydrous sodium sulfate to the top.** 

**11.9.2 Pre-elute the column with 100 mL of petroleu m ether. Discard the eluate** 

**and, just prior to exposure of the sodium sulfate l ayer to the air, quantitatively transfer the** 

**sample extract (Sec. 11.2) to the column by decanta tion and subsequent petroleum ether** 

**washings. Discard the eluate.**

**11.9.3 Just prior to exposure of the sodium sulfate layer to the air, begin eluting** 

**the column with 200 mL of petroleum ether and colle ct the eluate in flask (e.g., a 500-mL** 

**K-D flask equipped with a 10-mL concentrator tube). This fraction should contain the** 

**following chlorinated hydrocarbons:**

**2-Chloronaphthalene Hexachlorobenzene** 

**1,2-Dichlorobenzene Hexachlorobutadiene** 

**1,3-Dichlorobenzene Hexachlorocyclopentadiene** 

**1,4-Dichlorobenzene Hexachloroethane** 

**1,2,4-Trichlorobenzene**

**11.9.4 Concentrate the collected fraction to the vo lume listed in the determinative** 

**method, using the techniques described in the appro priate 3500 series method.** **SW-846 Update V 3620C - 15 Revision 4**

**October 2012 11.10 Cartridge procedure for chlorinated hydrocarb**  **ons**

**11.10.1 Using 1-g Florisil® cartridges, condition t he cartridges with 5 mL of** 

**acetone/hexane (10/90, v/v) as described in Sec. 11 .1.** 

**11.10.2 Transfer the extract (Sec. 11.2) to the car tridge. Open the cartridge valve** 

**to allow the extract to pass through the cartridge bed at approximately 2 mL/min.** 

**11.10.3 When the entire extract has passed through the cartridge, but before the** 

**cartridge becomes dry, rinse the sample vial with a n additional 0.5 mL of acetone/hexane** 

**(10/90), and add the rinse to the cartridge to comp lete the quantitative transfer.** 

**11.10.4 Close the cartridge valve and turn off the vacuum after the solvent has** 

**passed through, ensuring that the cartridge never g ets dry.** 

**11.10.5 Place a 5-mL vial or volumetric flask into the sample rack corresponding to** 

**the cartridge position. Attach a solvent rinsed st ainless steel solvent guide to the manifold** 

**cover and align it with the collection vial.**

**11.10.6 Add 10 mL of acetone/hexane (10/90, v/v) to the cartridge. Turn on the** 

**vacuum pump and adjust the pump pressure to 10 in ( 254 mm) of Hg. Allow the solvent to** 

**soak the sorbent bed for 1 min or less. Slowly op en the cartridge valve and collect the** 

**eluate into the collection vial.**

**11.10.7 Adjust the final volume of the eluant to th e volume listed in the** 

**determinative method, using the techniques describe d in the appropriate 3500 series** 

**method.**

**11.11 Column procedure for aniline and aniline deri vatives (see Table 8 for elution** 

**patterns)**

**11.11.1 Add a weight of activated Florisil® predete rmined by calibration (Sec.** 

**7.7.3.7) into a 20 mm ID chromatographic column. T ap the column to settle the Florisil®.** 

**11.11.2 Pre-elute the column with 100 mL of 2-propa nol/methylene chloride (5/95,** 

**v/v), followed by 100 mL of hexane/methylene chlori de (50/50, v/v), followed by 100 mL of** 

**hexane. Discard the eluate and leave a column of a bout 5 cm of hexane above the** 

**Florisil®.**

**11.11.3 Quantitatively transfer the 2-mL sample ext ract (Sec. 11.2) onto 2.0 g of** 

**activated Florisil® in a 50-mL beaker, using a smal l volume of methylene chloride, and dry** 

**under a gentle stream of nitrogen.**

**11.11.4 Place the dried Florisil® containing the sa mple extract onto the** 

**chromatographic column, and wash the beaker which c ontained the Florisil® with 75 mL of** 

**hexane, adding this wash to the reservoir.