Method for detecting pesticide residues in traditional Chinese medicine preparation based on UPLC-Q-TOF-MS analysis method

Abstract

The invention discloses a method for detecting pesticide residues in a traditional Chinese medicine preparation based on a UPLC-Q-TOF-MS (Ultra Performance Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry) analysis method, and provides a screening spectrum library of pesticide components in the traditional Chinese medicine preparation established based on the technology, and a method for detecting the pesticide residues in the traditional Chinese medicine preparation. The method provided by the invention can effectively identify the pesticide residue components in the traditional Chinese medicine preparation, is simple in pretreatment, less in component loss, more accurate and sensitive in detection and short in consumed time, can effectively separate 30 pesticide residue components within 15 minutes, realizes high-throughput detection and analysis identification of the pesticide residue components in the traditional Chinese medicine preparation, and has a wide application prospect. The method can rapidly and comprehensively analyze pesticide residues in the traditional Chinese medicine preparation raw materials, intermediates and finished products, overcomes the problems of long operation time, complex detection and difficult identification of the existing gas chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry, and is beneficial to quality control of the traditional Chinese medicine preparation raw materials, intermediates and finished products.

Description

Technical Field

[0001] The invention belongs to the technical field of analytical methods for chemical components in Chinese patent medicines, and specifically relates to a method for detecting pesticide residues in Chinese medicine preparations based on a UPLC-Q-TOF-MS analytical method. Background Art

[0002] Qingkailing preparations include Qingkailing oral liquid, Qingkailing tablets, Qingkailing soft capsules, Qingkailing effervescent tablets, Qingkailing injection, Qingkailing granules and other dosage forms. They have the effects of clearing away heat and detoxifying, calming the nerves, resolving phlegm and unblocking meridians, as well as awakening the brain and invigorating the mind, and are widely used in clinical practice. At present, pesticide residues in Chinese medicinal materials are a complex and widespread problem, mainly due to the large-scale use of pesticides in the cultivation of Chinese medicinal materials. In order to increase production and prevent and control pests and diseases, some farmers even use pesticides that are expressly prohibited by the state, resulting in excessive pesticide residues in Chinese medicinal materials, which seriously affects the quality and safety of Chinese medicinal materials.

[0003] In the actual production process of Qingkailing preparations, four plant-based medicinal materials, namely honeysuckle, gardenia, scutellaria and isatis root, are needed. These medicinal materials are crucial to the quality and efficacy of Qingkailing. Therefore, the detection of pesticide residues in these four medicinal materials is of great significance to the quality and efficacy control of the finished Qingkailing preparations.

[0004] In order to address this problem, the 2020 edition of the Chinese Pharmacopoeia added 33 prohibited pesticide detection items in the "0212 General Rules for the Verification of Medicinal Materials and Decoctions", and used gas chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry to conduct qualitative screening and quantitative determination of pesticide residues in Chinese medicinal materials. However, this method requires gas chromatography, tandem mass spectrometry, and liquid chromatography in the detection process, which makes the detection process too complicated, prolongs the detection time, and increases the detection cost. Moreover, this detection method can only detect a few of the pesticides, and the detection accuracy is low, which affects the quality of drug preparations to a certain extent. Summary of the invention

[0005] Based on this, the present invention provides a method for detecting pesticide residues in traditional Chinese medicine preparations based on the UPLC-Q-TOF-MS analysis method, which realizes high-throughput detection and analytical identification of pesticide residue components in traditional Chinese medicine preparations, can quickly and comprehensively analyze pesticide residue components, and overcomes the problems of long operation time, complex detection and difficult identification of existing gas chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] A method for detecting pesticide residues in traditional Chinese medicine preparations based on UPLC-Q-TOF-MS analysis method, comprising the following process:

[0008] S1. Preparation of test solution: If the sample to be tested is a liquid preparation of traditional Chinese medicine, the sample is filtered through an organic filter membrane; if the sample to be tested is a solid preparation of traditional Chinese medicine, the sample is dissolved in ethanol and then filtered through an organic filter membrane;

[0009] S2, separating the test solution obtained in step S1 by ultra-high performance liquid chromatography, and then detecting the separated pesticide components by quadrupole-time-of-flight mass spectrometry to obtain the retention time, molecular weight of the quasi-molecular ion and secondary mass spectrometry fragment ion data of the separated pesticide components, and further using the obtained data to establish a screening spectral library;

[0010] S3, identifying the pesticide components in the screening library established in step S2;

[0011] The ultra-high performance liquid chromatography conditions described in step S2 are as follows: the chromatographic column is Phenomenex C18; the column temperature is 28-32° C.; the mobile phase A is acetonitrile, and the mobile phase B is 0.1% formic acid aqueous solution; the gradient elution program is: 0-2 min, 0%-25% A; 2-5 min, 25%-40% A; 5-7 min 40-55% A; 7-8.5 min, 55%-70% A; 8.5-10.5 min, 70-95% A; the flow rate is 0.25 mL min -1 ; The injection volume was 0.2 μL; the detection wavelength was 254 nm.

