WO1998014768A2

WO1998014768A2 - Transdermal chemical monitoring device

Info

Publication number: WO1998014768A2
Application number: PCT/US1997/017856
Authority: WO
Inventors: Sorell L. Schwartz; Philip Witorsch; Robert R. Bies; Steven W. Collier; Arie Markus
Original assignee: Schwartz Sorell L; Philip Witorsch; Bies Robert R; Collier Steven W
Priority date: 1997-10-06
Filing date: 1997-10-06
Publication date: 1998-04-09
Also published as: WO1998014768A3

Abstract

A non-invasive chemical monitoring device that is placed on the surface of the skin collects and measures the concentration of various chemicals within the body by outward diffusion of the chemicals from the interior of the body, through the skin and into the device, is prepared with a molded polymeric collection medium (1), adhesive surface (3), and a skin to patch transfer link (2).

Description

TRANSDERMAL CHEMICAT. MONITORING DEVICE

This application is a continuation-m-part of U.S. provisional application serial number XXXXXX, (serial number of priority application unknown at time of filing) entitled A TRANSDERMAL

CHEMICAL MONITORING DEVICE FOR NON-INVASIVE TESTING OF THE BLOOD FOR CERTAIN DRUGS AND ENVIRONMENTAL CHEMICALS, filed by Sorrel L. Schwartz on October 4, 1996.

FIELD OF THE TNVF.NTTON

The invention relates to a non-invasive chemical monitoring device that is placed on the surface of the skin and collects ana measures the concentration of various chemicals within ne body by outward diffusion of the chemicals from the interior of the body, through the skin and into the device.

BACKGROUND DISCUSSION Uses for the TCMD include:

Environmental/Occupational Exposure Monitoring; Monitoring m the workplace for chemical exposure has taken on a number of different of different forms, mostly all of them appliea to those workplaces where the normal course of occupational activity involves chemicals (as opposed to office buildings, for example, where employees may be exposed to pesticides used by the building management) . This monitoring includes the periodic use of instruments to sample the air and the use of personal monitoring badges, which also monitor the air. This results in data presumptive of certain exposures, but not fully indicative of personal exposure. The most accurate representation of personal exposure is biological monitoring. Biological monitoring involves a measurement of the chemical or its surrogate in biological fluid or tissue. The most commonly used monitoring site is urine. This involves taking a urine sample at some specified time. The problem with this method is mat, because of pharmacokinetic factors, it does not give a full picture of exposure. It would be necessary to collect urine for 12 to ^π2 hours or longer to overcome this limitation. Such a procedure creates compliance problems. Another monitoring site is blood. It has tne αisadvantage of being invasive. It also has the same pharmacokinetic limitations that would make repeated sampling necessary to acnieve a full exposure picture. This presents obvious disadvantages and risks. The TCMD overcomes the aforementioned problems. It does not present compliance or risk problems, and tne collection period can be specifically tuned to the pharmacokinetic behavior of the chemicals under surveillance. Of course, the TCy will work only for chemicals that are transdermally transporteα. Based on physical chemical requirements, this includes a large fraction of the chemicals of interest such as therapeutic drugs, hormones, environmental toxins, pesticides, drugs of abuse and xenooiotics found in occupational settings.

A variety of diagnostic kits for monitoring an analyte IΓ sweat have oeen developed. For example, U.S. Pat. No. 3,552,929 to Fields, et a_. discloses a Dand-aid-type test patch suited for determining the cnloride ion concentration in perspiration as a method of diagnosing cystic fibrosis. The apparatus disclosed in Fields comprises an absorptive sweat collecting pad with an impermeable overlying layer for the purpose of preventing evaporation. When the absorptive pad is saturated, the patch is removed from the skin and exposed to a series of strips impregnated with incremental quantities of silver chromate or silver nitrate, the color of which undergoes a well known change upon conversion to the chloride salt. The Phillips U.S. Pat. No. 4,329,999 discloses a dermal patch for collecting sweat from a patient which includes a collecting pad for aDsorbmg sweat. Similarly, tne Phillips U.S. Pat. No. 4,595,011 dιsc_.cses a transoermal dosimeter device including a dermal contact bridge, a fluid collecting component for collecting and storing fluids collected from the skin and a process component for bmdinα or chemically converting the stored substances. The latter Phillips patent suggests that chemical conversion of collected substances to produce an observable color change in the device may be effected.

The Fogt et . al . U.S. Pat. No. 4,444,193 discloses an absoroent patch device for absorbing sweat, which patch device includes a chemical composition capable of reacting with chloride contained in the sweat.

The Pugliese U.S. Pat. No. 4,071,020 discloses an apparatus and metnoos for performing m-vivo measurements of enzyme activity wherem one or more reactants are placed directly on a predetermined area c: the skin surface and are free to migrate into the skin. Λhile the apparatus and methods of Pugliese are not dependent on collection of a substance such as sweat or interstitial fluid, they may be dangerous to the subject if the reactants have toxic effects upon mιgrat_on into the skin of the sub ect.

