Abuse-type drug test papers and methods of making and using same
This is a continuation-in-part of U.S. patent application Ser. No. 07/316,334 filed Feb. 24, 1989, now U.S. Pat. No. 4,992,296, which is a continuation-in-part of U.S. patent application Ser. No. 07/054,199, filed May 26, 1987, now abandoned, which is a continuation-in-part of U.S. patent application Ser. No. 808,378, filed Dec. 16, 1985, now abandoned, which is a continuation of U.S. patent application Ser. No. 520,463, filed Aug. 4, 1983, now abandoned. 
    
    
     TECHNICAL FIELD 
     This invention relates to a method and means for detecting the presence of certain drugs of abuse, namely amphetamines, cocaine, marijuana and morphine like compounds as a group as well as individual narcotics, specifically methadone and morphine glucorinide in physiological fluids. This invention relates to a combination of chemicals impregnated in a bibulous paper used as a carrier for said test chemicals reactive to indicate the presence of said drug abuse compound residues by use of said test paper. 
     BACKGROUND OF THE INVENTION 
     The increase of drug abuse has created a need for new methods of analyzing drug residues in physiological fluids that are accurate, rapid and cost effective. Most methods require large capital expenditures for equipment, are time consuming and are costly per unit test. 
     U.S. Pat. No. 3,915,639 issued Oct. 1975 to Friedenberg discloses a method for dipstick identification of drugs of abuse based upon the incorporation of an ion-exchange resin in bibulous support together with a hydrazine salt which acts as a stain intensifying agent. It is found that the preferred hydrazine, 2-4 dinitrophenyl, is not soluble in common solvents and is not functional in the manner disclosed. Applicant here uses neither the hydrazines or ion-exchange resins. Moreover, as noted below, the staining agents are composite salt solutions and are not used separately. 
     SUMMARY OF THE INVENTION 
     It is an object of this invention to provide a method and means, such as a chemically treated test paper, for the rapid determination of the above listed drug residues as they might exist in the physiological fluids of drug abusers. 
     Another object of this invention is to provide a means for rapid identification of particular species of drug residues when present in a concentration of over one microgram per milliliter of the test fluid. 
     It is a further object of this invention to provide a test paper of such high sensitivity to specific abuse type drugs in said quantity or range to provide a semi-permanent color as a record of the presence of such drug in the physiological fluid tested. 
     Still another object of the present invention is to provide a method and means for stabilizing addicts&#39; urine so that these urines may be used as biological controls for use with the test paper for an extended period of time without the need for refrigeration. 
     The term &#34;physiological fluid&#34; refers most commonly to urine. 
     The drug residues which are usefully identifiable are Amphetamine/broad deep grey band, Cocaine/orange band, Heroine/metabolite is morphine glucorindie--blue ring or band, Marijuana/THC metabolite-charcoal ring and Percodan/tan band. Rehabilitation patient urine will usually reveal Methadone/broad purple band. 
     The bibulous carrier is a chemically treated test paper which may be of any suitable shape or form and refers to paper, fibers or polymers having the requisite physical properties stated below. 
     It is found that the following indicators bismuth, usually as the subnitrate, iodide, usually potassium iodide, and various salts of platinum, applied as composite solutions to the test paper under specific physical conditions will yield a quick means for determining the presence of various drug residues in small urine samples. 
     An occasional addict patient may attempt to substitute water and this is quickly identified because one of the test papers will turn black immediately upon exposure to water instead of urine. 
     The urine is tested as follows: 0.05 ml is take up into a capillary tube and this is placed vertically directly in the center of a 11/4 inch square of chemically impregnated test paper. The urine is allowed to diffuse of its own accord into the paper--about 10 seconds. The paper is allowed to dry and can be read within five minutes. Better discrimination from a well defined chromatogram is achieved by introducing 0.1 mil of urine onto the 11/4 inch square of test paper by way of two drops delivered at 5 second intervals in the same locus from a vertically held medicine dropper of pasteur pipet. This is somewhat less sensitive than the first procedure but can be used as an adjunct. 
     Bibulous Test Paper 
     Cellulose filter paper will be insensitive if too thin and poor chromatograms will form if the paper is too thick. The paper should have a thickness a least in a range of 0.35 to 0.45 mm. A paper having about 0.4 mm thickness and a dry weight of about 185 g/m 2  is preferred. 
     TEST PAPER FORMULATIONS 
     Where Pt salt is shown in the below formulas plug in any high quality salt from the group: K 2  PtCl 4 , K 2  PtCl 6 , H 2  PtCl 6 , PtCl 3 , and PtCl 4 . The salt is put into solution by trituration in a glass mortar with a glass pestle using fresh saturated potassium iodide. This then washed into a flask with the water and acid and allowed to age for 48 hours prior to use. 
     The quality of a Pt salt can be determined by making an iodochloroplatinate test paper as for cocaine without the bismuth addition. A drop of fluid from a spiked urine specimen should reveal a faint grey ring if the concentration of the spike is one microgram per ml of caffeine or morphine sulfate. Two separate immersion procedures with a curing period of two weeks between the first and second dips are preferred. Four reagent solutions are required: 
     
