JP2003527613A - Systems, methods and devices for non-invasive sampling and analysis of body fluids - Google Patents

Abstract

(57) Abstract A system, method, and device for non-invasive body fluid sampling are provided. In one embodiment, the system includes a controller that controls the generation of ultrasound, an ultrasound applicator that applies ultrasound to the area of the biological membrane, and contacts the area of the biological membrane with fluid from the biological membrane. A receiving receptacle, and a meter for detecting the presence of at least one analyte of bodily fluid in the receptacle. The receiver may include a membrane and a medium, such as a hydrogel, fluid, liquid, etc., contained in the membrane. In one embodiment, the method comprises: (1) identifying a region of the biological membrane at a certain permeability level; (2) increasing the permeability level of the region of the biological membrane; Bringing the area of the membrane into contact with the receiver, (4) extracting body fluid through the biological membrane, (5) applying an external force to enhance body fluid extraction, (6) collecting body fluid in the receiver, 7) analyzing the collected bodily fluids to detect the presence of at least one analyte, and (8) providing a result of the bodily fluid analysis.

Description

Detailed Description of the Invention

CROSS REFERENCE TO RELATED APPLICATIONS The present invention is directed to US Provisional Patent Application No. 60/1 filed March 17, 2000.
Calls to enjoy the benefits of 89971. This disclosure is incorporated herein by reference in its entirety.

Further, the present invention is directed to US patent application Ser. No. 08/885931, entitled "Ultrasound Enhancement of Transdermal Transport,""Chemical and Physical Enhan."
US patent application Ser. No. 09 / 260,265 entitled "Cers and Ultrasound for Transdermal Drug Deivery" and "Method and Apparatus for Enhancement of
PCT International Patent Application No. PCT / US99 entitled "Transdermal Transport"
/ 30067. The entire disclosures of these disclosures are incorporated herein by reference.

BACKGROUND OF THE INVENTION 1. FIELD OF THE INVENTION The present invention relates to non-invasive sampling of body fluids, and more particularly to systems, methods and devices for non-invasive sampling and analysis of body fluids.

2. Description of the Related Art Diabetics frequently puncture their fingers and forearms to obtain blood in order to monitor blood glucose levels. Using blood for frequent monitoring is both painful and tedious. A new, pain-relieved bodily fluid sampling method is being considered,
It is disclosed. For example, these pain-free methods use small needles, iontophoresis, and ultrasound to sample body fluids such as blood and interstitial fluid.

It has been shown that it is possible to increase the permeability of the skin by applying ultrasound. Examples of such are US Pat. No. 4,767,402, US Pat. No. 594.
7921, and US Pat. No. 6,0029,61. The entire disclosures of these are incorporated by reference. Ultrasound is applied to the stratum corneum via a coupling medium to break the lipid bilayer structure by the action of cavitation and its bioacoustic effects. Disrupting the stratum corneum, which is a barrier to transport, enhances the diffusion of analytes such as glucose or drugs through and into and out of the skin.

Transport of analytes and body fluids can be further enhanced by the action of motive forces. These motive forces include sonophoretic, iontophoretic, electric, pressure, vacuum, electromagnetic, thermodynamic, magnetic, chemical, capillarity, and osmotic pressure, among others. The use of active force provides a means to obtain a fluid for subsequent analysis.

Motivating the skin before, during, and after making the skin permeable is described in US Pat. No. 5,279,543, US Pat. No. 5,722,397,
It is disclosed in US Pat. No. 5,947,921, US Pat. No. 6,0029,61, and US Pat. No. 6,093,343. The entire disclosures of these are incorporated by reference. The purpose of the motive force is to actively extract body fluids and their contents from the skin for use in analysis. As mentioned above, active forces such as vacuum, sonophoresis, and electroosmotic forces can form convection through the stratum corneum.
These forces can be used to extract body fluids, but certain limitations are imposed when they are applied to human skin. For example, a major limitation is the flow and volume of body fluids transported across the stratum corneum. High pressures are generally required to transport fluids across the highly permeable regions of the stratum corneum.
Vacuuming the skin for an extended period of time can cause the epidermis to physically separate from the dermis, resulting in scars and blisters.

[0008] Examples of other limitations include the amount of energy provided to the skin to form convection. Prolonged exposure to ultrasound can cause distress and skin damage when extracting usable quantities of body fluids. Similarly, electroosmotic extraction of body fluids through the stratum corneum requires the use of a high density of electrical current, which can damage the skin. When the above extraction method is applied to human skin, its use is obviously limited.

SUMMARY OF THE INVENTION Accordingly, there is a need for systems, methods and devices for non-invasive sampling and analysis of body fluids that overcome the above and other shortcomings of the related art.

Thus, methods for increasing the permeability of biological membranes such as the skin, mouth, and nails over time, as well as blood, interstitial fluid, lymph, in a non-invasive and practical discontinuous or continuous manner. Alternatively, a method for extracting body fluids for monitoring other body fluid analytes is needed.

A method for non-invasive sampling and analysis of body fluids is disclosed. According to one embodiment of the invention, the method comprises (1) identifying a region of the biological membrane having a certain permeability level, and (2) increasing the permeability level of the region of the biological membrane. A step, (3) contacting a region of the biological membrane with a receiver, (4) extracting body fluid from the biological membrane through the region of the biological membrane, (5) Applying an external force to enhance fluid extraction, (6) collecting fluid in a receiver, and (7) analyzing the fluid collected to detect the presence of at least one analyte. (8) providing the results of the step of analyzing the body fluid.

The area of the biological membrane is permeabilized using controlled ultrasonic waves of dosimetry. Extraction of body fluids is performed in the area exposed to ultrasound using osmotic transport. Body fluids are collected using a receiver. The receiver is a patch, wearable reservoir,
Attached to biological membranes in the form of membranes, absorbable strips, hydrogels, or equivalents. The receiver is analyzed to detect the presence of various analytes indicative of blood analytes. Electrochemical, biochemical, optical, fluorescence, absorbance (absorbance), reflectance, Raman, magnetism, mass spectrometry, infrared (IR) spectroscopy, and a combination thereof can be used for the analysis. The receiver can also be attached to the second receiver. The analyte concentration in the second receiver is continuously maintained to be substantially lower than the concentration in the body fluid, maximizing the chemical concentration driving force between the body fluid and the second receiver. To be. This is accomplished by chemical reaction or dilution volume or similar means. In one embodiment, the receiver and the second receiver may operate on different principles (eg, osmosis, dilution, etc.). In other embodiments, the receiver may operate on the same principle.

A system for non-invasive sampling and analysis of body fluids is disclosed. According to one embodiment of the invention, the system comprises a controller for controlling the generation of ultrasonic waves, an ultrasonic applicator for applying ultrasonic waves to the area of the biological membrane, and a biological contact for the area of the biological membrane. A receiver for receiving bodily fluid through the region of the biological membrane and from the biological membrane, and a meter that interacts with the receiver and detects the presence of at least one analyte in the bodily fluid within the receiver. The receiver can include a membrane and a medium such as a hydrogel, fluid, or liquid contained within the membrane.

A method for non-invasive sampling and analysis of body fluids is disclosed. According to one embodiment of the invention, the method comprises: (1) increasing the permeability level of the area of the biological membrane; (2) attaching a receiver to the area of the biological membrane;
) Extracting the analyte through and from the area of the biological membrane, (4) collecting the bodily fluid in a receiver, and (5) determining the concentration of at least one analyte in the bodily fluid. And a step of performing.

A device for non-invasive sampling and analysis of body fluids is disclosed. According to one embodiment of the present invention, a device is attached to a region of a biological membrane having enhanced permeability, a receiver for receiving body fluid from the biological membrane through the region of the biological membrane, and the received body fluid. A wearable meter for detecting the presence of at least one analyte in the and indicating the concentration of the analyte. The receiver includes a membrane and a medium such as a hydrogel, fluid, or liquid contained in the membrane. The meter has a processor and a device that detects the presence of the analyte. The detection device includes an electrochemical detector, a biochemical detector, a fluorescence detector, an absorbance (absorbance) detector, a reflectance detector, a Raman detector, a magnetic detector, a mass spectrometry detector, an IR spectroscopy detector, And combinations thereof.

According to one embodiment of the invention, the osmotic force is used to sample body fluid from a biological membrane through the biological membrane in an on-demand manner. Solutions, gels, hydrogels, or other forms of penetrants (osmotic agents) are used whenever the concentration of an analyte needs to be determined for diagnostic and monitoring purposes.
A receiver, such as a thin liquid reservoir, is used to apply the ultrasonically treated biological membrane. The receiver is attached to the biological membrane using an adhesive. The receiver is attached to the biological membrane for a short period of time. The solution in the receiver is then removed and analyzed to detect the presence of the analyte. In one embodiment, the receiver is configured in the form of a patch. The receiver may include a hydrogel and a penetrant. The receiver combines the penetrant and a chemical reagent to detect the presence of the analyte. Depending on the reagent (reactant), electrochemical, biochemistry, optics, fluorescence, absorbance, reflectance, Raman, magnetism, mass spectrometry, infrared (IR) spectroscopy methods, and combinations thereof are performed on the receiver. It will be possible.

In another embodiment, the osmotic force is used to sample body fluid from or through a biological membrane in a periodic or continuous manner. Solution penetrants can be applied to sonicated biological membranes using thin receivers, such as thin liquid reservoirs, when analyte concentration needs to be determined for diagnostics and monitoring. Given. The receiver is attached to the biological membrane using an adhesive. In one embodiment, the receiver is configured in the form of a patch. The receiver may include a hydrogel that includes a penetrant. The receiver may include means for manipulating the strength and duration of the osmotic force. The strength of the osmotic force is electric field force, magnetic field force, electromagnetic field force, biochemical reaction,
It can be operated using chemicals, molarity control, solvent control, pH control, ultrasonic field force, electroosmotic field force, ionic permeation field force, electroporation (electroporation) field force, and combinations thereof. The duration of osmotic force is electric field force, magnetic field force, electromagnetic field force, biochemical reaction, chemical product (drug), molar concentration adjustment, solvent adjustment, pH adjustment, ultrasonic field force, electroosmotic field force, ion infiltration field Can be operated using forces, electroporation field forces, and combinations thereof. The receiver may detect the presence of the analyte by combining the penetrant and the biochemical reagent. Reagents enable electrochemistry, biochemistry, optics, fluorescence, absorbance, reflectance, Raman, magnetism, mass spectrometry, infrared (IR) spectroscopy methods, and combinations of these on the receiver Become. The receiver can also be periodically removed for detection.

In one embodiment, the strength, duration, and frequency of exposing a biological membrane to osmotic forces are varied using electrical current to vary the concentration of osmotic agent in contact with the ultrasonically exposed biological membrane. Is operated. Penetrants are multiply charged agents that can dissociate into several charged species. These charged species are transported using electric field forces. Membranes are used to separate charged species. The charged species are free to diffuse and bind when the field force is removed.

In one embodiment, the strength, duration, and frequency of exposing a biological membrane to osmotic force is determined by determining the concentration of penetrant in contact with the ultrasonically exposed biological membrane using activating force. It is operated by changing it. The penetrant can be a neutral charge agent. Penetrants are transported using various field forces. The field force depends on the composition and cohesive nature of the penetrant selected. The field force produces the force necessary to move the penetrant towards and away from the biological membrane surface. Penetrant migration regulates the periodic and continuous extraction of body fluids through the stratum corneum.

In one embodiment, the strength, duration, and frequency of exposing a biological membrane to osmotic forces are manipulated by varying the concentration of penetrant that contacts the ultrasonically exposed biological membrane. It By manipulating the volume of solvent and the volume of hydrogel containing the penetrant, the concentration of penetrant is varied. The volume of the hydrogel can be varied by making the hydrogel sensitive to the concentration of molecules whose volume can diffuse into the gel. One example is a hydrogel configured to be sensitive to molecular glucose. The hydrogel volume can also be changed by manipulating its temperature and by changing the pH of the gel.

A receiver is disclosed that is attached to a region of a biological membrane with enhanced permeability and receives body fluid through and from the region of the biological membrane. According to one embodiment of the invention, the receiver comprises a first grid, a media layer containing at least one drug,
A membrane that induces a concentration gradient barrier for the at least one drug, an opposing grid, an oxidase layer, a detection layer, and a voltage source that provides a potential difference between the first grid and the opposing grid. Bodily fluids, including blood, interstitial fluid, analytes, and lymph, flow from or through the biological membrane, through the first grid, the opposing grid, and the oxidase layer to the detection layer.

A technical advantage of the present invention is the disclosure of systems, methods and devices for non-invasive sampling and analysis of body fluids. Another technical advantage of the present invention is that the concentration of the analyte is measured continuously or periodically.

For a more complete understanding of the present invention, objects and advantages of the present invention, reference is made to the following description with reference to the accompanying drawings. DETAILED DESCRIPTION OF THE INVENTION The preferred embodiments of the present invention and its advantages are best understood by referring to FIGS. 1-7 of the drawings. Like reference numerals are used to indicate like and corresponding parts in the various drawings.

The term “body fluid” as used herein includes blood, interstitial fluid, lymph fluid, and / or analytes. In addition, the term "biological membrane" as used herein includes tissues, mucous membranes, and keratinous tissues including skin, mouth and nails. Furthermore, the term "force" as used herein also includes a force gradient.

Although the present invention has been described in the context of human applications, veterinary applications are also within the spirit and scope of the present invention. Referring to FIG. 1, a flow chart is provided that illustrates a method for non-invasive sampling and analysis of body fluids according to one embodiment of the invention. In step 102, the permeability of the area of the biological membrane is increased. In one embodiment, the region of the biological membrane is located on the volar forearm of the mammalian subject. In another embodiment, the region of biological membrane is located on the thigh of a mammalian subject. In yet another embodiment, the area of biological membrane is located in the abdomen. In yet another embodiment, the area of biological membrane is located on the back. Other body positions may be used.

Generally, forming physical micropores to enhance the permeability of biological membranes,
Several techniques are available, such as physically disrupting the lipid bilayer structure, chemically modifying the lipid bilayer structure, physically destroying the stratum corneum, and chemically modifying the stratum corneum. Can be used. The formation of micropores (micropores) or its destruction can be performed by needles, microneedles, silicon microneedles, lasers, lasers combined with absorbing dyes, heat sources, ultrasonic needles, ultrasonic transducers, cryoablation, RF ablation, photoacoustic ablation, And accomplished by physical intrusion with a combination thereof.

In a preferred embodiment, ultrasound is applied to the area of the biological membrane to increase its permeability. Ultrasound is generally defined as sounds at frequencies above about 20 kHz. The therapeutic ultrasound is typically between 20 kHz and 5 MHz. Near ultrasound is typically about 10 kHz to about 20 kHz. It should be appreciated that in addition to ultrasound, near ultrasound can be used in embodiments of the invention.

In general, ultrasound or near ultrasound is preferably cavitation
Is applied to the region of the biological membrane at a frequency sufficient to cause and increase the permeability of the biological membrane. In one embodiment, the ultrasound is about 10 kHz to about 500 kHz.
given at the frequency of z. In another embodiment, the ultrasound is from about 20 kHz to about 1
Given at a frequency of 50 kHz. In yet another embodiment, the ultrasound is 50 kHz.
given by z. Ultrasound at other frequencies may be used to increase the permeability level of biological membranes.

In one embodiment, the ultrasound has an intensity in the range of about 0 to about 100 watts / cm ² , preferably 0 to about 20 watts / cm ² . Other suitable strengths are used as needed.

Techniques for increasing the permeability of biological membranes are disclosed in Kost et al., US Pat. No. 6,190,315. The entire disclosure of this is incorporated herein by reference.

In step 104, bodily fluid is extracted through or from the area of the biological membrane. In one embodiment, external forces such as osmotic forces aid extraction. In one embodiment, the osmotic force is controlled before, during, and after the permeability of the biological membrane is increased.

In one embodiment, the osmotic force is generated by applying an osmotic agent to a region of the biological membrane. Penetrants can be in the form of elements, molecules, macromolecules, chemical compounds, or combinations thereof. Penetrants can also be combined with liquid solvents, hydrogels, gels, or solvents having similar functions.

In step 106, the magnitude, strength, and duration of the external force may be adjusted by the at least one additional first energy and / or force. In one embodiment, the additional first energy and / or force is provided to control and regulate the movement and function of the osmotic agent for extracting body fluids through and from the biological membrane. The further first energy and / or force is heat, temperature force, pressure, electric force, mechanical vibration, ultrasound, iontophoresis,
It may be provided in the form of electromagnetic forces, magnetic forces, photothermal forces, photoacoustic forces, and combinations thereof. The effects of electric fields and ultrasound on transdermal drug delivery are described in US Pat. No. 6041.
No. 253. The entire disclosure of this disclosure is incorporated by reference.

In one embodiment, if the additional first energy and / or force is provided by ultrasound, the frequency of the ultrasound is different from the frequency used to increase the permeability of the biological membrane. Offered in frequency. In one embodiment, the frequency of the additional first energy / force ultrasonic waves may be higher than the frequency of the ultrasonic waves that enhance the transmittance.

In step 108, body fluid is collected in the receiver. In one embodiment, the receptacle contacts the biological membrane in the form of a patch, wearable reservoir, membrane, absorbent strip, hydrogel, or equivalent functioning structure. Other types and configurations of receivers may be used.

In one embodiment, the receiver is provided with a second receiver having an analyte concentration that is continuously maintained to be substantially lower than the concentration of the analyte in the body fluid. The chemical concentration driving force between the second receiver and the second receiver is maximized. This can be a chemical reaction or a volume for dilution.
dilution) or similar means.

In one embodiment, the second external energy / force is provided between the first receiver and the second receiver. In one embodiment, the second external energy / force may be different than the first external energy / force (eg, a different type of external force). In other embodiments, the second external energy / force can be the same as the first external energy / force (eg, the same type of external force). The first and second external energies / forces can be of various types, durations, and intensities and can be controlled through further different energies and / or forces.

At step 110, the collected body fluid is analyzed. In one embodiment, the analysis is suitable for electrochemistry, biochemistry, optics, fluorescence, absorbance, reflectance, Raman, magnetism, mass spectrometry, infrared (IR) spectroscopy measurement methods, and combinations thereof. Use of the method can be included.

In one embodiment, multiple analytes are analyzed simultaneously, in parallel, or sequentially. The results from these multiple analyzes can be used in combination with algorithms, for example, to improve the accuracy and / or precision of the analysis and measurement.

In one embodiment, the receiver is removed from contact with the biological membrane to analyze the collected bodily fluid. In other embodiments, the receiver is kept in contact with the biological membrane during the analysis of the collected body fluid.

Referring to FIG. 2, there is shown a device for the controlled application of ultrasonic waves to a biological membrane to enhance the permeability of the biological membrane, according to one embodiment of the present invention. The device 200 has a controller 202, which interfaces with the ultrasonic applicator 204 by any suitable means such as a cable. The controller 202 controls the application of ultrasound to the area of the biological membrane. In one embodiment, ultrasound or near ultrasound having an intensity in the range of about 0 to about 20 watts / cm ² is generated by controller 202 and ultrasound applicator 204. In one embodiment, the ultrasound waves have a frequency of about 20 kHz to about 150 kHz. In another embodiment, the ultrasound has a frequency of 50 kHz. Other ultrasonic frequencies can also be used.

In addition, the controller 202 uses the L
It may include a display such as a CD or LED display and the like. Controller 202 may also include user interfaces known in the art.

The ultrasonic applicator 204 includes a cartridge 2 containing an ultrasonic bonding solution 208.
06 is provided. The cartridge 206 may be formed of any material such as plastic that encapsulates the ultrasonic bonding solution 208. Suitable ultrasonic bonding solution 20
8 include, but are not limited to, water, brine, alcohols including ethanol and isopropanol (concentration range of 10 to 100% in aqueous solution), Triton X-
A surfactant such as 100, SLS, or SDS (preferably 0.
Concentration range between 001% and 10%), DMSO (preferably 1 in aqueous solution)
A concentration range between 0% and 100%), fatty acids such as linoleic acid (preferably,
Concentration range between 0.1% and 2% in ethanol-water (50:50) mixture)
Azone (preferably a concentration range between 0.1% and 10% in an ethanol-water (50:50) mixture), preferably polyethylene in a concentration range between 0.1% and 50% in an aqueous solution. Glycol, preferably 0.1 to 10 in aqueous solution
Histamine in a concentration range between 0 mg / ml, preferably EDTA in a concentration range between 1 and 100 mM, preferably sodium hydroxide, sodium octyl sulfate, N-lauroyl sarcosine, octyl trimethyl in a concentration range between 1 and 100 mM. Ammonium bromide, dodecyl trimethyl ammonium bromide, tetradecyl trimethyl ammonium bromide, hexadecyl trimethyl ammonium bromide, dodecyl pyridinium chloride hydrate, SPAN20, BRIJ30, glycolic acid ethoxylate 4-t-butyl phenyl ether, IGEPAL CO-21
Any material may be included such as 0, and combinations thereof.

In one embodiment, the binding medium also includes a chemical enhancer. Transport is enhanced by adding a capillary permeability enhancer, such as histamine, to the binding medium. The concentration of histamine in the binding medium ranges between 0.1 and 100 mg / ml. These agents are delivered across the biological membrane during the application of ultrasound, causing local edema which increases local fluid pressure and enhances transport of analytes through the biological membrane. In addition, the generation of free fluid by edema locally induces cavitation and enhances transport of analytes through biological membranes.

In one embodiment, the cartridge 206 is pierced when inserted into the ultrasonic applicator 204 and the ultrasonic binding solution 208 is transferred to the chamber (not shown).

A target identification device, such as target ring 210, is attached to the area of the biological membrane where the permeability is enhanced. The target ring 210 is attached to the area of the biological membrane by a transdermal adhesive (not shown). In one embodiment, the target ring 210 is pre-applied with a transdermal adhesive and treated after each use. In other embodiments, target ring 210 is reusable.

The target ring 210 is formed of any suitable material including plastic, ceramic, rubber, foam and the like. In general, the target ring 210 is
Identify areas of the biological membrane to enhance permeability and extract body fluids. In one embodiment, the target ring 210 is used to hold the receptacle 214 in contact with the biological membrane after the permeability of the biological membrane has been increased.

In one embodiment, the target ring 210 includes a “Method and Apparatus for
PCT International Patent Application No. P entitled "Enhancement of Transdermal Transport"
Used to monitor the permeability level of biological membranes as disclosed in CT / US99 / 30067. The entire disclosure of this disclosure is incorporated by reference. In such an embodiment, target ring 210 interfaces with ultrasonic applicator 204.

The ultrasonic applicator 204 is applied to the target ring 210 and activated to expose the ultrasonic binding solution 208 to the biological membrane. The controller 202
The ultrasonic applicator 204 is controlled to transmit ultrasonic waves through the ultrasonic coupling solution 208. During exposure to ultrasound, controller 202 monitors changes in the permeability of biological membranes and displays this information to the user.

The controller 202 stops or stops the application of ultrasound when the biological membrane permeability reaches a predetermined level. This level of transmission is pre-programmed or determined in real time while the ultrasound is being applied. Due to the differences in biological membranes between solids, a certain level of permeability for each solid is programmed.

After the transmittance reaches a predetermined level, the ultrasonic coupling solution 208 is discharged from the chamber (not shown) to the cartridge 206 and discarded. In other embodiments,
The ultrasonic bonding solution 208 is held in a holding area (not shown) within the ultrasonic applicator 204.
It will be discharged to and discarded later. The ultrasonic applicator 204 is then removed from the target ring 210.

Referring to FIG. 3, a device for analyzing bodily fluids is provided according to one embodiment of the present invention. The receiver 214 is disposed on the target ring 210 and may provide discontinuous or on-demand extraction of bodily fluids through and / or from the biological membrane. The receiver 214 may include a medium such as a hydrogel layer that incorporates a penetrant. In one embodiment, the hydrogel is phosphate buffered saline (PBS (phos
Phate buffered saline)) and the saline is sodium chloride having a concentration range of about 0.01M to about 10M. The hydrogel is buffered at pH 7. Other penetrants may also be used in place of or in addition to sodium chloride. Preferably, these penetrants are non-irritating, non-staining and non-immunogenic. Examples of such penetrants include lactate and magnesium sulfate, among others.

In other embodiments, the receiver 214 comprises a fluid or liquid medium such as water or a buffer contained within the semipermeable membrane. The receiver 214 may also include a spongy material such as foam.

The receiver 214 is applied to the biological membrane and contacts the ultrasonically exposed biological membrane. In one embodiment, the receiver 214 is contacted with the biological membrane for a period of time sufficient to collect a sufficient amount of bodily fluid to be detected. In another embodiment, the receiver 21
The 4 is contacted with the biological membrane for a period of time sufficient to collect a predetermined amount of body fluid. In yet another embodiment, the receiver 214 is contacted with the biological membrane for a period of time. In one embodiment, the contact between the receiver 214 and the biological membrane is 15 minutes or less. In other embodiments, the contact between the receiver 214 and the biological membrane is less than 5 minutes. In yet another embodiment, the contact between the receiver 214 and the biological membrane is less than 2 minutes. The actual duration of contact depends on the sensitivity of the detection method used in the analysis.

In one embodiment, the medium of the receiver 214 is at least 1 to detect the presence of a particular analyte in body fluid extracted from or through the biological membrane.
One reagent (not shown) may be included. In one embodiment, the hydrogel layer of the receiver 214 comprises a reagent, which is attached to the hydrogel by ionic and / or covalent means or immobilized by gel entrapment. The reagents are also arranged as a layer adjacent to the hydrogel, and the analyte from the body fluid extracted into the hydrogel can diffuse and react to produce a by-product. The by-products can then be detected using electrochemical, biochemical, optical, fluorescent, absorbance, reflectance, Raman, magnetic, mass spectrometry, IR spectroscopy measurement methods, and combinations thereof.

The detection method is accomplished by the meter 212. Meter 212 may include a processor (not shown) and a display such as an LCD display. Other suitable displays may also be provided.

In one embodiment, the meter 212 provides an interface that allows information to be downloaded to an external device such as a computer. Such an interface may be one that allows the connection of an interface cable or may be a wireless interface.

The meter 212 introduces glucose oxidase into the medium of the receiver 214 (in
corporate body) to determine the body fluid glucose concentration. In one embodiment, glucose from the extracted body fluid reacts with glucose oxidase to produce hydrogen peroxide. Hydrogen peroxide is detected by the oxidation of hydrogen peroxide on the surface of the electrode introduced into the receiver 214. Oxidation of hydrogen peroxide transfers electrons to the electrode surface, which produces a current flow that can be metered using a potentiostat introduced into meter 212. A glucose concentration proportional to the concentration of hydrogen peroxide is calculated and the result is reported to the user via the display. Various configurations of electrodes and reagents known to those skilled in the art can be introduced to perform the detection and analysis of glucose and other analytes.

The meter 212 is also expected to have analytes, such as glucose, that are expected to fluctuate in body fluid concentration, and body fluid concentrations to remain relatively stable over minutes, hours, or days. It can also be configured to simultaneously measure the concentration of an analyte such as creatinine or calcium. Analyte concentrations determined by an algorithm that considers relative concentrations of varying and stable analytes are reported to the user via a display.

In other embodiments, the meter 212 may analyze multiple analytes simultaneously, in parallel, or in series. The results from these multiple analyzes are used, for example, in combination with algorithms to improve the accuracy and / or precision of the analyzes and measurements.

The receiver 214 is discarded after the extraction and measurement steps. In other embodiments, the receiver 214 is reused. In one embodiment, the receiver 214 is cleaned, sterilized, etc. before being reused. Various configurations of electrodes and reagents known to those skilled in the art can be introduced to detect and analyze glucose and other analytes.

Referring to FIG. 4, in accordance with another embodiment of the present invention, there is provided a device for continuous extraction and analysis of body fluids for estimating analyte concentration. As shown, the site of the biological membrane on the forearm, abdomen, or thigh is exposed to ultrasound. Other biological membrane sites such as the back site can also be used. The receiver 402, which may be similar to the receiver 214, contacts the portion of the biological membrane that has been exposed to ultrasound and provides for continuous extraction of body fluids. In one embodiment, the receiver 402 is
It may include a medium such as a hydrogel layer into which a penetrant such as sodium chloride may be incorporated.
The hydrogel was prepared to include phosphate buffered saline (PBS), the saline solution at 0.
Sodium chloride having a concentration range of 01M to 10M. Hydrogel is p
Buffered with H7.

Other penetrants can be used in place of or in addition to sodium chloride. These penetrants are preferably non-irritating, non-staining and non-immunogenic. Examples of these other penetrants include lactate and magnesium sulfate, among others. The receptacle 402 is adapted to contact the ultrasonically exposed biological membrane. In one embodiment, the duration of this contact is 1
It can be 2 to 24 hours, or longer. In other embodiments, other contact durations, including substantially shorter durations and substantially longer durations, can be used as desired.

In other embodiments, the receptacle 402 may include a fluid or liquid medium such as water or a buffer contained within the semipermeable membrane. The receiver 402 may also include a spongy material such as foam.

In one embodiment, the medium of the receiver 402 includes at least one reagent (not shown) that detects the presence of the analyte through the biological membrane and in the body fluid extracted from the biological membrane. . In one embodiment, the hydrogel layer of the receiver 402 comprises reagents attached to the hydrogel by ionic and covalent means, or immobilized by gel entrapment. The reagent is also arranged as an adjacent layer of the hydrogel, and the analyte from the body fluid extracted into the hydrogel diffuses and reacts therewith to produce a by-product. By-products include electrochemistry, biochemistry, optics, fluorescence, absorbance, reflectance, Raman,
It can be detected using magnetic, mass spectrometry, IR spectroscopy measurement methods, and combinations thereof.

The detection method and result may be similar in function to the meter 212 described above.
04 and is presented to the user. In one embodiment, the meter 404 is wearable. For example, as shown, the meter 404 can be worn similar to wearing a wristwatch. The meter 404 can also be mounted on the belt, in a pocket, etc.

The meter 404 can introduce power and electronics, control periodic extraction of bodily fluids, detect analytes, and continuously present analyte concentrations.
Meter 404 includes electronics and software to obtain sensor signals,
Signal processing can be performed and analytical and trend information can be stored.

In one embodiment, meter 404 provides an interface for downloading information to an external device such as a computer. Such an interface allows connection of an interface cable or is a wireless interface.

Meter 404 is configured to determine body fluid glucose concentration by introducing glucose oxidase into the medium. In one embodiment, glucose from the extracted body fluid reacts with glucose oxidase to produce hydrogen peroxide. Hydrogen peroxide is detected by the oxidation of hydrogen peroxide on the surface of the electrodes introduced into the receiver 402. Oxidation of hydrogen peroxide carries electrons to the electrode surface, which produces a current flow that can be metered using a potentiostat introduced into meter 404. The glucose concentration, which is proportional to the concentration of hydrogen peroxide, is calculated and the result is reported to the user via the display. Various configurations of electrodes and reagents known to those of skill in the art can be introduced to detect and analyze glucose and other analytes.

[0070] In one embodiment, the meter 404 also provides an analyte, such as glucose, which is expected to have varying fluid concentrations, and a fluid concentration, such as creatinine or calcium, that is minutes, hours or It can also be configured to simultaneously measure the concentration of analyte that is expected to remain relatively stable over several days. Analyte concentrations that can be determined by an algorithm that considers relative concentrations of analytes that are variable and more stable can be reported to the user via a display.

In other embodiments, the meter 404 can analyze multiple analytes simultaneously, in parallel, or in series. The results from these numerous analyzes are
For example, it can be used in combination with an algorithm to improve the accuracy and / or precision of analysis and measurement.

In another embodiment, the receiver 402 is removed from contact with the biological membrane for analysis by the meter 404. The receptacle 402 is contacted with the biological membrane after such analysis.

The meter 404 provides the user with analyte readings periodically or continuously. For example, in one embodiment, in continuous monitoring of analyte glucose, glucose concentration is displayed to the user every 30 minutes, more preferably every 15 minutes, most preferably every 5 minutes, or even more frequently. obtain. In other embodiments, glucose concentration may be displayed continuously. The cycle depends on the sensitivity and method of analyte detection. In continuous glucose monitoring, in one embodiment, glucose detection is performed electrochemically with electrodes and reagents introduced into receiver 402 and detection and analysis is performed by meter 404. During the measurement period, the osmotic extraction of body fluid is continuously performed by the hydrogel layer of the receiver 402. Body fluid accumulates in the hydrogel of receiver 402. Glucose in body fluid diffuses, reacts with glucose oxidase, and is converted into hydrogen peroxide. Hydrogen peroxide is consumed by keeping the working electrode against the reference electrode. During the rest period, hydrogen peroxide accumulates and is consumed or destroyed before the measurement period. An operating potential large enough to quickly consume the accumulated hydrogen peroxide can be provided.

Referring to FIG. 5, an approach for periodic monitoring of analytes by performing periodic osmotic extraction of body fluids is shown, according to another embodiment of the invention. The strength and frequency of the osmotic extraction are manipulated with a penetrant that dissociates into multiple charged species, and the potential changes the concentration of charge towards the surface 550 of the biological membrane and of the biological membrane. Used to move from surface 550. The receiver 500 can be a grid, mesh or screen 504, a medium of hydrogel 5
06, membrane 508, opposing grid, mesh or screen 510, oxidase layer 512, and detection layer 514. Grid 504 and opposing grid 5
10 is connected to the voltage source 516. Membrane 508 is a semi-permeable membrane and media 506
It is used to induce a concentration gradient barrier to penetrants contained in. Preferred penetrants may include negative and positive species or counterions. By manipulating the concentration of charged species at the boundaries adjacent to the stratum corneum of biological membranes exposed to ultrasound, periodic extraction of body fluids can be performed.

In one embodiment, the receptacle 500 contacts the skin through the contact medium 502, which is a hydrogel or other suitable medium. The concentration of charged species is manipulated by applying a potential difference between grid 504 and opposing grid 510 using power supply 516. In one embodiment, the potential difference is large enough to manipulate the penetrant. Also, by changing the polarity of the grid,
Allows charge to be transported to and from the surface 550 of the biological membrane. The grid 504 and the opposing grid 510 are configured with optimal pores to transfer fluids and / or analytes from the stratum corneum, through the grid 504, the grid 510, and finally to the oxidase layer 512 and ultimately the detection layer 514. To be done. The oxidase layer 512 is used with a suitable catalyst or enzyme to provide specificity for analyte detection. The detection layer 514 includes a working electrode and a reference electrode (not shown) to allow detection of by-products of the oxidase layer 512 and quantify the desired analyte concentration of the assay.

EXAMPLES Examples are provided for a better understanding of the present invention. The examples are not intended to limit the invention in any way, but are intended to illustrate embodiments of the invention.

The following is a determination of body fluid glucose concentration in humans using a hyperosmotic extraction fluid and comparing this condition with an iso-osmotic extraction fluid according to one embodiment of the present invention. In order to do so, it is a description of experiments carried out to painlessly extract, collect, and analyze body fluids. Body fluid glucose concentration is used as an example to demonstrate feasibility, but other analytes are within the scope of the invention. Further, multiple analytes can be measured and / or analyzed simultaneously, in parallel, or in series, and the results from these multiple measurements can be improved, eg, in the accuracy and / or precision of the measurements. Can be used in combination with an algorithm. As will be appreciated by those skilled in the art, these steps can be automated and performed using the device described above.

Four sites on the volar forearm of volunteers were sonicated using the device shown in FIG. The ultrasonic transducer and its housing were placed on the volunteer's volar forearm with sufficient pressure to provide good contact between the skin and the outer transducer housing to prevent leakage. The area surrounding the transducer was then filled with a binding medium of silica particles and sodium dodecyl sulfate in phosphate buffered saline (PBS). Ultrasound was applied for a short period (5 to 30 seconds), the transducer device was removed from the biological membrane, and the skin was rinsed with tap water and allowed to dry.

FIG. 6 shows the components of a wearable brew chamber 600. Four extraction chambers were placed at each sonicated site in the volunteer. Form a thin circular foam chamber 602 with foam MED5636 Avery Dennison (7/1
6 ″ ID × 1 1/8 ″ OD) (1 ″ is about 2.54 cm). The foam chamber 602 was attached to one side of the element 602 with a double-sided adhesive (Adhese).
IVE Arcade 8570, 7/16 "ID x 7/8" OD)
It was attached concentrically to the site of the sonicated biological membrane. On the other side of the foam chamber 602, double-sided adhesive 604 (Adhesive Arcade 8570, 7/16)
“ID × 7/8” OD) was attached concentrically. The thin transparent lid 606 is 3M (
R) polyester 1012 (1 1/8 ″ × 1 1/8 ″). The double-sided adhesive 604 allowed a thin transparent lid 606 to be attached to the foam chamber 602 after the liquid was placed on the inner diameter of the foam chamber 602 when applied to the biological membrane. A thin transparent lid 606 served to prevent liquid from leaking from the brewing chamber and to allow the brewing chamber to be worn over time.

Each extraction chamber was alternately filled with 100 μl of extraction solution for 15 minutes and 100 μl of hydrate solution for 10-40 minutes. The extraction solution was PBS. At the two sites, PBS contained additional NaCl and the total concentration of NaCl was 1M. The hydrate solution was PBS for all sites.

The solution was collected and analyzed for glucose concentration using high pressure liquid chromatography. The HPLC concentration results were normalized for injection volume and total solution volume and reported as glucose flux (Q _g ), ie the mass of glucose across the sonicated site per unit area and unit time. Body fluid glucose concentrations (C _bg ) were obtained by testing capillary blood obtained from lanced fingers in a Bayer Glucometer Elite® meter. It was assumed that Q _g was linearly proportional to C _bg . FIG. 7 shows a graph of Q _g vs. C _bg . Unexpectedly, the Q _g from sonicated sites exposed to 1 M NaCl was much more strongly associated with C _bg than the Q _g from sonicated sites exposed to 0.15 M NaCl.

Other embodiments and uses of the invention will be apparent to those of ordinary skill in the art in view of the specification and practice of the invention disclosed herein. All references cited herein, including US and foreign patents and patent applications, are specifically incorporated herein by reference in their entireties. The specification and examples are intended for purposes of illustration only and the true scope and spirit of the invention is set forth in the following claims.

[Brief description of drawings]

FIG. 1 is a flow chart illustrating a method for non-invasive sampling of body fluids according to one embodiment of the present invention.

FIG. 2 illustrates a device for controlled application of ultrasound to a biological membrane to enhance permeability of the biological membrane, according to one embodiment of the invention.

FIG. 3 illustrates components for extracting and measuring discontinuities in body fluids to estimate analyte concentrations, according to one embodiment of the invention.

FIG. 4 illustrates components for performing continuous extraction and measurement of body fluids to estimate analyte concentration, according to one embodiment of the invention.

FIG. 5 illustrates an approach for periodic monitoring of analytes by performing periodic osmotic extraction of body fluids, according to one embodiment of the invention.

FIG. 6 illustrates components of a wearable brew chamber according to one embodiment of the present invention.

FIG. 7 is a graph of glucose flux versus blood glucose concentration according to one embodiment of the invention.

─────────────────────────────────────────────────── ─── Continued front page    (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, I T, LU, MC, NL, PT, SE, TR), OA (BF , BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, G M, KE, LS, MW, MZ, SD, SL, SZ, TZ , UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, B Z, CA, CH, CN, CR, CU, CZ, DE, DK , DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, J P, KE, KG, KP, KR, KZ, LC, LK, LR , LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, R O, RU, SD, SE, SG, SI, SK, SL, TJ , TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Elstrom, Tuan Ai             Ray, Illinois, United States 60044             Ku Bluff, West Sanctuary             Re-land 12780 (72) Inventor Kellogg, Scott Sea             02131, Massachusetts, United States             Boston, Sheffield Road 27 (72) Inventor Cost, Joseph             02141, Massachusetts, USA             Cambridge, Massachusetts Aveni             View 931, number 303 (72) Inventor Warner, Nicholas Ef             02178, Massachusetts, USA             Belmont, Concord Avenue             296 F term (reference) 2G045 BB60 CA30 DA31 FA25 FB05                       FB12 GC07 GC10 GC11 JA04                 4C038 TA10

Claims (71)

[Claims] 1. A method of non-invasive body fluid sampling and analysis, the method comprising: identifying a region of a biological membrane having a permeability level; and determining a permeability level of the region of the biological membrane. Increasing, contacting the area of the biological membrane with a receptacle, extracting body fluid through the area of the biological membrane, and providing an external force to enhance extraction of the body fluid Collecting bodily fluid in the receiver; analyzing the collected bodily fluid to detect the presence of at least one analyte; and providing the results of the step of analyzing the bodily fluid. A method comprising. 2. The method of claim 1, wherein identifying a region of a biological membrane having a permeability level provides a target identification device to the region of the biological membrane. A method comprising steps. 3. The method of claim 1, wherein the step of increasing the permeability level of a region of a biological membrane comprises ultrasonicating the biological waves having a frequency range of about 10 kHz to about 500 kHz. A method comprising applying to a region of a membrane. 4. The method of claim 1, wherein the step of increasing the permeability level of a region of a biological membrane comprises ultrasonicating the ultrasonic waves having a frequency range of about 20 kHz to about 150 kHz. A method comprising applying to a region of a membrane. 5. The method of claim 1, wherein the step of increasing the permeability level of a region of the biological membrane comprises ultrasonic waves having a frequency of about 50 kHz to the region of the biological membrane. A method comprising the step of applying. 6. The method of claim 1, wherein the step of increasing the permeability level of the region of the biological membrane increases the permeability level of the region of the biological membrane to a predetermined level. A method comprising the steps of: 7. The method of claim 1, wherein the step of increasing the permeability level of a region of a biological membrane comprises creating micropores in the region of the biological membrane, Physically disrupting the lipid bilayer structure in the region of the biological membrane, chemically altering the lipid bilayer structure in the region of the biological membrane, and stratum corneum in the region of the biological membrane Physically destroying the biological membrane and chemically altering the stratum corneum of the region of the biological membrane to increase the permeability level of the biological membrane in a method selected from the group: ,Method. 8. The method of claim 1, wherein said receiver is provided with a second receiver having a concentration of said at least one analyte, said second receiver. Maintaining the concentration of the at least one analyte at a level below the concentration of the at least one analyte in the body fluid. 9. The method of claim 8, further comprising the step of providing a second external force to the second receiver. 10. The method of claim 9, wherein the first external force and the second external force differ in at least one of type, duration, and intensity. 11. The method of claim 1, wherein the step of providing an external force to enhance extraction of the body fluid produces at least one osmotic force to enhance extraction of the body fluid. A method comprising steps. 12. The method of claim 11, wherein the step of generating at least one osmotic force to enhance extraction of the body fluid comprises at least one.
The method further comprising applying one osmotic agent to a region of the biological membrane to generate the at least one osmotic force. 13. The method of claim 11, wherein the step of generating at least one osmotic force to enhance extraction of the body fluid comprises heat, temperature force, pressure and electric force. Said at least one penetrating force with an external force selected from the group comprising: mechanical vibration, ultrasonic waves, iontophoresis, electromagnetic force, magnetic force, photothermal force, photoacoustic force, and combinations thereof. The method further comprising the step of adjusting. 14. The method of claim 11, wherein the step of generating at least one osmotic force to enhance extraction of the body fluid comprises the strength of the at least one osmotic force and the at least one The method further comprising manipulating at least one of one osmotic force period and said at least one osmotic force frequency. 15. The method according to claim 14, wherein applying an electric field force, applying a magnetic field force, applying an electromagnetic field force, applying a chemical substance, and Adjusting the molar concentration of at least one osmotic agent, adjusting the pH level of said at least one osmotic agent, applying an ultrasonic field force, and applying an electroosmotic field force , Applying an iontophoretic field force, applying an electroporation field force, and combinations thereof, the method further comprising the step of generating the external force. 16. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte comprises detecting the at least one analyte. The method further comprising providing at least one reagent to the receiver. 17. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte comprises electrochemical, biochemical, optical, fluorescent, absorption. Further comprising the step of detecting the presence of said at least one analyte using a method selected from the group consisting of methods of power, reflectance, Raman effect, magnetism, mass spectrometry, IR spectroscopy, and combinations thereof. ,Method. 18. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte detects the presence of the at least one analyte. For measuring using a method selected from the group comprising electrochemical, biochemistry, optics, fluorescence, absorbance, reflectance, Raman effect, magnetism, mass spectrometry, IR spectroscopy, and combinations thereof. The method further comprising the step of providing an instrument for analyzing a body fluid. 19. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte comprises: The method further comprising the step of periodically analyzing the collected body fluid. 20. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte comprises: The method further comprising the step of continuously analyzing the collected bodily fluid. 21. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte comprises the concentration of the at least one analyte in the bodily fluid. The method further comprising the step of determining 22. The method of claim 1, wherein the step of analyzing the collected bodily fluid to detect the presence of at least one analyte is based on the concentration of multiple analytes in the bodily fluid. And further determining the concentration of the at least one analyte in the body fluid. 23. The method of claim 1, further comprising removing the receiver from a region of the biological membrane after a predetermined condition. 24. The method of claim 23, wherein the step of removing the receiver from a region of the biological membrane after a predetermined condition comprises the step of receiving a sufficient amount of body fluid for analysis. Removing the receiver from the region of the biological membrane after being collected in a container. 25. The method of claim 23, wherein the step of removing the receiver from the area of the biological membrane after a predetermined condition comprises the area of the biological membrane of the receiver. Removing the receiver from the area of the biological membrane within 15 minutes of contacting the. 26. The method of claim 23, wherein the step of removing the receiver from the area of the biological membrane after a predetermined condition is such that the receiver is in the area of the biological membrane. Removing the receiver from the area of the biological membrane within 10 minutes after contacting. 27. The method of claim 23, wherein the step of removing the receiver from the area of the biological membrane after a predetermined condition is such that the receiver is in the area of the biological membrane. Removing the receiver from the area of the biological membrane within 5 minutes after contacting. 28. The method of claim 1, wherein providing the result of the step of analyzing the bodily fluid comprises displaying the result of the step of analyzing the bodily fluid. . 29. A system for non-invasively sampling and analyzing bodily fluids, the controller controlling the generation of ultrasonic waves, an ultrasonic applicator for applying the ultrasonic waves to a region of a biological membrane, A receptacle for contacting said region of a biological membrane and receiving bodily fluids therein from said region of said biological membrane, said receptacle comprising a membrane and a medium contained in said membrane; An instrument for interacting with the receiver to detect the presence of at least one analyte in the bodily fluid in the receiver. 30. The system according to claim 29, wherein the controller comprises a device for measuring the permeability level of the region of the biological membrane. 31. The system of claim 29, further comprising a cartridge containing an ultrasonic binding solution that is inserted into the ultrasonic applicator. 32. The system of claim 31, wherein the ultrasonic applicator comprises a cartridge chamber that receives the cartridge and a solution chamber that receives the ultrasonic coupling solution from the cartridge. 33. The system of claim 29, wherein the medium comprises at least one penetrant and at least one of a hydrogel layer, a fluid, and a liquid. 34. The system of claim 33, wherein the penetrant comprises at least one of sodium chloride, lactate, magnesium sulfide. 35. The system of claim 33, wherein the medium comprises at least one reagent. 36. The system of claim 29, wherein the instrument comprises a processor and an electrochemical detector, biochemical detector, fluorescent detector, absorbance detector, reflectance detector, Raman. A device for detecting the presence of said analyte selected from the group comprising a detector, a magnetic detector, a mass spectrometric detector, an IR spectroscopic detector and combinations thereof. 37. The system of claim 29, wherein the meter comprises a display that displays the concentration of the analyte. 38. The system of claim 29, further comprising a device that provides a first additional energy / force to the region of the biological membrane. 39. The system of claim 29, further comprising a target ring attached to the region of the biological membrane. 40. The system of claim 29, wherein the target ring is pre-applied with an adhesive. 41. The system of claim 38, further comprising a second receiver in communication with the receiver and having a concentration of the at least one analyte. The system wherein the concentration of the at least one analyte in is maintained at a lower level than the concentration of the at least one analyte in the body fluid. 42. The system of claim 41, further comprising a device that provides a second additional energy / force to the receiver. 43. The system of claim 42, wherein the first additional energy / force and the second energy / force differ in at least one of type, duration, and intensity. ,system. 44. The system of claim 29, wherein the instrument is wearable. 45. A method for non-invasively sampling and analyzing body fluids, comprising increasing the permeability level of a region of a biological membrane, and attaching a receiver to the region of the biological membrane. The step of: extracting the body fluid through the region of the biological membrane to the exterior thereof; collecting the body fluid in the receiver; and determining the concentration of at least one analyte in the body fluid. A method comprising steps and. 46. The method of claim 45, wherein the step of attaching a receiver to the region of the biological membrane comprises contacting the receiver with the region of the biological membrane using an adhesive. A method comprising the steps of: 47. The method of claim 45, wherein the step of determining the concentration of at least one analyte in the bodily fluid comprises electrochemical, biochemical, optical, fluorescent, absorbance, reflectance. Detecting the presence of said at least one analyte using a method selected from the group consisting of: Raman effect, magnetism, mass spectrometry, IR spectroscopy, and combinations thereof. 48. The method of claim 45, wherein the step of determining the concentration of at least one analyte in the bodily fluid is electrochemical to detect the presence of the at least one analyte. An instrument for analyzing the body fluid using a method selected from the group consisting of: biochemistry, optics, fluorescence, absorption, reflectance, Raman effect, magnetism, mass spectrometry, IR spectroscopy, and combinations thereof. Providing the method. 49. The method of claim 48, further comprising the step of mounting the instrument. 50. The method of claim 45, wherein the step of determining the concentration of at least one analyte in the body fluid comprises at least one of the body fluids.
A method comprising sequentially determining the concentrations of two analytes. 51. The method of claim 45, wherein the step of determining the concentration of at least one analyte in the body fluid comprises at least one of the body fluids.
A method comprising periodically determining the concentrations of two analytes. 52. The method of claim 45, wherein the step of determining the concentration of at least one analyte in the body fluid is based on the concentrations of multiple analytes in the body fluid. A method comprising determining a concentration of said at least one analyte in a bodily fluid. 53. The method of claim 45, further comprising displaying the concentration of the at least one analyte. 54. The method of claim 45, wherein the step of attaching a receiver to the area of the biological membrane attaches the receiver to the area of the biological membrane for at least 24 hours. A method comprising steps. 55. The method of claim 45, wherein the step of attaching a receiver to the area of the biological membrane attaches the receiver to the area of the biological membrane for at least 12 hours. A method comprising steps. 56. The method of claim 45, wherein the step of attaching a receiver to the area of the biological membrane attaches the receiver to the area of the biological membrane for at least 6 hours. A method comprising steps. 57. The method of claim 45, wherein the step of attaching a receiver to the area of the biological membrane attaches the receiver to the area of the biological membrane for at least 2 hours. A method comprising steps. 58. A device for non-invasively sampling and analyzing bodily fluids, the device being attached to a region of a biological membrane having increased permeability, passing through said region of the biological membrane and thereinto. A receiver for receiving bodily fluid from a body, the receiver comprising a membrane and a medium contained in the membrane, detecting the presence of at least one analyte in the received body fluid to determine the concentration of the analyte. A wearable instrument, as shown, comprising a processor, an electrochemical detector, a biochemical detector, a fluorescence detector, an absorbance detector, a reflectance detector, a Raman detector, and a magnetic detector. A mass spectrometric detector, an IR spectroscopic detector, and a combination thereof, the device detecting the presence of the analyte. 59. The device of claim 58, wherein the medium comprises at least one penetrant and at least one of a hydrogel layer, a fluid, and a liquid. 60. The device of claim 59, wherein the penetrant comprises at least one of sodium chloride, lactate, magnesium sulfide. 61. The apparatus of claim 59, wherein the medium is at least one.
An apparatus further comprising one reagent. 62. The apparatus of claim 58, wherein the wearable instrument comprises:
The device further comprising a display for displaying the concentration of the analyte. 63. The device of claim 58, wherein the receiver further comprises a transdermal adhesive. 64. A receptacle, attached to a region of a biological membrane having increased permeability, for receiving body fluid from the interior thereof through the region of the biological membrane, the first grid A medium layer containing at least one drug; a membrane that induces a concentration gradient barrier for the at least one drug; a counter grid; an oxidase layer; a detection layer; the first grid and the counter grid A voltage source that provides a potential difference between the first and second fluids, the bodily fluid being out of or passing through the biological membrane,
Flow through the grid of cells, the counter grid, and the oxidase layer to the detection layer. 65. The receiver of claim 64, wherein the body fluid comprises at least one of blood, interstitial fluid, analyte, and lymph. 66. The receiver of claim 64, wherein the media layer comprises at least one of a hydrogel and a liquid. 67. The receiver of claim 64, wherein the at least one drug comprises an osmotic drug. 68. The receiver of claim 67, wherein the osmotic agent comprises a negatively or positively charged nuclide. 69. The receiver of claim 68, wherein the concentration of the charged nuclide can be varied using the voltage source. 70. The receiver of claim 64, wherein the oxidase layer comprises at least one catalyst or enzyme that detects at least one analyte. 71. The receiver of claim 64, wherein the detection layer further comprises: at least one working electrode; and at least one reference electrode, the at least one working electrode and the at least one reference electrode. A receiver that enables the electrode to detect a by-product of the oxidase layer and quantify the concentration of a desired analyte to be detected.