Writing brush ionization probe device and application thereof in screening of key components of Chinese patent medicine

Abstract

The brush pen ionization probe device is constructed by taking a brush pen as a basic structure, and is used for directly and rapidly detecting complex samples with multiple shapes. The device has the advantages of more convenient sampling and ionization mode, stronger universality, less sample amount required by single analysis and low analysis cost, and can be coupled with various mass spectrometers to exert the qualitative and quantitative capabilities of high sensitivity, high specificity and high flux of mass spectrometry. In addition, the device is simple in structure, convenient to operate and high in analysis speed. According to the invention, 15 Chinese patent medicines in six dosage forms such as capsules, tablets, liquid preparations, pills, granules and powder are respectively taken as actual samples, acetaminophen, caffeine and chlorpheniramine maleate in anti-cold Chinese patent medicines and metformin, phenformin and glibenclamide in hypoglycemic Chinese patent medicines are taken as target molecules; an accurate and convenient rapid screening strategy for chemical medicine addition in the Chinese patent medicine is constructed, and the rapid detection device and method for the key components in the complex Chinese patent medicine based on the brush pen ionization probe are established.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine analysis, and particularly relates to a writing brush ionization probe device and an application thereof in screening key components of traditional Chinese medicines. Background Art

[0002] Chinese patent medicine is a Chinese medicine product made from Chinese medicine as raw materials, according to the prescribed prescriptions and preparation processes, for the prevention and treatment of diseases. As an indispensable part of traditional Chinese medicine, Chinese patent medicine has a long history, significant clinical efficacy, and is easy to carry and store. It has good therapeutic effects and a broad mass base. Most Chinese patent medicines are derived from ancient Chinese medicine prescriptions, and their dosage forms are diverse, including pills, tablets, powders, granules, liquid preparations, etc., and the prescriptions and processes are also complex and diverse. In recent years, Chinese patent medicine manufacturers have added chemical drug ingredients to Chinese patent medicines and health foods to solve the problems of slow onset, long course of treatment, and large dosage of Chinese patent medicines, so as to achieve the purpose of synergistic synergy. However, with the increasingly fierce competition in the pharmaceutical market, the phenomenon of illegal additions to Chinese patent medicine products has been repeatedly banned, posing a huge threat to public health and greatly hindering the high-quality development of the Chinese medicine industry. Due to the complexity of the dosage form and matrix of Chinese patent medicines and the diversity and uncertainty of the added ingredients, how to use modern technology to sensitively, accurately and quickly screen chemically added drugs in Chinese patent medicines has always been a bottleneck problem restricting the development of the quality control system of Chinese patent medicines.

[0003] At present, in the process of identifying the chemical additives in traditional Chinese patent medicines, most of them first use physical and chemical detection and thin-layer chromatography to screen positive samples, and then use chromatographic or mass spectrometry techniques with higher sensitivity and specificity for in-depth confirmation, which makes the inspection process rather cumbersome. With the progress of modern analytical techniques and the continuous improvement of the quality control system for traditional Chinese patent medicines, the rapid detection techniques and methods for traditional Chinese patent medicines have also developed rapidly. Currently, there are already various effective methods for detecting chemical additives in traditional Chinese patent medicines, such as vibrational spectroscopy detection techniques like Raman spectroscopy (RS), mid-infrared (MIR), and near-infrared spectroscopy (NIR), chromatographic techniques like thin-layer chromatography (TLC) and liquid chromatography (HPLC), and chromatographic-mass spectrometry coupling techniques like liquid chromatography-mass spectrometry (HPLC-MS) and gas chromatography-mass spectrometry (GC-MS). Spectral techniques are non-destructive analytical techniques, with simple and rapid operation, low analysis cost, and no pollution. However, they have limitations in terms of specificity, accuracy, sensitivity, and generality, and it is difficult to achieve highly sensitive and high-precision measurement of ultra-trace chemical additive drugs in traditional Chinese patent medicines. Chromatographic and chromatographic-mass spectrometry coupling techniques have the advantages of high sensitivity, good specificity, and the ability of accurate qualitative and quantitative analysis. However, the sample preparation is cumbersome, the analysis time is long, and the analysis cost is high. Moreover, during the long sample pretreatment stage, the target analytes are prone to loss and change. In addition, a large amount of organic chemical reagents are used in the analysis process, which does not conform to the green chemistry concept of rapid, economical, and environmental protection. With the development of the desorption electrospray ionization technique by Professor Cooks of Purdue University, direct ionization and mass spectrometry analysis of target molecules in complex matrix samples under atmospheric pressure have been achieved. However, such techniques still have problems such as complex sampling and sample loading processes, expensive consumables, high analysis cost, and it is difficult to solve prominent problems such as universality, handheld convenient sampling, and direct and rapid analysis for complex-shaped samples. Summary of the Invention

[0004] In order to improve the above technical problems, the present invention provides a brush ionization probe device and its application in screening key components of traditional Chinese patent medicines.

[0005] The present invention provides a brush ionization probe device, and the brush ionization probe device includes:

[0006] A brush pen shaft (1), an inert metal tube (2), and a polyamide fiber bundle (4);

[0007] The polyamide fiber bundle (4) is fixedly connected to the end of the pen shaft (1);

[0008] The inert metal tube (2) is wound around the end of the pen shaft (1) and is located at the connection between the pen shaft (1) and the polyamide fiber bundle (4).

[0009] According to an embodiment of the present invention, the tip of the polyamide fiber bundle (4) is adapted to the ion transfer tube (5) on the mass spectrometer; preferably, the tip of the polyamide fiber bundle (4) faces the ion transfer tube (5) on the mass spectrometer.

[0010] According to an embodiment of the present invention, the inert metal tube (2) is connected to the high-voltage power supply (3).

[0011] According to an embodiment of the present invention, the inert metal tube (2) is an inert metal tube such as a copper tube, a platinum tube, a silver tube, etc., for example, a copper tube.

[0012] According to an embodiment of the present invention, the fiber bundle length of the polyamide fiber bundle (4) is 3 mm - 50 mm, for example, 17 mm.

[0013] According to an embodiment of the present invention, the diameter of the writing brush pen shaft (1) is 0.7 mm - 10 mm, for example, 2.9 mm.

[0014] According to an embodiment of the present invention, the distance between the tip of the polyamide fiber bundle (4) and the ion transfer tube (5) is 1 mm - 20 mm, for example, 2 mm.

[0015] According to an embodiment of the present invention, the material of the polyamide fiber bundle (4) is an inert polyamide fiber, for example, Nylon 66.

[0016] According to an embodiment of the present invention, before using the device, the polyamide fiber bundle (4) is soaked and cleaned with a solvent (to remove the residues in the polyamide fiber bundle (4)); preferably, the solvent is selected from one or more of water, methanol, ethanol, dichloromethane, etc.

[0017] According to an embodiment of the present invention, by applying a high-voltage power supply (3) to the inert metal tube (2), an electric field is formed between the tip of the polyamide fiber bundle (4) and the ion transfer tube (5) on the mass spectrometer; preferably, the electric field can drive the sample (such as a sample solution) to form a Taylor cone at the tip, thereby enabling the analyte in the sample to be ionized.

[0018] According to an embodiment of the present invention, the device can be coupled with a mass spectrometer. According to an embodiment of the present invention, the mass spectrometer is selected from a linear ion trap mass spectrometer, an orbitrap mass spectrometer, a triple quadrupole mass spectrometer, and a time-of-flight mass spectrometer.

[0019] The present invention also provides a combined device, which includes the above-mentioned writing brush ionization probe device and a mass spectrometer; preferably, the mass spectrometer is selected from a linear ion trap mass spectrometer, an orbitrap mass spectrometer, a triple quadrupole mass spectrometer, and a time-of-flight mass spectrometer.

[0020] The present invention also provides a method for using the above-mentioned writing-brush ionization probe device and combined device, which is characterized in that the method comprises the following steps:

[0021] 1) Hold the pen shaft (1) of the writing brush and moisten the tip of the polyamide fiber bundle (4) with an extraction solvent, and then take a certain amount of sample with the moistened tip;

[0022] 2) Fix the device after sampling, with the polyamide fiber bundle (4) facing the ion transfer tube ⑤;

[0023] 3) Apply a high voltage (3) to the inert metal tube (2) to complete the ionization of the sample.

[0024] According to an embodiment of the present invention, in step 1), the sample is selected from solid samples or liquid samples.

[0025] According to an embodiment of the present invention, in step 1), brush the solid sample or dip the liquid sample with the tip of the polyamide fiber bundle (4).

[0026] According to an embodiment of the present invention, step 1) may alternatively be: dropping or spraying the sample to be tested onto the tip of the polyamide fiber bundle (4).

[0027] According to an embodiment of the present invention, the extraction solvent is selected according to the polarity and / or solubility of the component to be detected in the sample; for example, it is selected from methanol with different concentrations; for example, 25% methanol, 50% methanol, 75% methanol, etc.

[0028] The present invention also provides the application of the above-mentioned writing-brush ionization probe device and combined device in sample analysis and / or detection.

[0029] According to an embodiment of the present invention, the sample is selected from medicines (such as traditional Chinese medicine samples, traditional Chinese medicine samples added with chemical drugs), health products, etc.

[0030] According to an embodiment of the present invention, the sample is selected from solid samples, liquid samples, etc. The dosage forms of solid samples are, for example, traditional Chinese medicine pills, powders, capsules, granules, tablets, etc. The dosage forms of liquid samples are, for example, oral liquids, injections, etc.

[0031] According to an embodiment of the present invention, the sample is a cold medicine added with chemical drugs, a hypoglycemic drug added with chemical drugs; preferably, the chemical drug added to the cold medicine is selected from one or more of caffeine, paracetamol, chlorpheniramine (such as chlorpheniramine maleate) or its pharmaceutically acceptable salts; preferably, the chemical drug added to the hypoglycemic drug is selected from one or more of metformin, phenformin, pioglitazone (such as pioglitazone hydrochloride), glibenclamide or its pharmaceutically acceptable salts.

[0032] According to an embodiment of the present invention, an analyte is analyzed and / or detected based on the molecular ion peak information of the primary mass spectrum of the analyte in a sample, optionally the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum, optionally the fragmentation rule, optionally the standard quality spectrum information, etc.

[0033] According to an embodiment of the present invention, the molecular ion peak information [M+H] of the primary mass spectrum of caffeine, paracetamol, and chlorpheniramine + are m / z 195, m / z 152, and m / z 275 respectively.

[0034] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of caffeine are m / z 138 and m / z 110.

[0035] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of paracetamol are m / z 110.

[0036] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of chlorpheniramine are m / z 230.

[0037] According to an embodiment of the present invention, the molecular ion peak information [M+H] of the primary mass spectrum of metformin, phenformin, pioglitazone, and glibenclamide + are m / z 130, m / z 206, m / z 357, and m / z 494 respectively.

[0038] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of phenformin are m / z 189, m / z 164, m / z 105, and m / z 60.

[0039] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of pioglitazone are m / z 134 and m / z 119.

[0040] According to an embodiment of the present invention, the fragment ions of the secondary mass spectrum of the molecular ion peak information of the primary mass spectrum of glibenclamide are m / z 369 and m / z 395.

[0041] Beneficial effects

[0042] Based on a writing brush as the basic structure, the present invention constructs a Brush Ionization Probe (BIP) device for the direct and rapid detection of complex multi-morphology samples. This device has outstanding advantages in many aspects compared with existing devices / technologies:

[0043] 1) It has a more convenient sampling and ionization method. Soak the polyamide fiber bundle with an appropriate extraction solvent. Holding the pen shaft, one can sample the analyte to be tested with various morphologies (solid samples, liquid samples) (for example, directly brush or dip the sample to be tested, or directly drop / spray the sample to be tested onto the probe tip). The extraction solvent completes the in-situ extraction of the target molecule, and under the action of a high-voltage electric field, it forms a tip spray and enters the mass spectrometry ion transfer tube to complete the mass spectrometry analysis of the target molecule to be tested.

[0044] 2) It has stronger versatility. It can sample and analyze the analyte to be tested with various morphologies (solid samples, liquid samples), and is applicable to traditional Chinese medicine solid preparations such as pills, powders, capsules, granules, tablets, etc., as well as liquid samples such as oral liquids.

[0045] 3) The sample amount required for a single analysis is small. Samples as low as 1.0 μL or less of liquid and less than 0.1 mg of solid can be sampled in-situ. It is applicable to on-site sampling such as field sampling, in the fields, and archaeology.

[0046] 4) The analysis cost is low. The cost of a single brush pen probe device is less than 1.0 yuan in RMB, and it can be used once to avoid cross-contamination.

[0047] 5) It can be coupled and used with various types of mass spectrometry instruments (such as linear ion trap mass spectrometry, orbitrap mass spectrometry, triple quadrupole mass spectrometry, time-of-flight mass spectrometry, etc.) to exert the high-sensitivity, high-specificity, and high-throughput qualitative and quantitative capabilities of mass spectrometry.

[0048] In the present invention, the commonly used chemical additives in anti-cold traditional Chinese medicine preparations (paracetamol, caffeine, chlorpheniramine maleate) and the commonly used chemical additives in hypoglycemic traditional Chinese medicine (metformin, phenformin, glibenclamide) are used as target molecules respectively, and 15 traditional Chinese medicines in 6 dosage forms including capsules, tablets, liquid preparations, pills, granules, and powders are used as actual samples, realizing the accurate and convenient rapid screening of chemical additives in traditional Chinese medicines, and establishing a rapid detection device and method for chemical additives in complex traditional Chinese medicines based on a brush ionization probe. It is expected to provide a novel scientific tool for precise quality control and technological innovation in many fields such as pharmacy, traditional Chinese medicine, food, environment, and life science. Description of the Drawings

[0049] Figure 1 It is a schematic diagram of the overall structure of the brush ionization probe device. Figure 1 In it, 1 is the brush pen shaft, 2 is the inert metal tube, 3 is the high-voltage power supply, 4 is the polyamide fiber bundle, and 5 is the ion transfer tube.

[0050] Figure 2 It is the target ion intensity of the cold medicine group (a) and the hypoglycemic medicine group (b) under different solvents.

[0051] Figure 3Mass spectrometry analysis of chemical drugs easily added to Chinese patent medicines for cold relief. a) Full-scan mass spectrum of the mixed standard solution at the first level, b) Secondary mass spectrum of caffeine, c) Secondary mass spectrum of paracetamol, d) Secondary mass spectrum of chlorpheniramine maleate.

[0052] Figure 4 Mass spectrometry analysis of Chinese patent medicines for cold relief using a brush ionization probe. a) First-level mass spectrum of 999 Ganmaoling, b) First-level mass spectrum of Tankejing, c) First-level mass spectrum of Keterling, d) First-level mass spectrum of Vitamin C Yinqiao Tablets, e) First-level mass spectrum of Xiaochaihu Granules, f) First-level mass spectrum of Tongxuan Lifei Pills, g) First-level mass spectrum of 999 Oral Liquid.

[0053] Figure 5 Mass spectrometry analysis of chemical drugs easily added to Chinese patent medicines for blood sugar reduction. a) Full-scan mass spectrum of the mixed standard solution at the first level, b) Secondary mass spectrum of metformin, c) Secondary mass spectrum of phenformin, d) Secondary mass spectrum of pioglitazone, e) Secondary mass spectrum of glibenclamide.

[0054] Figure 6 Mass spectrometry analysis of Chinese patent medicines for blood sugar reduction using a brush ionization probe. a) First-level mass spectrum of Xiaoke Pills, b) First-level mass spectrum of Xiaotangling Capsules, c) First-level mass spectrum of Xiaoke Jiangtang Capsules, d) First-level mass spectrum of Jiangtang Tongmai Capsules, e) First-level mass spectrum of Shenqi Jiangtang Tablets, f) First-level mass spectrum of Yangyin Jiangtang Tablets, g) First-level mass spectrum of Beitushu Tangmaikang Granules, h) First-level mass spectrum of Huangqi Xuanshen Tea. Detailed implementation mode

[0055] The technical solution of the present invention will be further described in detail below in combination with specific embodiments. It should be understood that the following embodiments are only for illustrative explanation of the present invention and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection of the present invention.

[0056] Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods. For example, the brush used was purchased from Jiangxi Zifang Cultural Supplies Co., Ltd.

[0057] Example 1 Structure of the Brush Ionization Probe Device BIP

[0058] The overall structure of the brush ionization probe device is as Figure 1 shown.

[0059] The ionization probe body is a commercialized writing brush. Through preliminary experiments to screen the length of the polyamide fiber bundle at the front end, the optimized range of the fiber bundle length is 6.0 mm - 21 mm, and the diameter range is 0.8 mm - 3 mm. The results show that: the longer the polyamide fiber bundle and the larger the diameter, the greater the sample loading capacity and the longer the spraying time; however, as the polyamide fiber bundle continues to lengthen and the diameter increases, the spraying effect gradually weakens. Finally, a writing brush with a polyamide fiber bundle length of 17 mm and a diameter of 2.9 mm is selected for subsequent sample analysis. The probe material is inert polyamide fiber (Nylon 66), and polyamide fibers of different lengths are arranged orderly to form a polyamide fiber bundle with a tiny tip, constituting the ionization probe. Before use, the probe is soaked and ultrasonically cleaned with solvents such as deionized water, methanol, and dichloromethane.

[0060] During the experiment, the writing brush wetted with an appropriate extraction solvent can be held by hand, and the surface of the solid sample to be measured (such as traditional Chinese patent medicines in different dosage forms such as pills, powders, capsules, granules, tablets, etc.) can be directly brushed with the probe tip or a small amount of liquid sample (such as oral liquid, etc.) can be dipped, and the target analyte is adsorbed and extracted. Then, the probe tip is aligned in front of the ion transfer tube inlet of a linear ion trap mass spectrometer (LTQ XL, ThermoFisher, USA), with a distance of about 2 mm. An inert metal tube is sleeved at the connection between the end of the polyamide fiber bundle and the pen barrel, and a high voltage is applied to the inert metal tube to form a high electric field between the probe tip and the ion transfer tube, driving the extraction solution containing the analyte to be measured to form a Taylor cone at the probe tip, enabling the analyte to be measured to complete ionization ( Figure 1 ).

[0061] Example 2 Optimization of BIP-MS Conditions

[0062] Experimental Method

[0063] Commonly added chemical drugs in cold medicines (Caffeine, Paracetamol, Chlorpheniramine Maleate) and added chemical drugs in hypoglycemic drugs (Metformin, Phenformin, Pioglitazone, Glibenclamide) are selected as the detection objects. The above chemical drug standards are purchased from Beijing Beiterenkang Biotechnology Co., Ltd., with a purity of ≥98%. All standard products are configured into a stock solution of 1.0 mg / mL. Except for Phenformin using pure water as the solvent, other standard products are dissolved in methanol and stored in the dark at -20°C. Taking the moderately diluted mixed standard solution as the research object, the BIP device built in Example 1 is coupled with a linear ion trap mass spectrometer (LTQ) to optimize the mass spectrometry conditions such as the extraction solvent.

[0064] Five solutions of pure methanol, 75% methanol-water, 50% methanol-water, 25% methanol-water and pure water were selected for the investigation of extraction solvents. An appropriate amount of the standard stock solution was precisely pipetted and diluted into a mixed standard solution with the above extraction solvents respectively, so that the final concentration of each standard working solution was 1.0 μg / mL. A writing brush was held to dip into a trace amount of sample solution, the position of the BIP was fixed to align with the inlet of the mass spectrometer, and a voltage was applied to complete ionization and analysis.

[0065] Experimental results

[0066] Different extraction solvents will have different effects on the dissolution and ionization degree of target molecules. Using organic solvent methanol and its mixtures with different ratios of water as extraction solvents, solvent optimization was carried out, and the results are as Figure 2 shown.

[0067] Figure 2 a shows the ion intensities of each component in the cold medicine group under different solvents. Among them, the ion signal intensities of caffeine, paracetamol and chlorpheniramine maleate all first increased with the increase of methanol concentration. When the methanol ratio was 25%, the ion signal intensities of caffeine and paracetamol reached the maximum, and then decreased. The intensity of the ion signal of chlorpheniramine maleate reached the maximum when the methanol concentration increased to 75%, which was about 3 times the highest ion signal intensity of caffeine and paracetamol. Considering the ion signal intensities of the 3 compounds in the cold medicine group comprehensively, 25% methanol was the optimal extraction solvent.

[0068] Similarly, in the hypoglycemic drug group, although the ion signal intensities of metformin, phenformin and pioglitazone all reached the highest when the methanol concentration was 100%, the ion signal intensity of glibenclamide was extremely low at this time ( Figure 2 b), in order to expand the applicable range of the spray solvent in the quality control detection of hypoglycemic drugs, 25% methanol was selected as the extraction solvent.

[0069] Example 3 Screening for the addition of chemical drugs in traditional Chinese patent medicines for treating colds based on the BIP ionization probe device

[0070] Experimental method

[0071] Appropriately take appropriate amounts of the stock solutions of caffeine, paracetamol, and chlorpheniramine maleate standards, mix them, and dilute them to 1.0 μg / mL with a 25% methanol solution. Ionization and analysis are completed through the BIP device. Collect 7 kinds of proprietary Chinese medicines for colds, namely 999 Ganmaoling (granules, China Resources Sanjiu Medical & Pharmaceutical Co., Ltd.), Xiaochaihu Keli (granules, China Resources Sanjiu Pharmaceutical Co., Ltd.), Vitamin C Yinqiao Tablets (tablets, Guizhou Bailing Enterprise Group Pharmaceutical Co., Ltd.), Tankejing Powder (powder, Guangzhou Wanglaoji Pharmaceutical Co., Ltd.), Keterling Capsules (capsules, Tonghua Wantong Pharmaceutical Co., Ltd.), 999 Anti-viral Oral Liquid (solution, China Resources Sanjiu Pharmaceutical Co., Ltd.), and Tongxuan Lifei Pills (pills, Taiji Group Chongqing Traditional Chinese Medicine Factory No. 2 Co., Ltd.). Moisten the writing brush with a 25% methanol solution, directly brush the surface of the solid preparation to be tested with the tip of the probe or dip a small amount of the liquid preparation, and then perform ionization and analysis. In the Tune method, set Capillary Temp to 275 °C, Capillary Voltage to 35 V, Tube Lens to 140 V, and spray voltage to 4 V.

[0072] Experimental results

[0073] Figure 3 a is the full-scan first-order MS spectrum of the three chemical components easily added in cold medicines collected by this device under the optimized solvent conditions, namely paracetamol (m / z 152, [M+H] + ), caffeine (m / z 195, [M+H] + ), and chlorpheniramine maleate (m / z 275, [M+H] + ) (m / z 230 in the figure is the fragment ion generated by in-source cleavage of chlorpheniramine maleate).

[0074] Further, the second-order fragment ions of these molecular ion peaks are obtained through collision-induced dissociation (CID) experiments and their structures are annotated for the structural confirmation of the target compounds and to exclude possible interference from isomers. Figure 3 b - d are the MS / MS spectra and fragmentation assignments of caffeine, paracetamol, and chlorpheniramine maleate at the optimal CID energy, respectively. When the collision energy is 30 eV, caffeine (m / z 195) loses a fragment with a mass unit of 57 to form the m / z 138 [M+H - C2H3NO] + fragment peak, and continues to lose the CO group to generate the m / z 110 fragment peak; when the collision energy is 70 eV, paracetamol (m / z 152, [M+H] + ) loses the acetyl group (CH3CO—) to obtain a stronger fragment signal with m / z of 110; chlorpheniramine maleate loses a fragment with a mass unit of 45 (—NC2H7) at a collision energy of 9 eV to form the characteristic peak m / z 230.

[0075] The chemical components detected in 7 batches of Chinese patent medicines for treating colds were further analyzed by MS / MS, and identified by combining the quasi-molecular ions, secondary fragment ion information and fragmentation rules of the standard quality spectra. The results are as Figure 4 shown. Figure 4 (a) is the first-order and second-order mass spectra of Ganmaoling Granules. In the positive ion mode, 3 chemical drugs were mainly detected in Ganmaoling Granules, paracetamol (m / z 152), caffeine (m / z 195) and chlorpheniramine maleate (m / z 275); Tanke Jing Figure 4 (b) mainly detected caffeine (m / z 195); Keterling Figure 4 (c) detected chlorpheniramine maleate (m / z 275); Vitamin C Yinqiao Tablets Figure 4 (d) simultaneously detected paracetamol (m / z 152) and chlorpheniramine maleate (m / z 275); Xiaochaihu Granules Figure 4 (e), Tongxuan Lifei Pills Figure 4 (f) and 999 Oral Liquid Figure 4 (g) did not detect these 3 chemical components.

[0076] The chemical drug components screened out by the experiment were compared with the components indicated in the prescriptions of the actual drugs. The results showed that the chemical drug components screened above were indeed added to the prescriptions of Chinese patent medicines such as Ganmaoling Granules, Tanke Jing, Keterling and Vitamin C Yinqiao Tablets, while no chemical drugs were added to Xiaochaihu Granules, Tongxuan Lifei Pills and 999 Oral Liquid.

[0077] Example 4 Screening for Chemical Drug Additives in Antidiabetic Drugs Based on BIP Ionization Probe Device

[0078] Experimental Method

[0079] Appropriate amounts of metformin, phenformin, pioglitazone and glibenclamide standard stock solutions were mixed and diluted to 1.0 μg / mL with 25% methanol solution, and ionization and analysis were completed through the BIP device. 8 kinds of antidiabetic Chinese patent medicines in different dosage forms were collected from the market, Xiaoke Pills (pills, Guangzhou Baiyunshan Zhongyi Pharmaceutical Co., Ltd.), Xiaotangling Capsules (capsules, Liaoning Lvdan Pharmaceutical Co., Ltd.), Xiaoke Jiangtang Capsules (capsules, Luoyang Tiansheng Pharmaceutical Co., Ltd.), Jiangtang Tongmai Capsules (capsules, Qinghai Lukang Dadi Pharmaceutical Co., Ltd.), Shenqi Jiangtang Tablets (tablets, Guangdong Wannianqing Pharmaceutical Co., Ltd.), Yangyin Jiangtang Tablets (tablets, Shaanxi Fangzhou Pharmaceutical Co., Ltd.), Beitershutangmaikang Granules (granules, Chengdu Beitede Nuo Pharmaceutical Co., Ltd.), and the health product Huangqi Xuanshen Tea (granules, Xing'an Health Technology Co., Ltd.). Other experimental methods and conditions were the same as those in Example 3.

[0080] Experimental Results

[0081] All 4 hypoglycemic chemical drugs contain nitrogen atoms and are prone to form [M+H]+ peaks in the electrospray ionization source. Therefore, positive ion mode ionization is adopted. + Peaks, so positive ion mode ionization is used. Figure 5 a is the full-scan MS first-order spectrum of 4 chemical components that are easily added to hypoglycemic drugs. From left to right, they are metformin (m / z 130, [M+H]+ + ), phenformin (m / z 206, [M+H]+ + ), pioglitazone (m / z 357, [M+H]+ + ), and glibenclamide (m / z 494, [M+H]+ + ). Figure 5 b - e are the second-order mass spectra and fragmentation pathways of metformin, phenformin, pioglitazone, and glibenclamide, respectively. Biguanide drugs are prone to break at the carbon-nitrogen bond. However, when the collision energy is tuned up to 400 eV at most, the parent ion of metformin still remains stable and no fragment ions are produced. Therefore, the first-order mass spectrometry data (m / z 130) and the characteristics of its second-order mass spectrometry that are difficult to produce fragment ions are directly used to screen metformin in the sample; the fragment ions of phenformin at a collision energy of 25 eV are 189 [M+H - NH3]+ + , 164 [M + 2H - HNCNH2]+ + , 105 [M+H - C2H7N5]+ + and 60 [CN3H4 + 2H]+ + ; when the collision energy is 20 eV, the ethoxyphenyl ether bond of pioglitazone undergoes cleavage, and the characteristic fragment ions produced are m / z 134 and 119; the fragment ions m / z369 and m / z395 of glibenclamide are formed by the cleavage of two amide bonds respectively. Using the BIP device of the present invention to conduct mass spectrometry experiments on the standard product, the MS / MS fragments obtained can be used as the standard for identifying the analyte in the actual sample, and accurate and sensitive mass spectrometry characterization of the target analyte can be carried out.

[0082] The identification results of 4 common hypoglycemic chemical drug components in 8 batches of traditional Chinese patent medicines and health products for treating hyperglycemia are as Figure 6 shown. Except for glibenclamide (m / z 494) detected in Xiaoke Pills ( Figure 6 a) and Xiaotangling Capsules ( Figure 6 b), in Xiaoke Jiangtang Capsules ( Figure 6 c), Jiangtang Tongmai Capsules ( Figure 6 d), Shenqi Jiangtang Tablets ( Figure 6 e), Yangyin Jiangtang Tablets ( Figure 6 f), Beiteshutangmaikang Granules ( Figure 6 g) and Huangqi Xuanshen Tea ( Figure 6None of these four common antidiabetic drugs was detected in (h).

[0083] The chemical drug addition detected in the experiment was consistent with the chemical drug addition indicated in the antidiabetic drug prescription, which further confirmed the accuracy of the experimental results.

[0084] In summary, the brush ionization probe device provided by the present invention has the characteristics of simple device, convenient operation (handheld sampling), high sensitivity, fast analysis speed, strong universality, etc., can significantly reduce the analysis cost and the consumption of samples and reagents, and conforms to the modern analytical chemistry concept of environmental protection.

[0085] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A brush ionization probe device, the brush ionization probe device comprising: A brush pen shaft (1), an inert metal tube (2), a polyamide fiber bundle (4); The polyamide fiber bundle (4) is fixedly connected to the end of the pen shaft (1); The inert metal tube (2) is wound around the end of the pen shaft (1) and is located at the connection between the pen shaft (1) and the polyamide fiber bundle (4).

2. The device according to claim 1, characterized in that The tip of the polyamide fiber bundle (4) is adapted to the ion transfer tube (5) on the mass spectrometer; preferably, the tip of the polyamide fiber bundle (4) faces the ion transfer tube (5) on the mass spectrometer; Preferably, the inert metal tube (2) is connected to a high-voltage power supply (3); Preferably, the inert metal tube (2) is an inert metal tube such as a copper tube, a platinum tube, a silver tube, etc.; Preferably, the fiber bundle length of the polyamide fiber bundle (4) is 3 mm - 50 mm; Preferably, the diameter of the brush pen shaft (1) is 0.7 mm - 10 mm; Preferably, the distance between the tip of the polyamide fiber bundle (4) and the ion transfer tube (5) is 1 mm - 20 mm; Preferably, the material of the polyamide fiber bundle (4) is an inert polyamide fiber.

3. The device according to claim 1 or 2, characterized in that, Before using the device, soak and clean the polyamide fiber bundle (4) with a solvent; preferably, the solvent is selected from one or more of water, methanol, ethanol, dichloromethane, etc.; Preferably, by applying a high-voltage power supply (3) to the inert metal tube (2), an electric field is formed between the tip of the polyamide fiber bundle (4) and the ion transfer tube (5) on the mass spectrometer; Preferably, the device can be coupled and used in combination with a mass spectrometer.

4. A combined device, the combined device comprising the brush ionization probe device according to any one of claims 1 - 3 and a mass spectrometer; preferably, the mass spectrometer is selected from a linear ion trap mass spectrometer, an orbitrap mass spectrometer, a triple quadrupole mass spectrometer, a time-of-flight mass spectrometer.

5. The method of using the brush ionization probe device according to any one of claims 1-3 and the combined device according to claim 4, characterized in that, The usage method comprises the following steps: 1) Hold the brush pen shaft (1) and wet the tip of the polyamide fiber bundle (4) with an extraction solvent, and then take a certain amount of sample with the wetted tip; 2) Fix the device after sampling, with the polyamide fiber bundle (4) facing the ion transfer tube ⑤; 3) Apply a high voltage (3) to the inert metal tube (2), and the sample is ionized.

6. The usage method according to claim 5, characterized in that, In step 1), the sample is selected from a solid sample or a liquid sample; Preferably, in step 1), brush the solid sample or dip the liquid sample with the tip of the polyamide fiber bundle (4); Preferably, step 1) is alternatively: drop or spray the sample to be tested onto the tip of the polyamide fiber bundle (4); Preferably, the extraction solvent is selected according to the polarity and / or solubility of the component to be detected in the sample.

7. The application of the brush ionization probe device according to any one of claims 1 - 3 and the combined device according to claim 4 in sample analysis and / or detection.

8. The application according to claim 7, characterized in that The sample is selected from medicines (such as traditional Chinese medicine samples, traditional Chinese medicine samples added with chemical drugs), health products, etc.; Preferably, the sample is selected from solid samples, liquid samples, etc.; Preferably, the sample is a cold medicine added with a chemical drug or a hypoglycemic drug added with a chemical drug; preferably, the chemical drug added to the cold medicine is selected from one or more of caffeine, paracetamol, chlorpheniramine or a pharmaceutically acceptable salt thereof; preferably, the chemical drug added to the hypoglycemic drug is selected from one or more of metformin, phenformin, pioglitazone, glibenclamide or a pharmaceutically acceptable salt thereof; Preferably, based on the molecular ion peak information of the primary mass spectrum of the analyte in the sample, optionally the secondary mass spectrum fragments of the molecular ion peak information of the primary mass spectrum, optionally the fragmentation rules, optionally the standard quality spectrum information, etc., the analyte is analyzed and / or detected.

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