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(A)      Bismuth subnitrate 1.4    gm                                     
         Acetic acid, glacial                                             
                            14.0   ml                                     
         Water, qs          100    ml                                     
Allow 48 hours for complete dissolution.                                  
(B)      Potassium iodide   4.05   gm                                     
         Pt salt            0.45   gm                                     
         Water              100    ml                                     
         2N Hydrochloric acid                                             
                            100    ml                                     
(C)      Potassium iodide   6.0    gm                                     
         Pt salt            0.125  gm                                     
         Water              50     ml                                     
         2N Hydrochloric acid                                             
                            50     ml                                     
(C)      Alternative:                                                     
         Potassium iodide   7.2    gm                                     
         Pt salt            0.15   gm                                     
         Water              100    ml                                     
         2N Hydrochloric acid                                             
                            100    ml                                     
(D)      Potassium iodide   1.125  gm                                     
         Pt salt            0.125  gm                                     
         Water              100    ml                                     
         2N Hydrochloric acid                                             
                            100    ml                                     
(D)      Alternative                                                      
         Potassium iodide   1.35   gm                                     
         Pt salt            0.15   gm                                     
         Water              100    ml                                     
         2N Hydrochloric acid                                             
                            100    ml                                     
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     The first immersion using the (B) solution will receive the (C) solution two weeks later and the first immersion using the (D) solution will receive the (B) solution as a final dip two weeks later. 
     One day of use the (A)+(B) solutions are each warmed to 40° C. and mixed, the Pt solution is first poured into a 2 liter hand held beaker and swirled while the (A) solution is added slowly into the center of the swirl. The mixed composite reagent is then poured into a warm dish and held at this same temperature during the paper dipping process. This also applies when the (A)+(C) and (A)+(D) solutions are used. The bismuth, (A) solution, will tend to separate out of solution as black flakes if cold solutions are mixed. This may also occur if it is splashed on the sides of the mixing vessel. 
     Test paper made only from (B)+(D) solutions will show a grey ring for all alkaloids including caffeine. Methadone and water show nothing. When minute amounts of (A is added to the (B)+(D) solutions we find that the orange band occurring when cocaine is present is enhanced. Test paper made with only the (A) solution is insensitive to all drugs. But on addition of 5% KI, the (A) solution takes on a brilliant yellow color and paper dipped in that solution and then dried will only respond to water, revealing instantaneously a black spot. 
    
    
     Example 1 
     for all drugs in Table 1 but not cocaine. 
     First Immersion 
     10 ml of solution (A), prewarmed, is added to 100 ml of warm solution (B). Sheets of the bibulous paper, with splines affixed at one end, are immersed in the composite reagent held in a warm dish. The splines are then suspended from lines in an environmental room maintained at 30° C. for static drying, about 3 hours. The papers are then allowed to cure for two weeks in a holding area. They can be loosely boxed for this same period of time if preferred. 
     Second Immersion 
     100 ml prewarmed solution (A) is added slowly with continuous agitation to 100 ml of solution (C) also prewarmed. The mixed composite reagent is poured into a warm dish and the cured papers from the first dip are now immersed for the second time. The papers are again suspended in static air in the environmental room. The temperature for this drying cycle is 50° C. High humidity is also required. Drying is complete after 3 hours and the papers can then be cut to size and paced loosely in free air. The papers will remain effective for over six months if they are kept cool. 
     Example 2 
     a Cocaine Test Paper 
     First Immersion 
     1 ml of warm solution (A) is added to 100 ml of warm solution (D) Alternative. All conditions shown for first immersion in Example 1 are followed. 
     Second Immersion 
     1 ml of warm solution (A) is added to 100 ml of warm solution (B). All conditions shown for first immersion in Example 1 are followed. These papers will be ready for use upon drying. 
     Example 3 
     First Immersion 
     (for all drugs except cocaine) 
     Prewarmed (B) solution is poured into a warm dish and sheets of bibulous paper with splines attached at one end are immersed into the warm reagent. The splines are then suspended from lines in an environmental room where the temperature is maintained at about 30° C. The paper is dry in about 3 hours under static conditions after which they are loosely boxed for a curing period of 1-2 weeks. 
     Second Immersion 
     Papers from the first immersion are immersed in a composite reagent of 100 ml of warm solution (A) which has been mixed with 100 ml of warm solution (C) and then suspended in static air in the environmental room at temperature about 50° C., supplemental humidity being provided so that the relative humidity during the first hour is greater than about 90%. The relative humidity will diminish from that level gradually over the next two hours while drying is completed. The supplemental humidity is provided in any suitable manner, as known to those skilled in the art. 
     Example 4 
     First Immersion 
     Prewarmed solution (A) in the amount of 5 ml is added to 100 ml of prewarmed solution (B) and papers are immersed in it as described in Example 3. These papers then are dried and subjected to a second immersion, both steps being carried out using the procedure and the solution described in Example 3. 
     Example 5 
     for cocaine paper 
     First Immersion 
     Papers are immersed in warm solution (D) Alternative. These papers then are dried and subjected to a secondary immersion in warm solution (B)k, both steps being carried out using the drying procedure of the first immersion of Example 3. 
     
