PatentDe  


Dokumentenidentifikation EP1090301 05.09.2002
EP-Veröffentlichungsnummer 1090301
Titel NICHTINVASIVEN TRANSDERMAL DETEKTION VON ANALYTEN
Anmelder Transderm Technologies LLC, San Francisco, Calif., US
Erfinder BROWN, R., Larry, Newton, US;
EDELMAN, Elazer, Brookline, US;
TSENG, David, Santa Clara, US
Vertreter derzeit kein Vertreter bestellt
DE-Aktenzeichen 69902391
Vertragsstaaten AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LI, LU, MC, NL, PT, SE
Sprache des Dokument EN
EP-Anmeldetag 24.06.1999
EP-Aktenzeichen 999335698
WO-Anmeldetag 24.06.1999
PCT-Aktenzeichen PCT/US99/14288
WO-Veröffentlichungsnummer 0009967645
WO-Veröffentlichungsdatum 29.12.1999
EP-Offenlegungsdatum 11.04.2001
EP date of grant 31.07.2002
Veröffentlichungstag im Patentblatt 05.09.2002
IPC-Hauptklasse G01N 33/94
IPC-Nebenklasse G01N 33/84   G01N 33/52   

Beschreibung[en]
Background of the Invention

This application claims priority to "Non-Invasive Transdermal Detection of Heavy Metals" described in U.S. Serial Number 60/090,459 filed on June 24, 1998, by Larry R. Brown and Elazer R. Edelman.

This is generally in the area of non-invasive methods and apparatus for sampling of analytes present in body fluids, including sweat, such as glucose, heavy metals, and compounds of abuse.

The development of transdermal methods of delivering drugs through the skin has been made possible by the optimization of solvent conditions of individual drugs so that they solubilize and partition into the stratum corneum skin layer (Brown, L. and Langer, R., "Transdermal Delivery of Drugs." Ann. Rev. Med. 39: 221-229, 1988.). In addition, transdermal diffusion of drug compounds has been also enhanced using iontophoresis, electroporation and ultrasound. These observations have lead to the further investigation of using the transdermal route to detect metabolites in the skin. Virtually all of the work in the scientific literature has been focused in the area of glucose detection in the treatment of diabetes mellitus. However, large fluctuations in glucose concentrations occur can occur within minutes after the ingestion of a carbohydrate loaded meal. In contrast, glucose diffusion through the stratum corneum skin layer is relatively slow and lags behind these glycemic variations. Thus, there is a significant "lag time" between the skin measurement of glucose concentration through the stratum corneum skin layer and the actual blood concentration.

Previous attempts restricted to the detection of glucose have not and can not be extended to heavy metals. A more useful and uninvestigated application of transdermal diffusion is to non-invasively detect analytes whose concentration does not vary significantly over short time periods. This is especially important for the detection of chemicals such as heavy metals. For example, a heavy metals such as lithium used to treat manic depressives could be detected by psychiatrists to easily determine patient compliance. The detection of iron would be useful for the detection of iron overload diseases.

Lead (Pb) is an example of a heavy metal which is also a toxic substance. Public health organizations in the United States require all children to be tested several times before beginning school and during their early education. It is required for kindergarten grade entry in many school districts in the United States. Studies indicate that nearly 10% of the children in the United States ages six and under, or 1.7 million children, are victims of lead poisoning (Carolina Environment, Inc. 8/8/96). The United States Public Health Service estimates one out of six children under the age of six has enough lead in their blood to place them in what scientists consider the risky zone. Childhood lead poisoning has no predilection for socioeconomics or geography.

Although adults are susceptible to lead poisoning, children remain at the highest risk due to the natural instinct to introduce non-food items into their bodies. The effects of lead poisoning include learning, delinquent behavior, hyperactivity, decreased growth, kidney and heart disease, and even brain damage. Common potential lead-contaminated areas include older play equipment and chipping paint from window and door trim, and even walls.

The symptoms of lead poisoning include headaches, irritability, abdominal pain, vomiting, anemia, weight loss, poor attention span, noticeable learning difficulty, slowed speech development, and hyperactivity. The effects of lead poisoning include reading and learning disabilities, speech and language handicaps, lowered I. Q., neurological deficits, behavior problems, mental retardation, kidney disease, heart disease, stroke, and death.

Current test procedures for lead are traumatic for young children as they require venapuncture and extraction of a blood sample. The scientific literature does not describe non-invasive methodologies for detecting heavy metals via the dermal route.

There are numerous descriptions of the toxicology of Pb absorbed through the skin and then found to result in toxic effects or elevated levels in other tissues. This is especially true in the use of lead for topical cosmetic applications (Moore, et al. Food Cosmet. Toxicol. 18(4):399-405 (980)) and in industrial applications (Hine. et al., J. Occup. Med. 11(11):568-75 (1969)). In one citation in the scientific literature, sweat is used to assess the absorption of lead through the skin, but not from systemic blood supply and organs to the skin (Lilley, et al. Sci. Total Environ. 76(2-3):267-78 (1988)).

Other examples of plasma heavy metal concentrations caused by skin absorption include zinc (Morgan, et al. Br. J. Dermatol. 102(5):579-83 (1980)). Cadmium, chromium, and arsenic have also been detected in male reproductive organs (Danielsson, et al., Arch. Toxicol. Suppl. 7:177-80 (1984)).

Sweat lead detection was well studied by an Australian group at CSIRO, Menai, Australia. Stauber and colleagues worked on sweat lead detection for occupational lead absorption through skin. They found that even inorganic lead can be absorbed through skin and rapidly distributed through the body (Stauber et al., 1994). In one of their experiments (Lilley et al., 1988), lead powder was placed on the left arm of a healthy adult male volunteer, a certain region of skin of the right arm was induced to sweat for lead detection. The placing of 6 mg of lead as 0.5 M lead nitrate to the left arm resulted in the increase in lead concentration in pilocarpine-induced iontophoresis samples in the right arm. No changes were found in the blood or urine samples. However, Omokhodion and Crockford (1991) found that there is a good relationship between blood and sweat lead levels among non-occupationally exposed persons.

Many other substances must be measured on a frequent basis, resulting in trauma and pain to the patient. Examples include measuring blood glucose in diabetics and sampling drugs of abuse in cocaine addicts. Efforts for the non-invasive detection of glucose have focused on transdermal extraction of glucose using solvents, iontophoresis or using the penetration of infrared light through the skin. None of these glucose efforts have been commercialized. Among the scientific difficulties associated with glucose detection are that glucose concentrations change transiently and quickly throughout the day, and often do so at rates which exceed the permeability rate of the glucose molecule through the skin. Therefore the iontophoretic or solvent extraction routes have proven to yield irreproducible data. The use of infrared light has been plagued with interference from other substances of similar structure found in vivo and the inability to calibrate these devices reproducibly.

Transdermal detection of substrate other than lead from sweat has been applied to non-invasive devices. Several transdermal alcohol detection devices have been developed by Dermal Systems International to detect alcohol from sweat (Swift 1993). One of those methods is an alcohol dosimeter or "sweat patch" which is a portable, wearable, non-invasive, occlusive patch applied to the skin surface. However, it requires 7-10 day period for the detection. Another alcohol "Band-Aid" is a small strip applied to the skin that utilized enzymatic colorimetric detection to estimate blood ethanol concentration over several minutes (Roizman and Lichtor, 1990). However, estimating concentration of a drug (e.g. alcohol) across pharmacokinetic compartments (blood and skin) is not always straightforward. Sweat detection also involves the complexity of both passive diffusion through the skin (Scheuplein, and Blank, 1971) and active secretion by eccrine glands, primarily sweat glands (Brusilow and Gordis, 1966).

"Band-Aid" sweat pitch has also been tested in a human clinical study to monitor drug use, for example with Cocaine (Burns and Baselt, 1995). A cellulose based patch collected sweat in a seven day period and a substantial cocaine concentrations were detected in the patches. However, quantitative interpretation was beyond the "capabilities of the current state of the technology" (Burns and Baselt, 1995). This sweat patch was developed by Sudormed, Inc. which was later acquired by Pacific Biometrics, Inc. (Lake Forest, CA).

"Reverse" transdermal detection has been used for glucose detection. Because of the non-polar property of the glucose molecule, the "reverse" transdermal glucose detection requires certain enhanced methods, such as iontophoresis and ultrasound. Studies of "reverse" transdermal detection has demonstrated some results for the detection of glucose in the presence of current (Glikfeld et al., 1989; Rao et al., 1993). However, all of these methods require complex apparatus to extract the analytes.

. It is therefore an object of the present invention to provide a method and apparatus for non-invasively obtaining samples of body fluids for testing for the presence and/or amount of a particular analyte in the body fluid, which is easily and inexpensive.