U.S. Pat. No. 4,706,676 to Peck discloses a dermal collection device which comprises a binder to prevent reverse migration of an ana yte, a liquid transfer medium which permits transfer of an analyte from the dermal surface to the binder and an occlusive cover across the top of the liquid transfer medium and binder. Other U.S patents issued to Peck include No. 4,819,645, No. 4,960,467, No. 4,909,256 and No. 4,821,733.

Other devices are also known for measuring substances, particularly gases, in or emanating from the skin. For example, the Clark, Jr. U.S. Pat. Nos. 4,401,122 and 4,458,686 disclose apparatus and methods for measuring substances, particularly gases, which diffuse through the skin or are present underneath the skin in the blood or tissue using polarographic electrodes or enzyme electrodes. The v'esterager et al U.S. Pat. No. 4,274,418 discloses an apparatus for measuring gases, for example oxygen and carbon dioxide, whicn diffuse from blood vessels and through skin tissue wherein the gas is directed to a measuring chamber in wnich the partial pressure is measured.

The prior art discussed above deals only with the passive collection of sweat and the analysis of compounds within sweat. The compounds which appear in sweat are hydrophilic and thereby preclude the detection of a vast number of compounds of interest.

The collection of compounds which diffuse outwardly from the skin nas been described in U.S. patent 4,821,733 to Peck. The Peck devices are composed of a detector means including at least one detector chemical in solution and capable of chemically reacting with the analyte as the analyte migrates to the skin surface.

SUMMARY OF THE INVENTION

The transdermal chemical monitoring device (TCMD) s a patch applied to the skin and capable of providing data on the levels of specific drugs and/or chemicals in the blood. Transcutaneous chemical collection represents a relatively novel approach to the non-invasive sampling of certain blood components. The same principles that apply to the inward movement of drugs from transdermal dosing patches through the skin into the blood also apply to the outward movement to the skin surface. This outward movement of chemicals from blood to the s ιn provides the basis for transcutaneous monitoring of chemicals in the blood by use of a TCMD.

BRIEF DESCRIPTION OF THE PREFERRED EMBODIMENTS

The invention will be more fully appreciated from the following description, particularly when considered in conjunction with the attached drawings, wherein: Figure 1 shows a cross sectional view of the copo ymer paten;

Figure 2 shows a bottom section view of the copoiymer patch shown in figure 1; Figure 3 shows a cross sectional view of the copoiymer patch with me oil dispersion m the matrix;

Figure 4 shows a cross sectional view of the copoiymer patch consisting of several components, an outside copoiymer snell, an inner reservoir of uptake material, an inner binding compound and an adhesive attachment. The inner reservoir material can consist of polymers, oils, fats, fatty acids, fatty acid esters, waxes, and rubbers whicn act to trap the absorbed analyte in the transderma_. collection monitoring device;

Figure 5 shows a bottom sectional view of the copoiymer oatch device snown in figure 4;

F_gure 6 shows the forward flux through hairless rat skin and accumulation of estradiol in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y-axis vs time in hours on the x-axis.; Figure ^~ shows the reverse flux through hairless rat skin ana accumulation of estradiol in the receptor compartment. The axes snow cumulative amount in disintegrations per minute on the Y-axis vs time in hours on the x-axis.;

Figure 8 shows the forward flux through LSE and accumulation of estraαiol in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y-axis vs time in hours

Figure 9 shows the forward flux through LSE and accumulation of estradiol in the receptor compartment sampled every hour out to 24 hours . The axes show cumulative amount in disintegrations per minute in the receptor solution and remaining quantity of material in the donor solution on the Y-axis vs time in hours on the x-axis.; Figure 10 shows the rorward flux through SFLS LSE and accumulation of estradiol in the receptor compartment. The axes show cumulative amount m disintegrations per minute on the Y-axis vs time in hours on the x-axis.; Figure 11 shows the flux through CoTran #9270 polyethylene film membrane and accumulation of estradiol in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y-axis vs time in hours on the x-axis.;

Figure 12 shows the forward flux through hairless rat skin and accumulation of progesterone in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y-axis vs time m hours on the x-axis.;

Figure 13 shows the forward flux through LSE and accumulation of progesterone in the receptor compartment. The axes snow cumulative amount in disintegrations per minute on the .-axis "s time in hours on the x-axis.;

Figure 14 shows the forward flux through SFLS LSE and accumulation of progesterone in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y-axis vs time in hours on the x-axis.;

Figure 15 shows the forward flux througn LSE and accumulation of tritiated water in the receptor compartment. The axes show cumulative amount in disintegrations per minute in the receptor compartment and the remaining disintegrations per minute for the donor compartment on the Y-axis vs time in hours on the x-axis.;

Figure 16 shows the forward flux through SFLS LSE and accumulation of tritiated water in the receptor compartment. The axes show cumulative amount in disintegrations per minute on the Y- axis vs time in hours on tne x-axis .

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

TCMDs, when placed on the skin, collect a chemical by accumulating the chemical through thermodynamic factors. The patch is then analyzed to produce quantitative results on the accumulated chemical. On the basis of the applicable physical chemical principles, the device can be designed to accumulate a number of different molecules, which can be used as an advantage such as in the simultaneous monitoring of different chemicals.