**

**11.11.5 Elute the hexane from the column and discar d. Stop the column flow just** 

**prior to the exposure of the Florisil® to air.**

**11.11.6 Elute the column with 50 mL of methylene ch loride/hexane (50/50, v/v),** 

**using a drip rate of about 5 mL/min, and collect th e eluate in a flask (e.g., a 500-mL K-D flask** 

**equipped with a 10-mL concentrator tube). This is Fraction 1.** **SW-846 Update V 3620C - 16 Revision 4**

**October 2012 11.11.7 Elute the column with 50 mL of 2-propanol/h**  **exane (5/95, v/v), and collect**

**the eluate in a second flask. This is Fraction 2.**

**11.11.8 Elute the column a third time using 50 mL o f methanol/hexane (5/95, v/v).** 

**Collect the eluate in a third flask. This is Fract ion 3. Frequently, it will prove useful to** 

**combine the three fractions prior to analysis. How ever, in some situations, analysis of each** 

**separate fraction may be necessary. Refer to Metho d 8131.** 

**11.11.9 Concentrate the collected fractions to the volume listed in the determinative** 

**method, using the techniques described in the appro priate 3500 series method.** 

**11.12 Column procedure for organophosphates**

**11.12.1 Add a weight of activated Florisil®, predet ermined by calibration (Sec.** 

**7.7.3.7), to a 20 mm ID chromatographic column. Se ttle the Florisil® by tapping the** 

**column. Add anhydrous sodium sulfate to the top of the Florisil® to form a layer 1 to 2 cm** 

**deep.**

**11.12.2 Pre-elute the column with 50-60 mL of hexan e. Discard the eluate and** 

**just prior to exposure of the sulfate layer to air, quantitatively transfer the 10-mL sample** 

**extract (Sec. 11.2) onto the column using a hexane wash to complete the transfer.** 

**11.12.3 Just as the sample reaches the sodium sulfa te, elute the column with 100** 

**mL of diethyl ether/hexane (10/90, v/v). Discard t he eluate.** 

**11.12.4 Just prior to exposure of the sodium sulfat e to air, elute the column with 200** 

**mL of diethyl ether/hexane (30/70, v/v). This frac tion contains all of the target analytes** 

**except for tris(2,3-dibromopropyl) phosphate.**

**11.12.5 Elute the column with 200 mL of diethyl eth er/hexane (40/60, v/v). This** 

**fraction contains tris(2,3-dibromopropyl) phosphate**

**.**

**11.12.6 Concentrate the collected fraction to the v olume listed in the determinative** 

**method, using the techniques described in the appro priate 3500 series method.** 

**11.13 Column procedure for derivatized chlorophenox y acid herbicides** 

**11.13.1 Add a weight of activated Florisil® (nomina lly 4 g) predetermined by** 

**calibration (Sec. 7.7.3.7) into a 20 mm ID chromato graphic column. Tap the column to** 

**settle the Florisil® and add approximately 5 mm of anhydrous sodium sulfate to the top.** 

**11.13.2 Pre-elute the column with 15 mL of hexane. The rate for all elutions** 

**should be about 2 mL/min. Discard the eluate, and just prior to exposure of the sodium** 

**sulfate to air, quantitatively transfer the 2-mL sa mple extract (Sec. 11.2) onto the column,** 

**using an additional 2 mL of hexane to complete the transfer.** 

**11.13.3 Just prior to the exposure of the sodium su lfate layer to the air, elute the** 

**column with 35 mL of methylene chloride/hexane (20/ 80, v/v), collecting the eluate in a** 

**clean flask (e.g., a 500-mL K-D flask equipped with a concentrator tube). This is Fraction 1,** 

**and will contain any pentachlorophenyl methyl ester that is present.** 

**11.13.4 Elute the column with 60 mL of methylene ch loride/acetonitrile/hexane** 

**(50/0.35/49.65, v/v/v), collecting the eluate in a second flask. This is Fraction 2.** **SW-846 Update V 3620C - 17 Revision 4**

