Claims:WE CLAIM:
1. An apparatus 100 for detection of trehalose characterised in enabling quantitative detection of trehalose by using an optic-based sensor, the apparatus comprising:
one or more body parts of optic-based sensor 105, 106, wherein one of the body part of the optic-based sensor 106 comprises a light source 107, and a light detector 108, wherein the light source 107 comprises a laser diode for emitting laser light;
at least two optical fibres 111, 112;
a measurement cube 115;
a reflective mirror 114;
a sample container 117 comprising a liquid solution 116 containing trehalose, wherein the liquid solution 116 is used for testing;
a battery;
a signal processing circuit 102; and
a display screen 101;
wherein, when the battery is turned on, the light source 107 turns on, emitting a laser light such that the laser light passes through one of the optical fibre 112 to the measurement cube 115, wherein one side of the measurement cube comprises an aperture, wherein the reflective mirror 114 is placed at the bottom of the measurement cube 115, at a predefined distance from the end of the optical fibres 111, 112, wherein the reflective mirror 114 is configured to reflect the laser light passed through the liquid solution 116, wherein the liquid solution 116 is inserted in the aperture of the measurement cube 115;
wherein the reflected laser light is collected by another optical fibre 111 and guided towards the light detector 108 for detecting the reflected laser light with respect to viscosity of the liquid solution 116, further, wherein the detected laser light is in a form of a signal which is analysed by the signal processing circuit 102 and an analysed result is recorded and displayed on the display screen 101; and
wherein higher viscosity of the liquid solution 116 is indicative of a higher quantity of trehalose in the liquid solution 116, thereby leading to higher scattering of the laser light resulting in lesser detection of the laser light followed by a lower output.

2. The apparatus 100 as claimed in claim 1, wherein the optical fibres 111, 112 comprise one or more protective covers 110, 119, wherein the protective covers are of plastic pipes configured to keep the optical fibres 111, 112 parallel to each other and to prevent bending of said optical fibres 111, 112.

3. The apparatus 100 as claimed in claim 2, wherein the optical fibres are plastic with a core diameter of 488 µm, a length of 9cm each and a numerical aperture of 0.47 µm.

4. The apparatus 100 as claimed in claim 1, wherein the measurement cube 115 is made of plastic, wherein out of six sides of the measurement cube 115, five sides are covered and are leakage-proof, wherein a top side of the measurement cube 115 is made of transparent glass plate and one side is kept empty with an aperture for inserting the liquid solution 116 for testing.

5. The apparatus 100 as claimed in claim 1, wherein one or more body parts of optic-based sensor 105, 106 are detachable which can be separated and replaced when any component is damaged.

6. The apparatus 100 as claimed in claim 1, wherein the signal processing circuit 102 comprises and amplifier and like components.

7. The apparatus 100 as claimed in claim 1, wherein the apparatus 100 comprises a fully transparent glass plate 113 for protection of the optical fibres 111, 112.

8. A method of use 200 of an apparatus 100 for detection of trehalose, characterised in enabling quantitative detection of trehalose by using an optic-based sensor, the method comprising:
inserting a liquid solution 116 containing trehalose in an aperture comprised on one of the side of a measurement cube;
emitting via a light source 107, a laser light such that the laser light passes through one of the optical fibre 112 to the measurement cube 115;
reflecting via a reflective mirror 114 the laser light passed through the liquid solution 116 wherein the liquid solution 116 is inserted in the aperture of the measurement cube 115, wherein the reflective mirror 114 is placed at the bottom of the measurement cube 115, at a predefined distance from the ends of at least two optical fibres 111, 112;
collecting via an another optical fibre 111, the reflected laser light and guiding said reflected laser light towards a light detector 108;
detecting via the light detector 108, reflected laser light with respect to viscosity of the liquid solution 116, wherein the detected laser light is in a form of a signal;
analysing via a signal processing circuit 102 said signal; and
displaying via a display screen 101, an analysed result;
wherein a higher viscosity of the liquid solution 116 is indicative of a higher quantity of trehalose in the liquid solution 116, thereby leading to higher scattering of the laser light resulting in lesser detection of the laser light followed by a lower output.

Dated this 30th day of August, 2019

Priyank Gupta
Agent for the Applicant
IN/PA- 1454

, Description:FORM 2

THE PATENTS ACT, 1970
(39 of 1970)
&
THE PATENT RULES, 2003
COMPLETE SPECIFICATION

(See Section 10 and Rule 13)

Title of invention:
AN APPARATUS FOR DETECTION OF TREHALOSE AND A METHOD OF USE THEREOF

APPLICANT
Savitribai Phule Pune University,
An Indian Entity, having address as
Ganeshkhind, Pune 411007, Maharashtra India.

The following specification describes the invention and the manner in which it is to be performed.

CROSS-REFERENCE TO RELATED APPLICATIONS AND PRIORITY
The present application does not claim priority from any other patent application.
TECHNICAL FIELD
The present subject matter described herein, in general, relates to an apparatus for detection of trehalose and a method of use thereof.
BACKGROUND
Trehalose has wide applications in biomedical field and in food and cosmetics industry. Trehalose [C12H22O11; CAS 6138-23-4 (dihydrate); CAS 99-20-7 (anhydrous)] is a disaccharide sugar comprising two molecules of glucose bonded by an a,a-1,1 glycosidic linkage. Trehalose, a non-reducing disaccharide, is found in many organisms across the plant and animal kingdom including bacteria, algae, yeast, fungi, plants, nematodes and insects but is absent in vertebrates. Trehalose serves diverse functions in these organisms such as being a structural component of cell wall of bacterial, source of carbon and growth regulator in plants and fungi and as a source of energy and major blood sugar in insects and as a compatible solute to overcome environmental stresses. The most appreciated and well established role of trehalose in the field of life sciences is its ability to act as a protectant of organismal and cellular functions under environmental stress conditions such as drought and dehydration, global climate warming, hypoxia and chill or frost stress.

Few notable properties of biophysical and biochemical properties of trehalose include its high hydrophilicity, chemical stability, non-hygroscopic glass formation and the absence of internal hydrogen bond formation which has encouraged for efforts in its applications in DNA engineering, cryoprotection and food conservation. Among the other wide range of applications of trehalose are commercially available therapeutic products, biomedicine and healthcare, food and nutrition industry and for novel applications capitalizing on its promising potential as a bio-indicator of environmental stresses, especially in the current scenario of global climate change.
At present, there are only few available methods for trehalose detection and these methods are based on measurement of trehalose solution viscosity by dynamic light scattering and FRET sensor-based quantification of intracellular trehalose in mammalian cells. Although these approaches may be efficient, they are not free from limitations. The existing trehalose detecting methods are expensive and are labour intensive.

Given the growing use and applications of trehalose as a multifaceted biomolecule, there is a long standing need of a simple, cost effective and portable optics-based detection of trehalose.

SUMMARY

This summary is provided to introduce the concepts related to an apparatus for detection of trehalose and a method of use thereof and the concepts are further described in the detail description. This summary is not intended to identify essential features of the claimed subject matter nor it is intended to use in determining or limiting the scope of claimed subject matter.

In one implementation, the present subject matter describes an apparatus for detection of trehalose, in accordance to the present subject matter. In one embodiment, the apparatus is characterised in enabling quantitative detection of trehalose by using an optic-based sensor. The apparatus may comprise one or more body parts of optic-based sensor, wherein one of the body part of the optic-based sensor comprises a light source, and a light detector, wherein the light source comprises a laser diode for emitting laser light. The apparatus may comprise at least two optical fibers, a measurement cube, a reflective mirror, a sample container comprising a liquid solution containing trehalose, wherein the liquid solution is used for testing. The apparatus, may further comprise a battery, a signal processing circuit and a display screen. When the battery is turned on, the light source turns on emitting a laser light such that the laser light passes through one of the optical fibre to the measurement cube. One side of the measurement cube may comprise an aperture for inserting the liquid solution for testing. The reflective mirror may be placed at the bottom of the measurement cube, at a predefined distance from the end of the optical fibres, wherein the reflective mirror may be configured to reflect the laser light passed through the liquid solution, wherein the liquid solution is inserted in the aperture of the measurement cube. In one embodiment, the reflected laser light is collected by another optical fibre and guided towards the light detector for detecting the reflected laser light with respect to viscosity of the liquid solution. The detected laser light may be in a form of a signal which is analysed by the signal processing circuit and an analysed result is recorded and displayed on the display screen. A higher viscosity of the liquid solution is indicative of a higher quantity of trehalose in the liquid solution, thereby leading to higher scattering of the laser light resulting in lesser detection of the laser light followed by a lower output.
In another implementation, the present subject matter describes a method of use of an apparatus for detection of trehalose, in accordance to the present subject matter. The method may be characterised in enabling quantitative detection of trehalose by using an optic-based sensor. The method may comprise inserting a liquid solution containing trehalose in an aperture comprised on one of the side of a measurement cube. The method may further comprise emitting via a light source, a laser light such that the laser light passes through one of the optical fibre to the measurement cube. The method may further comprise reflecting via a reflective mirror the laser light passed through the liquid solution wherein the liquid solution is inserted in the aperture of the measurement cube, wherein the reflective mirror is placed at the bottom of the measurement cube, at a predefined distance from the ends of at least two optical fibres. The method may further comprise collecting via another optical fibre, the reflected laser light and guiding said reflected laser light towards a light detector. The method may further comprise detecting via the light detector, reflected laser light with respect to viscosity of the liquid solution, wherein the detected laser light is in a form of a signal. The method may furthermore comprise analysing via a signal processing circuit said signal. The method may comprise displaying via a display screen, an analysed result.
BRIEF DESCRIPTION OF DRAWINGS