                       TABLE 1                                                     
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Summary of color reactions with the test papers five minutes              
after placement of urine.*                                                
______________________________________                                    
Water adulterant  instant diffuse black *                                 
Normal            buff center, brown or thin                              
                  purple periphery                                        
Non specific normal                                                       
                  inner purple ring                                       
morphine sulfate SPIKE                                                    
Ammoniacal        green-grey ring inward of                               
                  the periphery                                           
Caffeine/Nicotine thin grey band against the                              
                  periphery                                               
Amphetamine       very broad deep grey band                               
                  against periphery                                       
Cocaine           broad orange band against                               
                  the periphery                                           
Heroin/Morphine   inner blue ring or broader                              
Glucorinine       blue band to the periphery                              
Marijuana/THC     inner thin charcoal ring                                
Methadone         broad deep purple band                                  
                  against the periphery                                   
Percodan          broad tan band against the                              
                  periphery                                               
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     I have found the sensitivity of the papers to coincide with thin layer chromatography results. It should be evident to one skilled in the art that greater sensitivity can be achieved by either molecular concentration techniques or extraction of urinary alkaloids and elution via ion exchange resins. 
     Urine specimens will be required for testing or control purposes. This could be an ongoing problem since urine decomposes rapidly unless frozen. As another invention of mine, however, I have found that urine can be preserved so that there is no change of pH or development of objectionable odor or other evidence of contamination when standing at room temperature for at least six months, by the addition of a small but effective amount of any ester of parahydroxybenzoic acid from the group comprising methyl, ethyl, propyl, butyl or benzyl esters. These esters are white powders and neutral and inert and present no interference with either the urinary drug or the test papers. Further, these esters are non-toxic and have neither odor nor taste, but have marked bacteriostatic and fungistatic effect upon a variety of microrganisms even in low concentrations. Thus, I have discovered that by incorporating these esters into urine this same urine will show no change of pH nor develope an objectionable odor for more than one year while standing at room temperature. I have also found that analytes or metabolites, if present in the urine, will not be affected and will remain measurably stable for at least six months under the same conditions. After this period, denaturation becomes a factor. This is consequently a valuable adjunct when added to addicts&#39; urine for use in a quality control program as stated herein. 
     Table II below shows the solvents which in accordance with this invention can be used with the esters of parahydroxybenzoic acid. The choice of a particular solvent will depend upon user preference or purpose and whether commercial shipping is involved. 
     
                       TABLE II                                                    
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Solvent    Time Required for Dissolution With Shaking                     
______________________________________                                    
Acetone    less than one minute                                           
Methanol   less than one minute                                           
95% Ethanol                                                               
           +/- one minute                                                 
70% Ethanol                                                               
           +/- five minutes                                               
Isopropyl alcohol                                                         
           two minutes                                                    
70% Isophypyl                                                             
           + five minutes                                                 
alcohol                                                                   
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     The use of two esters rather than one is desirable, as indicated in the following illustrative examples of the practice of this invention as I have carried it out on a number of occasions: 
     Example 6 
     
         ______________________________________                                    
Methyl Parahydroxybenzoic acid                                            
                         10.0   g                                         
Propyl Parahydroxybenzoic acid                                            
                         2.5    g                                         
Solvent (from Table I)   100    ml                                        
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     2 ml of the above is mixed with 100 ml of urine and the MPB will be at 0.2% concentration and the PPB will be at 0.05% concentration. 
     Example 7 
     
         ______________________________________                                    
Butyl Parahydroxybenzoic acid                                             
                        2.5    g                                          
Ethyl Parahydroxybenzoic acid                                             
                        7.5    g                                          
Solvent (from Table II) 100    ml                                         
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     2 ml of the above is mixed with 100 ml of urine and the BPB will be at 0.05% concentration and the EPB will be at 0.15% concentration. 
     As indicated above, taken in both its test paper aspect and its urine preservation aspect, has special utility in the detection of abuse type drugs in urine revealing visually, in a short interval, not only the fact that such a drug is present, but also identifying the particular drug involved. Additionally,, test papers of this invention will immediately expose a user&#39;s attempt to defeat the test by substituting water for urine specimen. Further, the art is hereby afforded a wide variety of choices of formulation as well as preferred embodiments of this invention incorporating special constituents for particular purposes as impregnants of celluosic papers of strength and thickness facilitating drug detection use to best advantage. Still further and also importantly, the concept of using parahydroxybenzoic acid esters, preferably the methyl or propyl esters, is a key novel feature for the reason that it solves the problem of urine preservation without interfering with the test purpose or diminishing the utility or sensitivity of the present novel test papers in such application. 
     In this specification and in the appended claims where ratio, proportion or percentage is stated, reference is to the weight basis unless otherwise expressly stated.