Summary of the Invention

A method and apparatus for in-invasive extraction and/or detection of chemicals such as heavy metals and cocaine and analytes such as blood glucose has been developed. A patch or hydrogel containing a reagent such as N-methyl pyrrole, or a similar pyrrolidone or mixture thereof, is used to extract the chemical or analyte to be measured through the epidermis and/or hair. This method is useful for detecting analytes which are generally present in relatively constant blood concentrations. The method is particularly useful for the detection of heavy metals such as lead, lithium, copper, iron, and has been demonstrated to be useful to extract both drugs and metabolic analytes. In the preferred embodiment, the method involves the use of the water miscible solvent N-methyl pyrrolidone (NMP) in an aqueous solution which is incorporated into an adsorbent pad or hydrogel. This adsorbent pad is placed on the skin for a defined time period, removed and then analyzed for the substance of interest. Analytical means can also be incorporated as part of the adsorbent pad in order to conduct both the extraction of the chemical of interest and the assay in situ. Examples demonstrate measurement of lead, cocaine, glucose, and acetominophen, demonstrating the very broad range of materials which can be extracted. Comparative examples demonstrate that the same results cannot be obtained using other reagents such as dimethylsulfoxide (DMSO).

Brief Description of the Drawings

  • Figure 1 is a schematic of
  • Figure 2 is a graph of lead extraction through skin at 3 hr (n=3).
  • Figure 3 is a graph of the lead flux (µg/cm2/hr)) through skin as a function of NMP concentration (0%, 25%, 50%, 75%NMP in water)
  • Figure 4 is a graph of the flux of lead (µg/cm2/hr)) through swine skin as a function of lead acetate concentration (µg/mL).
  • Figure 5 is a graph of lead concentration (µg/cm2) for control rat, water extracted rat, and NMP-hydrogel extracted rat.
  • Figure 6 is a graph of cadmium and mercury extraction through swine skin (n=3) over time (min).
  • Figures 7a-7f are graphs of extraction of lead: Figure 7a is a graph of blood lead lvels (µg/mL) in rats over time in days following administration of lead acetate in their drinking water. Figure 7b is a graph of total lead extraction (µg/cm2) using NMP-poloxamer over time in minutes. Figure 7c is a graph of lead extraction with NMP-poloxamer or agarose hydrogel patches. Figure 7d is a graph of total lead extraction (µg/cm2) through rodent skin with NMP-gauze patches, at four and 24 hours. Figure 7e is a graph of the correlation of the blood lead levels with lead extracted by the NMP-poloxamer hydrogel patches. Figure 7f isa graph showing extraction of lead from rodent fur using NMP-saline.
  • Figure 8a is a graph of acetaminophen extracted through human skin using 50%NMP:50%water-hydrogel. Figure 8b is a graph comparing extraction of acetaminophen through human skin with NMP-water versus 2-pyrrolidone-water.
  • Figure 9 is a graph comparing extraction of lead (µg/cm2/hr) through rat skin using either 50% NMP:50% water hydrogel or 25% DMSO-hydrogel.
  • Figure 10 is a graph of cumulated cocaine transported through skin (µg/cm2) over time in hours.
  • Figure 11 is a graph of cocaine extracted (µg/cm2/hr) by either polxamer or NMP-poloxamer.
  • Figure 12 is a graph of cumulated lead acetated extracted through swine skin over time, comparing 50% DMSO (circles) and 50% NMP (squares).
  • Figure 13 is a graph of the amount of lead extracted through rat skin (µg/cm2/hr) comparing 50%NMP-poloxamer with 25% DMSO-poloxamer.
  • Figure 14 is a graph comparing glucose concentration extracted using 50% NMP:50% 0.9% NaCl with capillary blood glucose (mg/dl) over time (min).

Detailed Description of the Invention

Compositions and methods for specifically extracting chemicals and chemical compounds, such as heavy metals through the skin, are disclosed. Glucose extraction through the skin with these compositions and methods is also disclosed.

Materials Used for Extraction of Chemical Compounds or Analytes Extraction Media

The general class of compounds that can be used to extract the compounds to be measured or detected include those pyrrolidones and alkyl pyrrolidones with the following chemical structure which can be co-solubilized with water in order to form a hydrogel using polymers such as poloxamer 407 (Plutonic™ F127, or Lutrol™ F127, BASF corporation) or polyvinyl alcohol, poly hydroxy methacrylate, or poly hydroxy ethyl methacrylate.

Examples of the general chemical structure for the class of compounds used are shown in the figure below. It should be noted that the preferred compounds are 2-pyrrolidone and N-methyl pyrrolidone and most preferred is the N-methyl -pyrrolidone (NMP).

  • R1 = H, CH3, HO-CH2-CH2-, CH3-CH2-, CH3-(CH2)n n=1 to 10
  • R3 = H, -OH, or an alkyl group such as dodecyl
  • R4 = H, methyloxycarbonyl
  • R5 = H, CH3, CH3-CH2-, HO-CH2-CH2

The following class of pyrrolidones are included: alkyl pyrrolidones and pyrrolidone derivatives; 2-pyrrolidone, 1 methyl-2-pyrrolidone, 1 ethyl 2 pyrrolidone, 1 hexyl-2-pyrrolidone, 1-lauyyl-2-pyrrolidone N-(2-hydroxyethyl)-2-pyrrolidone, 1, 5-dimethyl-2-pyrrolidone, 5-methyl-2-pyrrolidone, 1-hexyl-4-methyl oxycarbonyl-2-pyrrolidone, 1 lauryl-4-methyl oxycarbonyl-2―pyrrolidone, N-cyclohexyl-2-pyrrolidone, N-dodecyl-2-pyrrolidone, and 1-butyl-3-dodecyl-2-pyrrolidone. For ease of reference, this class of material is generally referred to as NMP. When referenced in the examples, NMP is N-methyl pyrrolidone.

Pharmaceutical grade NMP (Pharmasolve®) is available from International Specialty Products (Wayne, NJ). Pharmasolve® is presently used as a solvent in a gel in a FDA approved oral dental product developed by Atrix Laboratories (Fort Collins, Co.). The advantage of NMP is its water miscibility allowing the formation of hydrogels and its excellent safety and toxicity profile.

The examples demonstrate the extraction and solubilization of both water soluble and lipid soluble compounds from the skin. N-methyl Pyrrolidone (NMP) and 2-Pyrol (products of GAF) combined with 0.9% NaCl were used to extract and measure the amounts of heavy metals through the skin. Preferably, the NMP and 2-Pyrol are combined 50/50 with the 0.9% NaCl. The amounts of heavy metals thus extracted are indicative of circulating concentrations of these compounds.

N-methl-2-pyrrolide (NMP) has been used as an effective drug delivery skin penetration enhancer. Normal skin is capable of maintaining a reasonably constant level of hydration even when large changes in environment humidity occur. This is because of the presence of a natural moisture factor within the stratum corneum epidermal barrier. The natural moisture factor is believed to consist mainly of free fatty acids, urea, and pyrrolidone carboxylic acid and its sodium salt. Sodium pyrrolidone carboxylate appears to be the principal humectant and increases the water-binding capacity of the stratum corneum. NMP is one of the many analogs of sodium pyrrolidone carboxylate. NMP has shown to increase the transport of steroids, caffeine, ibuprofen, flurbiprofen, and aspirin across skin (Akhter and Barry, 1985; Walters, 1989). However, NMP has never been used to extract any chemical substances from skin.

Carriers

The extraction solvent can be administered as a solution, in an absorbant pad or other material, or in a hydrogel. Diluents are preferably aqueous, most preferably water or 0.9% NaCl. The NMP can also be mixed with, bound to, or incorporated into an absorbant material such as a cellulosic pad, agarose, acrylamide gel or beads, or other materials commonly used as absorbants. Alternatively, the NMP can be incorporated into a hydrogel formed of a preferably hydrophilic polymer or other water miscible materials such as a polaxamer (a polypropylene glycolpolypropyleneoxide block co-polymer) like poloxamer 407&peseta;(Plutonic™ F127, or Lutrol™ F127, BASF corporation), polyvinyl alcohol, poly hydroxy methacrylate, poly hydroxy ethyl methacrylate or propylene glycol. Surfactants such as Tween 20, Tween 80, Brij 30 and 36T, menthol, phospholipids, and azone.

The pyrrolidone in combination with carrier can be applied directly to the skin and/or hair from which the chemical compound is to be extracted. This will be left in place for a time effective to extract an amount of chemical compound to be detected and/or measured. The mixture may be applied as a device as simple as a bandaid type device, with the pyrrolidone absorbed into the cellulosic pad, or it may be packaged in a small metal or plastic "cup", which is strapped onto the appropriate site using an adhesive, tape, or an outer fabric or leather strap, similar to that worn as part of a watch. The entire device may be disposable or may be refillable.

Compounds to be Detected or Measured

A wide variety of compounds can be extracted using these materials. These include metals such as lead, cadmium, and mercury, organic compounds, especially drugs, such as cocaine, caffeine, and acetominophen, and biological analytes and/or metabolites, such as glucose.

