阅读说明：本技术 基于拉曼光谱的体液药物浓度的测量方法及检测装置 (Method and device for measuring body fluid drug concentration based on Raman spectrum ) 是由 郭爽 崔少博 王运锋 王同斌 于 2019-12-09 设计创作，主要内容包括：本发明涉及基于拉曼光谱的体液药物浓度的测量方法及检测装置,按照如下方法进行：体液提取：红细胞破碎处理：将破碎后的红细胞放入到离心机中进行离心处理,然后将离心处理后的红细胞的上清液进行提取；对将步骤三中收集到的上清液进行震动或摆动或旋转,然后在给上清液中加入纳米金或纳米银溶液均匀混合,然后通过离心浓缩；将步骤四中的液体进行取样,然后将取得的样品进行透射拉曼光谱检测,得到药物分子表面增强拉曼光谱图,通过模型计算得到血液中药物含量；通过该方法能够对服药后的人体血液浓度进行准确测量,确保不同的人在服用药量更加准确,对患者进行进一步的保护,对环境进行了有效的保护。(The invention relates to a method and a device for measuring body fluid drug concentration based on Raman spectroscopy, which comprises the following steps of extracting body fluid, namely breaking red blood cells, placing the broken red blood cells into a centrifuge for centrifugal treatment, extracting supernatant of the centrifuged red blood cells, vibrating or swinging or rotating the supernatant collected in the third step, adding a nano-gold or nano-silver solution into the supernatant for uniform mixing, carrying out centrifugal concentration, sampling liquid in the fourth step, carrying out transmission Raman spectroscopy detection on the obtained sample to obtain a drug molecule surface enhanced Raman spectrogram, and obtaining the drug content in blood through model calculation.)

1. The method for measuring the body fluid drug concentration based on the Raman spectrum is characterized by comprising the following steps of:

, extracting body fluid, namely extracting blood of a patient taking the medicine in a sterile environment, and diluting the extracted blood;

step two: and (3) red blood cell crushing treatment: placing the diluted blood into an ultrasonic power of 400-500W for acting for 25-30 min to carry out erythrocyte crushing treatment;

step three: placing the crushed red blood cells into a centrifuge for centrifugal treatment, and then extracting supernatant of the red blood cells after the centrifugal treatment;

step four: shaking or swinging or rotating the supernatant collected in the third step, adding a nano gold or nano silver solution into the collected supernatant, uniformly mixing, and then carrying out centrifugal concentration;

step five: and C, sampling the liquid in the step four, then carrying out transmission Raman spectrum detection on the sampled liquid to obtain a drug molecule surface enhanced Raman spectrogram, and calculating through a model to obtain the drug content in the blood.

2. A method for measuring a drug concentration in a body fluid based on Raman spectroscopy according to claim 1, wherein in the second step, the broken red blood cells are filtered by a filter membrane after the red blood cells are broken.

3. A method for measuring the concentration of a drug in body fluid based on raman spectroscopy according to claim 1, wherein the extracted blood is diluted with PBS buffer in step .

4. The method for measuring the drug concentration in body fluid based on Raman spectroscopy of claim 1, wherein the extracted blood is diluted with 1-2 times volume of double distilled water in step .

5. The method for measuring the concentration of a drug in a body fluid based on Raman spectroscopy according to claim 4, wherein the third step is performed as follows: adding anhydrous ethanol and chloroform into the crushed erythrocyte liquid sequentially, stirring while adding, standing for 30min after stirring, putting the mixed liquid into a centrifuge for centrifugation, and collecting the centrifuged supernatant; and then the ethanol and the chloroform are removed by a nitrogen blowing instrument.

6. A method for measuring body fluid drug concentration based on Raman spectroscopy according to claim 3, wherein said PBS buffer is composed of the following materials in parts by weight:

8-12% of sodium dihydrogen phosphate, 5-10% of disodium hydrogen phosphate, 1-3% of potassium chloride, 0.5-1.5% of pancreatin and the balance of water.

7. The device for detecting the concentration of a drug in a body fluid based on Raman spectroscopy according to claim 1, comprising:

an ultrasonic cell disruption/dispersion machine (1) for disrupting red blood cells in the extracted blood by the ultrasonic cell disruption/dispersion machine (1);

the centrifugal separation device (2) is used for injecting liquid into the centrifugal separation device (2) after the liquid is crushed by the ultrasonic cell crushing and dispersing machine (1), and the centrifugal separation device (2) is used for carrying out centrifugal separation on the crushed red blood cell liquid and separating out supernatant;

the mixed oscillation device (3) is used for feeding the supernatant separated by the centrifugal separation device (2) into the mixed oscillation device (3) to be mixed and oscillated with the nano-gold or nano-silver solution fed into the mixed oscillation device (3);

the centrifugal concentrator (4) extracts the liquid mixed by the mixing and oscillating device (3) and adds the liquid into the centrifugal concentrator (4) for centrifugal concentration treatment;

and the Raman spectrometer (5) is used for sampling the liquid centrifugally concentrated by the centrifugal concentrator (4) and putting the liquid into the Raman spectrometer (5) for transmission Raman spectrum detection to obtain the concentration of the medicine in the blood.

8. The device for detecting the drug concentration in body fluid based on Raman spectrum according to claim 7, wherein the centrifugal separation device (2) comprises a centrifugal outer cylinder (201), a centrifugal inner cylinder (202) with a plurality of centrifugal holes arranged on the circumference, an outer cylinder cover (203) is fixedly connected to the upper end of the centrifugal outer cylinder (201), an inner cylinder cover (204) is fixedly connected to the upper end of the centrifugal inner cylinder (202), a liquid inlet pipe (205) is fixedly connected to the inner cylinder cover (204), the upper end of the liquid inlet pipe (205) extends outwards through the outer cylinder cover (203), and the outer wall of the liquid inlet pipe (205) is fixedly connected with the outer cylinder cover (203) through a bearing; a check valve (206) is arranged on the liquid inlet pipe (205);

an independent sealing installation cavity (207) is formed in the bottom of the centrifugal outer cylinder (201), a centrifugal motor (208) is fixedly arranged in the independent sealing installation cavity (207), a rotating shaft of the centrifugal motor (208) penetrates through the top of the independent sealing installation cavity (207) to be fixedly connected with the bottom of the centrifugal inner cylinder (202), and the rotating shaft of the centrifugal motor (208) extending out of the independent sealing installation cavity (207) is located in a sealing rotating cylinder (209) movably connected with the bottom of the centrifugal inner cylinder (202) and the top of the independent sealing installation cavity (207);

a filtering membrane (210) is arranged between the outer side surface of the independent sealing installation cavity (207) and the inner side surface of the centrifugal outer cylinder (201), and a supernatant liquid outlet pipe (211) is communicated with the centrifugal outer cylinder (201) at the lower part of the filtering membrane (210).

9. The device for detecting the concentration of a body fluid drug based on a Raman spectrum according to claim 7 or 8, wherein the mixing oscillation device (3) comprises an outer sound-insulation box (301) and an oscillation box (302) arranged in the outer sound-insulation box (301), two sides of the oscillation box (302) are fixedly connected with the inner wall of the outer sound-insulation box (301) through a plurality of horizontal springs (303), the bottom of the oscillation box (302) is fixedly connected with the bottom in the outer sound-insulation box (301) through a plurality of vertical springs (304), and an oscillation driving device for driving the oscillation box (301) to oscillate up and down in the outer sound-insulation box (301) is fixedly arranged at the bottom in the outer sound-insulation box (301);

the top of the oscillation box body (302) is communicated with a liquid adding hose (313), and the end part of the liquid adding hose (313) penetrates through the top of the sound insulation outer box body (301) to extend outwards; the bottom of the oscillating box body (302) is communicated with a mixed liquid outlet hose (308), and the end part of the mixed liquid outlet hose (308) penetrates through the side wall of the lower part of the sound insulation outer box body (301) to extend outwards; the upper parts of the outer sound insulation box body (301) and the vibration box body (302) are respectively provided with an outer liquid inlet (305) and an inner liquid inlet (306), and the outer liquid inlet (305) and the inner liquid inlet (306) are communicated through a hose connecting assembly (307).

10. The device for detecting the drug concentration in body fluid based on Raman spectrum according to claim 9, wherein the oscillation driving device comprises a driving motor (312) fixedly arranged in the outer sound-proof box (301), a plurality of vertically downward push rods (309) fixedly connected to the oscillation box (302), and a driving shaft (314) fixedly connected to a rotating shaft of the driving motor (301), two ends of the driving shaft (314) are movably connected to a shaft connecting seat (310) fixedly arranged at the bottom of the outer sound-proof box (301), the driving shaft (314) is provided with a plurality of cams (311), and the circumferential surface of each cams (311) is in contact with the lower end of the push rod (309).

Technical Field

The invention relates to the technical field of body fluid drug concentration measurement, in particular to a body fluid drug concentration measurement method and a body fluid drug concentration detection device based on Raman spectroscopy.

Background

It is known that the amount of meal varies significantly among people and is not always proportional to body weight and body surface area. The same is true for the administration of standard doses, some patients may be under-dosed due to too rapid metabolism and some patients may be over-metabolized, resulting in substantial overdose and toxic side effects. If the concentration of the drug in blood is high or low, the curative effect is influenced, and the drug resistance of bacteria can be induced.

Therefore, monitoring of blood concentration is necessary, especially for drugs with narrow safety range and strong toxic and side effects, drugs with nonlinear pharmacokinetic properties, drugs with in vivo elimination rate constant dependent on dosage, drugs for patients with diseases of kidney, liver, heart, stomach and intestinal tract, drugs which need to be used for a long time, and drugs which are used in combination to cause the change of absorption, distribution or metabolism of the drugs due to the interaction of the drugs.

At present, phases are not accurate and feasible methods for detecting the content of the medicine in blood extracted after the medicine is taken by a human, so that the concentration content of the medicine in the blood cannot be accurately detected during detection, the medicine absorption rate of the human after the medicine is taken cannot be fully determined, and the risk of side effects is easily caused due to the fact that people with different physiques are not accurate enough during medicine taking.

Disclosure of Invention

In order to overcome the disadvantages of the prior art, the invention aims to provide body fluid drug concentration measuring methods and detecting devices based on raman spectroscopy, which can accurately measure the drug concentration in the body fluid of a human body after taking the drug.

In order to achieve the purpose, the invention adopts the following technical scheme, and the method for measuring the body fluid drug concentration based on the Raman spectrum is carried out according to the following method:

, extracting body fluid, namely extracting blood of a patient taking the medicine in a sterile environment, and diluting the extracted blood;

step two: and (3) red blood cell crushing treatment: placing the diluted blood into an ultrasonic power of 400-500W for acting for 25-30 min to carry out erythrocyte crushing treatment;

step three: placing the crushed red blood cells into a centrifuge for centrifugal treatment, and then extracting supernatant of the red blood cells after centrifugal treatment;

step four: shaking or swinging or rotating the supernatant collected in the third step, adding a nano gold or nano silver solution into the supernatant, uniformly mixing, and then carrying out centrifugal concentration;

step five: and C, sampling the liquid in the fourth step, carrying out transmission Raman spectrum detection on the obtained sample to obtain a drug molecule surface enhanced Raman spectrogram, and calculating through a model to obtain the drug content in the blood.

And in the second step, after the red blood cells are crushed, filtering the crushed red blood cells by a filter membrane.

The extracted blood was diluted with PBS buffer in step .

Diluting the extracted blood in step by using double distilled water with the volume of 1-2 times;

the third step is carried out according to the following method: adding anhydrous ethanol and chloroform into the crushed erythrocyte liquid sequentially, stirring while adding, standing for 30min after stirring, putting the mixed liquid into a centrifuge for centrifugation, and collecting the centrifuged supernatant; and then the ethanol and the chloroform are removed by a nitrogen blowing instrument.

The PBS buffer solution consists of the following substances in parts by weight:

8-12% of sodium dihydrogen phosphate, 5-10% of disodium hydrogen phosphate, 1-3% of potassium chloride, 0.5-1.5% of pancreatin and the balance of water.

Detection device of body fluid drug concentration based on raman spectroscopy includes:

an ultrasonic cell disruption and dispersion machine for disrupting red blood cells in the extracted blood;

the centrifugal separation device is used for injecting liquid after being crushed by the ultrasonic cell crushing and dispersing machine into the centrifugal separation device, and the centrifugal separation device is used for carrying out centrifugal separation on the crushed red blood cell liquid and separating out supernatant;

the mixed oscillation device is used for mixing and oscillating the supernatant separated by the centrifugal separation device with the nano gold or nano silver solution entering the mixed oscillation device;

a centrifugal concentrator, which extracts the liquid mixed by the mixing and oscillating device and adds the liquid into the centrifugal concentrator for centrifugal concentration treatment;

and the Raman spectrometer samples the liquid centrifugally concentrated by the centrifugal concentrator, and places the liquid into the Raman spectrometer for transmission Raman spectrum detection to obtain the concentration of the medicine in the blood.

The centrifugal separation device comprises a centrifugal outer barrel and a centrifugal inner barrel, wherein a plurality of centrifugal holes are formed in the centrifugal inner barrel in the circumferential direction, an outer barrel sealing cover is fixedly connected to the upper end of the centrifugal outer barrel, an inner barrel sealing cover is fixedly connected to the upper end of the centrifugal inner barrel, a liquid inlet pipe is fixedly connected to the inner barrel sealing cover, the upper end of the liquid inlet pipe penetrates through the outer barrel sealing cover to extend outwards, and the outer wall of the liquid inlet pipe is fixedly connected with the outer barrel sealing cover through a bearing; the liquid inlet pipe is provided with a one-way valve;

an independent sealing installation cavity is formed in the bottom of the centrifugal outer barrel, a centrifugal motor is fixedly arranged in the independent sealing installation cavity, a rotating shaft of the centrifugal motor penetrates through the top of the independent sealing installation cavity and is fixedly connected with the bottom of the centrifugal inner barrel, and the rotating shaft of the centrifugal motor, which extends out of the independent sealing installation cavity, is located in a sealing rotating barrel, wherein the bottom of the centrifugal inner barrel is movably connected with the top of the independent sealing installation cavity;

a filtering membrane is arranged between the outer side surface of the independent seal installation cavity and the inner side surface of the centrifugal outer cylinder, and a supernatant liquid outlet pipe is communicated with the centrifugal outer cylinder at the lower part of the filtering membrane;

the mixed oscillation device comprises an outer sound insulation box body and an oscillation box body arranged in the outer sound insulation box body, wherein two sides of the oscillation box body are fixedly connected with the inner wall of the sound insulation box body through a plurality of horizontal springs, the bottom of the oscillation box body is fixedly connected with the bottom in the outer sound insulation box body through a plurality of vertical springs, and an oscillation driving device for driving the oscillation box body to oscillate up and down in the outer sound insulation box body is fixedly arranged at the bottom in the outer sound insulation box body;

the top of the vibration box body is communicated with a liquid adding hose, and the end part of the liquid adding hose penetrates through the top of the sound insulation outer box body and extends outwards; the bottom of the vibration box body is communicated with a mixed liquid outlet hose, and the end part of the mixed liquid outlet hose penetrates through the side wall of the lower part of the sound insulation outer box body and extends outwards; the upper portions of the sound-insulation outer box body and the vibration box body are respectively provided with an outer liquid inlet and an inner liquid inlet, and the outer liquid inlet and the inner liquid inlet are communicated through a hose connecting assembly.

Vibrate drive arrangement including fixed establish driving motor, the fixed connection in the outer box that gives sound insulation in vibrate many perpendicular decurrent ejector pins of box, with driving motor's pivot fixed connection's drive shaft, the both ends of drive shaft and the fixed hub connection seat swing joint who is equipped with in outer box bottom gives sound insulation, be equipped with a plurality of cams in the drive shaft, and the circumference face of every cams contacts with the lower extreme of ejector pin.

The method has the advantages that the method can accurately measure the blood concentration of the human body after the medicine is taken, makes contribution to the absorption of different human bodies to the medicine, ensures that different people take more accurate medicine, protects patients from steps, prevents abuse of the medicine, and effectively protects the environment.

Drawings

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

FIG. 2 is a schematic view of the structure of the centrifugal separator of the present invention;

FIG. 3 is a schematic structural diagram of a hybrid oscillator device according to the present invention;

fig. 4 is a schematic structural view of the sealed rotary cylinder of the present invention.

Detailed Description

The invention is described in detail below with reference to the figures and examples.

In the description of the present invention, it is to be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplicity of description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention.

Thus, a feature defined as "", "second" may explicitly or implicitly include or more such features, and in the description of the invention, "plurality" means two or more unless otherwise indicated.