**October 2012 11.13.5 If picloram is to be determined, perform a**  **third elution with the volume of**

**diethyl ether determined from the Florisil® check i n Sec. 9.4.4, collecting this eluate in a** 

**third flask. This is Fraction 3, and will contain the picloram.** 

**11.13.6 The three fractions may be combined for ana lysis. Concentrate the** 

**combined fractions to the volume listed in the dete rminative method, using the techniques** 

**described in the appropriate 3500 series method.**

**12.0 DATA ANALYSIS AND CALCULATIONS**

**There are no calculations explicitly associated wit h this cleanup procedure. See the** 

**appropriate determinative method for the calculatio n of final sample results.** 

**13.0 METHOD PERFORMANCE**

**13.1 Performance data and related information are p rovided in SW-846 methods only as** 

**examples and guidance. The data do not represent r equired performance criteria for users of** 

**the methods. Instead, performance criteria should be developed on a project-specific basis,** 

**and the laboratory should establish in-house QC per formance criteria for the application of this** 

**method. These performance data are not intended to be and must not be used as absolute QC** 

**acceptance criteria for purposes of laboratory accr editation.** 

**13.2 Table 1 provides example recoveries of phthala te esters obtained from the Florisil®** 

**column procedure. These data are provided for guid ance purposes only.** 

**13.3 Table 2 provides example recoveries of phthala te esters obtained from the Florisil®** 

**cartridge procedure. These data are provided for g uidance purposes only.** 

**13.4 Table 3 provides an example distribution of or ganochlorine pesticides and PCBs** 

**from the Florisil® column procedure. These data ar e provided for guidance purposes only.** 

**13.5 Table 4 provides example recoveries of Aroclor s from the Florisil® cartridge** 

**procedure. These data are provided for guidance pu rposes only.** 

**13.6 Table 5 provides an example distribution of or ganochlorine pesticides from the** 

**Florisil® cartridge procedure, using 1-g cartridges . These data are provided for guidance** 

**purposes only.**

**13.7 Table 6 provides an example distribution of or ganophosphorus pesticides from the** 

**Florisil® column procedure. These data are provide d for guidance purposes only.** 

**13.8 Table 7 provides example recoveries of chlorin ated hydrocarbons obtained from the** 

**Florisil® cartridge procedure. These data are prov ided for guidance purposes only.** 

**13.9 Table 8 provides example elution patterns for aniline compounds from the Florisil®** 

**column procedure. These data are provided for guid ance purposes only.** **SW-846 Update V 3620C - 18 Revision 4**

**October 2012 14.0 POLLUTION PREVENTION** 

**14.1 Pollution prevention encompasses any techniqu e that reduces or eliminates the** 

**quantity and/or toxicity of waste at the point of g eneration. Numerous opportunities for pollution** 

**prevention exist in laboratory operation. The EPA has established a preferred hierarchy of** 

**environmental management techniques that places pol lution prevention as the management** 

**option of first choice. Whenever feasible, laborat ory personnel should use pollution prevention** 

**techniques to address their waste generation. When wastes cannot be feasibly reduced at the** 

**source, the Agency recommends recycling as the next best option.** 

**14.2 For information about pollution prevention th at may be applicable to laboratories** 

**and research institutions consult** Less is Better: Laboratory Chemical Management for Waste

Reduction, **a free publication available from the American Che mical Society (ACS), Committee on** 