[0012] The quadrupole-time-of-flight mass spectrometry conditions described in step S2 are as follows: using an electrospray ion source, scanning and analyzing in positive and negative ion modes; the ion source voltages are 5500V and -4500V, respectively; the ion source temperature is 500°C; the fragmentation voltage is 100V; the collision energy is ±30eV; the collision energy extension is 15eV; the nebulizing gas is nitrogen, the auxiliary gas 1 is 55psi, the auxiliary gas 2 is 55psi, and the curtain gas is 35psi; the primary mass spectrometry parent ion scanning range is 100 to 900Da, and the secondary mass spectrometry scanning daughter ion scanning range is 0 to 600Da.

[0013] Preferably, the traditional Chinese medicine preparation described in step S1 is a Qingkailing preparation, including Qingkailing preparation medicinal raw materials, intermediates, and finished products.

[0014] Preferably, the medicinal raw materials of the Qingkailing preparation include: honeysuckle, gardenia, isatis root, and scutellaria.

[0015] Preferably, the finished Qingkailing preparation includes: Qingkailing oral preparation, injection preparation, suppository, gel or ointment.

[0016] Preferably, the preparation method of the solid Chinese medicine preparation in step S1 is as follows: crush the medicinal materials or the solid Chinese medicine preparation, sieve, add 70-80% ethanol to mix, ultrasonically extract for 10-15 minutes, centrifuge to obtain the supernatant, and filter the supernatant with an organic filter membrane.

[0017] The present invention also provides a screening spectral library for pesticide components in traditional Chinese medicine preparations established using the method, wherein the screening spectral library contains retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide components; the Qingkailing preparation includes Qingkailing preparation medicinal raw materials, intermediates and finished products.

[0018] Preferably, the 30 pesticide ingredients include: aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusin, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethoprophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fonofos, chlorfenapyr, isocarbophos, methyl isofos, methamidophos, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate-methyl, phosphamidon, phosphamidon, terbufos sulfone, and terbufos sulfoxide.

[0019] Preferably, the retention time, molecular weight of the quasi-molecular ion and secondary mass spectrometry fragment ion data of the 30 pesticide components are as follows:

[0020] The retention time of aldicarb was 2.40min, the molecular weight of the quasi-molecular ion was 116.2761, and the secondary mass spectrometry fragment ion data was 70.6816;

[0021] The retention time of aldicarb sulfone was 0.94min, the molecular weight of the quasi-molecular ion was 223.2761, and the secondary mass spectrometry fragment ion data was 148.6816;

[0022] The retention time of aldicarb sulfoxide is 0.75min, the molecular weight of the quasi-molecular ion is 207.2761, and the secondary mass spectrometry fragment ion data is 89.6816;

[0023] The retention time of thiosulfate was 8.50min, the molecular weight of the quasi-molecular ion was 271.4065, and the secondary mass spectrometry fragment ion data was 131.3986;

[0024] The retention time of carbofuran was 4.08min, the molecular weight of the quasi-molecular ion was 222.2660, and the secondary mass spectrometry fragment ion data was 165.2581;

[0025] The retention time of 3-hydroxythiocarb is 1.32min, the molecular weight of the quasi-molecular ion is 238.1660, and the secondary mass spectrometry fragment ion data is 163.1581;

[0026] The retention time of chlordimeform was 1.08min, the molecular weight of the quasi-molecular ion was 197.18816, and the secondary mass spectrometry fragment ion data was 117.6802;

[0027] The retention time of chlorsulfuron is 4.28min, the molecular weight of the quasi-molecular ion is 358.0872, and the secondary mass spectrometry fragment ion data is 167.9792;

[0028] The retention time of coumaphos was 9.26min, the molecular weight of the quasi-molecular ion was 363.0819, and the secondary mass spectrometry fragment ion data was 334.7404;

[0029] The retention time of demeton was 6.69 min, the molecular weight of the quasi-molecular ion was 259.0528, and the secondary mass spectrometry fragment ion data was 61.0448;

[0030] The retention time of ethametsulfuron was 4.89 min, the molecular weight of the quasi-molecular ion was 411.5238, and the secondary mass spectrometry fragment ion data was 196.4159;

[0031] The retention time of ethoprophos was 7.24min, the molecular weight of the quasi-molecular ion was 243.1108, and the secondary mass spectrometry fragment ion data was 130.9028;

[0032] The retention time of fenamiphos was 7.08min, the molecular weight of the quasi-molecular ion was 304.1726, and the secondary mass spectrometry fragment ion data was 202.3468;

[0033] The retention time of fenamiphos sulfone was 3.92 min, the molecular weight of the quasi-molecular ion was 336.1726, and the secondary mass spectrometry fragment ion data was 308.3468;

[0034] The retention time of fenamiphos sulfoxide was 2.20min, the molecular weight of the quasi-molecular ion was 320.1726, and the secondary mass spectrometry fragment ion data was 233.3468;

[0035] The retention time of fofophos was 9.20min, the molecular weight of the quasi-molecular ion was 247.2271, and the secondary mass spectrometry fragment ion data was 109.3192;

[0036] The retention time of chlorfenapyr is 8.40min, the molecular weight of the quasi-molecular ion is 314.0559, and the secondary mass spectrometry fragment ion data is 120.7479;

[0037] The retention time of isocarbophos was 6.38min, the molecular weight of the quasi-molecular ion was 231.0029, and the secondary mass spectrometry fragment ion data was 109.2950;

[0038] The retention time of methyl isofenphos was 9.16min, the molecular weight of the quasi-molecular ion was 332.0836, and the secondary mass spectrometry fragment ion data was 272.7356;

