阅读说明：本技术 用于血糖检测纳米标志物、基于其的动态近红外光谱无损血糖仪及其制备方法 (Nano marker for blood glucose detection, dynamic near infrared spectrum nondestructive blood glucose meter based on nano marker and preparation method of dynamic near infrared spectrum nondestructive ) 是由 杨立峰 陈楚林 张恒静 田明睿 陈鹏 丁植 刘自强 张希仁 彭仁军 张靖 于 2019-11-21 设计创作，主要内容包括：本发明公开了用于血糖检测纳米标志物、基于其的动态近红外光谱无损血糖仪及其制备方法,包括激光器、激光聚焦单元、半透半反透镜,激光调制器、间质液吸附装置、微处理器以及显示模块,还包括设置有纳米标志物的纳米标志物盒、微型光谱仪一、缓冲反应皿一、分光系统、微型光谱仪二、缓冲反应皿二及差分电路模块,所述纳米标志物盒通过缓冲反应皿一与间质液吸附装置相连接,所述缓冲反应皿二与间质液吸附装置相连接,所述微型光谱仪一和微型光谱仪二与差分电路模块连接。使用纳米标志物具有荧光或磷光特性,增强检测效率且使用纳米标志物使得血糖检测更加直接,避免其他组织成分的影响。(The invention discloses a dynamic near infrared spectrum nondestructive blood glucose meter for blood glucose detection, which comprises a laser, a laser focusing unit, a semi-transparent semi-reflective lens, a laser modulator, an interstitial fluid adsorption device, a microprocessor, a display module, a nano marker box provided with a nano marker, a first micro spectrometer, a first buffer reaction vessel, a light splitting system, a second micro spectrometer, a second buffer reaction vessel and a differential circuit module, wherein the nano marker box is connected with the interstitial fluid adsorption device through the first buffer reaction vessel, the second buffer reaction vessel is connected with the interstitial fluid adsorption device, and the first micro spectrometer and the second micro spectrometer are connected with the differential circuit module. The nano marker has fluorescent or phosphorescent characteristics, the detection efficiency is enhanced, the blood sugar detection is more direct by using the nano marker, and the influence of other tissue components is avoided.)

1. The nanometer marker for blood sugar detection is characterized by comprising a resonance core layer and a peripheral coating layer, wherein the excitation excimer resonance core layer at least comprises a first gold nanosphere, a second gold nanosphere and substrate nanoparticles, and the peripheral coating layer comprises polypeptide molecules, sulfhydryl biological small molecules and polystyrene sulfonate joints.

2. The nano-marker for blood glucose detection according to claim 1, wherein the substrate nano-particle is a mesoporous silica nano-substrate.

3. The nano-marker for blood glucose detection according to claim 1, wherein the gold nanosphere I and the gold nanosphere II are conjugated on the substrate nanoparticle to form an excimer resonance core layer, and the distance between the gold nanosphere I and the gold nanosphere II is satisfied

4. The dynamic lossless glucometer based on the near infrared spectrum of the nano marker is characterized by comprising a laser, a laser focusing unit, a semi-transparent semi-reflective lens, a laser modulator, a interstitial fluid adsorption device, a microprocessor and a display module, wherein the semi-transparent semi-reflective lens is arranged between the laser and the laser focusing unit, the laser modulator is used for regulating and controlling the laser, the interstitial fluid adsorption device is used for regulating and controlling the laser, the dynamic lossless glucometer is characterized by further comprising a nano marker box, a first micro spectrometer, a first buffer reaction vessel, a light splitting system, a second micro spectrometer, a second buffer reaction vessel and a differential circuit module, the nano marker box is connected with the interstitial fluid adsorption device through the first buffer reaction vessel, the second buffer reaction vessel is connected with the interstitial fluid adsorption device, and the first micro spectrometer and the second micro spectrometer are connected with the differential circuit module.

5. The dynamic nondestructive blood glucose meter of near infrared spectrum based on nano-markers as claimed in claim 4, characterized in that the spectroscopic system comprises a total reflection mirror disposed between the semi-transparent semi-reflective lens and the second buffer reaction vessel.

6. A preparation method of a nano marker is characterized by comprising the following steps:

the method comprises the following steps: preparation of an excimer resonance core layer: fully dissolving the prepared gold nanosphere solution I and the prepared gold nanosphere solution II for 3-5 hours under ultrasonic stirring, adding the solution into the prepared substrate nanoparticle solution, fully stirring to form an excitation excimer resonance core layer, adding sulfhydryl biological micromolecules, and removing the precipitate formed by redundant gold nanospheres;

step two: and (3) adding the polypeptide molecule solution and the sodium polystyrene sulfonate solution into the solution obtained in the step one in sequence, and fully stirring to form a stable chemical bond, thereby obtaining the nano marker.

7. The method for preparing the nano marker according to claim 6, wherein the preparation of the gold nanosphere-one solution or the gold nanosphere-two solution comprises the following steps:

a) cetyl trimethyl ammonium bromide is taken as a surfactant, n-butyl alcohol is taken as a cosurfactant, n-octane is taken as an oil phase, and a chloroauric acid aqueous solution is taken as a water phase, namely 30 mu L of 0.02mol/L cetyl trimethyl ammonium bromide, 30 mu L of 0.1mol/L n-butyl alcohol, 120 mu L of 0.6mol/L n-octane and 100 mu L of 1% chloroauric acid aqueous solution are mixed together and stirred for 30 minutes to be uniformly mixed, so that uniform and transparent microemulsion is obtained;

b) adding 100 mu L of 1% sodium citrate into 10mL of ultrapure water to form a sodium citrate aqueous solution, dropwise adding the sodium citrate aqueous solution into the microemulsion under stirring, and then adding 0.05mL0.4mol/L of 1, 6-dimercaptohexane;

c) continuously stirring and reacting for 10-14 h to obtain a suspension of the gold nanosphere primary solution, standing for 23-25 h to completely precipitate suspended particles in the suspension, taking out the upper-layer solution part, and adding ethanol into the residual precipitate part to obtain a stable gold nanosphere primary solution; or continuously stirring and reacting for 20-24 h to obtain a suspension of the gold nanosphere two solution, standing for 23-25 h to completely precipitate suspended particles in the suspension, taking out the upper layer solution part, and adding ethanol into the rest precipitate part to obtain the stable gold nanosphere two solution.

8. The method for preparing the nano marker according to claim 6, wherein the base nanoparticle is prepared by a soft film plate method, and the diameter of the base nanoparticle is in a range of 58-200 nm.

9. The method as claimed in claim 6, wherein the radius r of the first gold nanosphere is larger than the radius of the second gold nanosphere₁Is 8nm to 12 nm.

10. The method for preparing the nano-marker according to claim 6, wherein the radius r of the second gold nanosphere₂Is 17nm to 26 nm.

Technical Field

The invention relates to a measuring instrument for medical detection of blood sugar level of a diabetic patient, in particular to a dynamic near infrared spectrum nondestructive blood sugar meter based on a nano marker.

Background

Diabetes is a metabolic disease characterized by chronic hyperglycemia due to various causes, and is accompanied by a series of metabolic disorders such as sugar, protein, fat, water, and electrolytes in the body, etc. due to insulin secretion or insulin action deficiency. The blood glucose monitoring can be implemented to better control the blood glucose change of the diabetic, and has important guiding significance on the rules of life, activities, sports, diet and reasonable medication, and the blood glucose meter is an electronic instrument for measuring the blood glucose level.

The dynamic blood sugar detector which is non-invasive, continuously measurable and portable has become the development trend of the current blood sugar meter. The diabetes map published by the international diabetes association shows that about 3.82 hundred million adults suffer from diabetes in 2013 globally, China becomes the most countries of diabetes patients worldwide, the total diabetes number is nearly one hundred million, and the number of people in the early stage of diabetes is about 1.5 hundred million. In order to control the development of a diabetic patient, it is necessary to monitor their blood glucose levels continuously in order to maintain their blood glucose levels within a normal range.

