Method for detection of analytes on a surface

The invention relates to a method for detecting an analyte on a surface by Wiping the analyte off the surface with a wiping member, elution of the analyte of the wiper member and detection of the analyte in the elution liquid by an immunological reaction.

In addition to the detection of analytes in test liquids, such as. For example, blood, urine, saliva, comes particularly in criminalistics the detection of analytes on solid surfaces, such. as to furniture, luggage, etc., of growing importance. Especially in the drug investigation unit, it is a desirable goal, even very low drug contamination from Objects quickly and easily be able to detect. The case sensitive a Nachweisver driving, the less analyte that contaminated a specific surface must, are thereby detected on this surface.

In DE 29 35 881 C2 discloses a device for the detection of traces of marijuana will be wrote. As detection reaction is a color reaction on the marijuana ingredient Cannabis used. The device consists of a cotton swab in a by account plastic tube is kept. In separate reagent chambers of Art Fabric tube required for the cannabis detection reagents are included. With the swab is the under investigation object or the person to be examined wiped, whereby optionally accumulate marijuana traces in cotton wool. This Cotton swab is then placed in a predetermined order in the reagent solutions be placed in storage tubes available, dipped. The detection reaction is based on a purely chemical reaction of the cannabis molecules with the device provided reagents at a defined pH. In the presence of Cannabis shows a color reaction in the final solution. Detection limits are in DE 29 35 881 C2 not specified. However, it is mentioned in the cited patent that a touch of cannabis containing samples is necessary. A detection of drugs in Body fluids on body surfaces is not mentioned.

EP-A-0229517 describes an absorbent, impregnated with reagent test paper for After setting of illegal drugs or their metabolites in biological fluid samples. The Sample liquid must be pretreated for the detection of drugs. Is given by a specially prepared syringe purified and concentrated by chromatography. After Pretreatment, the sample is applied to the sample application zone of the test paper. From there moves the sample liquid in a reagent impregnated region where the detection reaction takes place for the target analyte and is observable by means of a color reaction. here, too, is it a purely chemical detection reaction. It is known that the post weisempflindlichkeit and the specificity of such reactions is not sufficient to drugs in detect body fluids. Therefore, a step for concentrating and Reini movement of the sample upstream. The complicated nature of the sample preparation and the reaction lead in the detection reaction on the test paper can be that described in EP-A-0229517 Detection method unsuitable for use "on site" and in particular the application appear by "laymen".

To investigate any surfaces on analytes, especially drugs, are ver known various specific detection techniques, in which first the analyte by wiping collected a surface and after elution of the used wiping surface Immunology is proved logically. In the "Illicit Substance Detector" Westinghouse, Baltimore, USA (Security Management, Vol. 37/8, pages 12-15, 1993), is a nubbed plastic surface wipe a surface to be examined and the drug demonstrated an integrated in a disposable card process with three reagent solutions (DE-A-43 41 862). The test result is evaluated by an optical reader. The immunological detection principle is based on the inhibition of latex agglutination reaction by the detected drug. The lower limit of detection is using a micro program indicated. The disadvantage is in addition to the relatively high detection limit that Nachweisre action will be started by a mechanical pressing of the three liquid reservoirs have to be. In addition, the measurement result can only with an optical reading aid and not with the eye be evaluated.

The Roche Diagnostics provides a test kit for cocaine from urine samples by the same consuming to handle immunological detection principle. The detection sensitivity is in the range of 0.2 ug / ml.

The "AccuPress-Kit" of Thermetics, Wobuim, USA (Security Management, Vol. 37/8, Pages 12-15, 1993), consists of a specially coated reagent and three vessels with reagent. This test with a cotton swab to be tested wiped surface and the swab then washed with buffer. Next the comparable insensitive detection limit of not less than one ug is the hand dling of three different reagent cumbersome and also draws Error Possible opportunities to be.

In EP-A-0699906 describes a test kit is, wiped with the analytes of surfaces and to be rejected, which surfaces by direct contact with the hand dling were contaminated with such analytes.

The object of the invention to provide a method for the detection of Analytkontaminationen of Surfaces, particularly with drugs tracks to provide, in a simple manner and without the technical aids can be performed. In particular, the detection limit should apply the analyte absolutely, well below a ug possible below 100 ng.

The object is achieved by a method as characterized in the claims.