[0039] The retention time of methamidophos was 0.75min, the molecular weight of the quasi-molecular ion was 142.0404, and the secondary mass spectrometry fragment ion data was 125.1325;

[0040] The retention time of metsulfuron-methyl was 3.76 min, the molecular weight of the quasi-molecular ion was 382.0800, and the secondary mass spectrometry fragment ion data was 199.0721;

[0041] The retention time of monocrotophos was 0.87min, the molecular weight of the quasi-molecular ion was 224.0771, and the secondary mass spectrometry fragment ion data was 127.0691;

[0042] The retention time of phorate was 9.41min, the molecular weight of the quasi-molecular ion was 251.0228, and the secondary mass spectrometry fragment ion data was 47.0149;

[0043] The retention time of phorate sulfone was 6.34min, the molecular weight of the quasi-molecular ion was 293.0192, and the secondary mass spectrometry fragment ion data was 96.8839;

[0044] The retention time of phorate sulfoxide was 4.65min, the molecular weight of the quasi-molecular ion was 277.0919, and the secondary mass spectrometry fragment ion data was 142.9839;

[0045] The retention time of thiocyclic phosphorus was 1.77min, the molecular weight of the quasi-molecular ion was 256.1531, and the secondary mass spectrometry fragment ion data was 167.9451;

[0046] The retention time of phosphamide was 2.36min, the molecular weight of the quasi-molecular ion was 300.0028, and the secondary mass spectrometry fragment ion data was 127.0948;

[0047] The retention time of chlorpyrifos was 9.40min, the molecular weight of the quasi-molecular ion was 323.3474, and the secondary mass spectrometry fragment ion data was 96.8268;

[0048] The retention time of terbuthion sulfone was 7.49 min, the molecular weight of the quasi-molecular ion was 321.0456, and the secondary mass spectrometry fragment ion data was 96.3774;

[0049] The retention time of terbuthion sulfoxide was 6.19 min, the molecular weight of the quasi-molecular ion was 289.3796, and the secondary mass spectrometry fragment ion data was 96.9174.

[0050] The present invention also provides an application of the pesticide component screening spectral library in the detection of pesticide residues in traditional Chinese medicine preparations, wherein the screening spectral library contains retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide components.

[0051] The 30 pesticide ingredients are aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusin, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethoprophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fenofos, chlorfenapyr, isocarb, methyl isofos, methamidophos, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate, phosphamidon, chlorpyrifos, terbufos sulfone, terbufos sulfoxide;

[0052] The retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of the 30 pesticide components are as follows:

[0053] The retention time of aldicarb was 2.40min, the molecular weight of the quasi-molecular ion was 116.2761, and the secondary mass spectrometry fragment ion data was 70.6816;

[0054] The retention time of aldicarb sulfone was 0.94min, the molecular weight of the quasi-molecular ion was 223.2761, and the secondary mass spectrometry fragment ion data was 148.6816;

[0055] The retention time of aldicarb sulfoxide is 0.75min, the molecular weight of the quasi-molecular ion is 207.2761, and the secondary mass spectrometry fragment ion data is 89.6816;

[0056] The retention time of thiosulfate was 8.50min, the molecular weight of the quasi-molecular ion was 271.4065, and the secondary mass spectrometry fragment ion data was 131.3986;

[0057] The retention time of carbofuran was 4.08min, the molecular weight of the quasi-molecular ion was 222.2660, and the secondary mass spectrometry fragment ion data was 165.2581;

[0058] The retention time of 3-hydroxythiocarb is 1.32min, the molecular weight of the quasi-molecular ion is 238.1660, and the secondary mass spectrometry fragment ion data is 163.1581;

[0059] The retention time of chlordimeform was 1.08min, the molecular weight of the quasi-molecular ion was 197.18816, and the secondary mass spectrometry fragment ion data was 117.6802;

[0060] The retention time of chlorsulfuron is 4.28min, the molecular weight of the quasi-molecular ion is 358.0872, and the secondary mass spectrometry fragment ion data is 167.9792;

[0061] The retention time of coumaphos was 9.26min, the molecular weight of the quasi-molecular ion was 363.0819, and the secondary mass spectrometry fragment ion data was 334.7404;

[0062] The retention time of demeton was 6.69 min, the molecular weight of the quasi-molecular ion was 259.0528, and the secondary mass spectrometry fragment ion data was 61.0448;

[0063] The retention time of ethametsulfuron was 4.89 min, the molecular weight of the quasi-molecular ion was 411.5238, and the secondary mass spectrometry fragment ion data was 196.4159;

[0064] The retention time of ethoprophos was 7.24min, the molecular weight of the quasi-molecular ion was 243.1108, and the secondary mass spectrometry fragment ion data was 130.9028;

[0065] The retention time of fenamiphos was 7.08min, the molecular weight of the quasi-molecular ion was 304.1726, and the secondary mass spectrometry fragment ion data was 202.3468;

[0066] The retention time of fenamiphos sulfone was 3.92 min, the molecular weight of the quasi-molecular ion was 336.1726, and the secondary mass spectrometry fragment ion data was 308.3468;

[0067] The retention time of fenamiphos sulfoxide was 2.20min, the molecular weight of the quasi-molecular ion was 320.1726, and the secondary mass spectrometry fragment ion data was 233.3468;

