Method for detecting blood concentration in human body

Abstract

The invention provides a method for detecting blood concentration in a human body, which comprises the following steps of: A) placing a medicine standard substance on the surface of a substrate, and detecting by adopting Raman spectrum to obtain a standard fingerprint spectrum; and B) collecting the blood by adopting a micro-fluidic chip, detecting by adopting a Raman spectrum to obtain a Raman fingerprint spectrum in the blood, and comparing the Raman fingerprint spectrum with a standard spectrum to obtain the blood concentration. According to the application of the chiral film in blood concentration monitoring, the chiral nanostructure substrate provided by the invention can realize qualitative and quantitative monitoring of the blood concentration in a human body based on the surface enhanced Raman scattering effect, and clinical prediction can be carried out clinically according to metabolic levels of different blood concentrations of different patients.

Description

Technical Field

[0001] The present invention relates to the technical field of drug analysis, and in particular to a method for detecting blood drug concentration in a human body. Background Art

[0002] Nanobiodetection is a biological detection method and technology based on nanomaterials and nanostructures with special properties. Compared with traditional detection methods, nanobiodetection often has the advantages of high detection sensitivity, strong specificity, simultaneous detection of multiple targets, and rapid and instant detection. At the same time, chirality is a unique phenomenon in biological systems: from simple amino acids and sugars to polypeptides with multi-level structures to biomacromolecules such as proteins and nucleic acids, they all exhibit a single chiral structure. The chiral characteristics of biological systems not only determine the specificity of interactions between biomolecules, but also affect a series of processes in biological systems. In recent years, chiral nanomaterials have been widely used in the detection and identification of biomolecules due to their unique optical properties and stability. In addition, the biosafety effect of chiral nanomaterials is a prerequisite for their application in biological systems, and therefore they have gradually attracted people's attention and research. Chirality is widely present in organisms, and many disease markers are closely related to chirality, so chiral materials have broad application prospects in the biological field.

[0003] Monitoring blood drug concentrations is essential to ensure the safety and effectiveness of drug treatment. First, there may be significant differences in the absorption, distribution, metabolism and excretion of drugs between different individuals. By monitoring blood drug concentrations, doctors can adjust drug dosages to suit the specific needs of individuals. For example, individual genetic differences may affect the activity of drug-metabolizing enzymes, resulting in different responses to the same drug in different patients. Monitoring blood drug concentrations helps to take these genetic factors into account and optimize treatment plans. Secondly, because the therapeutic window of some drugs is narrow, that is, the gap between the effective concentration and the toxic concentration is small. Monitoring blood drug concentrations helps to avoid drug overdose and reduce potential toxic reactions and side effects. Ensuring that drug concentrations are maintained within the effective range can improve the treatment effect and reduce the risk of treatment failure. Moreover, when patients use multiple drugs at the same time, drug interactions may occur, affecting the blood concentration of drugs. Monitoring can help identify these interactions and adjust treatment plans in a timely manner. In addition, monitoring blood drug concentrations can reflect the patient's metabolic status, such as liver and kidney function, all of which affect the clearance rate of drugs. For patients who need long-term medication, regular monitoring of blood drug concentrations can help manage treatment plans in the long term and ensure continued efficacy and safety. At the same time, it is also considered that children, the elderly, pregnant women and patients with impaired liver and kidney function may need special dosage adjustments, and monitoring blood drug concentrations is particularly important for these special populations.

[0004] Clinical monitoring is to conduct real-time and continuous physical or chemical tests on patients' vital signs and physiological parameters through relevant equipment, and present them in the form of data or images to provide a basis for diagnosis and treatment. At present, there is a lack of direct and accurate methods to reflect the blood drug concentration in patients in clinical practice. In the past, common methods for monitoring body fluids using chemical materials included mass spectrometry, fluorescence detection, etc., but these methods are often accompanied by disadvantages such as long time consumption, high cost, and high sample requirements. In addition, traditional blood tests require centrifugation of blood samples after blood collection, which is very time-consuming and cannot be used for real-time monitoring of clinical blood drug concentrations.

[0005] Currently popular electrochemical-based sensors measure the current or potential on the surface of highly specific electrodes to convert analyte concentrations, and have been able to continuously track some electrolytes, metabolites (such as lactate and glucose), and drugs in body fluids; however, these sensors are usually limited to detecting one analyte at a time. In addition, the detection process is complicated and not integrated, and an integrated monitoring system cannot be established. In order to obtain dynamic and complete monitoring, it is necessary but challenging to combine several sensing mechanisms and detection methods into one sensing platform.

[0006] Therefore, the goal of the present invention is to develop a new sensor with universal target specificity rather than just one target, so that multiple targets can be tracked simultaneously. However, finding a more accurate, visual, and easier to put into clinical use blood drug concentration monitoring method is still very challenging and is also a hot topic of great concern to researchers in the field of life and health. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a method for detecting blood drug concentration in the human body. The method provided by the present invention can sensitively detect the substrate of low concentration of drugs in the blood, and has good signal reproducibility and uniformity, and at the same time has a low detection limit, and is integrated with a microfluidic chip that can separate plasma to form a mature monitoring system, thereby realizing its application in blood drug concentration detection. The present invention realizes rapid and accurate blood drug concentration monitoring by integrating a microfluidic chip that can separate plasma and a substrate with high chirality optical activity.

[0008] The present invention provides a method for detecting blood drug concentration in a human body, comprising the following steps:

[0009] A) placing a drug standard on the substrate surface and detecting it using Raman spectroscopy to obtain a standard fingerprint;

[0010] B) Blood is collected using a microfluidic chip and tested using Raman spectroscopy to obtain the Raman fingerprint of the blood, which is then compared with the standard spectrum to obtain the blood drug concentration.

[0011] The method for detecting blood drug concentration in human body provided by the present invention firstly prepares a substrate.

[0012] The substrate of the present invention is a nano-helical silver substrate with chiral optical activity.

[0013] In the present invention, preferably, the grazing angle deposition technique is used to prepare the nano-helical silver substrate having chiral optical activity.

[0014] In the present invention, the method for preparing the substrate comprises: using grazing angle deposition technology to prepare a nano-helical silver substrate with chiral optical activity.

[0015] The deposition rate is 0.3 nm / s; the theoretical pitch of the nano-helical silver substrate is 150 nm, the theoretical diameter is 70 nm, and the theoretical length is 856.64 nm.

[0016] The inventors creatively discovered that, compared with a rod-shaped nanosilver substrate under the same parameter conditions, a chiral substrate with a helical structure has a better surface enhanced Raman scattering effect, a lower detection limit and a better repeatability effect.

[0017] The drug standard is placed on the substrate surface and detected using Raman spectroscopy to obtain a standard fingerprint.

[0018] The pharmaceutical standard sample was dropped onto the surface of the chiral substrate. After drying, the chiral substrate was placed under a portable Raman laser light source and the power of the laser light source was adjusted to 10 mW.

[0019] The laser power of the Raman spectrum is 10 mW, the averaging time is 1 s, the wavelength of the laser light source is 785 nm, and the integration time is 3000.

[0020] The present invention has advantages in detecting complex samples that can resist matrix interface effects. Based on the surface enhanced Raman scattering effect of chiral nanomaterials and patient blood samples, the drug concentration in the patient's body is associated with the Raman signal intensity. The chiral nanomaterial with a special structure as a surface enhanced Raman scattering substrate can sensitively capture the low concentration of drug content in the patient's blood and display it as a spectrum signal. Since each drug molecule has a unique SERS spectrum, the target extracted from the plasma can be "fingerprinted" by our sensor. By using this sensor, as a proof-of-concept study, the changes in drug concentration in the human body can be tracked in real time to obtain an individual's drug metabolism profile. By observing the Raman signal intensity to directly reflect the different drug contents in the patient's body, the blood drug concentration can be monitored, which helps to achieve accurate metabolic assessment.

[0021] The present invention prepares a microfluidic chip.

[0022] According to the present invention, the microfluidic chip comprises a glass slide and a PDMS block disposed on the glass slide;

[0023] The PDMS block is provided with a blood inlet, a collecting end and a blood outlet; a thin pipeline connecting the blood inlet and the collecting end; a main pipeline connecting the blood inlet and the blood outlet; and the collecting end is provided with a base.

[0024] like Fig.10 As shown, 1 is the blood inlet, 2 is the glass slide, 3 is the PDMS block, 4 is the collecting end, and 5 is the blood outlet.

[0025] The main channel is 110 μm wide and 10 μm deep, and the thin channel is 2 μm wide and 1.2 μm deep.

[0026] The microfluidic chip of the invention is formed by bonding PDMS and a glass slide, and the PDMS block is provided with two capillary channels of different thicknesses and depths, a filter comb, a blood inlet and a plasma collection area.

[0027] Initially, a silicon master mold was made by photolithography. The channel pattern of the silicon mold was a main channel with a width of 110 μm and a depth of 10 μm, and a thin channel with a width of 2 μm and a depth of 1.2 μm. PDMS (the ratio of A glue and B in PDMS was 10:1 by volume) was poured on the silicon master mold and cured at 100 ° C for 60 minutes, and then carefully peeled off from the silicon master mold. Then the blood inlet, plasma collection and waste liquid collection areas were cut out. Finally, the glass slide and PDMS were placed in a plasma cleaner for 1 minute, and then the plasma-activated PDMS was placed on the glass. The bonded PDMS and glass slide were sealed and placed in a 60 ° C oven for 1 hour for reinforcement treatment, and then taken out to form a sealed and firm microfluidic chip.

[0028] The present invention uses a microfluidic chip to collect blood, specifically including: placing a blood sample into the blood inlet of the microfluidic chip, and after the blood flows through the main channel and the fine channel, the separated plasma enters the collection end, and the plasma can be detected after being adsorbed on the sample and coagulated.

[0029] The unique Raman fingerprint information of the drug is obtained by scattering the light source information after the excitation light source irradiated onto the drug on the chiral substrate integrated into the microfluidic chip is scattered back.

[0030] The solvent of the pharmaceutical standard product of the present invention is water; the concentration of the pharmaceutical standard product is the optimal concentration level of 1 mg / mL to 10 mg / mL.

[0031] Specifically, it can be 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL; or a range between any two of the above.

[0032] The medicine of the present invention comprises one of remifentanil, atracurium, methylprednisolone sodium succinate or rocuronium bromide.

[0033] According to the present invention, the characteristic peak in the Raman spectrum of remifentanil is located at 375.39 cm -1 、713.71cm -1 、835.09cm -1 、948.54cm -1 、1026.68cm -1 、1444.09cm -1 ;

[0034] The characteristic peak in the Raman spectrum of methylprednisolone sodium is located at 314.75 cm -1 、610.68cm -1 、688.06cm -1 、922.14cm -1 、1000.81cm -1 、1052.37cm -1 、1287.85cm -1 、1396.05cm -1 、1602.3cm -1 ;

[0035] The characteristic peak in the Raman spectrum of rocuronium bromide is located at 377.9 cm -1 、853.11cm -1 、1130.49cm -1 、1604.12cm -1 ;

[0036] The characteristic peak in the Raman spectrum of atracurium is located at 405.42 cm -1 、550.22cm -1 、666.96cm -1 、853.35cm -1 、957.3cm -1 、1140.92cm -1 、1378.58cm -1 、1585.92cm -1 .

[0037] The detection technology of the present invention can be used to obtain the spectrum information of various pure drugs, build a standard fingerprint spectrum information library, and use it as a standard sample to control the Raman spectrum of unknown drugs in the patient's blood sample. At the same time, the integrated microfluidic chip detection technology of the present invention can achieve rapid plasma separation, shorten the time from blood collection to plasma analysis, and speed up the diagnosis process. The detection principle of surface enhanced Raman scattering allows the use of trace blood samples for plasma separation, which is particularly beneficial for patients who need multiple blood draws, such as infants or anemic patients. Compared with the current traditional clinical blood drug concentration monitoring, the portable Raman detection based on chiral substrates has the advantages of small sample size, fast detection speed, strong visualization, and high accuracy. The peak width of the Raman spectrum is narrow, and the fingerprint spectrum information of different drugs will not interfere with each other, which can achieve accurate drug detection. Portable chip technology allows the processing and analysis of blood samples at clinical sites or remote areas, expanding the coverage of medical services. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 : The Raman substrate designed by the present invention has a lower detection limit for CV (crystal violet) material (proving the good detection performance of the substrate);

[0039] Figure 2 : It proves that the Raman substrate of the present invention has good reproducibility, and the difference is controlled within ±10%;

[0040] Figure 3 : Raman fingerprint of remifentanil;

[0041] Figure 4 : Raman fingerprint of methylprednisolone sodium;

[0042] Figure 5 : Raman fingerprint of rocuronium bromide;

[0043] Figure 6 : Raman fingerprint of atracurium;

[0044] Figure 7 : Integrated detection system to separate plasma;

[0045] Figure 8 : Schematic diagram of the pipeline structure of the microfluidic chip;

[0046] Fig. 9 :A real-shot structural diagram of the microfluidic chip;

[0047] Fig.10 : Microscope image of plasma separation using a microfluidic chip;

[0048] Fig.11 : Raman spectrum of patient plasma detected by the microfluidic chip of the present invention;

[0049] Fig.12 : Comparison of plasma spectra with drug spectra (proving that drugs can be detected in plasma);

[0050] Fig.13 : Raman spectra of blood collected from patients 30min, 60min, 90min, 135min, and 170min after injection of anesthetic drugs after being processed by the chip;

[0051] Fig.14 : The linear relationship between Raman intensity and time in the Raman spectrum of patient blood collected 30min, 60min, 90min, 135min, and 170min after injection of anesthetic drugs and processed by the chip. DETAILED DESCRIPTION

[0052] The present invention provides a method for detecting blood drug concentration in the human body. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar replacements and modifications are obvious to those skilled in the art, and they all fall within the scope of protection of the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0053] It should be understood that the expression "one or more of..." includes each of the items recited after the expression individually and in various different combinations of two or more of the recited items, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited items should be understood to have the same meaning, unless otherwise understood from the context.

[0054] The use of the terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, for example not excluding other unrecited elements or steps, unless otherwise specifically stated or otherwise understood from the context.

[0055] In this application, the term "and / or" describes the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural.

[0056] In this application, "at least one" means one or more, "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0057] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the present invention remains operable. In addition, two or more steps or actions may be performed simultaneously.

[0058] The use of any and all examples or exemplary language, such as "for example" or "including", herein is intended only to better illustrate the invention and does not limit the scope of the invention unless otherwise claimed. No language in this specification should be construed as indicating that any non-claimed element is essential to the practice of the invention.

[0059] In addition, the numerical ranges and parameters used to define the present invention are approximate values, and the relevant values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in this disclosure are modified by "about". Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0060] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0061] Some cases are recorded in the embodiments and comparative examples of the present invention, wherein the embodiments show certain implementation modes of the present invention. However, this does not mean that the effects of the present invention can only be achieved in these cases.

[0062] In order to further illustrate the present invention, a method for detecting blood drug concentration in human body provided by the present invention is described in detail below in conjunction with examples.

[0063] All raw materials of the present invention are not particularly limited in their sources and can be purchased from the market or prepared according to conventional methods known to those skilled in the art.

[0064] There is no particular restriction on the purity of all raw materials in the present invention. The present invention preferably uses analytically pure materials or materials of conventional purity used in preparing chiral nanomaterials.

[0065] The names and abbreviations of all process devices of the present invention are conventional names and abbreviations, and each name and abbreviation is clear and unambiguous in its relevant use. The technicians in this field can purchase them from the market or prepare them by conventional methods based on the names, abbreviations and corresponding uses.

[0066] The detection of the medicine in the present invention is carried out on a portable Raman spectrometer device. The device is purchased from Xia Rui Intelligent Technology Co., Ltd. and the model is XRaman-Easy. The wavelength of the laser light source is 785nm. During the detection, the sample to be tested is placed on a stable stage, the laser light source is aimed at the sample to be tested, and the appropriate focal length is selected by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. The computer is connected to the spectrometer, and the spectral information can be collected in real time and transmitted to the electronic device.

[0067] Taking methylprednisolone sodium and rocuronium bromide as examples, they are both commonly used anesthetics for clinical general anesthesia surgery, and their detection means, steps, and data processing methods are consistent. When preparing five kinds of anesthetic standard samples, the preparation method adopted is exactly the same, that is, the anesthetic is diluted to the optimal concentration. Depending on the sample concentration, the ratio of the drug to the solvent deionized water can be different. In the present invention, preferably, the concentration of the drug is controlled at a concentration of the order of mg / mL.

[0068] Example 1 Taking the gradient detection of CV (crystal violet) as an example to prove that the substrate has a lower detection limit

[0069] In the first step, take a crystal violet sample and prepare it with deionized water as solvent at a concentration of 10^ -9 M, 10^ -8 M, 10^ -7 M, 10^ -6 M, 10^ -5 Five solutions of M.

[0070] In the second step, use a pipette to take 10 μL of solution from five solutions of different concentrations, then drop them in the center of the substrate and leave them to dry at room temperature.

[0071] The third step is to wait for the sample droplet to dry, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the averaging time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal stabilizes, the computer collects the Raman fingerprint of crystal violet in the 0nm to 3000nm band. Figure 1 The characteristic spectra and characteristic peaks of crystal violet at different concentrations can be seen. The characteristic peak of crystal violet is: 725 cm -1 , 801cm -1 , 914 cm -1 , 1177cm -1 ,1392 cm -1 , and 1588 cm -1 At the same time, the lower detection limit of the substrate for crystal violet was demonstrated.

[0072] Example 2: Signal collection from different regions of the same sample in the detection of crystal violet proves that the sample has good signal uniformity

[0073] In the first step, take a crystal violet sample and prepare it with a concentration of 10 -9 The solution of M.

[0074] In the second step, use a pipette to take 10μL of the solution and drop it in the center of the substrate, and leave it to dry at room temperature. After the sample droplet is dry, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal is stable, the computer collects the Raman fingerprint of crystal violet in the 0nm to 3000nm band.

[0075] In the third step, the position of the substrate can be moved by adjusting the control knob on the stage, and 100 points are randomly selected in the substrate for signal detection. The results are as follows: Figure 2 As shown, it is proved that the substrate has good signal uniformity.

[0076] Example 3: Establishment of a standard curve for remifentanil as an example

[0077] In the first step, a standard sample of remifentanil was prepared. First, pure powder of remifentanil was taken and diluted to 1 mg / mL with deionized water as solvent.

[0078] The second step is to take 10μL of the standard drug solution with a pipette and drop it into the center of the chiral substrate. After the sample droplet is dry, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal is stable, the computer collects the Raman fingerprint of the standard drug remifentanil in the 0nm to 3000nm band. Figure 3 It can be seen that the characteristic spectrum and characteristic peak of remifentanil are located at 375.39 cm -1 、713.71cm -1 、835.09cm -1 、948.54cm -1 、1026.68cm -1 、1444.09cm -1 .

[0079] Example 4: Taking the standard curve of methylprednisolone sodium as an example

[0080] The first step is to prepare a standard sample of methylprednisolone sodium. First, take the pure powder of the drug methylprednisolone sodium and dilute it to the optimal concentration of 1 mg / mL with deionized water as solvent.

[0081] In the second step, take 10μL of the standard drug solution with a pipette and drop it into the center of the chiral substrate. After the sample droplet dries, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal stabilizes, the computer collects the Raman fingerprint of the standard drug methylprednisolone sodium in the 0nm to 3000nm band. Figure 3 The characteristic spectrum and characteristic peak of methylprednisolone sodium can be seen in the figure; the characteristic peak in the Raman spectrum of methylprednisolone sodium is located at 314.75 cm -1 、610.68cm -1 、688.06cm -1 、922.14cm -1 、1000.81cm -1 、1052.37cm -1 、1287.85cm -1 、1396.05cm -1 、1602.3cm -1 .

[0082] Example 5: Establishment of the standard curve of rocuronium bromide as an example

[0083] In the first step, a standard sample of rocuronium bromide is prepared. First, pure powder of the drug rocuronium bromide is taken and diluted to an optimal concentration of 1 mg / mL with deionized water as a solvent.

[0084] In the second step, use a pipette to take 10μL of the standard drug solution and drop it into the center of the chiral substrate. After the sample droplet dries, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal stabilizes, the computer collects the Raman fingerprint of the standard drug methylrocuronium bromide in the 0nm to 3000nm band. Figure 3 The characteristic spectrum and characteristic peaks of rocuronium bromide can be seen in the figure; the characteristic peak in the Raman spectrum of rocuronium bromide is located at 377.9cm -1 、853.11cm-1 、1130.49cm -1 、1604.12cm -1 .

[0085] Example 6: Taking the establishment of atracurium standard curve as an example

[0086] In the first step, a standard sample of atracurium was prepared. First, pure powder of atracurium was taken and diluted to 1 mg / mL with deionized water as solvent.

[0087] In the second step, take 10μL of the standard drug solution with a pipette and drop it into the center of the chiral substrate. After the sample droplet dries, place the substrate on the stage of the portable Raman spectrometer, aim the laser light source at the sample to be tested, set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal stabilizes, the computer collects the Raman fingerprint of the standard drug atracurium in the 0nm to 3000nm band. Figure 3 The characteristic spectrum and characteristic peak of atracurium can be seen in the figure; the characteristic peak in the Raman spectrum of atracurium is located at 405.42 cm -1 、550.22cm -1 、666.96cm -1 、853.35cm -1 、957.3cm -1 、1140.92cm -1 、1378.58cm -1 、1585.92cm -1 . .

[0088] Example 7: Taking the detection of actual patient blood samples as an example

[0089] The first step is to take samples from the patient at 30min, 60min, 90min, 135min, and 165min after the patient is injected with the drug. Take 10μL of the patient's real blood in the above time periods and put it into the microfluidic chip of the present invention. After the plasma is separated from the blood, it drips on the substrate of the collection area of ​​the chip. After the plasma is dried, immediately use a portable Raman laser light source to aim at the position where the plasma sample is dripped on the chiral substrate. Set the laser power to 10mW, the average time to 1s, the wavelength of the laser light source to 785nm, and the integration time to 3000. Select the appropriate focal length by adjusting the knob. When the focal length is appropriate, the Raman signal will be significantly enhanced. After the signal is stable, the computer terminal collects the Raman fingerprint spectrum of the patient's real blood in the 0nm to 3000nm band. Fig.11The characteristic spectrum and characteristic peaks of the patient's real blood in different time periods can be seen. The characteristic peaks are 375.39cm -1 (corresponding to remifentanil 375.39cm -1 characteristic peak), 853.11cm -1 (corresponding to rocuronium bromide 853.11cm -1 characteristic peaks), 1140.92cm -1 (corresponding to atracurium 1140.92cm -1 characteristic peaks), 1602.3cm -1 (corresponding to methylprednisolone sodium 1602.3cm -1 characteristic peaks) Fig.13 : Raman spectra of blood collected from patients 30min, 60min, 90min, 135min, and 170min after injection of anesthetic drugs after being processed by the chip

[0090] Example 8: Comparison of real blood samples from patients with standard drug profiles

[0091] The first step is to collect the Raman spectra of the patient's real blood at different time periods (30 minutes, 60 minutes, 90 minutes, 135 minutes, and 170 minutes after the injection of anesthetic drugs).

[0092] In the second step, by comparing the Raman characteristic peaks of remifentanil, methylprednisolone sodium, rocuronium, and atracurium collected in the above examples, the characteristic peak signals of different drugs can be found in the real blood of patients. Fig.12 As shown, since the Raman fingerprint patterns in the patient's blood samples at different times after drug injection are the same but with different intensities, the intensity will gradually decay with time. By correlating the drug content in the patient's body with the Raman signal intensity, a linear relationship between the intensity and time in the Raman spectrum can be established, thereby visually and accurately monitoring the patient's blood drug concentration.

[0093] Fig.14 : The linear relationship between Raman intensity and time in the Raman spectrum of patient blood collected 30min, 60min, 90min, 135min, and 170min after injection of anesthetic drugs and processed by the chip.

[0094] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for detecting blood drug concentration in human body, characterized in that: The steps include: A) placing a drug standard on the substrate surface and detecting it using Raman spectroscopy to obtain a standard fingerprint; B) Blood is collected using a microfluidic chip and tested using Raman spectroscopy to obtain the Raman fingerprint of the blood, which is then compared with the standard spectrum to obtain the blood drug concentration.

2. The method according to claim 1, characterized in that The power of the laser light source of the Raman spectrum is 30 mW, the averaging time is 1 s, the wavelength of the laser light source is 785 nm, and the integration time is 3000.

3. The method according to claim 1, characterized in that Step A) The substrate is a nano-helical silver substrate with chiral optical activity.

4. The method according to claim 3, characterized in that Step A) The preparation method of the substrate comprises: using grazing angle deposition technology to prepare a nano-helical silver substrate with chiral optical activity; the deposition rate is 0.3nm / s; the theoretical pitch of the nano-helical silver substrate is 150nm, the theoretical diameter is 70nm, and the theoretical length is 856.64nm.

5. The method according to claim 1, characterized in that The microfluidic chip comprises a glass slide and a PDMS block arranged on the glass slide; The PDMS block is provided with a blood inlet, a collection end and a blood outlet; A thin pipe connecting the blood inlet and the collecting end; a main pipe connecting the blood inlet and the blood outlet; and the collecting end is provided with a base.

6. The method according to claim 5, characterized in that The main channel is 110 μm wide and 10 μm deep, and the thin channel is 2 μm wide and 1.2 μm deep.

7. The method according to claim 5, characterized in that The blood collection using the microfluidic chip specifically includes: placing a blood sample into the blood inlet of the microfluidic chip, and after the blood flows through the main channel and the thin channel, the separated plasma enters the collection end, and the plasma can be detected after being adsorbed on the sample and coagulated.

8. The method according to claim 1, characterized in that The solvent of the pharmaceutical standard is water; the concentration of the pharmaceutical standard is 1 mg / mL to 10 mg / mL.

9. The method according to claim 1, characterized in that: The drug includes one of remifentanil, atracurium, methylprednisolone sodium succinate or rocuronium bromide.

10. The method according to claim 9, characterized in that The characteristic peak in the Raman spectrum of remifentanil is located at 375.39 cm -1 、713.71cm -1 、835.09cm -1 、948.54cm -1 、1026.68cm -1 、1444.09cm -1 ; The characteristic peak in the Raman spectrum of methylprednisolone sodium is located at 314.75 cm -1 、610.68cm -1 、688.06cm -1 、922.14cm -1 、1000.81cm -1 、1052.37cm -1 、1287.85cm -1 、1396.05cm -1 、1602.3cm -1 ; The characteristic peak in the Raman spectrum of rocuronium bromide is located at 377.9 cm -1 、853.11cm -1 、1130.49cm -1 、1604.12cm -1 ; The characteristic peak in the Raman spectrum of atracurium is located at 405.42 cm -1 、550.22cm -1 、666.96cm -1 、853.35cm -1 、957.3cm -1 、1140.92cm -1 、1378.58cm -1 、1585.92cm -1 .

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