It has been found, surprisingly, that the test kit, which is described in EP-A-0699906, can be successfully used for the analysis of body regions that are not contaminated when handled with the substance to be detected by direct contact were. Essentially there are those parts of the body, the body fluids such as excrete sweat, saliva, tears, urine, etc., or carry, such as mouth, oral mucosa skin, tongue, armpits, eyes, genital area, etc.

The invention provides a method for detecting an analyte in body fluid with a test kit which comprises

<ul><li>a) a test strip comprising one or more capillary-active chromatographiefähigen sheet-like materials in fluid contact with each other </li><li>- An eluting agent at one end and a target zone at the other end,</li><li>- A capture zone between the eluting agent and target zone in which an ex capture reagent is immobilized, which is capable either the analyte, a specific specifically bind analyte or a labeled binding partner and</li><li>- A conjugate zone between the eluting agent application and capture zone, the one migratable labeled contains binding partner, capable of either the analyte specifically binding or the specific analyte or the capture reagent, wherein the binding of the labeled binding partner to a detectable signal in the Capture zone or into the target zone, indicating the presence of the analyte,</li><li>b) a wiping element separately for test strip surface</li><li>c) a pressing device which causes the wiping member with the test strip surface between Elutionsmittelauftragszone and conjugate zone or in the conjugate zone itself can be contacted,characterized in that the wiper element body fluid from a body is taken and examined.</li></ul>

This process comprises wiping the analyte from a body surface with a Wiping member of a wiping material, elution of the analyte from the wiping element and Detection of the analyte in the elution liquid by an immunological detection reaction, characterized in that

<ul><li>a) the wiped with the wiping member to the analyte to be examined surface,</li><li>b) the wiping member with the flat surface of a capillary chromatographiefähigen test strip at one end of an eluting agent and containing a target zone at the other end of the strip, in a region between two zones will be contacted,</li><li>c) elution liquid is applied to the eluting agent application, by capillary power to the target zone at the contact point with the wiping surface moves past, said analyte is recorded on the wiping element of the elution liquid and</li><li>d) the analyte measured in the target zone due to an immunological binding reaction is.</li></ul>

The test strip for the present process may consist of a single chromatographic graphiefähigen striped material or preferably from several to a base layer substantially juxtaposed capillary-active surfaces of the same or ver different materials, which are in fluid contact with each other, so that they a make liquid transport path along which a liquid by capillary forces ge exaggerated, flows from the elution agent to the target zone.

A chromatographic material are all liquid-porous or capillary-active materials in question, eg., cellulose and derivatives thereof, glass fibers and fleeces and woven fabrics of synthetic or natural materials.

Various in the inventive immunoassay guides be applied to those of the analyte due to one or more immunological Bin forming reactions is demonstrated. In a preferred embodiment (<b>Fig.</b> 1) contains A suitable for the invention chromatography test strip optionally on a Carrier film (<b>5</b>) Between the eluting agent application (<b>1</b>) And the target zone (<b>4</b>) An interception zone (<b>3</b>) Which contains a capture reagent in immobilized form, which is capable of either Analyte, a specific binding partner of the analyte or a labeled binding partner specifically binding. Before the capture of the test strip preferably contains a conjugate (<b>2</b>) Containing a migratable labeled binding partner, capable of is either the analyte, a specific binding partner of the analyte or the Ab capture reagent in the capture of specifically binding.

The "specific binding partner of the analyte" is a trek enabled, unlabeled analyte with a binding site for the capture reagent. When such a binding partner is used in the immunoassay, it can before up or in the conjugate zone or preferably between conjugate zone and capture zone be introduced. In addition, after the target zone still further liquid absorbent Connecting material liquid after a hike through the individual zones takes.

Depending on the type of immunoassay which can be applied in the process according to the invention should lie in the different zones, different binding partners: With a Sandwich immunoassay are preferably present in the conjugate, a labeled Analytbindungs partner non-immobilized form. This forms with the analyte a complex which either is bound by the capture reagent through its binding to the analyte, or a second, unlabeled, free hiking capable Analytbindepartner with a specific bin binding site for the capture reagent first forms a sandwich complex with the analyte, the TIALLY using the specific binding site of the binding partner in the capture zone the will. Preferably, the labeling of the complex is measured in the capture zone. In this case, capture zone and target zone are identical.