TCMDs can be designed according to the methods described m a variety of ways so as to accommodate various detection techniques and application configurations.

Preparation of the TCMD

An embodiment of the device are shown in Figure 1. The device shown is comprised of a single-piece of molded polymer (1) . The molded polymer is treated on its skin contact surface with a tacky material that serves as the skm-to-patch transfer link (SPTL) (2). The components of the SPTL can vary, depending upon the physical chemical properties of the compounds for which monitoring is oeing done. The components of the polymer are specifically dictated by physical chemical properties needed for transferring the analyte from the skin into the collection medium. In some situations the polymer can be formulated in a tacky consistency such that it provides its own SPTL. This active unit can then be attached to the outer anchoring adhesive (3) . The bottom view of this embodiment is shown in Figure 2.

The TCMD can be provided with a thermodynamic sink in the form of an area within the molded polymer in which an additional substance is provided wnicn nas a difference nydrophilicity or lipophilicity from tne molded polymer. The additional substance can also be a polymeric material which has been selected and formulated for it physical chemical properties. Figure 3 shows such an embodiment in which αroplets of oil (4) are added to the poiymer in order to enhance tne tnermodynamic sink for certain lipophilic chemicals.

Figure 4 shows the fabrication of an embodiment that contains a center well that can be used for reagents that will react with the analyte such that the patch can consist of a self-contained analytical system to quantify the amount of analyte. The pre-pc_ —eπzed mixture is poured into a mold so that it polymerizes in a for™ containing a well (1). The reagent material is then aαdeo to the veil (5). Alternatively, the well can contain material to further concentrate the analyte, e.g., mineral oil. A film material such as commercially available paraffin film (6) is then placed over the e__, and a layer of pre polymerized mixture poured over that cover, .'.nen the added mixture polymerizes (7), a single piece device (8) results. The SPTL material is added (2) and the unit attached to the outer anchoring adhesive.

F_gure 5 shows the bottom view of this embodiment of the TCMD: the tac<y polymeric surface (shown as a section) (2) for bonding tne patch tc the skin; an outline of the reservoir (5) can oe seen trough the translucent polymer (8); the anchoring outer adhesive (3).

Components used for TCMD polymer preparation are listed below:

PVP: Polyvmylpyrrolidone PLASDONE K-90 is 2-pyrrolιdιnone, 1- ethenyl-, homopolymer (ISP Technologies, Wayne NJ) witn a molecular formula of (C₆ ^H ₉ ^NO, _x Also known as Polyvmylpyrrolidone. CAS#9003-39-8. PVA: Poly Vinyl Alcohol (Sigma Chemical Company, St. Louis MO) with molecular weight of 70,000-100,000 Daltons . Also known as vinyl alcohol polymer. CAS#9002-89-5.

GANEX P-904: 2-pyrrolιdιnone, 1-ethenyl-, polymer with 1-butene (ISP Technologies, Wayne NJ) with a molecular weight of 16000 Daltons. Also known as Butylated polyvmylpyrrolidone. CAS#26160-96- 3.

Desmophen 1800: Polyester Polyol (Polydiethylene Adipate) . (Bayer, Pittsburgh PA)

Desmophen 2200B: Polyester Polyol CAS#28183-09-7 Comprises acrylic acid/ethenediol repeats (Bayer, Leverkusen, Germany) .

Multranol 9111: Bayer, Pittsburgh PA. Polyether Polyol (Poly- oxyalkylene) Polyol) .

Isonate 181: Dow, Midland MI. Pre-polymer diphenylmethylenedi- isocyanate.

Isonate M301: Dow Benelux, 4 , 4-dιphenyl, methyl-diisocyanate prepolymer .

Isonate M342: Dow Benelux, 4, 4-dιphenyl, methyl-diisocyanate prepolymer

Mineral Oil

CoTran membrane #9270 Polyethylene film. (3M, St. Paul, MN)

Bayflex 0549B: Polyester Polyol Bayer, Leverkusen, Germany Bayflex 0681: Polyester Polyol CAS#26570-73-0' Bayer, Leverkusen, Germany

Voranate M580: Polymeric methylene oiphenyl dusocyanate, Dow, Beneiix

PEG 1300: Carbowax Sentry Polyethylene Glycol 1000NF, FCC Grace (Union Carbide, Danbury CT) . Polyethylene Glycol, an oxyalkylene Polymer with the formula ^HO"ICH ₂ ^CH ₂ ^01I1"H with a molecular weignt range of

950-IC50 Daltons. Also known as polyoxyethylene 1000 and polytoxy- 1, 2emaneoιyI) a-hydro-w-hyoroxy~. CAS# 25322-68-3

PEG 3350: Carbowax Sentry Polyethylene Glycol 3350 Flake NF, FCC Grace 'Union Carbide, Danbury CT) . Polyethylene Glycol, an oxyaκ.lene polymer with tne formula ^{H0 ICH},^{CH 0ln_H} with a molecular weigr range of 3000-3600 Daltons. Also known as polyoxyethylene 3350 and poly (oxy-1, 2ethanedιyl) a-nydro-w-nydroxy- . CAS#25322-68-3

