Method of using an optical scanning device and a cartridge to collect light reflected by a reagent in the cartridge that is exposed to human breath, and kit for same

BACKGROUND OF THE INVENTION

Handheld devices for detecting the presence of a preselected substance (biomarker) in the breath are well-known. For example, U.S. Patent No. 7,285,246 (Martin) discloses a deformable housing that forms a test chamber for allowing a reagent to interact with breath. A rupturable ampoule positioned in the housing holds the reagent. After the ampoule is broken and exposed to breath, color changes in the reagent are visually detected and are compared to color codes imprinted directly on the housing.

There is an unmet need to provide an improved process and a computerized platform to allow for testing a wide variety of different biomarkers in the breath, including certain biomarkers that have not previously been tested using breath. Furthermore, while the housing in U.S. Patent No. 7,285,246 inhibits the user's risk of being cut by glass shards from the ruptured ampoule and of inhaling the reagent, further improvements in reducing these risks are desirable. It is further desirable to provide a housing for the reagent having an improved form factor. The present invention fulfills such needs.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

The following detailed description of preferred embodiments of the invention will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown.

In the drawings:

Fig. 1 is a front and left side perspective view of one preferred embodiment of the optical scanning device (breath analyzer);

Fig. 2 is a front elevational view thereof;

Fig. 3 is a rear elevational view thereof;

Fig. 4 is a left side elevational view thereof; a right side elevational view being a mirror image thereof;

Fig. 5 is a top plan view thereof; Fig. 6 is a bottom plan view thereof;

Figs. 7A, 7B and 8 show internal components of the optical scanning device;

Figs 9A-9K, taken together, show electrical components of the optical scanning device;

Figs. 10A-10D, taken together, show a listing of the electrical components illustrated in Figs. 9A-9K;

Fig. 1 1 is a front and left side perspective view of first preferred embodiment of hand-held cartridge for a breath analyzer in accordance with the present invention;

Fig. 12 is a front elevational view of the hand-held cartridge of Fig. 1 1 ;

Fig. 13 is a rear elevational view of the hand-held cartridge of Fig. 1 1 ;

Fig. 14 is a top plan view of the hand-held cartridge of Fig. 1 1 ;

Fig. 15 is a bottom plan view of the hand-held cartridge of Fig. 1 1 ;

Fig. 16 is a left side elevational view of the hand-held cartridge of Fig. 1 1 ;

Fig. 17 is a right side elevational view of the hand-held cartridge of Fig. 1 1 ;

Fig. 18 is a cross-sectional view in elevation of the hand-held cartridge of Fig. 16 taken along the line 8-8;

Fig. 19 is a left side elevational view of the lens cap of the hand-held cartridge of Fig. 1 1 ;

Fig. 20 is a top plan view of the lens cap of Fig. 19;

Fig. 21A-21 G graphic illustrations of the steps for using the hand-held cartridge of Fig. 1 1 ;

Figs 22A-22G show user interface display screens for use with an app associated with the optical scanning device; and

Fig. 23 shows a schematic diagram of the computer environment for using the app associated with the optical scanning device.

DETAILED DESCRIPTION OF THE INVENTION

Certain terminology is used herein for convenience only and is not to be taken as a limitation on the present invention.

Certain terminology is used in the following description for convenience only and i not limiting. The words "right," "left," "lower" and "upper" designate directions in the drawings to which reference is made. The words "inwardly" and "outwardly" refer to directions toward and away from, respectively, the geometric center of hand-held cartridge and designated parts thereof. The terminology includes the words noted above, derivatives thereof and words of similar import.

Preferred embodiments of the present invention are described in the context of the following products commercialized by Akers Biosciences Inc., Thorofare, New Jersey: i. BreathScan Lync™ Reader (reader), also referred to herein as a "BreathScan device" or "optical scanning device," and a related BreathScan™ mobile application (app). The reader is also referred to generically as a "breath analyzer."

ii. KetoChek™ rapid breath test for measuring the level of ketones.

iii. OxiChek™ rapid breath test for measuring the level of oxidative stress/free radicals.