In a competitive assay is preferred in the conjugate, a labeled Analyte analogue, the analyte in the presence of either the binding sites of the ex konicurriert capture reagent in the capture zone (in this case, the capture reagent is a Ana lytbindungspartner) or, if an additional hike capable analyte is present to compete the binding site. Complex of labeled analyte analogue and wanderungsfähigem binding partner are then a specific bin binding site, eg., biotin, bound in the capture zone. In this case, the capture reagent a binding partner of the analyte binding migratable, eg., streptavidin. Be the marking is preferably in this method, not in the capture zone, but in the Target zone in the form of uncomplexed analyte analogs as a measure of the presence of Analyte measured.

Preferably, the inventive method is for a 1EMA-analog test principle (Also "immunenzymometrisch-analog" Test principle) performed. The performance of IEMA tests on test strips, for example, in EP-A-0407904, EP-A-0353570 or DE OS No. 40 24 919th

In the conjugate are labeled binding partner for the analyte in excess. After chromatographic migration are not bound to analyte labeled Bin binding partner immobilized by solid phase-bound analyte analogues in the capture, weiterchroma while complexes of analyte and labeled binding partner in the target zone tographieren. The entered the target zone marker can be used as a measure of the presence or also the concentration of the analyte to be measured.

As analyte used in the present invention, all immunologically detectable Substan zen particular antigens and haptens. Is very particularly suitable the present inven tion for the detection of drugs, eg. as cocaine, morphine, heroin. In this case, the analyte to the surface be wiped as a molecule or particle-particle or adsorbed ago come.

As the labeling are customary labels such as enzyme labels, fluorescence, color fabric marker in question. Preference is direct label, in particular metal label, most preferably gold label. This has the advantage that directly to the eye The test result can be read.

The binding partner for the analyte in particular antibodies and antibody Ask coming elements into consideration.

If a hike capable unlabeled analyte in addition to trapping reagent is used, it transmits a binding site for the capture reagent. For this I binding site are all specific binding partner of a specific binding pair in Purpose, for. Example, lectins, antibodies, antigens, preferably biotin (which binds with streptavidin). The label in the capture zone or target zone can have conventional detection methods he follow, z. B. by reflection or visually. Particularly with metal label, z. B. Gold mark, the measurement result can be read visually in a simple manner.

To carry out the inventive process with a wiping member on a ge to be examined surface advantageously using a slight pressure wipes. The body region to be examined usually carries moisture. It is advantageous to when the wiping operation is repeated several times. For better handling, the Wiping member (<b>13</b>) Preferably on a support (<b>11</b>) Attached (<b>Fig.</b> 3). A good Wiping performance results in particular at a high mechanical load of Wiping member. Particularly advantageously, it has been demonstrated, when the material of Wiping member with the support (<b>11</b>) Is ultrasonically welded.

When wiping members substrates such as plastics, fabrics, non-wovens can be used, stay on those analytes, particularly drugs and adhere to accumulate by wiping leave. On the other hand should the adherent analytes in fluid contact slightly be desorbed. The skilled person may select such materials without effort. Before given to have absorbent materials such as nonwovens, tissue or porous matrices such as membranes or sponges. Very particularly suitable are webs of fibrous materials, wherein the Fibers are assembled disorderly generally, z. B. paper or glass fiber webs. The Wiping surface is preferably dry, but can also be moistened.

If rough surfaces such. B. anodized aluminum surfaces or other roughened Metal surfaces investigated is preferably used a moistened wipe surface. Suitable liquids to moisten the wiping surface are primarily water and aqueous Buffer systems which may be adapted to the subsequent immune response. the water or aqueous buffers may be added to detergents or organic solvents. As detergents Tween 20, Tween 80, octylglucoside, polidocanol, Synperonic, z. B. F can 68, or zwitterionic detergents such Cholamidopropansulfonat in Konzen concentration below 5 wt .-%, preferably 0.01 wt .-% to 1 wt .-%, alone or in mixtures be used. Organic solvents which can eg., Dimethyl sulfoxide, glycerol or Ethanol can be used alone or in mixtures. The proportion of solvent in the Fluid can clearly be less than 30 wt .-%. but it is also possible to use organic to use solvents or mixtures of organic solvents without water.

The moistening of the wiping surface is per the task of 1-50 ul of liquid cm², Preferably 3-20 ul / cm² z. B. by task with a pipette or automatic achieved matic dosing. Alternatively, the humidification by brief Contact the wiping surface with a damp sponge or another feuchtigkeitsabge inputting surface occur.