Lead (Pb) is a particularly important toxic metal to measure and is presently measured in all young children. Other metals which can be extracted include lithium, copper, iron, mercury, and cadmium.

Many drugs are tested, either to establish that a person is adhering to a program to stop a particular addiction (caffeine, cocaine, smoking), or to monitor drug levels (ex. rheumatoid arthritis patients taking high doses of non-steroidal antiinflammatories such as acetominophen). These can all be detected by extracting the chemical compounds into the polypyrrole.

Other materials to be tested for include metabolic analytes or chemical compounds such as glucose.

The device can include means for detection for these chemical compounds. Detection means may be qualitative or quantitative. Examples include enzyme chromatographic means, such as an enzyme-substrate assay that results in a color change in the presence of substrate; antibody assays, where the antibodies are labelled and migrate to a region where reaction with substrate can be detected. Alternatively, the extracted chemical compound can be removed, for example, using a pipette, capillary tube or syringe type means, and put into a separate detection device such as an HPLC or spectrophotometer.

Kits containing these devices will typically include one or more devices, each of which contains the pyrrolidone/carrier mixture, optionally including devices for removing the extracted chemical compounds from the pyrrolidone/carrier mixture for measurment or detection of the chemical compounds and/or means for measurement or detection of the chemical compounds.

The present invention will be further understood by reference to the following non-limiting examples.

Example 1. Extraction of lead through skin skin in a diffusion cell.

All of the results from the in vitro experiments were conducted using flow teflon diffusion cells of the type described by Bronaugh and Steward (J. Pharm. Sci. 1985, 74: 64-67). The cross section area of a single diffusion cell was 0.64cm2. Figure 1 is a schematic diagram of a typical diffusion cell setup.

Solutions containing 75% N-methyl pyrrolidone (Pharmasolve, ISP, Wayne, NJ) (NMP):25% deionized water, 50% NMP:50% deionized water, 25% NMP:75% deionized water and 0% NMP: 100% deionized water were used as extraction solvents and placed in the receptor compartment of the diffusion cell shown below. The donor solution contained 5, 10, or 20µg/ml lead acetate. Samples were measured on an Atomic Absorption Spectrometer (Perkin-Elmer AAnalyst 300).

Donor solutions containing either 10µg/ml or 5µg/ml lead acetate were introduced on the surface of pig skin. The receptor solutions contained either 50% NMP-water or water alone. Figure 2 shows that there was insignificant diffusion of lead into receptor solutions containing water alone, while those receptor solutions containing 50% NMP:50% water extracted significant amounts of lead at values in excess of 0.02µg/ml (p<0.01, Figure 2).

Example 2. Determination of Optimal NMP Concentration for Lead Extraction.

In an experiment designed to show that there is an optimal mixture of NMP and water or aqueous buffer, the NMP:deionized H2O solutions were varied between 0 and 75% NMP. Solutions containing 75% N-methyl pyrrolidone (Pharmasolve, ISP, Wayne, NJ) (NMP):25% deionized water, 50% NMP:50% deionized water, 25% NMP:75% deionized water and 0% NMP:100% deionized water were used as extraction solvents and placed in the receptor compartment of the diffusion cell shown in Figure 1. The donor solution contained 20µg/ml lead acetate. The data in Figure 3 demonstrates the ability of NMP to enhance the extraction of Pb significantly when compared to H2O alone, and that 50% NMP:50% H2O results in the maximum flux when compared to other mixtures of NMP and water.

Example 3. Extraction of Lead over Time Using NMP.

Experiments were conducted over a 4 hour period. The donor solutions contained lead acetate at 20µg/ml, 10µg/ml, and 5µg/ml, and 0µg/ml. The extraction solution contained 50% NMP:50% water.

As shown by Figure 4, the 50% NMP-water solution was capable of extracting lead acetate through the pig skin for up to a 4 hour period, linearly with time and at concentrations as low as 5µg/ml.

Example 4. In vivo demonstration of the detectability of lead extracted through the skin of rats.

A clear hydrogel was formed containing 1 gram of the copolymer Pluronic™ F127 (BASF) and 2.5 mL of dionized water and 2.5 mL of N-methyl pyrrolidone (Pharmasolve, ISP, Wayne, NJ). The hydrogel is easily molded into any shape or container. It also allows and maintains excellent skin surface contact. Lead is introduced into the blood of the rats by feeding the rats with 750 to 1,000 ppm lead acetate drinking water.

Figure 5 shows that the 50% NMP:50% water Pluronic™ hydrogel extracts lead from rats in an in vivo experiment. Each bar represents the mean of 5 rats.

Example 5. In vitro extraction of the heavy metals mercury (Hg) and cadmium (Cd).

The goal of the in vitro experiments was to prove that NMP could enhance the diffusion of cadmium acetate (Cd) and mercury (Hg) through swine skin. The experiments were conducted in a continuous diffusion cell system. Three diffusion cells contained cadmium acetate (50µg/ml) in donor chambers and NMP:dH2O (50:50) solution in receptor chambers. Three diffusion cells contained mercury (50µg/ml) in donor chambers and NMP:dH2O (50:50) solution in receptor chambers. Two diffusion cells contained cadmium acetate (50µg/ml) in donor chambers and dH2O solution in receptor chambers. Initial samples were taken at 5min time points. Subsequent samples were taken at 15min intervals thereafter.

The cadmium acetate concentrations from receptor chambers were measured on graphite Atomic Absorption Spectrometer (Perkin-Elmer AAnalyst 300). The mercury concentrations from receptor chambers were measured on flame Atomic Absorption Spectrometer (Perkin-Elmer AAnalyst 300). The results are shown in Figure 6. The results were summarized in and shown as X±SD.

The large error bars in Hg extraction was a result of flame AA measurements which generated larger data fluctuations than those of graphite AA measurement which were used for Pb and Cd assays.

The experimental results showed that NMP/dH2O solution enhances the diffusion of cadmium acetate (Cd) through swine skin; NMP/dH2O solution enhances the diffusion of mercury (Hg) through swine skin; and dH2O (control) solution agent is not able to enhance the diffusion of target chemicals (e.g. Cd) through swine skin.

The experimental results showed that NMP:dH2O solution enhances the diffusion of cadmium acetate (Cd) through pig skin; NMP:dH2O solution enhances the diffusion of mercury (Hg) through pig skin, and the extraction solvent containing only deionized H2O as a control did not result in significant cadmium extraction from pig skin.

Example 6 In vivo correlation of blood lead and lead extracted through in of rats in vivo.

Five male Sprague Dawley rats (approximately 400 gms) were fed a diet which contained 750-1,000 ppm lead acetate in their water supply. Determinations of blood lead (Pb) concentrations were conducted by obtaining a blood sample from the tail vein of the animals and analyzing them using an ESA brand LeadCare Analyzer (Chelmsford, MA). After the blood lead levels were determined to be in the 50-60 µg/dL range for several weeks , the leaded drinking water supply was removed (Figure 7a).

At that time, a clear hydrogel formulation containing 1 gram of the copolymer Pluronic F127 (BASF), and 2.0 mL of deionized water and 2.0 mL of N-methyl pyrrolidone (Pharmasolve, ISP, Wayne, NJ) was prepared. Approximately 1.2 grams of hydrogel was applied to a 2.4 cm diameter polypropylene plastic disk. The disks were attached to a jacket which was affixed to the backs of the rats. The disks were removed after 1 hour. The NMP - hydrogel was removed and the lead content was determined by Atomic Absorption Spectroscopy (Perkin-Elmer AAnalyst 300). A similar study was conducted using agarose instead of the Poloxamer. The data is shown in Figure 7b as the µg of lead per cm2 over time in minutes and in Figure 7c as total lead extracted.

Similar extractions were made using gauze patches, as shown in Figure 7d.

Figure 7e shows the correlation between blood lead concentrations and lead extraction. The data representing 60 blood lead and skin extracted lead determinations clearly demonstrates the NMP-hydrogel's ability to extract significant quantities of lead, which correlate with blood lead concentrations.

Figure 7f demonstrates that lead can also be extracted using the NMP-hydrogel from rodent fur.

Example 7. The detection of Acetaminophen in human skin.

Two human subjects ingested approximately 5 grams of acetaminophen over a 26 hour period. The acetaminophen was detected through human skin by applying a two different hydrogels on the forearm of each subject. One hydrogel contained 25% Lutrol™ F127 (Pluronic™ F127, BASF) formed with water alone. The second hydrogel was formed by combining 1 gram of the copolymer Pluronic™ F127 (Lutrol™, BASF,) AND 2.5 mL of deionized water and 2.5 mL of N-methyl 2-pyrrolidone (Pharmasolve™, ISP, Wayne, NJ) to form 50% NMP:50% H2O, a clear hydrogel.

The hydrogel is easily molded into any shape or container. It also allows and maintains excellent skin surface contact. Approximately 1.44 gram of the hydrogel was applied to a 10.5 cm2 polypropylene plastic frame. The gel was secured to the test subjects' forearms using surgical tape. The gel was removed after 40 minutes and analyzed using the Sigma Diagnostics Acetaminophen Assay (catalog #430-A). Samples containing significant concentrations of acetaminophen turn yellow in color. The color intensity was also measured spectrophotometrically. The results are graphed in Figure 8a, which demonstrates the ability of the NMP:water Pluronic™ hydrogel to effectively extract acetaminophen from humans treated with the oral drugs of the drug by extracting small quantities of acetaminophen through the skin. However, significantly more acetaminophen was extracted when the F127 hydrogel contained 50% NMP:50% water.

A second trial with two subjects yielded total acetaminophen values that were approximately 36% higher for the NMP hydrogel for test subject one and 17% higher for test subject two compared to water only Pluronic hydrogel.

Example 8. Extraction of Acetaminophen in human skin using alternate hydrogels.

Using the techniques in Example 7, 50% NMP:50% water was loaded into Pluronic™ 127, polyvinyl alcohol, Sephadex™ and Agarose. The gels were placed on a test subjected who had ingested 1 gram of acetaminophen for 45 minutes. The gels were analyzed for acetaminophen using the Sigma Diagnostics Acetaminophen Assay (catalog #430-A).

Yellow color development was greatest with the Pluronic™ F127 formulation. The Poly Vinyl Alcohol formulation also showed the extraction of acetaminophen, however the yellow color development was less than the Pluronic F127 formulation. Little or no yellow color change was seen with the agarose or Sephadex™ gels.

Example 9. Extraction of Acetaminophen in human skin using alternate pyrrolidone derivatives.

4 grams of acetaminophen was orally ingested at a dose equal to 1000 mg every 6 hours over a 26 hour period by a human subject.

1 gram Pluronic™ F-127 was dissolved in either:

4 mL. of water (water hydrogel), or

4 mL. of a 50% by volume solution ofN-methyl-2-pyrrolidone-water (NMP - hydrogel), or

4 mL. of a 50% by volume solution of 2-pyrrolidone- water (2-pyrrolidone hydrogel).

Approximately 1.25 gram of the hydrogel was applied to a 10.5 cm2 polypropylene plastic disk. The plastic disk containing the hydrogel was secured to the test subject's forearm using surgical tape. The gel was removed after 40 minutes and analyzed using the Sigma Diagnostics Acetaminophen Assay (catalog # 430-A). Samples containing significant concentrations of acetaminophen turn yellow in color. The color intensity was also measured spectrophotometrically.

The results are shown in Figure 8b. Yellow color development was greatest with the NMP-hydrogel. A significant amount of acetaminophen was also recovered using the 2-pyrollidone hydrogel. Only very small quantities of acetaminophen was detected using the water only hydrogel. Thus, it appears that the transdermal extraction of acetaminophen can be accomplished with the general class of alkyl (N-methyl) and H (2-pyrrolidone) substituted pyrrolidone transdermal enhancing agents.

Example 10. The detection of [14C]-caffeine administered to rats through skin.

Rats were shaven and fasted for 24 hours prior to caffeine administration by intraperitoneal injection of 14C labeled caffeine to the 600 gram rats. Each animal received approximately 16.7 µCi of 14C caffeine via intraperitoneal injection of 0.5 mL of ethanol and water.

A 50% NMP:50% water - Pluronic™ F127 hydrogel was applied to a 2.4 cm diameter plastic disks. The disks were attached to a jacket which was affixed to the backs of the rats. The disks were removed after 2.5 hours The gel was removed and counted. The rats were sacrificed and blood and tissue were removed for analysis from each animal. The tissue samples were placed in liquid scintillation vials with 1.5 hours. The gel was removed and counted. The rats were sacrificed and blood and tissue were removed for analysis from each animal. The tissue samples were placed in liquid scintillation vials with 1.5mL of soluene (Packard Instruments, Downers Grove, IL) in order to dissolve the tissues for valid liquid scintillation counting.

The table below shows the relative amounts of radioactivity found in each of the samples. Sample Radioactivity (mean dpm ± SD) Skin 52.15 ± 0.07 dpm/mg Muscle 64.30 ± 8.73 dpm/mg Blood 87.24 dpm/µL NMP-Pluronic F127 Hydrogel 58.82 dpm/cm2/hr

The data demonstrates that the tissue, skin and blood concentration of 14C labeled caffeine are of the same order of magnitude. This suggests that the ability to extract drugs from the skin should result in a value which is representative of the animal's steady state concentrations of the drug, molecule or chemical. Indeed, the data in the table shows that the NMP containing hydrogel was shown to extract a significant quantity of radioactive caffeine through the rat.

Example 11. In vitro Extraction of Cocaine.

3H-cocaine transport through swine skin was conducted with diffusion cells (4.5 cm diameter). 3H-cocaine donor solution (20µCi/ml) was loaded in the upper chamber. The receptor solution (50% NMP:H2O) was loaded in lower chamber. Samples from lower chamber (100µl) were taken every hour and counted.

The results are shown in Figure 10. The flux of 3H-cocaine through pig skin in vitro was determined to be 0.00317 µg/cm2/hr.

Example 12. In vivo Extraction of Cocaine.

Rats (500-600 g) were shaven prior to cocaine administration by intraperitoneal injection of 3H-cocaine (20µCi/ml). Each animal received a dose of approximately 40 µCi. Using the techniques described in Example 8, both NMP:Poloxamer and Water:Poloxamer patches were applied to the shaved rat skin. The hydrogel patches were removed after two hours. The hydrogel was collected and the amount of 1H-cocaine extracted in hydrogel was counted by liquid scintillation counting.

The results are shown in Figure 11.

Example 13. Comparison of Extraction using either NMP or DMSO from Swine Skin (in vitro) or Rats (in vivo).

In vitro extraction of lead acetate by NMP and DMSO through swine skin was conducted with diffusion cells (4.5cm diameter). Lead acetate donor solution (200 µg/ml) was loaded in upper chamber. The receptor solution (50% NMP:H2O was loaded in lower chamber. Samples from lower chamber (1ml) were taken every hour and measured by Atomic Absorption Spectroscopy.

The results are shown in Figure 12.. The fluxes for lead acetate through swine skin for NMP and DMSO as extraction agents are 0.279µg/cm2/hr and 0.040µg/cm2/hr respectively.

Rats fed with lead acetate were used in the experiment. Poloxamer dydrogel patches contained 50% NMP or 25% DMSO were applied to the shaved rat skin. The hydrogel patches were removed after one hour. Hydrogel was collected and the amount of lead extracted in hydrogel was measured on AA.

The results are shown in Figure 13.

Example 14. Extraction of glucose from human skin using NMP or 2-Pyrol Aqueous Solutions.

N-methyl 2-Pyrrolidone (NMP) and 2-Pyrol combined in 50% by volume and 50% by 0.9% NaCl in aqueous solution have been shown to allow significant glucose concentrations measurement through the skin.

NMP or 2-Pyrrol were mixed in equal volumetric proportions with 0.9% NaCl solution in water. In addition, a solution containing 0.9% NaCl alone was prepared. The skin on the forearm was thoroughly washed with soap and water and swabbed clean with 70% isopropyl alcohol. Approximately 25 µl of a 50:50 solution of NMP:0.9% NaCl solution in H2O or the 0.9% NaCI solution were placed on the forearm for a 5-10 minute period contained within a foil backed adhesive with a circular opening in the center at the end of the sampling period, an additional 10 µl was then placed on the skin and used to recover the test solutions. An approximate 5 to 10 µl drop was applied to a Medisense Exactech (Cambridge, MA) glucose test strip. The normal glucose determination procedure described in the Exactech brochure was followed for the determination of glucose concentration using the Exactech meter.

Simultaneously, capillary blood samples were obtained from a diabetic subject and glucose concentrations were measured with the Medisense Exactech meter as described in their device brochure. A small drop of blood was obtained and placed on the Exactech strip and measured for glucose concentration.

The results are tabulated below. Measurements of Glusoe in Blood compared to Skin Extraction Blood glucose (mg/dl) Skin Glucose (mg/dl) 0.9% NaCl in H2O 159 42 (5 min skin application of test solution) LO (below detection) 54 (10 min skin application of test solution) 207 66 LO 49

The results demonstrat that the NMP:0.9% NaCl solution was effective in enhancing the ability to detect glucose on the skin compared to the 0.9% NaCI control, and that the formulation appears to enable one to detect skin glucose. However, the skin glucose values measured were approximately 1/3 those of the blood glucose values. The NaCl in the water control indicates the importance of the penetration enhancing qualities offered by the NMP used to "extract" the glucose from the skin.

Example 15. Effect of time on glucose concentration collection from skin.