[0068] The retention time of fofophos was 9.20min, the molecular weight of the quasi-molecular ion was 247.2271, and the secondary mass spectrometry fragment ion data was 109.3192;

[0069] The retention time of chlorfenapyr is 8.40min, the molecular weight of the quasi-molecular ion is 314.0559, and the secondary mass spectrometry fragment ion data is 120.7479;

[0070] The retention time of isocarbophos was 6.38min, the molecular weight of the quasi-molecular ion was 231.0029, and the secondary mass spectrometry fragment ion data was 109.2950;

[0071] The retention time of methyl isofenphos was 9.16min, the molecular weight of the quasi-molecular ion was 332.0836, and the secondary mass spectrometry fragment ion data was 272.7356;

[0072] The retention time of methamidophos was 0.75min, the molecular weight of the quasi-molecular ion was 142.0404, and the secondary mass spectrometry fragment ion data was 125.1325;

[0073] The retention time of metsulfuron-methyl was 3.76 min, the molecular weight of the quasi-molecular ion was 382.0800, and the secondary mass spectrometry fragment ion data was 199.0721;

[0074] The retention time of monocrotophos was 0.87min, the molecular weight of the quasi-molecular ion was 224.0771, and the secondary mass spectrometry fragment ion data was 127.0691;

[0075] The retention time of phorate was 9.41min, the molecular weight of the quasi-molecular ion was 251.0228, and the secondary mass spectrometry fragment ion data was 47.0149;

[0076] The retention time of phorate sulfone was 6.34min, the molecular weight of the quasi-molecular ion was 293.0192, and the secondary mass spectrometry fragment ion data was 96.8839;

[0077] The retention time of phorate sulfoxide was 4.65min, the molecular weight of the quasi-molecular ion was 277.0919, and the secondary mass spectrometry fragment ion data was 142.9839;

[0078] The retention time of thiocyclic phosphorus was 1.77min, the molecular weight of the quasi-molecular ion was 256.1531, and the secondary mass spectrometry fragment ion data was 167.9451;

[0079] The retention time of phosphamide was 2.36min, the molecular weight of the quasi-molecular ion was 300.0028, and the secondary mass spectrometry fragment ion data was 127.0948;

[0080] The retention time of chlorpyrifos was 9.40min, the molecular weight of the quasi-molecular ion was 323.3474, and the secondary mass spectrometry fragment ion data was 96.8268;

[0081] The retention time of terbuthion sulfone was 7.49 min, the molecular weight of the quasi-molecular ion was 321.0456, and the secondary mass spectrometry fragment ion data was 96.3774;

[0082] The retention time of terbuthion sulfoxide was 6.19 min, the molecular weight of the quasi-molecular ion was 289.3796, and the secondary mass spectrometry fragment ion data was 96.9174.

[0083] The Chinese medicine preparation is a Qingkailing preparation, including Qingkailing preparation medicinal raw materials, intermediates and finished products.

[0084] The present invention also provides a method for detecting pesticide residues in traditional Chinese medicine preparations. The method comprises: using the UPLC-Q-TOF-MS technology to detect and identify pesticide residues in traditional Chinese medicine preparations of different batches or in the same batch at different time points. The pesticide residues include any one or several of aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusafos, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethoprophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fonofos, chlorfenapyr, isocarbophos, methyl isofos, methamidophos, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate, phosphamidon, chlorpyrifos, terbufos sulfone and terbufos sulfoxide; the traditional Chinese medicine preparation is a Qingkailing preparation, including medicinal raw materials, intermediates and finished products of the Qingkailing preparation.

[0085] The present invention provides a method for analyzing and identifying pesticide residues in traditional Chinese medicine preparations based on UPLC-Q-TOF-MS technology, provides a screening spectral library for pesticide components in traditional Chinese medicine preparations established based on the technology, and provides a method for detecting pesticide residue components in traditional Chinese medicine preparations.

[0086] Compared with the prior art, the present invention has the following advantages:

[0087] (1) When analyzing the pesticide residue components in traditional Chinese medicine preparations according to the method of the present invention, the pretreatment is simple, the component loss is small, the detection is more accurate and sensitive, and the time consumption is short. 30 kinds of pesticide residue components in traditional Chinese medicine preparations can be effectively identified within 15 minutes;

[0088] (2) The present invention realizes high-throughput detection and analytical identification of pesticide residue components in traditional Chinese medicine preparations, and can quickly and comprehensively analyze the pesticide residue components in traditional Chinese medicine preparations, overcoming the problems of long operation time, complex detection and difficult identification of existing gas chromatography-tandem mass spectrometry and liquid chromatography-tandem mass spectrometry, which is beneficial to the quality control of traditional Chinese medicine preparations, Chinese medicinal materials, intermediates and finished products.

[0089] (3) Compared with the existing pharmacopoeia detection methods, the detection sensitivity, accuracy and efficiency have been greatly improved, the detection cost is lower, and it is more environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0090] Figure 1 This is a chromatogram of the chemical components of Qingkailing Injection detected based on UPLC-Q-TOF-MS technology. DETAILED DESCRIPTION

[0091] The present invention is further explained below in conjunction with specific examples, but it should be noted that the following examples are only used to explain the present invention, and cannot be used to limit the present invention, and all technical solutions that are the same or similar to the present invention are within the scope of protection of the present invention. If no specific technology or conditions are specified in this embodiment, the operation is carried out according to the conventional technical methods and instrument instructions in the art; if the manufacturer of the reagents or instruments is not specified, they are all conventional products that can be purchased commercially.