The moistening of the wiping surface can also be advantageously achieved so that the liquid in microencapsulated or blister packaged form on the due date for the wiping part is already kept. When used as liquid water or aqueous buffer systems be that encapsulation is advantageous by inclusion in waxy substances, such as z. B. Paraffin, made. The release of the liquid can by mechanical means, eg. B. by pressing on the surface to be examined, done.

Preferably, the wiping member is a sheet-like product, in particular a fleece before given to from fibers based on cellulose and / or polyester fibers. In addition, the Fibers of an organic binder, the preferred hydroxyl and / or ester groups contains, are held together. Such fabrics are described in German patent application DE-OS 38 02 366 described.

Preferably used cellulose fibers rayon, wood pulp or cotton linters. As Zellwolle is designated a material to by alkalization of cellulose to alkali cellulose, closing treatment with carbon disulphide and the formation of cellulosic Xantho genaten, dissolution of cellulose xanthate in caustic and spinning of viscose filament yarn was obtained. Pulp, by a complete chemical digestion cellulosic materials and subsequent bleaching are obtained. As linters short, not spinnable cotton fibers referred to, which are obtained from seed cotton. The cellulose fibers have preferably a fiber density of 1.7 to 4.5 dtex and have Lengths of 1 to 20 mm, preferably 3 to 12 mm. Particularly preferred polyester fibers are Fibers and having a specific gravity of about 1.17 g / cm a length of 3 to 6 mm a fiber fineness from 1.7 to 3.3 dtex.

As a further constituent, the webs may comprise an organic binder which OH and / or having ester groups include. this are preferably polyvinyl alcohol and epichloro hydrinharze used. The polyvinyl alcohol is preferably used as fiber material with a Length of 3-5 mm and a specific gravity 1.26 to 1.30 g / cm applied. Out These components and demineralized water is a non-woven on a inclined wire produced by the conventional method in papermaking. Particularly preferred non-woven materials are also described in DE-OS 38 02 366th

The thickness of the material of the wiping member is not critical. Advantageously, it should have the flattest possible surface, the thickness is usually between 0.1 and 3 mm lies. The wiping surface should be on the dimensions of the chromatography test strip be fit, ie the width of the wiping surface should not substantially the width of the test strip exceed. Preferred dimensions of the wiping surface are between 0.3 and 2 cm, special IDEM preferably from 0.5 to 0.8 cm in length and in width from 0.3 to 1 cm, particularly preferably from 0.4 to 0.8 cm.

After wiping a contaminated surface with a wiping element is the Wiping surface of this element with an area of ​​the test strip surface between the contacted eluting agent and the target zone, preferably lightly pressed. This Region is preferably between the eluting agent and the conjugate or on the conjugate zone itself. It is particularly preferred also if the zone where the is pressed wiping member from one of the suitable for the wiping element materials consists, in particular the non-woven materials described in DE-OS 38 02 366th All It is particularly preferred if the wiping member is pressed onto the conjugate is. Preferably, therefore, is that the wiping element between 25% and 150% of the area, particularly preferably has approximately the same area as the conjugate zone.

The pressure at which the wiping member is pressed, should be at least so great that a planar fluid contact between the two surfaces is possible.

In order to facilitate the user pressing, in one embodiment of the test carriers (<b>5</b>) In a housing (<b>6</b>. <b>8th</b>) be housed (<b>Fig.</b> 2a: housing without cover, <b>Fig.</b> 2b: Case Closed). The housing has an opening (<b>9</b>) For printing the wiper element (<b>13</b>) on. The wiping element (<b>13</b>) On a support (<b>11</b>) As a hinge (<b>14</b>) fixed the housing that this vehicle is expanded for wiping and for pressing of the wiping member on the opening (<b>9</b>) Flipped the test strip housing and given if can be locked. The wiper member may also hand or a clamp with will contact the test strip.

In the next step elution to the elution agent application (<b>1</b>) Applied. The liquid may be applied to the eluting agent or the test strips are dipped in an elution. If the liquid is applied, there is the Elutionsmittelaufnahmezone preferably made of a particularly absorbent material at least as much liquid absorbs that the liquid until the end of the chromatogram phy strip travels. If the test carrier in a housing, the housing preferably has an opening (<b>7</b>) For supplying the liquid.