The detailed description is described with reference to the accompanying figures. In the Figures, the left-most digit(s) of a reference number identifies the Figure in which the reference number first appears. The same numbers are used throughout the drawings to refer like features and components.

Figure 1 illustrates, an apparatus 100 for detection of trehalose, in accordance with an embodiment of a present subject matter.

Figure 2 illustrates, a method of use 200 of an apparatus for detection of trehalose, in accordance with an embodiment of a present subject matter.

DETAILED DESCRIPTION

Reference throughout the specification to “various embodiments,” “some embodiments,” “one embodiment,” or “an embodiment” means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, appearances of the phrases “in various embodiments,” “in some embodiments,” “in one embodiment,” or “in an embodiment” in places throughout the specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner in one or more embodiments.

The present disclosure comprising a trehalose sensor is based on the concentration-dependent clustering tendency of trehalose aqueous solutions resulting in changes in viscosity. The varying concentrations of trehalose solutions (%) tested indicated variations in output using signal processing system. This change in the output with respect to concentrations of trehalose solution is due to viscosity change of the solution. With rising concentrations of trehalose, there was increase in viscosity of the trehalose solution which in turn, led to an increase in light scattering resulting in reduction in the amount of light received at the detector. Thus, the working mechanism of the apparatus for detecting trehalose is based on the principles of viscosity and scattering of light. Moreover, the apparatus 100 may be configured for quantitative detection of trehalose in a liquid solution. The detection mechanism is primarily based on viscosity of trehalose solution and scattering of light.

Referring, to figure 1, an apparatus 100 for detection of trehalose, is illustrated in accordance with an embodiment of a present subject matter. In an embodiment, the apparatus 100 for detection of trehalose may be characterised in enabling quantitative detection of trehalose by using an optic-based sensor. The optic-based sensor may comprise one or more body parts 105, 106. The apparatus 100 may comprise an optic-based sensor comprising a light source 107, a light detector 108 and at least two optical fibres 112, 111. The light source 107 may comprise a laser diode for emitting laser light. Further, the apparatus 100 may comprise a measurement cube 115, a reflective mirror 114, a sample container 117 comprising a liquid solution 116 containing trehalose, wherein the liquid solution is used for testing. The apparatus 100 may further comprise a battery, a signal processing circuit 102 and a display screen. In one embodiment, when the battery is turned on, the light source 107 turns on emitting a laser light such that the laser light passes through one of the optical fibre 112 to the measurement cube 115. One side of the measurement cube comprises an aperture for inserting the liquid solution 116 for testing. In one embodiment, the reflective mirror 114 may be placed at the bottom of the measurement cube 115, at a predefined distance from the end of the optical fibres 111, 112. The reflective mirror 114 may be configured to reflect the laser light passed through the liquid solution 116 wherein the liquid solution 116 is inserted in the aperture of the measurement cube 115. Further, the reflected laser light may then be collected by another optical fibre 111 and guided towards the light detector 108 for detecting the reflected laser light with respect to viscosity of the liquid solution 116. In one embodiment, the changes in the viscosity may depend on varying concentrations of trehalose in the liquid solution 116. In one embodiment a sample container 117 may comprise a liquid solution 116 containing trehalose. Further, the detected laser light may be in a form of a signal which may be analysed by the signal processing circuit 102 and an analysed result is recorded and displayed on the display screen 101. In one embodiment, a higher viscosity of the liquid solution 116 is indicative of a higher quantity of trehalose in the liquid solution 116, thereby leading to higher scattering of the laser light resulting in lesser detection of the laser light followed by a lower output. In one embodiment, a change in temperature of the liquid solution 116 may create disturbance in reading of the optic-based sensor, thus the viscosity of the liquid solution 117 containing trehalose may increase with temperature drop.
In one embodiment, one or more body parts of optic-based sensor 105, 106 may be made of plastic or polymer. The said parts may be detachable two parts. The parts may be separated to replace any damaged part of the sensor like light source, detector, fibers. The body of the sensor has provision to keep light source and detector inside it. In one embodiment, the optical fibres 112, 111 comprise one or more protective covers 110, 119, wherein the protective covers are of plastic pipes configured to keep the optical fibres 111, 112 parallel to each other and to prevent bending of said optical fibres 111, 112. The optical fibres 111, 112 may be configured to carry light from the light source 107 to the measurement cube 115 and further from the measurement cube 115 to the light detector 112. In one embodiment, the optical fibres may be made of plastic with a core diameter of 488 µm, a length of 9cm each and a numerical aperture of 0.47 µm. in one embodiment, both the transmitting and receiving fibres may be kept inside the plastic pipe and placed parallel to each other at 0º angle between them. In one embodiment, the sample container 117 is used to hold the liquid solution 116. The sample container 117 are chosen such way that, liquid solution in sample container cannot go beyond the protecting glass of the optic-based sensor. In one embodiment, the signal processing circuit 102 may comprise necessary circuit components like amplifier and display or measuring equipment used to display the measured readings. In one embodiment, the apparatus 100 comprises a fully transparent glass plate 113 for protection of the optical fibres 111, 112.
The apparatus 100 for detection of trehalose is cost-effective, portable, easy to operate, sensitive and simple battery operated device. The apparatus 100 may be configured to measure trace levels of trehalose as a part of its applications process.
The apparatus 100 may be used in nutritional health intervention, biomedical applications, environmental and climate change studies, food industry, cosmetics industry, neutraceuticals industry and such like.

Referring to figure 2, illustrates a method of use 200 of an apparatus for detection of trehalose, in accordance with an embodiment of a present subject matter. The method 200 is characterised in enabling quantitative detection of trehalose by using an optic-based sensor.
At step 201, a liquid solution 116 may be inserted. In one embodiment, the method may comprise inserting a liquid solution 116 containing trehalose in an aperture comprised on one of the side of a measurement cube.
At step 202, a laser light may be emitted. In one embodiment, the method may comprise emitting via a light source 107, a laser light such that the laser light passes through one of the optical fibre 112 to the measurement cube 115.
At step 203, the laser light may be reflected. In one embodiment, the method may comprise reflecting via a reflective mirror 114 the laser light passed through the liquid solution 116 wherein the liquid solution 116 is inserted in the aperture of the measurement cube 115. In one embodiment, the reflective mirror 114 may be placed at the bottom of the measurement cube 115, at a predefined distance from the ends of at least two optical fibres 111, 112.
At step 204, the reflected laser light may be collected. In one embodiment, the method may comprise collecting via another optical fibre 111, the reflected laser light and guiding said reflected laser light towards a light detector 108.
At step 205, the reflected laser light may be detected. In one embodiment, the method may comprise detecting via the light detector 108, reflected laser light with respect to viscosity of the liquid solution 116, wherein the detected laser light is in a form of a signal.
At step 206, the method may comprise analysing via a signal processing circuit 102 said signal.
At step 207, the method may comprise displaying via a display screen 101, an analysed result.
In one embodiment, the measurement cube 115 may be made of plastic, wherein out of six sides of the measurement cube 115, five sides are covered and are leakage-proof, wherein a top side of the measurement cube 115 is made of transparent glass plate and one side is kept empty with an aperture for inserting the liquid solution 116 for testing.
In one embodiment, a higher viscosity of the liquid solution 116 is indicative of a higher quantity of trehalose in the liquid solution 116, thereby leading to higher scattering of the laser light resulting in lesser detection of the laser light followed by a lower output.

The embodiments, examples and alternatives of the preceding paragraphs or the description and drawings, including any of their various aspects or respective individual features, may be taken independently or in any combination. Features described in connection with one embodiment are applicable to all embodiments, unless such features are incompatible.

Although implementations for the computer implemented an apparatus for detection of trehalose and a method of use thereof have been described in language specific to structural features and/or methods, it is to be understood that the appended claims are not necessarily limited to the specific features or methods described. Rather, the specific features and methods are disclosed as examples of implementations for an apparatus for detection of trehalose and a method of use thereof.