Many patents have been published on non-invasive blood glucose meters, such as "a wearable noninvasive dynamic blood glucose monitor based on photoacoustic spectrum characteristics" (CN105559794A), which uses a photoacoustic spectrum multi-array to enhance the signals of each sensor in an infrared sensor array to measure the blood glucose level; a composite photoacoustic nondestructive dynamic blood sugar detector (CN104706363A) improves the detection of photoacoustic signals by modulating signals and a piezoelectric transducer array; a noninvasive self-test blood glucose meter (CN1271562A) measures blood glucose level by using an infrared light emitting tube as an infrared light source (wavelength: 1000 to 2900nm) and adopting a transmission type.

The photoacoustic spectrums adopted by the method for detecting the blood glucose signals are seriously interfered and have high requirements on the environment, and the problem of low photoacoustic conversion efficiency cannot be thoroughly solved. The nondestructive blood glucose meter can not be successfully researched and developed in time, and the main reasons are as follows: (1) the bottlenecks of stability and accuracy are difficult to break through; (2) the photoacoustic signal is weak, and the detection difficulty is high.

Disclosure of Invention

The invention aims to: aiming at the problems that the blood glucose signals detected by a photoacoustic spectrum are seriously interfered and have high requirements on the environment, and the problem of low photoacoustic conversion efficiency cannot be thoroughly solved, the invention provides the nano marker for detecting the blood glucose.

It is another object of the present invention to provide a dynamic non-destructive glucometer based on near infrared spectrum of nano-markers.

It is still another object of the present invention to provide a method for preparing a nano-tag.

The technical scheme adopted by the invention is as follows:

the nanometer marker for blood sugar detection comprises a resonance core layer and a peripheral coating layer, wherein the excitation excimer resonance core layer at least comprises a gold nanosphere I, a gold nanosphere II and substrate nanoparticles, and the peripheral coating layer comprises polypeptide molecules, sulfhydryl biological small molecules and polystyrene sulfonate joints. The nano-marker of the present invention may be linked to the plasma glucose protein of interstitial fluid via a covalent bond, thereby forming plasma glucose protein carrying the nano-marker.

Preferably, the substrate nanoparticles are mesoporous silica nanomatrix.

Preferably, the first gold nanosphere and the second gold nanosphere are conjugated on the substrate nanoparticle to form an excimer resonance core layer, and the distance between the first gold nanosphere and the second gold nanosphere satisfies the requirement

The near infrared spectrum dynamic lossless glucometer based on the nano marker comprises a laser, a laser focusing unit, a semi-transparent semi-reflective lens, a laser modulator, a interstitial fluid adsorption device, a microprocessor and a display module, wherein the semi-transparent semi-reflective lens is arranged between the laser and the laser focusing unit and is used for regulating and controlling the laser, the nano marker box is provided with the nano marker according to any one of claims 1 to 4, a micro spectrometer I, a buffer reaction vessel I, a light splitting system, a micro spectrometer II, a buffer reaction vessel II and a differential circuit module, the nano marker box is connected with the interstitial fluid adsorption device through the buffer reaction vessel I, the buffer reaction vessel II is connected with the interstitial fluid adsorption device, and the micro spectrometer I and the micro spectrometer II are connected with the differential circuit module.

The nano marker box is characterized in that a box for containing a nano marker is connected with an interstitial fluid adsorption device through a first buffer reaction vessel, the interstitial fluid adsorption device adsorbs interstitial fluid with blood sugar from skin, the interstitial fluid enters the first buffer reaction vessel, and the nano marker in the first buffer reaction vessel is linked to blood glucose protein of the interstitial fluid through a covalent bond to form blood glucose protein carrying the nano marker; the laser is connected with the laser modulator, the laser is controlled by the laser modulator, corresponding wavelength and pulse width signals can be output, the laser can work under different wavelengths and pulse widths, after light energy is absorbed by the first buffer reaction vessel carrying the nano marker blood glucose protein, the signal carrying the nano marker blood glucose protein after being enhanced is detected by the first micro spectrometer, on the other hand, light emitted by the laser passes through the second buffer reaction vessel after the semi-transparent semi-reflective lens and the fully reflective lens, the liquid spectrum without the nano marker can be detected by the second micro spectrometer, and finally, the microprocessor calculates the information containing the blood glucose protein content from the differential circuit module.