As elution water and the common in immunoassays, buffer solutions comes in Question. The liquid travels along the strip towards the target zone (<b>4</b>) And happened while the zone with the printed wiping member. Surprisingly it on was added to the wiping element adhering analyte molecules of the liquid stream and in the other zones transported. , Is preferred when the zone of the test strip, the the wiping member is pressed ( "receiving zone") from one of the wiping element Preferred non-woven materials. It is particularly preferable if the receiving zone ( "Up Fleece") by the conjugate (<b>2</b>) Itself is formed.

In a preferred test variant analyte molecules as they pass the conjugate complexed by labeled analyte binding partner, non-complexed labeled binding partners are in the capture by immobilized analyte analogs such. B. polyhaptens, detained while labeled binding partners that have an analyte bound to Ab pass, catch zone and reach the target zone. In the target zone then the marked Complexes are detected. To the signal in the target zone optically better of the mark to distinguish in the capture, the capture can abge advantageously be covered. If the test carrier is accommodated in a housing, this has over the target zone preferably an opening (<b>10</b>) For observing the signal.

The analysis on a Analytkontamination the examined surface is positive if at least a portion of the target zone having a coloration. Staining can easily visually are detected or photometry.

It is also possible to detect a test Roaming Vice multiple analytes. To this end, , the conjugate zone and the capture and optionally also the chromatographic Material parallel of the conjugate to the target zone in several, in test strip direction, advantageously from each other divided part separate strips, in each part konjugatzonen and Teilabfangzonen binding partner for the different evidence the analyte or different analyte analogues are included. After contacting the Wiping element with a common receiving zone off the Konjugatzonen and task the elution, is divided into the various Teilkonjugatzonen the analyte various Teilchromatographiestrecken on with another in any part of the target zone Analyte can be detected.

The sensitivity of the method according to the invention is surprisingly much higher than that of the processes from the prior art. The wiping and transfer efficiency the wiping member of analyte on the test strip is so great that with unexpectedly the procedure absolute amounts of up to 10 ng analyte, in particular drugs, on surfaces can be detected. The process requires very few steps and handling the result can be determined very quickly and with simple means. particularly surprisingly it is that analyte particular drug successfully nachge in body fluids can be recognized, which are only present in small amounts on or in regions of the body, such as especially sweat and saliva. By the sampling of body regions which are not in handling with the analyte to be determined may have come into contact, can be decided after the presentation of the result whether a recording of Substance was present in the body or not.

example 1

a. Preparation of Benzoylecgoninmaleimidoethylamid

2.4 g Benzoylecgoninhydrochlorid be in 200 ml dry acetonitrile with 1 g of N-hydroxysuccinimide added and 1.8 g of dicyclohexylcarbodiimide and stirred for 3 h.. From Precipitate is filtered off, the filtrate was evaporated, taken up in nitromethane and filtered again. After evaporation of the solvent is triturated with ether. You get 1:13 g Benzoylecgoninsuccinimidylester. This product is used together with 0:47 g Maleimi doethylaminhydrochlorid (s. WO 90/15798) was added to 100 ml of dry acetonitrile. Are added to 1.1 g triethylamine and stirred for 12 hours at room temperature. The reaction mixture is evaporated, in 50 ml of ethyl acetate was added and three times with sodium bicarbonate shaken. The ethyl acetate phase is evaporated and the Product converted by recording in 10 ml dioxane saturated with HCl into the hydrochloride. It is filtered, washed with ether and obtains 1 g Benzoylecgoninmaleimidoethylamid hydrochloride.