However, the scope of the invention is not limited to these specific products.

The BreathScan Lync™ Reader delivers test results directly to Bluetooth ^® -enabled iOS ^® or Android ^® devices via the app. Optics in the reader measures chemical reactions via color changes when breath mixes with the reagents in the test devices. The results are displayed as a score having a number range or a quantifiable metric {e.g., equivalent mmol/liter). The results may be tracked and graphed to monitor changes over time in response to diet, exercise, nutritional supplements and other regimens.

1. Summary

One preferred embodiment is a kit for collecting light reflected from a visible, powder-like reagent exposed to human breath. The kit includes one or more cartridges and an optical scanning device (reader).

Each cartridge includes (i) a transparent bottom surface, (ii) an ampoule positioned above the transparent bottom surface that initially holds the reagent, and (iii) a breathing path that allows human breath to enter, flow through, and exit the cartridge.

The optical scanning device includes (i) a cartridge holder, (ii) a light source that faces and is in close proximity to the transparent bottom surface of the cartridge when the cartridge is placed in the slot so as to illuminate the reagent, and (iii) a color sensor {e.g., RGB color sensor) that also faces and is in close proximity to the transparent bottom surface of the cartridge when the cartridge is placed in the cartridge holder. The color sensor is configured to collect light reflected by the illuminated reagent. The reflected light is comprised of different wavelengths which are subsequently analyzed to determine whether . In operation, the optical scanning device and cartridge are used as follows:

(a) The ampoule is broken to release the reagent within the breathing path of the cartridge.

(b) A user breathes into the breathing path of the cartridge for a predefined period of time, thereby allowing the human breath to pass through the reagent that is in the breathing path of the cartridge.

(c) Upon completion of the predefined period of time, the cartridge is placed into the cartridge holder so that the transparent bottom surface faces and is in close proximity to the light source and the color sensor. Gravity causes the reagent within the breathing path to collect on and around the transparent bottom surface.

(d) The reagent is then illuminated with the light source and light reflected by the reagent is detected by the color sensor.

2. Detailed disclosure of optical scanning device

Fig. 1 is a front and left side perspective view of one preferred embodiment of the optical scanning device.

Fig. 2 is a front elevational view thereof;

Fig. 3 is a rear elevational view thereof;

Fig. 4 is a left side elevational view thereof; a right side elevational view being a mirror image thereof;

Fig. 5 is a top plan view thereof; and

Fig. 6 is a bottom plan view thereof.

Referring to Fig. 1, the optical scanning device 100 includes a housing 102, a slot 104 for receiving and holding the cartridge, and LED indicator lights 106 and 108. Figs. 2- 6 show different views of these parts. The cartridge is described in more detail below with respect to Figs. 1 1 - 18. In these figures, the cartridge is labeled as element 10 having a lens 20. The lens 20 is interchangeably referred to as the "transparent bottom surface."

Figs. 7A and 7B show internal components of the optical scanning device 100, and also show how a cartridge 10 inserts into the optical scanning device 100. In Figs. 7A and 7B, a sidewall of the optical scanning device 100 has been schematically removed to show the internal components. Referring to Fig. 7B, the cartridge 10 is placed into the optical scanning device 100 so that a transparent bottom surface (lens) of the cartridge 10 faces and is in close proximity to (e.g., adjacent to) light source 1 12 and color sensor 1 14 mounted on daughterboard 1 10. (In Figs. 7A and 7B, the light source 1 12 is not visible.) Fig. 8 and 9A-9 (taken together) show electrical components of the optical scanning device 100. In one assembly process, the electrical components include a main board 109 and the daughterboard 1 10. A portion of the main board 109 is shown physically in Figs. 7A and 7B, and is shown schematically in Figs. 9A-9J. The daughterboard 1 10 is shown physically in Fig. 8 and schematically in Fig. 9 . A portion of the daughterboard 1 10 is also visible in Figs. 7A and 7B. The daughterboard 1 10 is attached perpendicular to the main board 109. The LED indicator lights 106 and 108 are mounted off of the main board 109 and are bent 90 degrees to the main board 109. Referring to Fig. 8, the daughterboard 1 10 includes the light source 1 12 and the color sensor 1 14 mounted thereon. Unless otherwise specified, the following notes apply to the electrical components in Figs. 9A-9K:

1. All resistor values are in ohms and have values of 1% tolerance at 1/10 Watt.

2. All capacitor values are in microfarads and have values of 10% tolerance at 16V.

3. All inductor values are in microhenrys.

A sample listing of the electrical components shown in Figs. 9A-9K is provided in

Figs. 10A-10D. However, the scope of the present invention is not limited to any specific electrical components, and includes any equivalent components that can perform similar functions.

Figs. 1 OA- I OC, taken together, is a bill of materials for the main board. Fig. 10D is a bill of materials for the daughterboard 1 10. As shown in the parts listing for the daughterboard 1 10, one suitable color sensor 1 14 is an RGB digital color sensor commercially available from Hamamatsu Photonics, having part number S9706. One suitable light source 1 12 is a Surface-Mounted-Device Light-Emitting Diode (SMD LED) commercially available from LITE-ON Technology Corp, having part number LTW- 150T . Other types of suitable light sources include high luminescence COB (chip on board) and MCOB (multi-COB) LEDs, and low luminescence Dual In Line Package (DIP) LEDs.

3. Detailed disclosure of cartridge and manner of using same

Referring to the drawings in detail, where like numerals indicate like elements throughout, there is shown in Figs. 1 1 -20 a first preferred embodiment of a hand-held cartridge generally designated 10 for insertion in a breath analyzer able to detect the presence of biomarkers in a breath condensate sample exhaled into the cartridge 10. The cartridge 10 comprises a housing 12 having the general shape of a hollow, elongated tube with a generally rectangular cross section with rounded corners as shown in Figs. 1 1 , 14 and 15. Although a generally rectangular cross-sectional shape is preferable, the cross-sectional shape of the housing 12 is not limited to a rectangular shape and may have any geometrical shape such as circular or polygonal that allows insertion of the cartridge 10 in a corresponding slot in the breath analyzer. A mouthpiece 14 by which breath condensate may be exhaled into the cartridge is provided at the proximal end 12a of the housing 12. An end cap 16 having a cavity 18 therein is provided at the distal end 12b of the housing 12. A transparent lens 20 is centrally located in the distal end 16b of the end cap 16. A partition 22 extending substantially the axial length of the housing 12 creates two passageways 24, 26 within the housing 12. The first passageway 24 has a proximal end 24a in fluid communication with the mouthpiece 14 and an open distal end 24b in fluid communication with the cavity 18 in the end cap 16. The second passageway 26 has a closed proximal end 26a adjacent to or abutting the mouthpiece 14 and an open distal end 26b in fluid communication with the cavity 18 in the end cap 16. In some embodiments, the cavity 1 8 may be formed in the distal end 12b of the housing 12 and may not be in the end cap 16. An outwardly extending vent hole 28 in a proximal end portion of the housing 12 is in fluid communication with the second passageway 26. (See, Figs. 6 and 8) The fluid flow path through the cartridge 10 is shown in Fig. 8 a dotted line with arrows indicating the direction of flow.

A rupturable ampoule 30 containing a chemical indicator which reacts with biomarkers in breath condensate exhaled into the cartridge 10 is positioned in the first passageway 26 preferably in the distal end portion thereof. However, in some

embodiments, the ampoule 30 may be positioned in the central portion or proximal portion of the first passageway 26. The diameter of the ampoule 30 is sized for a tolerance fit in the first passageway 26 and therefore has a diameter only slightly less than the diameter of the first passageway 26.