PEG 4600: Carbowax Sentry Polyethylene Glycol 4600 Flake NF, FCC Graαe (Union Caroide, DanDury CT) . Polyethylene Glycoi, an oxyai-c/lene polymer with the formula ^H0"ICH ₂ ^CH ₂ ^01n_H with a molecular weight range of 4400-4800 Daltons. Also known as polyoxyethylene 4600 and poly(oxy-l,2ethanedιyl) , 1-hydro-w-hydroxy- . CAS#25322-68-3

PEG 8000: Carbowax Sentry Polyethylene Glycol 8000 Flake NF, FCC Grade (Union Carbide, Danbury CT) . Polyethylene Glycol, an oxyalkylene polymer with the formula ^{H0~ ,CH} ₂ ^CH ₂ ⁰¹"-" with a molecular weight range of 7000-9000 Daltons. Also known as polyoxyethylene 8000 and polyιoxy-l,2ethanedιyl) , 1-hydro-w-hydroxy- . CAS#25322-68-3. Proceαures for the preparation of me polymer formulations are described below: PVP/PVA

Polymers were prepared in 100 gram batches. 4g of Polyvmylalcohol (PVA) and 24g of glycerol were added to 85mL of solvent consisting of a 50/50 mixture of absolute ethanol and H20 contained in a stainless steel beaker. The mixture was stirreα on low speed setting #2 using a Caframo RZR1 stirrer (Wharton Ontario Canada). 28grams of PVP K-90 is placed in a beaker with 115mL of a 50/50 ethanol/H20 solvent mixture. When the oil bath reached 90°C the PVP solution mentioned previously was added to the PVA/Glycerol mixture. The mixture was boiled at 130°C with stirring and heat continued for five minutes beyond this point. The beaker was removed from the heat (oil bath) and 3 grams of the copoiymer was poureα into each of 10 wells of a nonstick mold. Approximately 48 hours later, a good film had formed. The disks removed from the mold had a weignt of 1.1-1.3 grams each

PVA/GANEX 4g of PVA was mixed with 85mL of 50% Ethanol at 90°C (in an oil bath) . 12 grams of glycerol was added and this mixture was heated to 110 degrees Celsius. At that point 14.5 grams of GA EX V516(ISP Technologies, Wayne NJ) was added to the mixture. The GANEX was not dissolved in any solvent prior to its addition to the PVA/Glycerol/Ethanol mixture. The final solution was mixed and heated to 130°C. The mixture was then poured into a nonstick mold and a non-stick sheet. The film took approximately 48 hours to form.

Polyurethane Patches with Incorporated Oil Isonate M342 & Desmophen 2200B 17 grams of Desmophen 2200B (Bayer, Leverkusen, Germany) and 3 grams of Isonate M342 (Dow, Benelux) and 4 ml of heavy mineral oι_ were ccmoineα as stirred over heat until a homogeneous mixture was oDtameα. This mixture was men poured into a nonstick mold and placed in an oven (Blue M Electric, Blue Island ILL) overnight at 60°C and subsequently removed.

Isonate M342 & Bavflex 0549

17 grams of Bayflex 0549 (Bayer, Leverkusen, Germany) was combined with 3 grams of Isonate M342 (Dow, Benelux) and 4 ml of heavy mineral oil and heated with stirring until a homogeneous mixture was obtained. This mixture was then poured into nonstick molαs and placed in an oven overnight at 60°C.

Isonate M342 & Bavflex 0681 17 grams of Bayflεx 0681 (Bayer, Leverkusen, Germany) was combined with 3 grams of Isonate M342 (Dow, Benelux) and 4 ml of heavy mineral oil (source as above) and heated with stirring unt_l a homogeneous mixture was obtained. The mixture was then poured into nonstick molds and placed in an oven overnight at 60°C.

Polvurethane Patches Isonate M342 & Demsophen 2200B

As outlined above including the proportions of the mixture except that mineral oil was not incorporated

Isonate M342 & Bavflex 0681

As outlined above including the proportions of the mixture except that mineral oil was not incorporated.

Isonate M342 & Bayflex 0549

As outlined above including the proportions of the mixture except that mineral oil was not incorporated. Isonate M301 & Desmophen 2200B

These components were combined in a 3:17 ratio and heated with stirring until a homogenous mixture was obtained. The mixture was then poured into a nonstick mold and polymerized in a dry oven at 60 degrees centigrade overnight.

Isonate M301 & Bayflex 0681

Isonate M301 & Bayflex 0549

Voranate M580 & Multranol 9111 These components were combined in a 3:17 ratio and heated with stirring until a homogenous mixture was obtained. The mixture was then poured into a nonstick mold and polymerized in a dry oven at 60 degrees centigrade overnight.

Voranate M580 with PEG 8000

19.5 g of PEG 8000 (Union Carbide, Danbury CT) was combined with 0.5 g of Voranate M580 (Dow, Midland MI) with heat and stirring until a homogeneous mixture was obtained. Once obtained, the mixture was poured into nonstick molds and placed in an oven overnight at 60°C. Voranate M580 with PEG 1000

16 g of PEG 1000 (Union Carbide, Danbury CT) and 4g of Voranate M580 were combined with heat and stirring until a homogeneous mixture was ODtaineα. The mixture was poured into nonstick oiαs and placed m an oven overnight at 60 degrees Celsius .