b. Preparation of a biotinylated cocaine polyhapten for the capture

Rabbit IgG is at a concentration of 25 mg / ml in phosphate buffer pH 8 with the 6x molar amount S Acetylthiopropionsäuresuccinimidylester dissolved in dimethyl sulfoxide implemented. After 1 hour at 25 ° C the reaction is stopped by adding a solution of 1 mol / l Lysine stopped. It follows dialysis against 0.1 mol / l potassium phosphate buffer, pH 6 with 1 mmol / l EDTA. Subsequently, the pH adjusted to 7.8 is adjusted with 1 mol / l hydroxylamine solution, pH 7.5 ad 20 mmol / l at 25 ° C incubated for 1 hour. For coupling a 5-fold molar shot Benzoylecgoninmaleimidoethylamidhydrochlorid dissolved in dimethyl sulfoxide and added to the stirred solution of the modified Sulffiydrylgruppen rabbit IgG. After Incubation at 25 ° C for 2 hours, the reaction is stopped by the successive addition of of 0.1 mol / l cysteine ​​solution ad 1 mmol / l and 0.5 mol / l Jodacetamidlösung ad 5 mmol / l. Of the Batch is dialyzed overnight against 0.1 mol / l potassium phosphate buffer, pH 8.5 and over Membrane filtration is concentrated to a protein concentration of 10 mg / ml. Thereafter, the obtained cocaine Polyhapten with a 8-fold molar excess Biotinylcapronsäure succinimidyl ester, dissolved in dimethyl sulfoxide, biotinylated. The mixture is dialyzed against 20 mmol / l Sodium acetate, pH 4.3, dialyzed, and purified by FPLC.

c. Preparation of morphine-3-O-essigsäuremaleimidoethylamidhydrochlorid

Analog Example 1a. is morphine-3-O-acetic acid with Maleimidoethylaminhydrochlorid to Morphine-3-O-essigsäuremaleimidoethylamidhydrochlorid implemented.

d. Preparation of a biotinylated morphine Polyhaptenes

Analogously to Example 1b. is modified with sulfhydryl rabbit IgG with morphine-3- O essigsäuremaleimidoethylamidhydrochlorid and Biotinylcapronsäuresuccinimidylester to a biotinylated morphine Folyhapten implemented.

e. Preparation of a gold conjugate of an anti-cocaine antibody

Gold sol with a specific by photon correlation spectroscopy particle diameter of 20 nm was by standard methods (Frens, Nature Vol. 241, p 20-22, 1973) produced. The conjugation with the antibody, the cocaine and benzoyl ecgonine recognizes is according to the prior art performed (Geoghegan et al., J. Immunol. Meth. Vol. 34, P 11-31, 1980)

f. Preparation of a gold conjugate of an anti-morphine antibody

Analogously to Example 1e, an antibody, morphine and heroin was seen to gold particles bound.

example 2

a. Test carrier for the determination of cocaine

Construction of the test carrier see <b>Fig.</b> 1.

Eluting agent (breather) (1)

Polyester fleece of the firm Binzer, Hatzfeld, Federal Republic of Germany. It is a pure polyester fleece, which is solidified with 10% Kuralon. The thickness of 1.0-1.2 mm, which Absorbent Capacity 1800 ml / m²,

Conjugate (conjugate fleece) (2)

A mixed fleece of 80 parts polyester and 20 parts of viscose staple fiber, solidified with 20 parts Kuralon, in a thickness of 0.32 mm and with a suction capacity of 500 ml / m² will with impregnated following solution and then dried: 100 mmol / l HEPES buffer pH 7.5, 100 mol / l NaCl, conjugate of gold particles and an anti-cocaine antibody, which also Benzyl ecgonine binds in a concentration to an optical density of 10 at 520 nm has.

Capture zone (3)

A nonwoven made of 100% cotton linters, solidified with 2% Etadurin having a thickness 0.35 mm and a Absorptive capacity of 372 ml / m² is impregnated with the following solution and then getrock net: 10 mmol / l sodium phosphate pH 7.5, polymerized streptavidin 200 mg / l (Preparation according to Example 1c, EP A 0331127).

The pre-impregnated fabric is then soaked again and then dried: 10 mmol / l sodium phosphate pH 7.5, 200 mg / l biotinylated cocaine polyhapten from Example 1b.

Detection field (target zone) (4)

It is a non-woven 100% cotton linters, solidified with 2% Etadurin having a thickness 0.35 mm and a suction capacity of 372 ml / m² used.

All fabrics have a width of 5 mm. The conjugate fleece has a size of 5 × 5 mm. The Fleeces are under <b>Fig.</b> 1 on a carrier film (<b>5</b>) 5 mm wide glued.

b. Test carrier for the detection of heroin

Analogously, from the gold conjugate of the anti-heroin antibody and the biotinylated Morphine Polyhapten a test carrier for the detection of heroin produced.

example 3

On a polyethylene surface in each case 10 .mu.l of a dilute heroin hydrochloride solution applied in methanol and allowed to dry. This gives each with 10, 20, 40, 60 and 80 ng heroin hydrochloride contaminated areas of about 1 cm² Area. From each lot be prepared three test fields.

A fleece was prepared consisting of 80 parts of polyester fibers with a fiber fineness of 3.3 dtex and a fiber length of 4 mm is, 20 parts viscose staple having a fiber fineness of 1.7 dtex and a cut length of 3 mm and 20 parts of polyvinyl alcohol a Average length of 4 mm. The fibers of polyester, viscose and polyvinyl alcohol with pitched demineralized water at a consistency of 0.3% in mixing vats or sporadically. Of the Pulp was then pumped to a rotating sieve. While the fiber mixture dewatered and the water is sucked out by vacuum, the fibers are oriented on the Wire side and a non-woven having a solids content of about 20% over the drying cylinders contact dried. There is obtained a nonwoven fabric having a basis weight of 80 g / m² and a Thickness 0.32 mm.

On a support (<b>1</b>) Are 5 × 5 mm pieces of this fabric (<b>2</b>) glued (<b>Fig.</b> 3). With the mounted on the carrier fleece wiping be contaminated with heroin Test fields dusted with light pressure.

The mounted to the support wiping fleece is the conjugate fleece of according to Example 2b. positioned test carrier prepared and pressed on under gentle pressure and with a Clamp fixed.

The Elutionsmittelsaufgabezone is 5 sec. In a chromatography buffer (150 mmol / l NaCl, 50 mmol / l potassium phosphate buffer pH 7.2) immersed. You put on a non-absorbent position from, and after 2 minutes with a Chroma Fa. Minolta the chroma (C-value). Thereafter, the detection field is checked for the presence of pink color visually.

example 4

With a dilute solution of cocaine in water were analogously to Example 3 on a polyethylene film test areas with 5, 10, 25, 50, 75 and 100 ng cocaine applied. Analogously to Example 3 With a device according to Scheme 2, 2a each test carriers according to example. to Be contains humor of cocaine, wiped the test areas. The wiping fleeces are around at a diameter of 4 mm. The detection field is each visually checked on pink coloration.

<tgroup cols="2"><tbody><entry align="left">Volume cocaine / test field [ng]</entry><entry align="left">visual reading²</entry><row><entry align="center">0</entry><entry align="left">0</entry></row><row><entry align="center">5</entry><entry align="left">+/-</entry></row><row><entry align="center">10</entry><entry align="left">+/-</entry></row><row><entry align="center">25</entry><entry align="left">+</entry></row><row><entry align="center">50</entry><entry align="left">+ / +</entry></row><row><entry align="center">75</entry><entry align="left">++</entry></row><row><entry align="center">100</entry><entry align="left">++</entry></row></tbody></tgroup>

2

0: no pink coloration in the detection field visible+/- Faint color detectable+ Colors detectable++ Strong color detectable

example 5

Some cocaine is mixed with 1000 parts of lactose. 5 mg of this mixture are on an area of ​​220 cm² a cotton cloth spread. Approximately 10 centimeters² these range wiped off as in Example 4 and analyzed. In all experiments, a pink coloration in the Detection field detected visually. Analogously, a polyethylene film of 2 cm² with cocainhal contaminated term particles and analyzed by wiping. It is also in all experiments a pink coloration in the detection field detected visually.

example 6

On a rough anodized aluminum plate are each 10 ul diluted accordingly given solutions Cocainhydrochlorid in water and on an area of ​​1 cm² ver Splits. After drying at 37 ° C for 15 min. be contaminated surfaces with a round fleece abge according to Example 3 with a Durchinesser of 5 mm with light pressure wipes. It is both without humidification (dry) and after previous moistening the wiping fleece with 2 ul water gewüscht. The wiping fabrics are then each of the placed conjugate of a test carrier for the detection of cocaine in Example 3 and with a flat tweezers detained. The breather of the test carrier is in half for 10 sec. In Water immersed. Then, you insert the test carrier on a flat non-absorbent Un terlage from and after 2 minutes in the detection field of the color.

example 7

On examination of the body surface of people the following picture:In the examination of the body surface under the arm is connected to the test carrier in Individuals who had consumed opiates or cocaine (as evidenced by a positive Findings in the urine), a positive result obtained. includes the location on the body surface out that it concerns with the analyte detected by drug molecules through external contamination are reached on the body surface. Proven Drugs molecules and their metabolites have been deposited by means of welding, that is, they must have been consumed.