Preferably, the light splitting system comprises a total reflection mirror, and the total reflection mirror is arranged between the semi-transparent semi-reflective lens and the second buffer reaction vessel.

Further, the laser may be a multi-wavelength tunable laser, or may be a laser array formed by a plurality of lasers with different wavelengths.

Further, the interstitial fluid adsorption device may be an electrical or guided device that allows for the adsorption of dermal interstitial fluid to the nano-marker cassette junction.

A preparation method of a nano marker comprises the following steps:

the method comprises the following steps: preparation of an excimer resonance core layer: fully dissolving the prepared gold nanosphere solution I and the prepared gold nanosphere solution II for 3-5 hours under ultrasonic stirring, adding the solution into the prepared substrate nanoparticle solution, fully stirring to form an excitation excimer resonance core layer, adding sulfhydryl biological micromolecules, and removing the precipitate formed by redundant gold nanospheres;

step two: and (3) adding the polypeptide molecule solution and the sodium polystyrene sulfonate solution into the solution obtained in the step one in sequence, and fully stirring to form a stable chemical bond, thereby obtaining the nano marker.

Preferably, the substrate nanoparticles are prepared by a soft membrane plate method, and the diameter range is 58-200 nm

Preferably, the preparation of the gold nanosphere-one solution or the gold nanosphere-two solution comprises the following steps:

the preparation of the gold nanosphere solution I or the gold nanosphere solution II comprises the following steps:

a) cetyl trimethyl ammonium bromide is taken as a surfactant, n-butyl alcohol is taken as a cosurfactant, n-octane is taken as an oil phase, and a chloroauric acid aqueous solution is taken as a water phase, namely 30 mu L of 0.02mol/L cetyl trimethyl ammonium bromide, 30 mu L of 0.1mol/L n-butyl alcohol, 120 mu L of 0.6mol/L n-octane and 100 mu L of 1% chloroauric acid aqueous solution are mixed together and stirred for 30 minutes to be uniformly mixed, so that uniform and transparent microemulsion is obtained;

b) adding 100 mu L of 1% sodium citrate into 10mL of ultrapure water to form a sodium citrate aqueous solution, dropwise adding the sodium citrate aqueous solution into the microemulsion under stirring, and then adding 0.05mL of 0.4 mol/L1, 6-dimercaptohexane;

c) continuously stirring and reacting for 10-14 h to obtain a suspension of the gold nanosphere primary solution, standing for 23-25 h to completely precipitate suspended particles in the suspension, taking out the upper-layer solution part, and adding ethanol into the residual precipitate part to obtain a stable gold nanosphere primary solution; or continuously stirring and reacting for 20-24 h to obtain a suspension of the gold nanosphere two solution, standing for 23-25 h to completely precipitate suspended particles in the suspension, taking out the upper layer solution part, and adding ethanol into the rest precipitate part to obtain the stable gold nanosphere two solution.

Preferably, the substrate nanoparticles are prepared by a soft membrane plate method, and the diameter range is 58-200 nm

Preferably, the radius r of the gold nanosphere I₁Is 8nm to 12 nm.

Preferably, the radius r of the gold nanosphere II₂Is 17nm to 26 nm.

Compared with the prior art, the invention has the beneficial effects that:

(1) the nano-markers have fluorescent or phosphorescent characteristics, and the detection efficiency is enhanced.

(2) The use of the nano-markers enables blood glucose detection to be more direct, and avoids the influence of other tissue components.

(3) And a reference light path is adopted, and data is detected in a differential mode without being influenced by the surrounding environment.

Drawings

FIG. 1 is a schematic structural view of the present invention;

FIG. 2 gold nanospheres-TEM scan results;

FIG. 3 two TEM scan of gold nanospheres.

Labeled as: the system comprises an 11-laser, a 12-laser focusing unit, a 13-nano marker box, a 14-micro spectrometer I, a 15-buffer reaction vessel I, a 16-interstitial fluid adsorption device, a 17-laser modulator, an 18-microprocessor, a 19-display module, a 110-differential circuit module, a 111-semi-transparent semi-reflective lens, a 112-total reflective mirror, a 113-micro spectrometer II and a 114-buffer reaction vessel II.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.

Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.

It is noted that relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.