Voranate M580 with PEG 3350

18 g of PEG 3350 (Union Carbide, Danbury CT) and 2 g of Voranate M580 were combined as for other Voranate PEG mixtures.

Voranate M580 with PEG 4600

19 g of PEG 4600 (Union Carbide, Danoury CT) ana 1 g of Voranate M580 were combined as for other Voranate PEG mixtures.

Isonate M342 with PEG 8000

19.5 g of PEG 8000 was mixed with 0.5 g of IsonateM342 with heat and stirring until a homogeneous mixture was obtained. The mixture was poured into nonstick molds and and placed in an oven overnight at °60 C.

Uptake into Polymer Patches

Uptake was assessed for estradiol botn througn nairless rat skin and CoTran membrane using a Franz Cell Apparatus.

Direct uptake assessments into Copoiymer Patch formulations Franz Cells were filled with HHBSS. The copoiymer patch formulations were then mounted on the Franz Cells. The patches were allowed to equilibrate with the donor solution for a period of 24 hours at 32°C using a recirculating water bath prior to dosing. The C14 labelled test compound was then introduced via the side arm of the Franz cell and the system was run for an additional 24 hours. At the end of the 24 hour period the patch was removed from the franz cell and digested for analysis . The donor solution was also collected and counted in its entirety in addition to the first rinse of the franz cell. The copoiymer patch was digested using concentrated sulphuric acid and then quenched using Safety Solve scintillation cocktail and IN NaOH. The resulting mixture was counteα by scintillation counting. Skin Assessments

Hairless rat skin was cut using a Padgett Dermatome (Padgett Instruments, Kansas City MO) to a thickness of between 200 and 250 microns. The skin was mounted on a franz cell containing HBSS as the donor solution facing the papillary dermis . The HBSS was stirred and maintained at a temperature of 32°C using a circulating water batn. The skin was equilibrated with the HBSS prior to dosing for a period of 24 hours. After the equilibration period, [4-¹C] -Estradiol was introduced into the donor solution and a polymer collection patch applied using CoTran #9871 Pharmaceutical Grade Transfer Adhesive (3M, St. Paul MN) . After an additional 24 hours, the skin and the patch were removed and the donor solution collected for counting along with two rinses of tne Franz cell. The skin was solubilized using the tissue solubilizer TS-2 (RPI, Mount Prospect IL) . The resulting material was brought to a neutral pH using 0.01 N sooiurr hydroxide. The patch was extracted using ethanol (95%) for a period of 24 hours. The donor solution and aforementioned patch materials and solubilized skin were quenched using scintillation cocktail and counted by scintillation counting. CoTran Assessments

Carried out as mentioned for skin, except that the CoTran was not cut or modified in any way, it was mounted directly onto the Franz cell apparatus in tne same fashion as the hairless rat skin. Measurement of Chemical Flux into TCMD

The flux of parathion, chlorpyrifos, estradiol and progesterone a variety of membranes including dermatome cut hairless rat skin, CoTran f9270 (3M, St. Paul MN) , Living Skin Equivalent iLSE), ana serum free lipid supplemented living skin equivalent (SFLS LSE) . The Kp values describing tne movement of these comoounαs are shoii^r Table 1 .

Table 1 : Kp values for the Movement of test compounds tnrough various membranes. (All values are listed as the Mean ± Standard Deviation of the Kp, units are cm/hr) .

Test Hairless Rat Skin LSE SFLS LSE Cmod FWD REV FWD REV FWD REV

PTN 0.1310.0 3

CLP 0.02310. 008

EST 0.025 0.00210. 0.9+0.2 2+0.3 0.3710.11 0006

PRG 0.05±0.0 0.07±0.0 0.2 0.2 0.07±0.01 12 1

^JH 0 0.5 0.5 0.4210.11 0.54r 0.15

Once a steady state flux was observed (figure 6) as evidenceα by a l_near portion of the cumulative movement vs time plot values m this range for donor and receptor solution concentrations, skin thickness and time period were considered to determine the flux and the permeation constant (Kp) .