NMP or 2-Pyrrol were mixed in equal volumetric proportions with 0.9% NaCl solution in water. In addition, a solution containing 0.9% NaCl alone was prepared. The skin on the forearm was thoroughly washed with soap and water and swabbed clean with 70% isopropyl alcohol. Approximately 25 µl of the 50:50 solution of NMP:0.9% NaCl solution in H2O or the 0.9% NaCl solution was placed on the forearm and samples were measured over a 75 minute period from within a foil backed adhesive with a circular opening in the center to contain the test solution. An approximate 5 to 10 µl drop was applied to a Medisense Exactech glucose test strip. The normal glucose determination procedure described in the Exactech brochure was followed for the determination of glucose concentration using the Exactech meter.

Simultaneously, capillary blood samples were obtained from a diabetic subject and glucose concentrations were measured with the Medisense Exactech meter as described in their device brochure. A small drop of blood was obtained and placed on the Exactech strip and measured for glucose concentration. The results of the time course experiment are shown in Figure 14.

The data indicates that glucose extraction from the skin is not measurable using the 0.9% NaCl test solution. In contrast, and after a 15 minute lag time, the sampling of the 50% NMP:50% 0.9% NaCl solution resulted in significant and measurable skin glucose concentrations over the course of the experiment.


Anspruch[de]
  1. Vorrichtung für die transdermale Detektion von Analyten, die in einem Subjekt vorliegen,

       mit einer wirksamen Menge an Pyrrolidon der folgenden Formel in einem wässrigen Beimischung,
       wobei
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2- oder CH3-(CH2)n-,

    wobei n = 1 bis 11,
    R3 =
    H, -OH, CH3- oder CH3-(CH2)n-, wobei n = 1 bis 11,
    R4 =
    H oder Methyloxycarbonyl und
    R5 =
    H, CH3, CH3-CH2 oder HO-CH2-CH2; und
       mit Mitteln zum Detektieren der Anwesenheit oder der Menge des Analyts, das in die wässrige Pyrrolidon-Mischung extrahiert wurde.
  2. Vorrichtung nach Anspruch 1, wobei das Pyrrolidon aus der Gruppe ausgewählt ist, die aus N-Methyl-2-pyrrolidon, 2-Pyrrolidon, 1-Methyl-2-pyrrolidon, 1-Ethyl-2-pyrrolidon, 1-Hexyl-2-pyrrolidon, 1-Lauryl-2-pyrrolidon, N-(2-Hydroxyethyl)-2-pyrrolidon, 1,5-Dimethyl-2-pyrrolidon, 5-Methyl-2-pyrrolidon, 1-Hexyl-4-methyloxycarbonyl-2-pyrrolidon, 1-Lauryl-4-methyloxycarbonyl-2-pyrrolidon, N-Zyklohexyl-2-pyrrolidon, N-Dodecyl-2-pyrrolidon und 1-Butyl-3-dodecyl-2-pyrrolidon.
  3. Vorrichtung nach Anspruch 2, wobei das Pyrrolidon N-Methyl-2-pyrrolidon (NMP) oder 2-Pyrrolidon ist.
  4. Vorrichtung nach Anspruch 3, wobei das Pyrrolidon N-Methyl-2-Pyrrolidon (NMP) ist.
  5. Vorrichtung nach einem der Ansprüche 1 bis 4, wobei dem Pyrrolidon Wasser beigemischt ist.
  6. Vorrichtung nach einem der Ansprüche 1 bis 4, wobei dem Pyrrolidon eine Salzlösung beigemischt ist.
  7. Vorrichtung nach einem der Ansprüche 1 bis 6, wobei das Pyrrolidon in einer wässrigen Konzentration von mindestens 25 % vorliegt.
  8. Vorrichtung nach einem der Ansprüche 1 bis 7, wobei das Pyrrolidon in einer wässrigen Konzentration von nicht mehr als 75 % vorliegt.
  9. Vorrichtung nach einem der Ansprüche 1 bis 8, wobei das Pyrrolidon NMP ist.
  10. Vorrichtung nach Anspruch 9, wobei das NMP in einer wässrigen Konzentration von etwa 50 % vorliegt.
  11. Vorrichtung nach einem der Ansprüche 1 bis 10 und weiterhin mit:
    • einem Hydrogel,
       wobei die wässrige Pyrrolidon-Mischung in dem Hydrogel vorliegt.
  12. Vorrichtung nach Anspruch 11, wobei das Hydrogel ein Polymer aufweist, das aus der Gruppe ausgewählt ist, welche besteht aus: Poloxameren, Polyvinylalkohol, Polyhydroxymethacrylat, Polyhydroxyethylmethacrylat, Propylenglycol und Acrylamid.
  13. Vorrichtung nach Anspruch 12, wobei das Hydrogel ein Poloxamer aufweist.
  14. Vorrichtung nach Anspruch 13, wobei das Poloxamer ein Poloxamer 407 ist.
  15. Vorrichtung nach einem der Ansprüche 1 bis 10, die weiterhin aufweist:
    • ein Absorptionsmittel,
       wobei die wässrige Pyrrolidon-Mischung innerhalb des Absorptionsmittels vorliegt.
  16. Vorrichtung nach Anspruch 15, wobei das Absorptionsmittel ein Cellulosekissen oder ein Wattebausch ist.
  17. Vorrichtung nach einem der Ansprüche 1 bis 16, wobei die Detektionsmittel einen Antikörpertest umfassen, der spezifisch für das Analyt ist.
  18. Vorrichtung nach Anspruch 17, wobei die Antikörper in der Lage sind, innerhalb der Vorrichtung zu wandern.
  19. Vorrichtung nach Anspruch 17 oder Anspruch 18, wobei der Antikörper spezifisch für ein Analyt ist, das aus der Gruppe ausgewählt ist, welche aus Glukose, Metaboliten, Missbrauchsdrogen, Metaboliten von Drogen und pharmazeutischen Verbindungen besteht.
  20. Vorrichtung nach einem der Ansprüche 1 bis 16, wobei das Analyt ein Schwermetall ist.
  21. Vorrichtung nach Anspruch 20, wobei das Schwermetall aus der Gruppe ausgewählt ist, die aus Blei, Cadmium, Quecksilber, Lithium und Kupfer besteht.
  22. Vorrichtung nach Anspruch 21, wobei das Schwermetall Blei ist.
  23. Vorrichtung nach einem der Ansprüche 1 bis 22, wobei die Detektionsmittel enzymchromatographische Mittel umfassen.
  24. Vorrichtung nach einem der Ansprüche 1 bis 23, die weiterhin ein Oberflächeneigenschaften modifizierendes Mittel aufweist, das aus der Gruppe ausgewählt ist, welche aus Detergenzien, Menthol, Phosphorlipiden und Azone® besteht.
  25. Verfahren für die nichtinvasive transdermale Detektion der Anwesenheit oder der Menge eines Analyts, das in einem Subjekt vorliegt, mit den Schritten:
    • Kontaktieren der Haut des Subjekts mit einer effektiven Menge an Pyrrolidon der folgenden Formel in einer wässrigen Beimischung,
       wobei
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2- oder CH3-(CH2)n-,

    wobei n = 1 bis 11,
    R3 =
    H, -OH, CH3- oder CH3-(CH2)n-, wobei n = 1 bis 11,
    R4 =
    H oder Methyloxycarbonyl, und
    R5 =
    H, CH3, CH3-CH2 oder HO-CH2-CH2,
    für eine Zeit, die ausreichend ist, um es nachweisbaren Mengen des Analyts zu ermöglichen, aus dem Subjekt in die wässrige Pyrrolidon-Mischung extrahiert zu werden; und dann

       Detektieren des extrahierten Analyts.
  26. Verfahren nach Anspruch 25, wobei das Pyrrolidon aus der Gruppe ausgewählt ist, die aus N-Methyl-2-pyrrolidon, 2-Pyrrolidon, 1-Methyl-2-pyrrolidon, 1-Ethyl-2-pyrrolidon, 1-Hexyl-2-pyrrolidon, 1-Lauryl-2-pyrrolidon. N-(2-Hydroxyethyl)-2-pyrrolidon, 1,5-Dimethyl-2-pyrrolidon, 5-Methyl-2-pyrrolidon, 1-Hexyl-4-Methyloxycarbonyl-2-pyrrolidon, 1-Lauryl-4-methyloxycarbonyl-2-pyrrolidon, N-Zyklohexyl-2-pyrrolidon, N-Dodecyl-2-pyrrolidon, und 1-Butyl-3-dodecyl-2-pyrrolidon.
  27. Verfahren nach Anspruch 25 oder Anspruch 26, wobei das Pyrrolidon N-Methyl-2-pyrrolidon (NMP) oder 2-Pyrrolidon ist.
  28. Verfahren nach einem der Ansprüche 25 bis 27, wobei das Pyrrolidon N-Methyl-2-pyrrolidon (NMP) ist.
  29. Verfahren nach einem der Ansprüche 25 bis 28, wobei dem Pyrrolidion Wasser beigemischt ist.
  30. Verfahren nach einem der Ansprüche 25 bis 28, wobei dem Pyrrolidon eine Salzlösung beigemischt ist.
  31. Verfahren nach einem der Ansprüche 25 bis 30, wobei das Pyrrolidon in einer wässrigen Konzentration von mindestens 25 % vorliegt.
  32. Verfahren nach einem der Ansprüche 25 bis 31, wobei das Pyrrolidon in einer wässrigen Konzentration von nicht mehr als 75 % vorliegt.
  33. Verfahren nach einem der Ansprüche 25 bis 32, wobei das Pyrrolidon NMP ist.
  34. Verfahren nach Anspruch 33, wobei das NMP in einer wässrigen Konzentration von etwa 50 % vorliegt.
  35. Verfahren nach einem der Ansprüche 25 bis 34, wobei die wässrige Pyrrolidon-Mischung in einem Hydrogel vorliegt.
  36. Verfahren nach einem der Ansprüche 25 bis 34, wobei die wässrige Pyrrolidon-Mischung in einem Absorptionsmittel vorliegt.
  37. Verfahren nach Anspruch 36, wobei das Absorptionsmittel aus der Gruppe ausgewählt ist, die aus Cellulosekissen und Wattebäuschen besteht.
  38. Verfahren nach einem der Ansprüche 25 bis 37, wobei die Zeit nicht mehr als 24 Stunden beträgt.
  39. Verfahren nach einem der Ansprüche 25 bis 37, wobei die Zeit nicht mehr als sechs Stunden beträgt.
  40. Verfahren nach einem der Ansprüche 25 bis 37, wobei die Zeit nicht mehr als 60 Minuten beträgt.
  41. Verfahren nach einem der Ansprüche 25 bis 37, wobei die Zeit nicht mehr als 30 Minuten beträgt.
  42. Verfahren nach einem der Ansprüche 25 bis 41, wobei das Analyt innerhalb der wässrigen Pyrrolidon-Mischung detektiert wird.
  43. Verfahren nach einem der Ansprüche 25 bis 41, das weiterhin vor dem Detektieren des Analyts den Schritt aufweist:
    • Entfernen des extrahierten Analyts von der Pyrrolidon-Mischung.
  44. Verfahren nach einem der Ansprüche 25 bis 43, wobei die Analytdetektion qualitativ ist.
  45. Verfahren nach einem der Ansprüche 25 bis 43, wobei die Analytdetektion quantitativ ist.
  46. Verfahren nach einem der Ansprüche 25 bis 45, wobei das Analyt kolorimetrisch detektiert wird.
  47. Verfahren nach einem der Ansprüche 25 bis 45, wobei das Analyt spektralphotometrisch detektiert wird.
  48. Verfahren nach einem der Ansprüche 25 bis 45, wobei das Analyt enzymchromatographisch detektiert wird.
  49. Verfahren nach einem der Ansprüche 25 bis 45, wobei das Analyt durch einen Antikörpertest detektiert wird.
  50. Verfahren nach einem der Ansprüche 25 bis 47, wobei das Analyt ein Schwermetall ist.
  51. Verfahren nach Anspruch 50, wobei das Schwermetall aus der Gruppe ausgewählt ist, die aus Blei, Cadmium, Quecksilber, Lithium und Kupfer besteht.
  52. Verfahren nach Anspruch 51, wobei das Schwermetall Blei ist.
  53. Verfahren nach einem der Ansprüche 25-49, wobei das Analyt Glukose ist.
  54. Verfahren nach einem der Ansprüche 25-49, wobei das Analyt ein Metabolit ist.
  55. Verfahren nach einem der Ansprüche 25 bis 49, wobei das Analyt eine Missbrauchsdroge oder ein Abbauprodukt derselben ist.
  56. Verfahren nach Anspruch 55, wobei die Missbrauchsdroge aus der Gruppe ausgewählt ist, die aus Kokain, Nikotin und Koffein besteht.
  57. Verfahren nach einem der Ansprüche 25-49, wobei das Analyt eine pharmazeutische Verbindung ist.
  58. Verfahren nach Anspruch 57, wobei die pharmazeutische Verbindung ein nichtsteroidales entzündungshemmendes Mittel ist.
  59. Set für die transdermale Detektion von Analyten, die in einem Subjekt vorliegen,

       mit einer Vorrichtung zur transdermalen Extraktion von detektierbaren Mengen des Analyts, wobei die Vorrichtung eine effektive Menge an Pyrrolidon der folgenden Formel in einer wässrigen Beimischung aufweist,
    wobei
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2- oder CH3-(CH2)n-,

    wobei n = 1 bis 11,
    R3 =
    H, -OH, CH3- oder CH3-(CH2)n-, wobei n = 1 bis 11,
    R4 =
    H oder Methyloxycarbonyl, und
    R6 =
    H, CH3, CH3-CH2 oder HO-CH2-CH2; und
       Mit Mittel zum Detektieren des in die wässrige Pyrrolidon-Mischung extrahierten Analyts.
Anspruch[en]
  1. A device for the transdermal detection of analytes present within a subject, comprising:
    • an effective amount of pyrrolidone of the following formula in aqueous admixture,
    wherein
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2-, or CH3-(CH2)n-, wherein n=1 to 11,
    R3 =
    H, -OH, CH3-, or CH3-(CH2)n-, wherein n=1 to 11,
    R4 =
    H, or methyloxycarbonyl, and
    R5 =
    H, CH3-, CH3-CH2-, or HO-CH2-CH2; and
       means for detecting the presence or amount of analyte extracted into said aqueous pyrrolidone mixture.
  2. The device of claim 1, wherein said pyrrolidone is selected from the group consisting of N-methyl 2-pyrrolidone, 2-pyrrolidone, 1-methyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, 1-hexyl-2-pyrrolidone, 1-lauryl-2-pyrrolidone, N-(2-hydroxyethyl)-2-pyrrolidone, 1,5-dimethyl-2-pyrrolidone, 5-methyl-2-pyrrolidone, 1-hexyl-4-methyloxycarbonyl-2-pyrrolidone, 1- lauryl-4-methyloxycarbonyl-2-pyrrolidone, N-cyclohexyl-2-pyrrolidone, N-dodecyl-2-pyrrolidone, and 1-butyl-3-dodecyl-2-pyrrolidone.
  3. The device of claim 2, wherein said pyrrolidone is N-methyl 2-pyrrolidone (NMP) or 2-pyrrolidone.
  4. The device of claim 3, wherein said pyrrolidone is N-methyl 2-pyrrolidone (NMP).
  5. The device of any one of claims 1 - 4, wherein said pyrrolidone is admixed with water.
  6. The device of any one of claims 1 - 4, wherein said pyrrolidone is admixed with saline.
  7. The device of any one of claims 1 - 6, wherein said pyrrolidone is at an aqueous concentration of at least 25%.
  8. The device of any one of claims 1 - 7, wherein said pyrrolidone is at an aqueous concentration of no more than 75%.
  9. The device of any one of claims 1 - 8, wherein said pyrrolidone is NMP.
  10. The device of claim 9, wherein said NMP is present at an aqueous concentration of 50%.
  11. The device of any one of claims 1 - 10, further comprising:
    • a hydrogel,
    wherein said aqueous pyrrolidone mixture is within said hydrogel.
  12. The device of claim 11, wherein said hydrogel comprises a polymer selected from the group consisting of: poloxamers, polyvinyl alcohol, polyhydroxymethacrylate, polyhydroxyethyl methacrylate, propylene glycol and acrylamide.
  13. The device of claim 12, wherein said hydrogel comprises a poloxamer.
  14. The device of claim 13, wherein said poloxamer is poloxamer 407.
  15. The device of any one of claims 1 - 10, further comprising:
    • an absorbant,
    wherein said aqueous pyrrolidone mixture is within said absorbant.
  16. The device of claim 15, wherein said absorbant is a cellulosic pad or gauze patch.
  17. The device of any one of claims 1 - 16, wherein said detection means include antibody assays specific for said analyte.
  18. The device of claim 17, wherein said antibodies are capable of migrating within said device.
  19. The device of claim 17 or claim 18, wherein said antibody is specific for an analyte selected from the group consisting of glucose, metabolites, drugs of abuse, metabolites of drugs of abuse, and pharmaceutical compounds.
  20. The device of any one of claims 1-16, wherein said analyte is a heavy metal.
  21. The device of claim 20, wherein said heavy metal is selected from the group consisting of lead, cadmium, mercury, lithium and copper.
  22. The device of claim 21, wherein said heavy metal is lead.
  23. The device of any one of claims 1 - 22, wherein said detection means include enzyme chromatographic means.
  24. The device of any one of claims 1 - 23, further comprising a surface property modifying agent selected from the group consisting of surfactants, menthol, phospholipids, and Azone®.
  25. A method for the noninvasive transdermal detection of the presence or amount of an analyte present within a subject, comprising:
    • contacting the skin of said subject with an effective amount of pyrrolidone of the following formula in aqueous admixture,
       wherein
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2-, or CH3-(CH2)n-, wherein n= 1 to 11,
    R3 =
    H, -OH, CH3-, or CH3-(CH2)n-, wherein n= 1 to 11,
    R4 =
    H, or methyloxycarbonyl, and
    R5 =
    H, CH3-, CH3-CH2-, or HO-CH2-CH2,
    for a time sufficient to permit detectable amounts of said analyte to be extracted from said subject into said aqueous pyrrolidone mixture; and then detecting said extracted analyte.
  26. The method of claim 25, wherein said pyrrolidone is selected from the group consisting of N-methyl 2-pyrrolidone, 2-pyrrolidone, 1-methyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, 1-hexyl-2-pyrrolidone, 1-lauryl-2-pyrrolidone, N-(2-hydrbxyethyl)-2-pyrrolidone, 1,5-dimethyl-2-pyrrolidone, 5-methyl-2-pyrrolidone, 1-hexyl-4-methyloxycarbonyl-2-pyrrolidone, 1- lauryl-4-methyloxycarbonyl-2-pyrrolidone, N-cyclohexyl-2-pyrrolidone, N-dodecyl-2-pyrrolidone, and 1-butyl-3-dodecyl-2-pyrrolidone.
  27. The method of either of claims 25 or 26, wherein said pyrrolidone is N-methyl 2-pyrrolidone (NMP) or 2-pyrrolidone.
  28. The method of any one of claims 25 - 27, wherein said pyrrolidone is N-methyl 2-pyrrolidone (NMP).
  29. The method of any one of claims 25 - 28, wherein said pyrrolidone is admixed with water.
  30. The method of any one of claims 25 - 28, wherein said pyrrolidone is admixed with saline.
  31. The method of any one of claims 25 - 30 wherein said pyrrolidone is at an aqueous concentration of at least 25%.
  32. The method of any one of claims 25 - 31, wherein said pyrrolidone is at an aqueous concentration of no more than 75%.
  33. The method of any one of claims 25 - 32, wherein said pyrrolidone is NMP.
  34. The method of claim 33, wherein said NMP is present at an aqueous concentration of 50%.
  35. The method of any one of claims 25 - 34, wherein said aqueous pyrrolidone mixture is included within a hydrogel.
  36. The method of any one of claims 25 - 34 wherein said aqueous pyrrolidone mixture is included within an absorbant.
  37. The method of claim 36, wherein said absorbant is selected from the group consisting of cellulosic pad and gauze patch.
  38. The method of any one of claims 25 - 37, wherein said time is no more than 24 hours.
  39. The method of any one of claims 25 to 37, wherein said time is no more than 6 hours.
  40. The method of any one of claims 25 to 37, wherein said time is no more than 60 minutes.
  41. The method of any one of claims 25 to 37, wherein said time is no more than 30 minutes.
  42. The method of any one of claims 25 - 41, wherein said analyte is detected within said aqueous pyrrolidone mixture.
  43. The method of any one of claims 25 - 41, further comprising the step, prior to detecting said analyte, of:
    • removing said extracted analyte from said pyrrolidone mixture.
  44. The method of any one of claims 25 - 43, wherein said analyte detection is qualitative.
  45. The method of any one of claims 25 - 43, wherein said analyte detection is quantitative.
  46. The method of any one of claims 25 - 45, wherein said analyte is detected colorimetrically.
  47. The method of any one of claims 25 - 45, wherein said analyte is detected spectrophotometrically.
  48. The method of any one of claims 25 - 45, wherein said analyte is detected enzyme chromatographically.
  49. The method of any one of claims 25 - 45, wherein said analyte is detected by antibody assay.
  50. The method of any one of claims 25 - 47, wherein said analyte is a heavy metal.
  51. The method of claim 50, wherein said heavy metal is selected from the group consisting of lead, cadmium, mercury, lithium and copper.
  52. The method of claim 51, wherein said heavy metal is lead.
  53. The method of any one of claims 25 - 49, wherein said analyte is glucose.
  54. The method of any one of claims 25 - 49, wherein said analyte is a metabolite.
  55. The method of any one of claims 25 - 49, wherein said analyte is a drug of abuse or metabolite thereof.
  56. The method of claim 55, wherein said drug of abuse is selected from the group consisting of cocaine, nicotine, and caffeine.
  57. The method of any one of claims 25 - 49, wherein said analyte is a pharmaceutical compound.
  58. The method of claim 57, wherein said pharmaceutical compound is a nonsteroidal antiinflammatory agent.
  59. A kit for the transdermal detection of analytes present within a subject, comprising:
    • a device for transdermal extraction of detectable amounts of said analyte, said device comprising an effective amount of pyrrolidone of the following formula in aqueous admixture,
       wherein
    R1 =
    H, CH3-, HO-CH2-CH2-, CH3-CH2-, or CH3-(CH2)n-, wherein n=1 to 11,
    R3 =
    H, -OH, CH3-, or CH3-(CH2)n-, wherein n=1 to 11,
    R4 =
    H, or methyloxycarbonyl, and
    R5 =
    H, CH3-, CH3-CH2-, or HO-CH2-CH2; and
       means for detecting analyte extracted into said aqueous pyrrolidone mixture.
Anspruch[fr]
  1. Dispositif pour la détection transdermique d'analytes présents chez un sujet, comprenant :
    • . une quantité efficace de pyrrolidone de formule suivante en mélange aqueux :
         dans laquelle
      R1 =
      H, CH3-, HO-CH2-CH2-, CH3-CH2- ou CH3-(CH2)n-, où n=1 à 11 ;
      R3 =
      H, -OH, CH3- ou CH3-(CH2)n-, où n=1 à 11 ;
      R4 =
      H ou un groupe méthyloxycarbonyle ; et
      R5 =
      H, CH3-, CH3-CH2- ou HO-CH2-CH2- ; et
    • un moyen pour détecter la présence ou la quantité d'analyte extrait dans ledit mélange aqueux de pyrrolidone.
  2. Dispositif selon la revendication 1, dans lequel ladite pyrrolidone est choisie dans le groupe formé par
    • la N-méthyl-2-pyrrolidone, la 2-pyrrolidone,
    • la l-méthyl-2-pyrrolidone, la 1-éthyl-2-pyrrolidone,
    • la 1-hexyl-2-pyrrolidone, la 1-lauryl-2-pyrrolidone,
    • la N-(2-hydroxyéthyl)-2-pyrrolidone,
    • la 1,5-diméthyl-2-pyrrolidone, la 5-méthyl-2-pyrrolidone,
    • la 1-hexyl-4-méthyloxycarbonyl-2-pyrrolidone,
    • la 1-lauryl-4-méthyloxycarbonyl-2-pyrrolidone,
    • la N-cyclohexyl-2-pyrrolidone, la N-dodécyl-2-pyrrolidone et
    • la 1-butyl-3-dodécyl-2-pyrrolidone.
  3. Dispositif selon la revendication 2, dans lequel ladite pyrrolidone est la N-méthyl-2-pyrrolidone (NMP) ou la 2-pyrrolidone.
  4. Dispositif selon la revendication 3, dans lequel ladite pyrrolidone est la N-méthyl-2-pyrrolidone (NMP).
  5. Dispositif selon l'une quelconque des revendications 1 à 4, dans lequel ladite pyrrolidone est mélangée à de l'eau.
  6. Dispositif selon l'une quelconque des revendications 1 à 4, dans lequel ladite pyrrolidone est mélangée à du sérum physiologique.
  7. Dispositif selon l'une quelconque des revendications 1 à 6, dans lequel ladite pyrrolidone est à une concentration aqueuse d'au moins 25%.
  8. Dispositif selon l'une quelconque des revendications 1 à 7, dans lequel ladite pyrrolidone est à une concentration aqueuse ne dépassant pas 75%.
  9. Dispositif selon l'une quelconque des revendications 1 à 8, dans lequel ladite pyrrolidone est NMP.
  10. Dispositif selon la revendication 9, dans lequel ladite NMP est présente à une concentration aqueuse d'environ 50%.
  11. Dispositif selon l'une quelconque des revendications 1 à 10, comprenant de plus :
    • un hydrogel,
    dans lequel ledit mélange aqueux de pyrrolidone est dans ledit hydrogel.
  12. Dispositif selon la revendication 11, dans lequel ledit hydrogel comprend un polymère choisi dans le groupe formé par : les poloxamères, le poly(alcool vinylique), le polyhydroxyméthacrylate, le poly(méthacrylate d'hydroxyéthyle), le propylèneglycol et l'acrylamide.
  13. Dispositif selon la revendication 12, dans lequel ledit hydrogel comprend un poloxamère.
  14. Dispositif selon la revendication 13, dans lequel ledit poloxamère est le poloxamère 407.
  15. Dispositif selon l'une quelconque des revendications 1 à 10, comprenant de plus :
    • un absorbant,
    dans lequel ledit mélange aqueux de pyrrolidone est dans ledit absorbant.
  16. Dispositif selon la revendication 15, dans lequel ledit absorbant est un tampon cellulosique ou une pièce de gaze.
  17. Dispositif selon l'une quelconque des revendications 1 à 16, dans lequel ledit moyen de détection inclut des analyses d'anticorps spécifiques pour ledit analyte.
  18. Dispositif selon la revendication 17, dans lequel lesdits anticorps sont capables de migrer à l'intérieur du dispositif.
  19. Dispositif selon la revendication 17 ou 18, dans lequel ledit anticorps est spécifique pour un analyte choisi dans le groupe formé par le glucose, les métabolites, les drogues dont on abuse, les métabolites de drogues et les composés pharmaceutiques.
  20. Dispositif selon l'une quelconque des revendications 1 à 16, dans lequel ledit analyte est un métal lourd.
  21. Dispositif selon la revendication 20, dans lequel ledit métal lourd est choisi dans le groupe formé par le plomb, le cadmium, le mercure, le lithium et le cuivre.
  22. Dispositif selon la revendication 21, dans lequel ledit métal lourd est le plomb.
  23. Dispositif selon l'une quelconque des revendications 1 à 22, dans lequel ledit moyen de détection inclut un moyen chromatographique enzymatique.
  24. Dispositif selon l'une quelconque des revendications 1 à 23, comprenant de plus un agent modifiant les propriétés de surface choisi dans le groupe formé par les tensio-actifs, le menthol, les phospholipides et l'Azone®.
  25. Procédé pour la détection transdermique non invasive de la présence ou de la quantité d'un analyte présent chez un sujet, comprenant :
    • la mise en contact de la peau dudit sujet avec une quantité efficace de pyrrolidone de formule suivante en mélange aqueux :
      dans laquelle
      R1 =
      H, CH3-, HO-CH2-CH2-, CH3-CH2- ou CH3-(CH2)n-, où n=1 à 11 ;
      R3 =
      H, -OH, CH3- ou CH3-(CH2)n-, où n=1 à 11 ;
      R4 =
      H ou un groupe méthyloxycarbonyle ; et
      R5 =
      H, CH3-, CH3-CH2- ou HO-CH2-CH2- ;
      pendant une durée suffisante pour permettre à des quantités détectables dudit analyte d'être extraites dudit sujet dans ledit mélange aqueux de pyrrolidone ; puis
    • la détection dudit analyte extrait.
  26. Procédé selon la revendication 25, dans lequel ladite pyrrolidone est choisie dans le groupe formé par
    • la N-méthyl-2-pyrrolidone, la 2-pyrrolidone,
    • la 1-méthyl-2-pyrrolidone, la 1-éthyl-2-pyrrolidone,
    • la 1-hexyl-2-pyrrolidone, la 1-lauryl-2-pyrrolidone,
    • la N-(2-hydroxyéthyl)-2-pyrrolidone,
    • la 1,5-diméthyl-2-pyrrolidone, la 5-méthyl-2-pyrrolidone,
    • la 1-hexyl-4-méthyloxycarbonyl-2-pyrrolidone,
    • la 1-lauryl-4-méthyloxycarbonyl-2-pyrrolidone,
    • la N-cyclohexyl-2-pyrrolidone, la N-dodécyl-2-pyrrolidone et
    • la 1-butyl-3-dodécyl-2-pyrrolidone.
  27. Procédé selon la revendication 25 ou 26, dans lequel ladite pyrrolidone est la N-méthyl-2-pyrrolidone (NMP) ou la 2-pyrrolidone.
  28. Procédé selon l'une quelconque des revendications 25 à 27, dans lequel ladite pyrrolidone est la N-méthyl-2-pyrrolidone (NMP).
  29. Procédé selon l'une quelconque des revendications 25 à 28, dans lequel ladite pyrrolidone est mélangée à de l'eau.
  30. Procédé selon l'une quelconque des revendications 25 à 28, dans lequel ladite pyrrolidone est mélangée à du sérum physiologique.
  31. Procédé selon l'une quelconque des revendications 25 à 30, dans lequel ladite pyrrolidone est à une concentration aqueuse d'au moins environ 25%.
  32. Procédé selon l'une quelconque des revendications 25 à 31, dans lequel ladite pyrrolidone est à une concentration aqueuse ne dépassant pas environ 75%.
  33. Procédé selon l'une quelconque des revendications 25 à 32, dans lequel ladite pyrrolidone est NMP.
  34. Procédé selon la revendication 33, dans lequel ladite NMP est présente à une concentration aqueuse d'environ 50%.
  35. Procédé selon l'une quelconque des revendications 25 à 34, dans lequel ledit mélange aqueux de pyrrolidone est inclus à l'intérieur d'un hydrogel.
  36. Procédé selon l'une quelconque des revendications 25 à 34, dans lequel ledit mélange aqueux de pyrrolidone est inclus à l'intérieur d'un absorbant.
  37. Procédé selon la revendication 36, dans lequel ledit absorbant est choisi dans le groupe formé par un tampon cellulosique et une pièce de gaze.
  38. Procédé selon l'une quelconque des revendications 25 à 37, dans lequel ladite durée ne dépasse pas environ 24 heures.
  39. Procédé selon l'une quelconque des revendications 25 à 37, dans lequel ladite durée ne dépasse pas environ 6 heures.
  40. Procédé selon l'une quelconque des revendications 25 à 37, dans lequel ladite durée ne dépasse pas environ 60 minutes.
  41. Procédé selon l'une quelconque des revendications 25 à 37, dans lequel ladite durée ne dépasse pas environ 30 minutes.
  42. Procédé selon l'une quelconque des revendications 25 à 41, dans lequel ledit analyte est détecté à l'intérieur dudit mélange aqueux de pyrrolidone.
  43. Procédé selon l'une quelconque des revendications 25 à 41, comprenant de plus l'étape, précédent la détection dudit analyte, consistant à :
    • retirer ledit analyte extrait dudit mélange de pyrrolidone.
  44. Procédé selon l'une quelconque des revendications 25 à 43, dans lequel ladite détection de l'analyte est qualitative.
  45. Procédé selon l'une quelconque des revendications 25 à 43, dans lequel ladite détection de l'analyte est quantitative.
  46. Procédé selon l'une quelconque des revendications 25 à 45, dans lequel ledit analyte est détecté colorimétriquement.
  47. Procédé selon l'une quelconque des revendications 25 à 45, dans lequel ledit analyte est détecté spectrophotométriquement.
  48. Procédé selon l'une quelconque des revendications 25 à 45, dans lequel ledit analyte est détecté chromatographiquement par enzyme.
  49. Procédé selon l'une quelconque des revendications 25 à 45, dans lequel ledit analyte est détecté par analyse d'anticorps.
  50. Procédé selon l'une quelconque des revendications 25 à 47, dans lequel ledit analyte est un métal lourd.
  51. Procédé selon la revendication 50, dans lequel ledit métal lourd est choisi dans le groupe formé par le plomb, le cadmium, le mercure, le lithium et le cuivre.
  52. Procédé selon la revendication 51, dans lequel ledit métal lourd est le plomb.
  53. Procédé selon l'une quelconque des revendications 25 à 49, dans lequel ledit analyte est le glucose.
  54. Procédé selon l'une quelconque des revendications 25 à 49, dans lequel ledit analyte est un métabolite.
  55. Procédé selon l'une quelconque des revendications 25 à 49, dans lequel ledit analyte est une drogue dont on abuse ou un de ses métabolites.
  56. Procédé selon la revendication 55, dans lequel ladite drogue est choisie dans le groupe formé par la cocaïne, la nicotine et la caféine.
  57. Procédé selon l'une quelconque des revendications 25 à 49, dans lequel ledit analyte est un composé pharmaceutique.
  58. Procédé selon la revendication 57, dans lequel ledit composé pharmaceutique est un agent anti-inflammatoire non stéroïdien.
  59. Trousse pour la détection transdermique d'analytes présents chez un sujet, comprenant :
    • un dispositif pour l'extraction transdermique de quantités détectables dudit analyte, ledit dispositif comprenant une quantité efficace de pyrrolidone de formule suivante en mélange aqueux :
      dans laquelle
      R1 =
      H, CH3-, HO-CH2-CH2-, CH3-CH2- ou CH3-(CH2)n-, où n=1 à 11 ;
      R3 =
      H, -OH, CH3- ou CH3-(CH2)n-, où n=1 à 11 ;
      R4 =
      H ou un groupe méthyloxycarbonyle ; et
      R5 =
      H, CH3-, CH3-CH2- ou HO-CH2-CH2- ; et
      un moyen pour détecter l'analyte extrait dans ledit mélange aqueux de pyrrolidone.






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