[0092] Example 1 Establishment of a screening library for 30 pesticide components in traditional Chinese medicine preparations based on UPLC-Q-TOF-MS technology

[0093] The present invention establishes a screening spectral library for pesticide components in a traditional Chinese medicine preparation based on UPLC-Q-TOF-MS technology, wherein the screening spectral library contains retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide components, and the traditional Chinese medicine preparation is a Qingkailing preparation.

[0094] 1. Experimental methods

[0095] The instruments and reagents used in the embodiments of the present invention are: Triple-TOF TM 5600 + The instrument was a triple quadrupole time-of-flight mass spectrometer equipped with an electrospray ion source. The data acquisition software was Analyst TF 1.7 software. The data processing software was Peakview 2.0. LC-MS grade acetonitrile was purchased from Merck KGaA (Darmstadt, Germany). Chromatographic grade formic acid was purchased from Tianjin Komiou Chemical Reagent Co., Ltd. (Tianjin, China). Purified water was purchased from Watsons Group Co., Ltd. (Guangzhou, China). 30 pesticide reference substances were purchased from the National Institutes for Food and Drug Control.

[0096] The ultra-high performance liquid chromatography conditions were as follows: the chromatographic column was Phenomenex C18; the column temperature was 28-32°C; the mobile phase A was acetonitrile and the mobile phase B was 0.1% formic acid aqueous solution; the gradient elution program was: 0-2 min, 0%-25% A; 2-5 min, 25%-40% A; 5-7 min, 40-55% A; 7-8.5 min, 55%-70% A; 8.5-10.5 min, 70-95% A; the flow rate was 0.25 mL min -1 ; The injection volume was 0.2 μL; the detection wavelength was 254 nm.

[0097] The conditions of quadrupole-time of flight mass spectrometry are as follows: using an electrospray ion source, scanning and analysis in positive and negative ion modes; the ion source voltages are 5500 V and -4500 V, respectively; the ion source temperature is 500°C; the fragmentation voltage is 100 V; the collision energy is ±30 eV; the collision energy extension is 15 eV; the nebulizing gas is nitrogen, the auxiliary gas 1 is 55 psi, the auxiliary gas 2 is 55 psi, and the curtain gas is 35 psi; the primary mass spectrometry parent ion scanning range is 100-900 Da, and the secondary mass spectrometry scanning daughter ion scanning range is 0-600 Da.

[0098] The data processing method is as follows: the present invention uses Peakview 2.0 software to process the data. First, the chemical structure of each pesticide component is analyzed by mass spectrometry to summarize the fragmentation rules and neutral loss rules; secondly, the target compound is oriented by structural characteristics and structural correlation, and a target precursor ion list is constructed; thirdly, the correctness of the structure of the target precursor ion is verified by secondary mass spectrometry, and the pesticide component information is comprehensively and quickly analyzed by comprehensively analyzing the mass spectrum information, retention time and other data.

[0099] 2. Experimental results

[0100] The retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide residue components are as follows:

[0101] The retention time of aldicarb was 2.40min, the molecular weight of the quasi-molecular ion was 116.2761, and the secondary mass spectrometry fragment ion data was 70.6816;

[0102] The retention time of aldicarb sulfone was 0.94min, the molecular weight of the quasi-molecular ion was 223.2761, and the secondary mass spectrometry fragment ion data was 148.6816;

[0103] The retention time of aldicarb sulfoxide is 0.75min, the molecular weight of the quasi-molecular ion is 207.2761, and the secondary mass spectrometry fragment ion data is 89.6816;

[0104] The retention time of thiosulfate was 8.50min, the molecular weight of the quasi-molecular ion was 271.4065, and the secondary mass spectrometry fragment ion data was 131.3986;

[0105] The retention time of carbofuran was 4.08min, the molecular weight of the quasi-molecular ion was 222.2660, and the secondary mass spectrometry fragment ion data was 165.2581;

[0106] The retention time of 3-hydroxythiocarb is 1.32min, the molecular weight of the quasi-molecular ion is 238.1660, and the secondary mass spectrometry fragment ion data is 163.1581;

[0107] The retention time of chlordimeform was 1.08min, the molecular weight of the quasi-molecular ion was 197.18816, and the secondary mass spectrometry fragment ion data was 117.6802;

[0108] The retention time of chlorsulfuron is 4.28min, the molecular weight of the quasi-molecular ion is 358.0872, and the secondary mass spectrometry fragment ion data is 167.9792;

[0109] The retention time of coumaphos was 9.26min, the molecular weight of the quasi-molecular ion was 363.0819, and the secondary mass spectrometry fragment ion data was 334.7404;

[0110] The retention time of demeton was 6.69 min, the molecular weight of the quasi-molecular ion was 259.0528, and the secondary mass spectrometry fragment ion data was 61.0448;

[0111] The retention time of ethametsulfuron was 4.89 min, the molecular weight of the quasi-molecular ion was 411.5238, and the secondary mass spectrometry fragment ion data was 196.4159;

[0112] The retention time of ethoprophos was 7.24min, the molecular weight of the quasi-molecular ion was 243.1108, and the secondary mass spectrometry fragment ion data was 130.9028;

[0113] The retention time of fenamiphos was 7.08min, the molecular weight of the quasi-molecular ion was 304.1726, and the secondary mass spectrometry fragment ion data was 202.3468;

[0114] The retention time of fenamiphos sulfone was 3.92 min, the molecular weight of the quasi-molecular ion was 336.1726, and the secondary mass spectrometry fragment ion data was 308.3468;

[0115] The retention time of fenamiphos sulfoxide was 2.20min, the molecular weight of the quasi-molecular ion was 320.1726, and the secondary mass spectrometry fragment ion data was 233.3468;

[0116] The retention time of fofophos was 9.20min, the molecular weight of the quasi-molecular ion was 247.2271, and the secondary mass spectrometry fragment ion data was 109.3192;

[0117] The retention time of chlorfenapyr is 8.40min, the molecular weight of the quasi-molecular ion is 314.0559, and the secondary mass spectrometry fragment ion data is 120.7479;

[0118] The retention time of isocarbophos was 6.38min, the molecular weight of the quasi-molecular ion was 231.0029, and the secondary mass spectrometry fragment ion data was 109.2950;

[0119] The retention time of methyl isofenphos was 9.16min, the molecular weight of the quasi-molecular ion was 332.0836, and the secondary mass spectrometry fragment ion data was 272.7356;

[0120] The retention time of methamidophos was 0.75min, the molecular weight of the quasi-molecular ion was 142.0404, and the secondary mass spectrometry fragment ion data was 125.1325;

[0121] The retention time of metsulfuron-methyl was 3.76 min, the molecular weight of the quasi-molecular ion was 382.0800, and the secondary mass spectrometry fragment ion data was 199.0721;

[0122] The retention time of monocrotophos was 0.87min, the molecular weight of the quasi-molecular ion was 224.0771, and the secondary mass spectrometry fragment ion data was 127.0691;

[0123] The retention time of phorate was 9.41min, the molecular weight of the quasi-molecular ion was 251.0228, and the secondary mass spectrometry fragment ion data was 47.0149;

[0124] The retention time of phorate sulfone was 6.34min, the molecular weight of the quasi-molecular ion was 293.0192, and the secondary mass spectrometry fragment ion data was 96.8839;

[0125] The retention time of phorate sulfoxide was 4.65min, the molecular weight of the quasi-molecular ion was 277.0919, and the secondary mass spectrometry fragment ion data was 142.9839;

[0126] The retention time of thiocyclic phosphorus was 1.77min, the molecular weight of the quasi-molecular ion was 256.1531, and the secondary mass spectrometry fragment ion data was 167.9451;

[0127] The retention time of phosphamide was 2.36min, the molecular weight of the quasi-molecular ion was 300.0028, and the secondary mass spectrometry fragment ion data was 127.0948;

[0128] The retention time of chlorpyrifos was 9.40min, the molecular weight of the quasi-molecular ion was 323.3474, and the secondary mass spectrometry fragment ion data was 96.8268;

[0129] The retention time of terbuthion sulfone was 7.49 min, the molecular weight of the quasi-molecular ion was 321.0456, and the secondary mass spectrometry fragment ion data was 96.3774;

[0130] The retention time of terbuthion sulfoxide was 6.19 min, the molecular weight of the quasi-molecular ion was 289.3796, and the secondary mass spectrometry fragment ion data was 96.9174.

[0131] Example 2 A method for detecting pesticide residues in a traditional Chinese medicine preparation

[0132] The present invention also provides a method for detecting pesticide residues in traditional Chinese medicine preparations, using the method described in Example 1 to separate and detect pesticide residues in traditional Chinese medicine preparations of different batches or at different time points in the same batch, obtaining the retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of the separated components; comparing the obtained data with the screening spectral library described in Example 1; judging the pesticide residues in traditional Chinese medicine preparations of different batches or at different time points in the same batch according to the comparison results. The traditional Chinese medicine preparation is a Qingkailing preparation, including Qingkailing preparation medicinal material raw materials, intermediates and finished products.

[0133] Specifically, the criteria for judging the results are as follows:

[0134] If the retention time deviation of the compared components is ±0.2 min, the accurate mass deviation is ±5 mDa, and two or more fragment ions can be matched, it indicates that the compared components are the same substance;

[0135] 2. Application of the detection method in Qingkailing injection

[0136] The present invention uses the above detection method to detect pesticide residues in different batches of Qingkailing injection (batch numbers: 1243001, 1243002, 1243003), and uses the screening library for identification. Preparation of Qingkailing injection test solution: Qingkailing injection is filtered using a 0.22 μm PTFE filter membrane. The chromatogram of Qingkailing injection under the chromatographic conditions of Example 1 is shown in FIG. Figure 1 As shown, Figure 1 The vertical axis in the figure is the peak ultraviolet absorption intensity of the components in the sample, and the peak is the peak substance of different components in the actual sample (non-pesticide residue). Figure 1 It can be seen that the samples of Qingkailing injection from different batches tested do not contain the above 30 pesticide components. The results show that there is no pesticide residue in different batches of Qingkailing, which also shows that this detection method can be used for the detection of pesticide residues in Qingkailing preparations.