Tie forward flux of estrad±oi across dermatome cut hairless rat skin is shown in Figure 6 as a cumulative amount of material mov_.rg across the membrane versus time graph. It appears as tnough a steady state or consistent flux condition is approacned at approximatel_y four hours after the start of the experiment. Figure 7 shows the movement of estradiol through dermatome cut hairless rat skin but this time in the reverse direction. The reverse direction in this case is the movement from the papillary dermal face of the skin through to the stratum corneum face of the skin. The movement cf estradiol across the LSE is shown in figure 8. This graph shows the forward flux - i.e. from the stratum corneum side to the papillary dermis side - of the material as a cumulative movement vs time piot. The cnambers were sampled in this case hourly out to 8 hours and then at 18,20,22 and 24 hours. Once again, a steady state condition appears to be established by four hours into the experiment. The forwarα movement of estradiol is shown again m figure 9 with samplirg every hour out to twenty four hours. During the period from hour 8 through to hour 18, the receptor side solution was not repiaceα allowing the material to accumulate and significantly reduce tne concentration gradient observed across the membrane as well as the rate of movement. In addition, this graph shows the decline m the donor fluid concentrations. They decrease quite rapidly up to approximately 8 hours where they stabilize after the receptor fluid is left to accumulate material. The reverse flux of estradiol througn LSE occurs in a similar manner to the forward flux shown in figure 9. The forward and reverse fluxes in these last two cases are virtually identical (table 1) . Subsequent to the LSE experiments the SFLS LSE protocol was attempted and the forward flux of estradiol througn the SFLS LSE is shown in figure 10. The flux of estradiol througn the CoTran membrane was also assessed. This was undertaken as the CoTran was used as a "skin substitute" in several of the copoiymer patch formulation uptake experiments. A cumulative amount vs time graph of this data is shown in figure 11.

The forward flux of progesterone through hairless rat skin is shown in figure 12. This figure shows the cumulative amount of progesterone taken up on the receptor side. It also appears that the steady state is approached much faster in this case - by 3 hours it appears that there is a constant flux across the membrane from that point on to eight hours. The receptor fluid was sampled hourly out to eight hours and then again at 18,20,22 and 24 hours. The reverse flux of progesterone through hairless rat skin is similar to the forward flux shown m figure 12. Once again, there appears to be a steady state condition approached at 3 - 4 hours. The forward flux of progesterone through living skin equivalent is shown m figure 13. The flux is quite rapid as it was for estradiol and there is quite a lot of variability when compared to the data for nairless rat s ιn. As for estradiol, tne forward movement of progesterone through the SFLS LSE was also assessed. The flux was considerably less for progesterone through this new formulation than that seen with the regular LSE. A cumulative amount vs time for eight hours of movement through the membrane is shown in figure 14.

The movement of tritiated water was also assessed through the LSE and SFLS LSE membrane. The data for LSE is represented in plots whicn snow the cumulative amount of tritiated water that has moved across the membrane and into the receptor solution as well as the amount of tπtiateα water remaining in the donor solution compartment. The forward movement through LSE is shown in figure 15. The αata for the SFLS LSE is shown in a plot of cumulative amount in receptor fluid vs time in figure 16 for forward flux. The reverse flux in the two previous cases was very similar to the forward flux.

Patch Uptake Direct uptake of Parathion into Copoiymer Formulation

The direct uptake of parathion into one of the copoiymer formulations was assessed. The movement of parathion from the αonor solution consisting of HHBSS into the Isonate 181/Desmophen 1800 formulation resulted in the uptake of 69.9121.2% of the administered dose of parathion.

Parathion Uptake through CoTran Membrane The ability of a series of patch formulations to take up

Paratnion was assessed with the use of a "skin substitute" - CoTran #9270 polyethylene membrane from 3M. There was a general trend of increasing hydrophobicity in the uptake device resulting in a greater amount of parathion taken up. The Isonate M342/Desmophen 2200B formulation consistently took up a significant portion of the dose (up to 70%) and also resulted in good total dose recoveries, (table Table 2 - Uptake of Parathion through CoTran Membrane

Copoiymer Paten % Copoiymer patch Formulation uptake

Voranate M580 34.5_z2.41 Bayflex 0549

Isonate M301 57.516.4 Desmophen 2200B

Isonate M301 Bayflex 0681 69.3+7.8

Isonate M301 Bayflex 0549 47.3+18.08

Isonate M342 75.5+3.1 Desmophen 2200B

Isonate M342 Bayflex 0681 78.9+1.81

Isonate M342 Bayflex 0681 71.2+0.22

Isonate M342 Bayflex 0549 72.5+14.8

Uptake of Parathion through Hairless Rat Skin into Copoiymer Patches

The ability of several of these formulations to take up parathion was then assessed but through hairless rat skin instead of the CoTran membrane (table 3) . The uptake levels were significantly lower in these cases. Only relatively hydrophilic PVP/PVA and

PVA/GANEX formulations were assessed in this case.

Table 3 - Uptake of Parathion through Hairless Rat Skin

Direct Uptake of Chlorpyrifos into Copoiymer Patch Formulation The direct uptake of chlorpyrifos into the I181/D1800 copoiymer formulation was assessed and found to take up 6314.4% of the administered dose to the Franz cell. Uptake of Chlorpyrifos through CoTran membrane into Copoiymer Patch Formulations

Tne aoility of these copoiymer patch uptake devices to take up Chlorpyrifos tnrough the CoTran Membrane was then assessed (table 4). Both Isonate formulations tested provided a high level of uptake with the IM342/D2200B formulation being better than the other isonate combination tested - the IM342/BF0549 formulation, (table 4)

Table 4 - UptaKe of Chlorpyrifos tnrough CoTran Membrane

Copoiymer Patch % CoPolymer Patch Formulation Uptake

Isonate M342 51.4+6.34

_.:. Desmophen 2200B

Isonate M342 31.5112.36 Bayflex 0549

20 Uptake of Chlorpyrifos through Hairless Rat Skin into Copoivmer Patch Formulations

The ability of several formulations collect chlorpyrifos througn hairless rat skin was then assessed with a relatively 25 hydrophilic patch formulation - pvp/pva. The results are shown in table 5. The level of uptake is rather low and there is quite a wide range m the amount of material collected even within the same experiment .