The material from which ampoule 30 is formed is substantially inert to and insoluble with respect to the chemical indicator or reagent(s) contained therein. ("Reagent" and "chemical indicator" are used interchangeably herein.) The ampoule 30 may be capable of containing a solid, liquid or gaseous reagent and is preferably of thin-walled, easily rupturable construction. Suitable materials include, but are not limited to, glass, plastic and the like. The ampoule 30 may also be segmented or compartmentalized through the use of dividers, or some other means to separate plural reagents from one another. The dividers may be porous or non-porous. When plural reagents are used, each may be dedicated to the detection of a different biomarker. There may also be plural ampoules within a single housing, where each ampoule includes a different indicator reagent dedicated to detection of a biomarker.

The ampoule 30 is thin-walled and may be formed of glass that has been slightly scored at or near the center. A relatively small pressure applied to the portion of the housing 12 adjacent the ampoule 30 should be sufficient to rupture the walls of ampoule 30 without shattering the ampoule 30. The deformable character of the housing 12 in the vicinity of the ampoule 30 presupposes some elasticity so that housing 12 returns substantially to its original shape after a user squeezes it to rupture the glass ampoule 30. In a preferred embodiment, housing 12 is formed from a polymeric material such as polypropylene, polychloroprene or other inert, flexible material that can be shaped into tubing of the desired cross-sectional shape and size.

To identify the location of the ampoule 30 in the housing 12 and to facilitate rupture of the ampoule 30, opposed portions of the outer surface of the housing 12 between which the ampoule 30 is located are preferable marked by raised concave finger grips 32 which when squeezed toward each other cause the ampoule 30 to rupture.

A fin 36 extends outwardly from the surface of the distal portion of the housing 12 and is used to index the cartridge 10 in the slot in the breath analyzer.

Filters 34, made from a material such as crystal polystyrene, through which exhaled breath may pass but through which contents released from the ruptured ampoule 30 may not pass are located at the proximal end 24a of the first passageway 24 and the distal end 26b of the second passageway 26. In addition to preventing the reagents released from the ruptured ampoule 30 from leaving the housing 12, the filters 34 also reduce the variation in the velocity of breath condensate and the fluid pressure in the first passageway 24 and the cavity 18 due to the wide range of force exerted by a respiratory system in exhaling breath into the mouthpiece 14. The range of exerted force varies depending on the size, strength, age and health of a user's respiratory system. The degree to which fluid flow through the filters is attenuated also depends, in part, on the porosity of the filters. The reduction in the range of exerted forces improves test result analysis.

Referring to FIGS. 21 A-21 G, in use, the user holds the mouthpiece (or open end) 14 of the cartridge 10 upward and squeezes the finger grips (or designated pinch point) 32 between the thumb and forefinger to rupture the ampoule 30. This releases a chemical indicator or reagent in the first passageway 24 of the cartridge 10. While continuing to hold the cartridge 10 in the vertical position with the mouthpiece 14 up, the user forcefully exhales (or blows) their breath into the mouthpiece 14. The breath passes through the filter 34 in the proximal end 24a of the first passageway 24, past the ruptured ampoule 30, into the cavity 18 in the end cap 16, through the filter 34 in the distal end 26b of the second passageway 26, up the second passageway 26 and out the vent hole 28 and the breath condensate remains between the two filters 34.

After exhaling multiple consecutive breaths into the cartridge 10, the cartridge 10 is shaken vigorously up and down to mix the breath condensate with the reagent. Thereafter, the side of the cartridge is flicked several times to allow the reagent which has bonded with the biomarkers in the breath condensate to settle in the cavity 18 above the lens 20. The cartridge 10 is then inserted distal end 12b first into the breath analyzer and irradiated to determine the level of biomarkers in the cartridge through spectral analysis techniques.

The following detailed user instructions are provided for the OxiChek™ Cartridge:

Fig. 21A: Wait 15 minutes after last food or drink.

Fig. 21 B: Hold the open end of the OxiChek™ Cartridge upward and squeeze the designated "pinch point" between thumb and forefinger to break the inner glass capsule. You should be able to feel the capsule break - squeeze only once. This releases a yellow powder (the Reagent) within the Cartridge.

Fig. 21 C: Hold the cartridge vertically so the open end is facing up, and flick it with your finger several times, to ensure the yellow powder settled on the clear bottom window.