[0137] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for detecting pesticide residues in traditional Chinese medicine preparations based on UPLC-Q-TOF-MS analysis method, characterized in that: The process includes the following: S1. Preparation of test solution: If the sample to be tested is a liquid preparation of traditional Chinese medicine, the sample is filtered through an organic filter membrane; if the sample to be tested is a solid preparation of medicinal materials, the sample is dissolved in ethanol and then filtered through an organic filter membrane; S2, separating the test solution obtained in step S1 by ultra-high performance liquid chromatography, and then detecting the separated pesticide components by quadrupole-time-of-flight mass spectrometry to obtain the retention time, molecular weight of the quasi-molecular ion and secondary mass spectrometry fragment ion data of the separated pesticide components, and further using the obtained data to establish a screening spectral library; S3, identifying the pesticide components in the screening library established in step S2; The ultra-high performance liquid chromatography conditions described in step S2 are as follows: the chromatographic column is Phenomenex C18; the column temperature is 28-32°C; the mobile phase A is acetonitrile, and the mobile phase B is 0.1% formic acid aqueous solution; the gradient elution program is: 0-2 min, 0%-25% A; 2-5 min, 25%-40% A; 5-7 min 40-55% A; 7-8.5 min, 55%-70% A; 8.5-10.5 min, 70-95% A; the flow rate is 0.25 mL min -1 ; The injection volume was 0.2 μL; the detection wavelength was 254 nm; The quadrupole-time-of-flight mass spectrometry conditions described in step S2 are as follows: using an electrospray ion source, scanning and analyzing in positive and negative ion modes; the ion source voltages are 5500V and -4500V, respectively; the ion source temperature is 500°C; the fragmentation voltage is 100V; the collision energy is ±30eV; the collision energy extension is 15eV; the nebulizing gas is nitrogen, the auxiliary gas 1 is 55psi, the auxiliary gas 2 is 55psi, and the curtain gas is 35psi; the primary mass spectrometry parent ion scanning range is 100 to 900Da, and the secondary mass spectrometry scanning daughter ion scanning range is 0 to 600Da.

2. The method according to claim 1, characterized in that The traditional Chinese medicine preparation described in step S1 is a Qingkailing preparation, including Qingkailing preparation medicinal raw materials, intermediates, and finished products.

3. The method according to claim 2, characterized in that The medicinal raw materials of Qingkailing preparation include: honeysuckle, gardenia, isatis root and scutellaria baicalensis.

4. The method according to claim 2, characterized in that The finished products of Qingkailing preparations include: Qingkailing oral preparations, injection preparations, suppositories, gels or ointments.

5. The method according to claim 1, characterized in that The preparation method of the medicinal solid preparation in step S1 is as follows: crush the medicinal solid preparation, sieve it, add 70-80% ethanol to mix it, ultrasonically extract it for 10-15 minutes, centrifuge it to obtain the supernatant, and filter the supernatant with an organic filter membrane to obtain the solid preparation.

6. A screening library for pesticide components in traditional Chinese medicine preparations established by the method according to any one of claims 1 to 5, characterized in that: The screening spectral library contains the retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide components; the traditional Chinese medicine preparation is Qingkailing preparation, including Qingkailing preparation medicinal raw materials, intermediates or finished products.

7. The pesticide component screening library according to claim 6, characterized in that: The 30 pesticide ingredients include aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusin, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethanophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fonofos, chlorpyrifos, isocarb, methyl isofos, methyl parathion, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate-methyl, phosphamidon, phosphamidon, terbufos sulfone, and terbufos sulfoxide.