30 Table 5 - Uptake of Chlorpyrifos through Hairless Rat Skin

Copoiymer Patch % Copoiymer Formulation Patch Uptake

PVP/PVA 11.66±5.8

35 PVP/PVA 2.2+1

Direct Uptake of Estradiol - various embodiments 40 The amount absorbed over 24 hrs 15-77 % depending on the formulation assessed. This uptake increased with increasing hydrophobicity to a certain extent but when the optimal hydrophobicity was exceeded this uptake decreased. This may be an artifact of the extraction procedure. Perhaps the ethanol extraction proceαure was unable to remove the estradiol from the patch in these cases. The ideal uptake polymer was a combination of Isonate M342 and Desmophen D2200B providing a good recovery of the total dose ana estraaiol uptake from 35-57%. Table 6 outlines the uptake observed. The uptake into the copolymers was very consistent (approximately 35%) wnen the material had to pass through the 3M pharmaceutical grade adhesive. Without the adhesive the uptake varied from 21.8% to 77.8%

Table 6 - Uptake of Estradiol into Copoiymer formulations directly from Donor solution

Formulation % Uptake (mean+sd)

Isonate M301 Bayflex 0549 (PGTA) 34.2

Isonate M301 Bayflex 0681 (PGTA) 34.5

Isonate M301 Desmophen 2200B (PGTA) 34.5

Isonate M342 Desmophen 2200B (PGTA) 34.2

Voranate M580 Multranol 9111 21.8

Bayflex 0681 Isonate M301 77.8

Isonate 181 Desmophen 1800 70.6+14.7

Isonate M342 Desmophen 2200B 57.1

PGTA = Pharmaceutical Grade Transfer Adhesive applied in these experiments .

Uptake of Estradiol through CoTran membrane into Copoiymer Patch Formulations

The ability of a series of copoiymer formulations to collect or take up estradiol through the CoTran membrane was assessed. There was quite a wide range of uptake values. The percent of dose taken up by the patches ranged from 0.09 to 5. The most material (5% of the αose) was collected in the IM342/D2200B patch (table 7) . Table 7 - UptaKe of Estradiol through CoTran Membrane

Copoiymer Patch %CoTran % Patch Formulation Uptake Uptake (mean+SD) (mean+SD)

Isonate M342 Desmophan 4.3+2.23 0.134+0.086 2200B with oil

Isonate M342 Bayflex 0549 4.6+1.37 0.21+0.1

Isonate M342 Bayflex 0681 2.7+0.53 0.0910.015 with oil

Voranate M580 PEG 1000 5.1+1.2 2.27+0.77

Voranate M580 PEG 3350 6.1+1.1 1.62+0. Si

Voranate M580 PEG 4600 5.611.3 0.6±0.015

Voranate M580 PEG 8000 4.8310.002 0.910.035

Isonate M342 4.1+0.9 5+0.5 Desmophen 2200B

Isonate M342 PEG 8000 4.710.5 0.99+0.04

Uptake of Estradiol through Hairless Rat Skin into Copoiymer Patch Formulations

The ability of a series of formulations to take up estradiol througn hairless rat skin was then assessed. The greatest level of uptake occurred with the IM342/D2200B patches. In addition, the ability of CoTran to take up estradiol through hairless rat skin was assessed and determined to be approximately 2%, although quite variable (table 8).

Another method of detection that has been utilized with the TCMD is acousto-optic tunable filter near infrared spectroscopy (AOTF-NIR) . With AOTF-NIR the concentration of the analyte can be measured nondestructively from directly within the TCMD. AOTF-NIR measurements with the TCMD are typically sensitive enough to measure subnanogram quantities of estradiol and other analytes. Other methods of detection that can be utilized include laser desorption mass spectrometry as well as techniques sucn as liquid chromatography and gas chromatography of extracted TCMDs. An advantage of the TCMD is that it can be cast as a solid one piece device and can incorporate chemical sensors directly into the body of the TCMD. The sensors may also be quite specialized and include sensitive and selective detectors for specific volatile or nonvolatile chemical or biological compounds such pesticides or otner anti-acetylcholmesterase compounds. The TCMD may also incorporate several types of sensors into one integrated device and the sensors may oe used to perform quantitative analyte measurements while tne device is still attached to the skin or after it has been removed. Typically, the sensors can be integrated into the structure of the TCMD. Such sensors can utilize different methods of detection. There are many examples of miniaturized detectors such as electrochemical sensors including chemical selective electrodes, immunosensors and biosensors which may contain specific antibodies or other ligands that will selectively bind the chemical compound of interest. Biosensors utilizing specific antibodies or ligands typically are bound to piezo crystals or electrodes or optical waveguides for fluorescent, photometric, or surface plasmon spectroscopy. Miniature photoacoustic detectors may also be employed. Because it is possible to produce TCMDs which are transparent or near transparent, any method of optical analytical measurement technique utilizing visible, infrared, or ultraviolet light my be utilized. Waveguides can be cast into the body of the TCMD with optically active ligands bound to the surface of the wave guide for the detection and measurement of collected analyte.