**Chemical Safety, http://portal.acs.org/portal/fileF etch/C/WPCP_012290/pdf/WPCP_012290.pdf.** 

**15.0 WASTE MANAGEMENT**

**The Environmental Protection Agency requires that l aboratory waste management** 

**practices be conducted consistent with all applicab le rules and regulations. The Agency urges** 

**laboratories to protect the air, water, and land by minimizing and controlling all releases from** 

**hoods and bench operations, complying with the lett er and spirit of any sewer discharge permits** 

**and regulations, and by complying with all solid an d hazardous waste regulations, particularly the** 

**hazardous waste identification rules and land dispo sal restrictions. For further information on** 

**waste management, consult** The Waste Management Manual for Laboratory Personne l **available**

**from the American Chemical Society at the address l isted in Sec. 14.2.** 

**16.0 REFERENCES**

**1. A. J. Gordon and R. A. Ford, The Chemist's Compa nion: A Handbook of Practical Data,** 

**Techniques , and References, New York: John Wiley & Sons, Inc**  **., pp. 372, 374, and 375,**

**1972.**

**2. Floridin of ITT System, Florisil: Properties, Ap plication, Bibliography**  **, Pittsburgh,**

**Pennsylvania, 5M381DW.**

**3. P. A. Mills, "Variation of Florisil Activity; Si mple Method for Measuring Absorbent Capacity** 

**and its use in Standardizing Florisil Columns,"** Journal of the Association of Official

Analytical Chemists **, 51, 29, 1968.**

**4. U.S. Food and Drug Association, Pesticides Analy tical Manual**  **(Volume 1), July 1985.**

**5. V. Lopez-Avila, J. Milanes, N. S. Dodhiwala, an d W.F. Beckert, "Cleanup of Environmental** 

**Sample Extracts Using Florisil Solid-Phase Extracti on Cartridges,"**  J. Chrom. Sci. **27,**

**209-215, 1989.**

**6. U.S. EPA, "Evaluation of Sample Extract Cleanup Using Solid Phase Extraction Cartridges,"** 

**Project Report, December 1989.**

**7. U.S. EPA, Method 650, "Aniline and Selected Subs tituted Derivatives."** **SW-846 Update V 3620C - 19 Revision 4**

**October 2012 8. W. F. Beckert and V. Lopez-Avila, "Evaluation of**  **SW-846 Method 8060 for Phthalate**

**Esters," Proceedings of the Fifth Annual Waste Test ing and Quality Assurance Symposium,** 

**1989, pp. 144-156.**

**9. U.S. EPA Method 608, Organochlorine Pesticides a nd PCBs, 40 CFR 136, October 26,** 

**1984.**

**17.0 TABLES, DIAGRAMS, FLOW CHARTS, AND VALIDATION DATA** 

**The following pages contain the tables referenced b y this method.