Abstract:
A hybrid spectrophotometric instrument utilizing absorption and fluorescence observations requires different cell parameters for optimal performance. The ability to have an instrument with a variable optical path length with a single set of optical surfaces and one detector to reduce system variance is unique. A Zero Angle Photon Spectrometer with the additional hardware and software to support a variable path length instrument is described.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
       [0001]    This application is related to and claims priority to U.S. Provisional Patent Application number 61/912,153 filed on Dec. 5, 2013. 
     
    
     BACKGROUND 
       [0002]    Waters have been monitored spectrophotometrically for many years. However, generally only one chemical or physical parameter was measured at a time, and measurements were generally made using a single optical technique, such as light scattering for turbidity or fluorescence for chlorophyll-a. Typically, water monitoring occurs sporadically by occasional field sampling, which is taken to a laboratory for analysis. Some specialized sensors have been used on water flowing through a pipe, which provide real-time measurement of one component of the water. Generally it is desirable to move laboratory instruments to the field to perform continuous on-line water monitoring but such monitoring has been for a very limited number of water components. Typically, measurement of multiple components using more than one optical technique has required multiple sensors. This approach requires a skilled operator capable of performing multiple calibration and maintenance procedures. Multiple sensors also propagate errors from each device, which limits the monitoring system&#39;s ability to accurately detect anomalies or events using a combination of readings. 
         [0003]    Several relationships have been established in the art. The Beer-Lambert Law relates to absorbance readings. This is most often used in a quantitative way to determine concentrations of an absorbing species in solution: 
         [0000]      A=log 10 (I 0 /I)=ε1c
   where A is the measured absorbance, I 0  is the incident light intensity at a given wavelength, I is the transmitted light intensity, 1 the path length through the sample, and c the concentration of the absorbing species. For each species and wavelength, ε is a constant known as the molar absorptivity or extinction coefficient. This constant is a fundamental molecular property in a given solvent, at a particular temperature and pressure, and has units of 1/M*cm or often AU/M*cm.   
 
         [0005]    The Beer-Lambert Law predicts a linear relationship between absorption and concentration and is useful for characterizing many compounds but does not hold as a universal relationship. A second order polynomial relationship between absorption and concentration is sometimes encountered for very large, complex molecules or simpler compounds at relatively high concentration. The Beer-Lambert law has implicit assumptions that must be met experimentally for it to apply. For instance, the chemical makeup and physical environment of the sample can alter its extinction coefficient. The chemical and physical conditions of a test sample therefore must match reference measurements for conclusions to be valid. The Beer-Lambert law also only applies to pure solutions and unencumbered absorbance. In the real world, scattering from particles and non-specific absorption contribute to measured values. The ability to measure the sample at multiple optical path lengths allows the molecular absorptivity to be calculated directly for the sample being evaluated at the wavelength being used. 
         [0006]    One apparatus for measuring the purity of fluids is known from the disclosure of U.S. Pat. No. 8,102,518 to Haught et al. Devices used for measuring fluid purity in general, and for identifying and quantifying the amount of impurities in particular, commonly use light as a probing mechanism. Such devices are generally referred to as photometers. A specific type of photometer is the spectrophotometer, which permits adjustment of the light frequency (i.e., Wavelength), for making measurements at multiple frequencies. An optical sample cell contains a portion of fluid being analyzed at any given moment. 
         [0007]    Electromagnetic energy that is used to irradiate the aqueous stream may either be rejected by material in the aqueous stream, transmitted through the aqueous stream and its material load, or absorbed by the aqueous stream material. In the instance where the electromagnetic energy is absorbed by the aqueous stream material, the aqueous stream material may also fluoresce. In devices used to measure purity, one of three basic measurement methodologies following from these potential interactions of the electromagnetic energy with the aqueous stream is generally employed. These methodologies measure the parameters absorption, reflectance, and fluorescence of the aqueous stream in the optical sample cell. In accomplishing the various methodologies, an electromagnetic energy detector has been disposed with respect to an electromagnetic energy transmitter so that the detector is optimally positioned to be responsive to the associated parameter. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0008]      FIG. 1A  is a cross-sectional view of one example of an apparatus which can provide a continuously variable path length over a specified distance. 
           [0009]      FIG. 1B  is a cross-sectional side view the apparatus shown in  FIG. 1A . 
           [0010]      FIG. 2  is a chart showing the correlation between optical path length and the transmitted light for several wavelengths. 
           [0011]      FIG. 3  is a block diagram of one example of a hybrid spectrophotometric instrument with a variable optical path length. 
           [0012]      FIG. 4A  is a chart showing the relationship of the counts for a Ultra Violet electromagnetic signal, in photon counts and the optical path length in centimeters. 
           [0013]      FIG. 4B  is a chart which shows the calculated absorbance using Beer-Lambert equation, using the values from  FIG. 4A  as input values. 
           [0014]      FIG. 5A  is a chart showing the relationship of the counts for an ultra violet electromagnetic signal at a wavelength different from  FIG. 4A . 
           [0015]      FIG. 5B  is a chart which shows the calculated absorbance using Beer-Lambert equation, using the values from  FIG. 5A  as input values. 
           [0016]      FIG. 6A  is a chart showing the relationship of the counts for a visible electromagnetic signal, in photon counts and the optical path length in centimeters. 
           [0017]      FIG. 6B  is a chart which shows the calculated absorbance using Beer-Lambert equation, using the values from  FIG. 6A  as input values. 
           [0018]      FIG. 7  is a chart showing the cysteine fluorescence signal, when there is no cysteine material in the fluid. 
       