The preceding examples are set forth to illustrate specific embodiments of the invention and are not intended to limit the scope of the apparatus and method of the present invention. Additional emboαiments and advantages within the scope of the claimed invention will be apparent to one of ordinary skill in the art. Table S - Uptake of Estradiol through Hairless Rat Skin

Copoiymer Patch Formulation % Skin Uptake % Patch (mean±SD) Uptake

(mean±SD)

PVP/PVA High Dose 16.0414 3.9±0.404

PVP/PVA Low Dose 19.9914.1 4±0.95

PVA/GANEX - 4.03±1.46

Isonate M342 Desmophen 2200B 7.9210.57 14.6110.2

Isonate M342 Desmophen 2200B 5.714.6 6.9111.6

Isonate M342 Desmophen 2200B 16±9.75 6.116.3

Isonate M342 Desmophen 2200B 10.913.26 4.3115.13

Isonate M342 Desmophen 2200B 19.516.3 1.2710.824

CoTran 7.2±5 2.311.3

Table 9 - Direct Uptake of Progesterone into Copoiymer Patch Form.

Copoiymer Patch % Uptake

Formulation (Mean±SD)

Isonate M342/Desmophen 77.5113.1

2200B

IM342/D2200B with 61.7121

CoTran Adhesive #9871

IM342/D2200B with 76.4124

National Starch

Adhesive #87-2516

IM342/D2200B with 84.817.1

MacTac#545 Adhesive

IM301/Bayflex 0549 67.5110.9

IM342/Bayflex 0681 57.6132

I181/D1800 (mold 1) 90.3110

I181/D1800 (mold 2 71.4117.9

1.5mm)

Claims

WHAT IS CLAIMED IS:

1. A transdermal collection monitoring device comprising a molded polymeric collection medium.

2. A transdermal collection monitoring device as in claim 1 wherein the molded polymeric collection medium has a skm-to- patch transfer link.

3. A transdermal collection monitoring device as in claim 1, further comprising a reservoir medium.

4. A transdermal collection monitoring device as in claim 2 further comprising a reservoir medium.

5. A transdermal collection monitoring device as in claim 2 wherein the skin-to-patch transfer link is a polyacrylate adhesive.

6. A transdermal collection monitoring device as in claim 4 wherein the skm-to-patch transfer link is a polyacrylate adhesive .

7. A transdermal collection monitoring device as in claim 1 wherein the polymeric collection medium is a copoiymer.

8. A transdermal collection monitoring device as m claim 7 wherein the copoiymer is made from an isocyanate pre-polymer and a polyester polyol.

9. A transdermal collection monitoring device as in claim 1 wherein the polymeric collection medium is transparent or translucent .

10. A transdermal collection monitoring device as in claim 1 further comprising a reservoir medium.

11. A transαermal collection monitoring device as in claim 10 wnerein the reservoir medium is selected from the group consisting of polymers, oils, fats, fatty acids, fatty acid esters, waxes, and rubbers.

12. A transdermal collection monitoring device as in claim 1 further comprising an chemical sensor.

13. A transdermal collection monitoring device as in claim 12 wnerein the chemical sensor is selected from the group consisting of optical sensors, electrochemical sensors, biosensors and immunosensors.

14. A method of measuring a chemical in the blood of an animal comprising the steps of; applying a transdermal collection monitoring device as in claim 1 to the surface of the skin of an animal needing blood chemical measurements, collecting a chemical in the transdermal collection monitoring device and analyzing the chemical collected in the transdermal collection monitoring device.

15. A method of producing the transdermal collection monitoring device as in claim 1 comprising the steps of; producing a polymeric collection medium and casting the polymeric collection medium to form a molded polymeric collection medium.

16. A method of producing the transdermal collection monitoring device as in claim 15 wherein the step of producing the polymeric collection medium comprises reacting an isocyanate pre-polymer and a polyester polyol.

17. A method of measuring a chemical in the blood of an animal as in claim 14 wherein the chemical collected in the transdermal monitoring device is analyzed by acousto-optic tunable filter near infrared spectroscopy or laser desorption mass spectrometry.

18. A method of measuring a chemical m the blood of an animal as in claim 14 wherein the chemical collected in the transdermal monitoring device is analyzed by a chemical sensor wherein the sensor is integrated into the polymeric collection medium.

19. A Method of measuring a chemical in the blood of an animal as in claim 14 wherein the chemical collected in the transdermal collection monitoring device is analyzed by a method selected from the group consisting of infrared spectroscopy, visible spectroscopy and ultraviolet spectroscopy.

20. A transdermal collection monitoring device as in claim 1 wherein the polymeric collection medium is a form for collecting chemicals selected from the group consisting of therapeutic drugs, endogenous hormones, drugs of abuse, environmental toxins and industrial chemicals.

21. A transdermal collection monitoring device as in claim 11 wherein the reservoir medium is dispersed within the polymeric collection medium.