** **SW-846 Update V 3620C - 20 Revision 4**

**October 2012 TABLE 1** 

**EXAMPLE AVERAGE RECOVERIES OF 16 PHTHALATE ESTERS FROM THE** 

**Florisil® COLUMN PROCEDURE a** 

**Compound Average Recovery (%)** 

**Dimethyl phthalate 40** 

**Diethyl phthalate 57** 

**Diisobutyl phthalate 80** 

**Di-** n **-butyl phthalate 85** 

**Bis(4-methyl-2-pentyl) phthalate 84** 

**Bis(2-methoxyethyl) phthalate 0** 

**Diamyl phthalate 82** 

**Bis(2-ethoxyethyl) phthalate 0** 

**Hexyl 2-ethylhexyl phthalate 105** 

**Dihexyl phthalate 74** 

**Benzyl butyl phthalate 90** 

**Bis(2-** n **-butoxyethyl) phthalate 0** 

**Bis(2-ethylhexyl) phthalate 82** 

**Dicyclohexyl phthalate 84** 

**Di-** n **-octyl phthalate 115** 

**Dinonyl phthalate 72** 

**a Average recovery from two determinations, data from**  **Reference 8. SW-846 Update V 3620C - 21 Revision 4** 

**October 2012 TABLE 2** 

**EXAMPLE AVERAGE RECOVERIES OF 16 PHTHALATE ESTERS**

**FROM Florisil® CARTRIDGES a** 

**Compound Average Recovery (%)** 

**Dimethyl phthalate 89** 

**Diethyl phthalate 97** 

**Diisobutyl phthalate 92** 

**Di-** n **-butyl phthalate 102** 

**Bis(4-methyl-2-pentyl) phthalate 105** 

**Bis(2-methoxyethyl) phthalate 78** 

**Diamyl phthalate 94** 

**Bis(2-ethoxyethyl) phthalate 94** 

**Hexyl 2-ethylhexyl phthalate 96** 

**Dihexyl phthalate 97** 

**Benzyl butyl phthalate 99** 

**Bis(2-** n **-butoxyethyl) phthalate 92** 

**Bis(2-ethylhexyl) phthalate 98** 

**Dicyclohexyl phthalate 90** 

**Di-** n **-octyl phthalate 97** 

**Dinonyl phthalate 105** 

**a Average recovery from two determinations, data fr**  **om Reference 6. SW-846 Update V 3620C - 22 Revision 4** 

**October 2012 TABLE 3** 

**EXAMPLE DISTRIBUTION OF ORGANOCHLORINE PESTICIDES AND PCBs** 

**IN Florisil® COLUMN FRACTIONS**

**Percent Recovery by Fraction a** 

**Compound Fraction 1**  **Fraction 2 Fraction 3** 

**Aldrin 100** 

α **-BHC 100** 

β **-BHC 97** 

δ **-BHC 98** 

γ **-BHC 100** 

**Chlordane 100** 

**4,4'-DDD 99** 

**4,4'-DDE 98** 

**4,4'-DDT 100** 

**Dieldrin 0**  **100**

**Endosulfan I 37**  **64**

**Endosulfan II 0**  **7 91** 

**Endosulfan sulfate 0**  **0 106** 

**Endrin 4**  **96**

**Endrin aldehyde 0**  **68 26** 

**Heptachlor 100** 

**Heptachlor epoxide 100** 

**Toxaphene 96** 

**Aroclor 1016 97** 

**Aroclor 1221 97** 

**Aroclor 1232 95**  **4**

**Aroclor 1242 97** 

**Aroclor 1248 103** 

**Aroclor 1254 90** 

**Aroclor 1260 95** 

**a Eluant composition Fraction 1 -- 200 mL of 6% ethyl**  **ether in hexane**

**Fraction 2 -- 200 mL of 15% ethyl ether in hexane**

**Fraction 3 -- 200 mL of 50% ethyl ether in hexane**

**Data from Reference 9. SW-846 Update V 3620C - 23 Revision 4** 

**October 2012 TABLE 4** 

**EXAMPLE AVERAGE RECOVERIES OF AROCLORS FROM Floris il® CARTRIDGES** 

**Compound Average Recovery (%)** 

**Aroclor 1016 105** 

**Aroclor 1221 76** 

**Aroclor 1232 90** 

**Aroclor 1242 94** 

**Aroclor 1248 97** 

**Aroclor 1254 95** 

**Aroclor 1260 90** **SW-846 Update V 3620C - 24 Revision 4**

**October 2012 TABLE 5** 

**EXAMPLE ELUTION PATTERNS AND RECOVERIES OF ORGANOC HLORINE PESTICIDES** 

**FROM Florisil® CARTRIDGES a** 

**Fraction 1 Fraction 2**  **Fraction 3**

**Compound % Rec.**  **RSD % Rec.**  **RSD % Rec.**  **RSD**

α **-BHC**

**- -**  **111 8.3** 

**- -** 

β **-BHC**

**- -**  **109 7.8** 

**- -** 

γ **-BHC**

**- -**  **110 8.5** 

**- -** 

δ **-BHC**

**- -**  **106 9.3** 

**- -** 

**Heptachlor 98**  **11**

**- -**  **- -** 

**Aldrin 97**  **10**

**- -**  **- -** 

**Heptachlor epoxide**

**- -**  **109 7.9** 

**- -** 

**Chlordane**

**- -**  **105 3.5** 

**- -** 

**Endosulfan I**

**- -**  **111 6.2** 

**- -** 

**4,4'-DDE 104**  **5.7**

**- -**  **- -** 

**Dieldrin**

**- -**  **110 7.8** 

**- -** 

**4,4'-DDD**

**- -**  **111 6.2** 

**- -** 

**Endosulfan II**

**- -**  **- -**  **111 2.3** 

**Endrin aldehyde**

**- -**  **49 14**  **48 12** 

**4,4'-DDT b**  **40 2.6**  **17 24**  **63 3.2** 

**Endosulfan sulfate b** 

**- -**  **- -**  **- -** 

**Methoxychlor**

**- -**  **85 2.2**  **37 29** 

**a 1-g Florisil® cartridges spiked with 0.5 μg of each**  **compound.**

**b These two compounds coelute on the DB-5 capillary c**  **olumn.**

**Eluant composition: Fraction 1 -- 3 mL of hexane**

**Fraction 2 -- 5 mL of methylene chloride/hexane (26 /74, v/v)** 

**Fraction 3 -- 5 mL of acetone/hexane (10/90, v/v) SW-846 Update V 3620C - 25 Revision 4** 

**October 2012 TABLE 6** 

**EXAMPLE DISTRIBUTION OF ORGANOPHOSPHORUS PESTICIDE S** 

**IN Florisil® CLEANUP FRACTIONS**

**Percent Recovery by Fraction**

**Compound Fraction 1**  **Fraction 2 Fraction 3**  **Fraction 4**

**Azinphos methyl 20**  **80**

**Bolstar (Sulprofos) ND**  **ND ND**  **ND**

**Chlorpyrifos >80** 

**Coumaphos NR**  **NR NR** 

**Demeton 100** 

**Diazinon 100** 

**Dichlorvos NR**  **NR NR** 

**Dimethoate ND**  **ND ND**  **ND**

**Disulfoton 25-40** 

**EPN >80** 

**Ethoprop V**  **V V** 

**Fensulfothion ND**  **ND ND**  **ND**

**Fenthion R**  **R**

**Malathion 5**  **95**

**Merphos V**  **V V** 

**Mevinphos ND**  **ND ND**  **ND**

**Monochrotophos ND**  **ND ND**  **ND**

**Naled NR**  **NR NR** 

**Parathion 100** 

**Parathion methyl 100** 

**Phorate 0-62** 

**Ronnel >80** 

**Stirophos (Tetrachlorvinphos) ND**  **ND ND**  **ND**

**Sulfotepp V**  **V**

**TEPP ND**  **ND ND**  **ND**

**Tokuthion (Prothiofos) >80** 

**Trichloronate >80** 

**a Eluant composition: Fraction 1 -- 200 mL of 6% ethy**  **l ether in hexane**

**Fraction 2 -- 200 mL of 15% ethyl ether in hexane**

**Fraction 3 -- 200 mL of 50% ethyl ether in hexane**

**Fraction 4 -- 200 mL of 100% ethyl ether**

**R = Recovered (no percent recovery data provided by U.S. FDA)** 

**NR = Not recovered (U. S. FDA)**

**V = Variable recovery (U. S. FDA)**

**ND = Not determined SW-846 Update V 3620C - 26 Revision 4** 

**October 2012 TABLE 7** 

**EXAMPLE PERCENT RECOVERIES AND ELUTION PATTERNS FO R 22** 

**CHLORINATED HYDROCARBONS FROM 1-g Florisil® CARTRI DGES**  **a**

**Fraction 2**

**Compound Average Recovery (%)**  **RSD**

**Hexachloroethane 95**  **2.0**

**1,3-Dichlorobenzene 101**  **2.3**

**1,4-Dichlorobenzene 100**  **2.3**

**1,2-Dichlorobenzene 102**  **1.6**

**Benzyl chloride 101**  **1.5**

**1,3,5-Trichlorobenzene 98**  **2.2**

**Hexachlorobutadiene 95**  **2.0**

**Benzal chloride 99**  **0.8**

**1,2,4-Trichlorobenzene 99**  **0.8**

**Benzotrichloride 90**  **6.5**

**1,2,3-Trichlorobenzene 97**  **2.0**

**Hexachlorocyclopentadiene 103**  **3.3**

**1,2,4,5-Tetrachlorobenzene 98**  **2.3**

**1,2,3,5-Tetrachlorobenzene 98**  **2.3**

**1,2,3,4-Tetrachlorobenzene 99**  **1.3**

**2-Chloronaphthalene 95**  **1.4**

**Pentachlorobenzene 104**  **1.5**

**Hexachlorobenzene 78**  **1.1**

α **-BHC 100**  **0.4**

γ **-BHC 99**  **0.7**

β **-BHC 95**  **1.8**

δ **-BHC 97**  **2.7**

**a Florisil® cartridges (Supelco, Inc.) were condition**  **ed with 4 mL of hexane. Five replicate**

**experiments were performed.**

**The cartridges were spiked with 1.0 μg per cartridg e for hexachloroethane, hexachlorobutadiene,** 

**hexachloropentadiene, pentachlorobenzne, and hexach lorobenzene. The trichlorobenzenes,** 

**tetrachlorobenzenes, benzal chloride, benzotrichlor ide, and the BHCs were spiked at 10 μg per** 

**cartridge. The dichlorobenzenes and benzyl chlorid e were spiked at 100 μg per cartridge, and** 

**2-chloronaphthalene was spiked at 200 μg per cartri dge.** 

**The cartridges were eluted with 5 mL of acetone/hex ane (10/90, v/v).** **SW-846 Update V 3620C - 27 Revision 4**

**October 2012 TABLE 8** 

**EXAMPLE DISTRIBUTION OF ANILINES IN Florisil® CLEA NUP FRACTIONS** 

**Percent Recovery by Fraction a** 

**Compound Fraction 1**  **Fraction 2 Fraction 3** 

**Aniline 41**  **52**

**2-Chloroaniline 71**  **10**

**3-Chloroaniline 78**  **4**

**4-Chloroaniline 7**  **56 13** 

**4-Bromoaniline 71**  **10**

**3,4-Dichloroaniline 83**  **1**

**2,4,6-Trichloroaniline 70**  **14**

**2,4,5-Trichloroaniline 35**  **53**

**2-Nitroaniline 91**  **9**

**3-Nitroaniline 89**  **11**

**4-Nitroaniline 67**  **30**

**2,4-Dinitroaniline 75** 

**4-Chloro-2-nitroaniline 84** 

**2-Chloro-4-nitroaniline 71**  **10**

**2,6-Dichloro-4-nitroaniline 89**  **9**

**2,6-Dibromo-4-nitroaniline 89**  **9**

**2-Bromo-6-chloro-4-nitroaniline 88**  **16**

**2-Chloro-4,6-dinitroaniline 76** 

**2-Bromo-4,6-dinitroaniline 100** 

**Eluant composition: Fraction 1 -- 50% methylene chl oride in hexane** 

**Fraction 2 -- 5% 2-propanol in hexane**

**Fraction 3 -- 5% methanol in hexane SW-846 Update V 3620C - 28 Revision 4** 

**October 2012 Appendix A** 

**Summary of Revisions to Method 3620C (From Revision 3, February 2007)** 

**1. Improved overall method formatting for consisten cy with new SW-846 methods style** 

**guidance.**

**2. Minor editorial and technical revisions were mad e throughout to improve method clarity.** 

**3. The unit for trichlorophenol in Sec. 7.1.1 was c orrected to 0.1 mg/L.** 

**4. The ratio specified in Sec. 11.3.4 was corrected to methylene choride/acetonitrile/hexane** 

**(50/0.35/49.65).**