8. The pesticide component screening library according to claim 6, characterized in that: The retention time, molecular weight of quasi-molecular ions and secondary mass spectrometry fragment ion data of 30 pesticide components are as follows: The retention time of aldicarb was 2.40min, the molecular weight of the quasi-molecular ion was 116.2761, and the secondary mass spectrometry fragment ion data was 70.6816; The retention time of aldicarb sulfone was 0.94min, the molecular weight of the quasi-molecular ion was 223.2761, and the secondary mass spectrometry fragment ion data was 148.6816; The retention time of aldicarb sulfoxide is 0.75min, the molecular weight of the quasi-molecular ion is 207.2761, and the secondary mass spectrometry fragment ion data is 89.6816; The retention time of thiosulfate was 8.50min, the molecular weight of the quasi-molecular ion was 271.4065, and the secondary mass spectrometry fragment ion data was 131.3986; The retention time of carbofuran was 4.08min, the molecular weight of the quasi-molecular ion was 222.2660, and the secondary mass spectrometry fragment ion data was 165.2581; The retention time of 3-hydroxythiocarb is 1.32min, the molecular weight of the quasi-molecular ion is 238.1660, and the secondary mass spectrometry fragment ion data is 163.1581; The retention time of chlordimeform was 1.08min, the molecular weight of the quasi-molecular ion was 197.18816, and the secondary mass spectrometry fragment ion data was 117.6802; The retention time of chlorsulfuron is 4.28min, the molecular weight of the quasi-molecular ion is 358.0872, and the secondary mass spectrometry fragment ion data is 167.9792; The retention time of coumaphos was 9.26min, the molecular weight of the quasi-molecular ion was 363.0819, and the secondary mass spectrometry fragment ion data was 334.7404; The retention time of demeton was 6.69 min, the molecular weight of the quasi-molecular ion was 259.0528, and the secondary mass spectrometry fragment ion data was 61.0448; The retention time of ethametsulfuron was 4.89 min, the molecular weight of the quasi-molecular ion was 411.5238, and the secondary mass spectrometry fragment ion data was 196.4159; The retention time of ethoprophos was 7.24min, the molecular weight of the quasi-molecular ion was 243.1108, and the secondary mass spectrometry fragment ion data was 130.9028; The retention time of fenamiphos was 7.08min, the molecular weight of the quasi-molecular ion was 304.1726, and the secondary mass spectrometry fragment ion data was 202.3468; The retention time of fenamiphos sulfone was 3.92 min, the molecular weight of the quasi-molecular ion was 336.1726, and the secondary mass spectrometry fragment ion data was 308.3468; The retention time of fenamiphos sulfoxide was 2.20min, the molecular weight of the quasi-molecular ion was 320.1726, and the secondary mass spectrometry fragment ion data was 233.3468; The retention time of fofophos was 9.20min, the molecular weight of the quasi-molecular ion was 247.2271, and the secondary mass spectrometry fragment ion data was 109.3192; The retention time of chlorfenapyr is 8.40min, the molecular weight of the quasi-molecular ion is 314.0559, and the secondary mass spectrometry fragment ion data is 120.7479; The retention time of isocarbophos was 6.38min, the molecular weight of the quasi-molecular ion was 231.0029, and the secondary mass spectrometry fragment ion data was 109.2950; The retention time of methyl isofenphos was 9.16min, the molecular weight of the quasi-molecular ion was 332.0836, and the secondary mass spectrometry fragment ion data was 272.7356; The retention time of methamidophos was 0.75min, the molecular weight of the quasi-molecular ion was 142.0404, and the secondary mass spectrometry fragment ion data was 125.1325; The retention time of metsulfuron-methyl was 3.76 min, the molecular weight of the quasi-molecular ion was 382.0800, and the secondary mass spectrometry fragment ion data was 199.0721; The retention time of monocrotophos was 0.87min, the molecular weight of the quasi-molecular ion was 224.0771, and the secondary mass spectrometry fragment ion data was 127.0691; The retention time of phorate was 9.41min, the molecular weight of the quasi-molecular ion was 251.0228, and the secondary mass spectrometry fragment ion data was 47.0149; The retention time of phorate sulfone was 6.34min, the molecular weight of the quasi-molecular ion was 293.0192, and the secondary mass spectrometry fragment ion data was 96.8839; The retention time of phorate sulfoxide was 4.65min, the molecular weight of the quasi-molecular ion was 277.0919, and the secondary mass spectrometry fragment ion data was 142.9839; The retention time of thiocyclic phosphorus was 1.77min, the molecular weight of the quasi-molecular ion was 256.1531, and the secondary mass spectrometry fragment ion data was 167.9451; The retention time of phosphamide was 2.36min, the molecular weight of the quasi-molecular ion was 300.0028, and the secondary mass spectrometry fragment ion data was 127.0948; The retention time of chlorpyrifos was 9.40min, the molecular weight of the quasi-molecular ion was 323.3474, and the secondary mass spectrometry fragment ion data was 96.8268; The retention time of terbuthion sulfone was 7.49 min, the molecular weight of the quasi-molecular ion was 321.0456, and the secondary mass spectrometry fragment ion data was 96.3774; The retention time of terbuthion sulfoxide was 6.19 min, the molecular weight of the quasi-molecular ion was 289.3796, and the secondary mass spectrometry fragment ion data was 96.9174.

9. An application of the pesticide component screening library as claimed in any one of claims 6 to 8 in the detection of pesticide residues in traditional Chinese medicine preparations, characterized in that: The screening library contains retention time, molecular weight of quasi-molecular ion and secondary mass spectrometry fragment ion data of 30 pesticide components, and the 30 pesticide components are aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusin, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethoprophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fonofos, chlorfenapyr, isocarbophos, methyl isofos, methamidophos, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate, phosphamidon, chlorpyrifos, terbufos sulfone and terbufos sulfoxide; the traditional Chinese medicine preparation is Qingkailing preparation, including Qingkailing preparation medicinal material raw materials, intermediates or finished products.

10. A method for detecting pesticide residues in traditional Chinese medicine preparations, characterized in that: The method according to any one of claims 1 to 5 is used to detect and identify pesticide residues in different batches of traditional Chinese medicine preparations or in the same batch at different time points, wherein the pesticide residues include any one or more of aldicarb, aldicarb sulfone, aldicarb sulfoxide, cadusafos, carbofuran, 3-hydroxycarbofuran, chlordimeform, chlorsulfuron, coumaphos, demeton, ethametsulfuron, ethoprophos, fenamiphos, fenamiphos sulfone, fenamiphos sulfoxide, fonofos, chlorfenapyr, isocarbophos, methyl isofos, methamidophos, metsulfuron, monocrotophos, phorate, phorate sulfone, phorate sulfoxide, thiophanate, phosphamidon, chlorpyrifos, terbufos sulfone and terbufos sulfoxide; the traditional Chinese medicine preparation is a Qingkailing preparation, including raw materials, intermediates and finished products of the Qingkailing preparation.

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