Fig. 2 I D: While continuing to hold the Cartridge in a vertical position with the open end up, blow forcefully through the Cartridge for a total of 30 seconds. This may be done in multiple consecutive breaths (e.g., two 15-second breaths, etc.) in quick succession. Do not cover the vent hole with a finger while blowing. Do not inhale through the Cartridge.

Fig. 2 I E: While keeping the open end of the Cartridge facing upward, shake vigorously up and down for 5 seconds. This helps mix the reagent powder.

Fig. 21 F: Keeping the cartridge in its original vertical position with the open end facing up, flick the side of the cartridge several times. The reagent powder should be settled evenly on the clear bottom window.

Fig. 21 G: Line up the fin on the Cartridge with the slot in the BreathScan Lync™ reader, and insert the Cartridge into the device reader with the clear bottom window down. Make sure the Cartridge is fully seated within the reader. The reader will automatically power "On" and the green light will illuminate.

Similar instructions are provided for a KetoChek™ Cartridge, except that the color of the reagent may be different.

4. App

Figs. 22A-22E show sample user interface display screens for use with the optical scanning device 100 in an iOS app environment. The user's mobile device must initially make a Bluetooth connection with a BreathScan device via conventional Bluetooth pairing. In these sample display screens, the BreathScan device is in a professional office, such as a physician's office or wellness center, and there are a plurality of clients (e.g., patients) associated with the office or center. Accordingly, the user in this scenario is a clinician of the office or center who administers or oversees taking a particular test sample from a client. In one embodiment, only the clinician can log into client accounts to upload test samples and view charts. However, in an alternative embodiment, the clients may have access to their test results and charts which they can view on a website that they can log into with their own login credentials.

Upon opening the app, the user logs into their account (not shown). The following explanations are provided for the app's user interface screens:

Fig. 22A: Client "Steve C" has been selected from the client list.

Fig. 22B: The specific breath test to be taken is selected. Here, the two available choices are OxiChek and KetoChek. In this example, OxiChek was selected.

The display screens then prompt the client through the seven steps described above with respect to Figs. 21 A-21 G.

Fig. 22C shows a template of the actual user interface display screens which highlight the current step and provide forward and backward buttons for the client to move forward or backwards through the steps. The explanatory icons are shown in Figs. 21 A- 21G and the textual instructions are also described above in the explanation of Figs. 21 A- 21 G. When the last step is completed (not shown), the client selects a button that reads "I've completed all steps as advised."

Next, the Bluetooth pairing is confirmed and data is received from the BreathScan device. Fig. 22D shows a sample test result record. The value ranges from 0 to 1 ,000 for OxiCheck, with lower being better. The values are a proprietary score. In this example, the client's latest test result shows a value of 208. For KetoChek, the values are displayed in equivalent mmol/liter from 0.0 to 1.5.

Fig. 22E shows a display screen of "What my Result means?" Different score ranges identify different levels of improvement recommended (e.g., slight, moderate, significant) and different respective recommendations. The most common known treatment for oxidative stress is to consume substances (e.g., foods and/or supplements) that have antioxidant effects on the body.

Fig. 22F shows a sample test record for KetoCheck, wherein the values are displayed in equivalent mmol/liter from 0.0 to 1.5.

Fig. 22G shows a display screen of "What my Result means?" for KetoChek.

Different score ranges indicate whether the client is or is not in ketosis, with different recommendations for each score range.

The BreathScan device may be in a user's home, and the user may be the same entity as the client. For example, an individual may choose to buy a kit for their personal use, in which case the client and clinician may be the same entity.

5. Hardware/software for app hosting

Fig. 23 shows one preferred embodiment for implementing the app hosting. A plurality of user devices 1 16 connect to a cloud-based server 1 18 via an electronic network 120 such as the internet. The user devices 1 16 may be mobile devices (e.g., smartphones, tablets) or they may be personal computers that run apps. As discussed above, the user devices 1 16 are preferably used by clinicians. However, in an alternative, embodiment, a client may have access to their test data via their own user device 1 16 and client credentials provided by the clinician.