    
    
     DESCRIPTION 
       [0019]    For the purposes of promoting an understanding of the principles of the claimed technology and presenting its currently understood best mode of operation, reference will now be made to the embodiments illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the claimed technology is thereby intended, with such alterations and further modifications in the illustrated device and such further applications of the principles of the claimed technology as illustrated therein being contemplated as would normally occur to one skilled in the art to which the claimed technology relates. 
       FIG. 1 Key 
       [0000]    
       
           101  fiber optic bundle—common end 
           102  sample outlet—flow cell 
           103  cleaning nozzles 
           104  flow cell 
           105  reflective target 
           106  target support 
           107  sample inlet—flow cell 
           108  fluid seal 
           109  flow cell motor adaptor 
           110  motor 
           111  position transducer 
       
     
       FIG. 3 Key 
       [0000]    
       
           201  flow cell 
           203  sample inlet, flow cell 
           205  sample outlet, flow cell 
           208  isolation valve 
           209  electronics enclosure 
           211  fiber optic bundle, common end 
           215  illuminated fluid 
           217  power supply and broad spectrum light source 
           219  illumination computer controlled optical filter wheel 
           221  fiber optic bundle, illumination branch 
           225  reflective target, moonbeam lens 
           227  fiber optic bundle, detector branch 
           229  detector computer controlled optical filter wheel 
           231  detector 
           233  digital computer controlled counter 
           235  embedded computer 
           237  local storage (optional) 
           239  local display (optional) 
           241  computer controlled stepper motor 
           243  trigger control for light source 
           245  window/enable control for optical detector 
           248  LAN, wireless communication device, modem, or other communication means (optional) 
       
     
         [0053]    The present invention is an enhancement of an integrated spectrophotometric system for monitoring changes in a fluid system such as that described in U.S. Pat. No. 8,102,518 to Haught, et al. It is understood that the disclosed technology could also be adapted to work with other examples of spectrophotometric systems. For example, the disclosed system could also be adapted for use with the zero angle photon spectrometers described in U.S. Pat. Nos., 5,304,492; 7,411,668 B2; 8,102,518 B2; and U.S. Patent Application 2012/0205547. The disclosed technology could also be adapted and used with other suitable spectrophotometric systems. 
         [0054]    In one example, the disclosed technology can be used as an in-line, on-line instrument for monitoring various chemical parameters in a fluid system on a continuous or intermittent basis, such as for water flowing by a monitoring station containing the disclosed technology. Such a system may be used for monitoring natural, drinking, purified, or wastewaters from industrial or municipal facilities for routine screening and as an indicator of changes in the water. In one example the instrument has an on board computer  235  which runs through a programmed sequence which: cycles through a set of optical filter pairs and records the values, performs calculations using those values to generate statistical information of those readings, uses calculated results to generate client results, sends all or a subset of this data to the client by using a local area network, a 4 to 20 rnA current signal, or a cellular modem to a remote, secure server which a user can access. Optionally, all raw counts, calculated results, and statistical information may be saved on a local hard drive or other storage device to be collected later. 
         [0055]    In this particular example, the fixed optical path length base, mirror support, and mirror have been removed from the new flow cell and replaced with a optical reflective target  105 , reflective target support  106 , fluid seal  108 , flow cell/motor adapter  109 , linear servo motor  110 , and a linear position transducer  111 , as one example of how to implement a variable path length hybrid spectrophotometric instrument. The servo motor is controlled by the embedded computer  235  and the position transducer is monitored by the computer  235 . In one embodiment there are hard stops at the minimum and maximum distance for the lens assembly. The instrument tracks the position of the lens assembly by maintaining a record of the number of steps the motor has taken from the home position and checks this position with the output of the linear position transducer. The transducer is repeatable to  0 . 1  mm. If the calculated position and the measured position do not match, the computer  235  will re-home the lens assembly. The home position may also be checked on a periodic basis. 
         [0056]    Another embodiment of an analytical instrument for the determination of impurities in an aqueous stream which may employ the present invention will now be described. The aqueous stream to be analyzed could be a municipal water supply, industrial effluent, groundwater, and the like. In one embodiment, an aqueous sample is caused to flow through an optical sample cell from an inlet to an outlet. Valves may be controlled to regulate the flow rate of aqueous stream through the optical sample cell, particularly if the stream is to be intermittent, reduced, or combined at certain times. Optical components and electronics are optically connected to the optical sample cell through an optical fiber bundle. A narrowband electromagnetic energy source for supplying an illumination probe to the aqueous stream, in an embodiment utilizes a source of electromagnetic energy such as a flash lamp. Light (a portion of electromagnetic radiation occurring within wavelengths including infrared, visible light, and ultraviolet light) is directed by the system through a bandpass optical filter to select a desired wavelength and passed into and through a portion of an optical fiber assembly to the optical sample cell. The light is then directed, as the illumination probe, through the aqueous sample in the optical sample cell and reflected from a light reflector disposed in the optical sample cell, through the aqueous sample again, and back into the optical fiber bundle. A separate portion of the optical fiber bundle directs the returning light through another bandpass optical filter and is detected by an optical detector. 
         [0057]    Signals from detector are coupled to a counter and then to a computer. The computer may be, alternatively, a programmed general-purpose computer or a specialized computer controller to control the movement of a positioning motor (capable of repositioning light reflector), valves (if any), optical filters, etc. Signals from the optical detector are received, recorded, and analyzed by the computer and the results are stored in a memory, reported on a display, and/or output to external devices, or signals are fed back to affect appropriate change in the sensor system or fluid control system as a feedback loop. Further, algorithms may be stored in memory to be recalled and executed by the computer to extract desired information from the data collected. 
       EXAMPLES 
       [0058]    Several test runs were generated using a spectrophotometric system according to the disclosed technology. The path length was varied from 3.0 cm to 5.0 cm in 0.5 cm increments. The instrument was set up to measure 15 sequential sets of data for the 10 different filter pairs at each path length. Seven filter pairs established an absorbance measurement condition and three established a fluorescent measurement condition. Each of the 150 represented measurements in this 20 minute block represents an average of 400 individual measurements. Only four of the 10 different measurements are shown in  FIGS. 4A through 7 . The measured photon counts were then used to calculate the absorbance for each of the absorbance filter pairings using the counts and the path length. The raw counts were used as an indication of background counts in the fluorescent filter pairings. Treated city drinking water was routed through the instrument while this data was generated. This source is constant and typically has low levels of material which show an optical signature over this time interval. 
         [0059]    In  FIGS. 4A ,  5 A,  6 A, and  7  the solid lines indicate the programmed path length (scale on the right) at which measurements took place. The dashed line indicates the photon count values (scale to the left) measured for various wavelengths and spectrophotometric techniques at each programmed path length. In  FIGS. 4B ,  5 B,  6 B the line is the calculated absorbance at a particular point in time using obtained values for path length and photon counts. Over an eight hour test period the spectrophotometric system recorded approximately 330 results for each of 10 wavelengths (filter pairs), with each of those being the average value for 400 measurements. 
         [0060]    The absorption measurements shown in  FIGS. 4B ,  5 B, and  6 B demonstrate the ability of the disclosed technology to accurately reposition a spectrophotometric instrument&#39;s optics at a desired path length. The diagrams also demonstrate the disclosed technology&#39;s ability to accurately and precisely track zero values at various wavelengths against which low ( FIG. 4 ), very low ( FIG. 5 ) and near detection ( FIG. 6 ) values of absorption are determined. The ability to optimize the path length for individual wavelengths enhances the disclosed technology&#39;s ability to analyze at the most appropriate path length for each wavelength as dictated by a matrix of interest (short path lengths at wavelengths with strong absorptive properties and longer path lengths at wavelengths with weak absorptive properties). 
         [0061]    The fluorescence measurements shown in  FIG. 7  illustrate the ability of the disclosed technology to enhance fluorescence measurements by allowing a device to operate at maximum path length thus reducing the background values used during fluorescence measurements. The reduction of background (or scattered) light in fluorescence measurements is directly proportional to the signal to noise ratio, a critical limitation of such measurements. These cysteine fluorescence measurements were made with no fluorescent material in the fluid being measured. 
         [0062]    Variable path length technology allows hybrid spectrophotometric analyzers to be optimized by allowing the system to work at optimized path length depending on the absorptive properties of the matrix of interest while also allowing background light to be minimized during fluorescence measurements. These data demonstrate several advantages continuous path length technology has over both fixed and multiple path length devices. 
         [0063]    While the claimed technology has been illustrated and described in detail in the drawings and foregoing description, the same is to be considered as illustrative and not restrictive in character. It is understood that the embodiments have been shown and described in the foregoing specification in satisfaction of the best mode and enablement requirements. It is understood that one of ordinary skill in the art could readily make a nigh-infinite number of insubstantial changes and modifications to the above-described embodiments and that it would be impractical to attempt to describe all such embodiment variations in the present specification. Accordingly, it is understood that all changes and modifications that come within the spirit of the claimed technology are desired to be protected.