In one preferred embodiment, the server 1 18 is hosted within a Microsoft ^® Azure ^® cloud hosting platform using .NET as the development platform and Microsoft SQL Server for the user data. The data preferably resides in HIPPA-compliant databases. A cloud server-based secure REST API web application endpoint is used for the client mobile application. 6. Other considerations

The chemical composition (constituents) of the reagents will vary depending upon the biomarker being tested. One suitable biomarker for measuring the level of ketones using etoChek is disclosed in U.S. Patent No. 8,871 ,521 (Akers, Jr.), which is incorporated by reference herein.

In the preferred embodiment, reagents are used that undergo a chemical change when exposed to the biomarkers of interest, and the chemical change causes the reagents to change color, thereby allowing a digital color sensor to detect the color by measuring light reflected off of the chemically changed reagent. This spectral signature can then be used to detect the presence and amounts of the biomarker in the breath, which, in turn, may be correlated to blood levels using known relationships. If the biomarker of interest is not present in the breath, the reagent will not experience the chemical change or color change, and the spectral signature of the reflected light will be unchanged from a test sample of a reagent that has never been exposed to breath.

The algorithms used in the app to convert the data from the color sensor 1 14 into a number range or a quantifiable metric will depend upon the specific reagent being used and the calibrations that must be performed for the particular biomarker. The algorithms will specify the weightings given to the different wavelengths detected by the color sensor 1 14. The preferred embodiment uses an RGB color sensor. However, other types of color sensors may be used, such as a CMYK color sensor. The algorithms for wavelength analysis must be adjusted accordingly with a different type of color sensor.

The digital color sensor 1 14 significantly improves sensitivity and accuracy in detecting color changes in the reagent after exposure to breath compared to using printed color codes, such as disclosed in U.S. Patent No. 7,285,246. The digital color sensor 1 14 is thus not simply a substitute for printed color codes, but provides functionality that is not available when using printed color codes.

As described above, the cartridge 10 has a transparent bottom surface defined by a transparent lens 20 that is centrally located in the distal end 16b of the cartridge's end cap 16. The transparent bottom surface is preferably flat so as to minimize distortions caused by reflections of light from the light source 1 12 as it impinges on, and is reflected by, the reagent that settles on the inside surface of the lens 20. If light was directed towards a sidewall of the cartridge 10 so as to reflect off of the reagent, there would be increased distortion. A similar disadvantage would occur using the housing disclosed in U.S. Patent No. 7,285,246. As described above, one suitable use for the present invention is to measure the level of oxidative stress/free radicals in a person. In chemistry, a free radical is an atom, molecule, or ion that has unpaired valence electrons. These unpaired electrons make free radicals highly chemically reactive towards other substances, or even towards themselves. Low and moderate concentrations of free radicals are not associated with any health risks, and are considered a normal part of human biology. However, high concentrations of free radicals leads to oxidative stress which has been shown to have harmful effects on the human body. Oxidative stress can damage cell structures and DNA, thereby causing a large number diseases.

Conventional tests for measuring the level of oxidative stress/free radicals include measuring TBARS (thiobarbituric acid reactive substance) levels and performing an ELISA (enzyme linked immunosorbent) assay using absorbent spectrophotometry. These conventional methods require taking blood or urine samples and also require expensive test equipment. The kit described herein may be used to measure the level of oxidative stress/free radicals in a person using a simple breath test. Since free radicals are unstable in breath, the kit preferably measures one or more byproducts of free radicals.

The scope of the present invention also includes the use of a reactant (i.e., a substance consumed in the course of a chemical reaction) instead of a reagent provided that the reactant changes in a way that is detectable by a color sensor.

It will be appreciated by those skilled in the art that changes could be made to the embodiments described above without departing from the broad inventive concept thereof. It is understood, therefore, that this invention is not limited to the particular embodiments disclosed, but it is intended to cover modifications within the spirit and scope of the present invention.

What is claimed is: