CN113155986A

CN113155986A - Method for detecting perchlorate and chlorate in panax notoginseng

Info

Publication number: CN113155986A
Application number: CN202011567975.4A
Authority: CN
Inventors: 胡佳哲; 赖宇红; 方继辉
Original assignee: Guangdong Institute For Drug Control (guangdong Institute For Drug Quality
Current assignee: Guangdong Institute For Drug Control (guangdong Institute For Drug Quality
Priority date: 2020-12-25
Filing date: 2020-12-25
Publication date: 2021-07-23
Anticipated expiration: 2040-12-25
Also published as: CN113155986B

Abstract

The invention relates to a method for detecting perchlorate and chlorate in panax notoginseng, belonging to the technical field of traditional Chinese medicine detection. The invention takes precious Chinese medicinal material pseudo-ginseng as a research object, establishes an HPLC-MS/MS detection method for detecting perchlorate and chlorate in Chinese medicinal materials such as pseudo-ginseng and the like, fills the research blank of perchlorate which is an environmental pollutant in the field of Chinese medicinal materials, provides technical support for the research of perchlorate in Chinese medicinal materials, protects the medication safety of consumers, and promotes and develops great economic and social benefits of Chinese medicinal materials. The detection method of the invention has strong practicability, simple and convenient operation, accurate result and good reproducibility.

Description

Method for detecting perchlorate and chlorate in panax notoginseng

Technical Field

The invention relates to a method for detecting perchlorate and chlorate in panax notoginseng, belonging to the technical field of traditional Chinese medicine detection.

Background

The record of the compendium of materia Medica is Shi Yi: ginseng, radix Ginseng and Notoginseng are the most valuable Chinese herbs because they tonify qi the first and tonify blood the first, and both the flavor and action of Notoginseng are the same. Since ancient times, China has a statement that ginseng exists in the north and pseudo-ginseng exists in the south, and pseudo-ginseng is a famous and precious Chinese medicinal material and is a large variety next to ginseng. At present, more than 400 Chinese patent medicine varieties using pseudo-ginseng as raw materials exist in China, and 1300 manufacturers involved need 800 kilo of pseudo-ginseng every year, and the sales of pseudo-ginseng-containing Chinese patent medicine preparations reaches 320 billion yuan. The pseudo-ginseng has good hemostatic effect and remarkable hematopoietic function; can strengthen and improve coronary microcirculation, and dilate blood vessel; relieving pain, and has effects of relieving fatigue, and improving learning and memory ability; anti-inflammatory effects; anti-tumor effect; anti-aging and anti-oxidation effects; reducing blood lipid and cholesterol; has liver protecting effect. Notoginseng radix is dry root and rhizome of Panax notoginseng belonging to Araliaceae, and is prepared by cleaning Notoginseng radix in soil with water, sun drying, and pulverizing into powder for administration. At present, most middle-aged and elderly people paying attention to health preservation take the pseudo-ginseng powder for a long time, and take the pseudo-ginseng powder with warm boiled water once in the morning and at night, but the pollution of perchlorate in tap water, irrigation water, soil, ore, surface water, sludge and other environments is serious. Perchlorate is a persistent environmental pollutant, is extremely difficult to degrade under natural conditions, can interfere the synthesis and secretion of thyroxine, influences normal metabolism, and has serious harm to nerves, reproduction, heredity and the like.

At present, the research on perchlorate pollution in the fields of environment, food, water quality and the like is widely regarded, but the research reports in the field of traditional Chinese medicinal materials are few. The method aims to establish an HPLC-MS/MS detection method for detecting perchlorate and chlorate in traditional Chinese medicinal materials such as pseudo-ginseng by taking a precious traditional Chinese medicinal material pseudo-ginseng as a research object, fills the research blank of perchlorate which is an environmental pollutant in the field of traditional Chinese medicinal materials, provides technical support for the research of perchlorate in traditional Chinese medicinal materials, protects the medication safety of consumers, and promotes and develops great economic and social benefits of traditional Chinese medicinal materials.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides the method for detecting the perchlorate and the chlorate in the panax notoginseng, which has the advantages of strong practicability, simple and convenient operation, accurate result and good reproducibility.

In order to achieve the purpose, the invention adopts the technical scheme that: a method for detecting perchlorate and chlorate in pseudo-ginseng, the method comprising the steps of:

(1) preparation of control solutions:

preparing external standard solution from perchlorate solution and chlorate solution¹⁸Marking the perchlorate intermediate solution by O to prepare an internal standard solution, and then preparing a standard series working solution by using the external standard solution and the internal standard solution;

(2) preparation of a test solution:

drying Notoginseng radix, pulverizing, and sieving to obtain Notoginseng radix powder; mixing Notoginseng radix powder, alkaline aluminum oxide, graphite carbon black powder,¹⁸Adding the O-labeled perchlorate intermediate solution and diatomite into an extraction tank, taking water as an extraction solvent, carrying out rapid extraction, transferring an extraction solution after extraction and purification are finished, carrying out constant volume with water, shaking up, filtering, and measuring by a liquid chromatography-tandem mass spectrometer;

(3) and (3) detection: precisely measuring the test solution and the reference solution, and injecting into a liquid chromatogram-tandem mass spectrometer for qualitative detection or quantitative detection.

As a preferred embodiment of the method of the present invention, in the step (1), the preparation of the reference solution is specifically as follows:

preparation of external standard solution: preparing a mixed solution, wherein the concentrations of a perchlorate solution and a chlorate solution in the mixed solution are both 10 mu g/mL; then preparing the mixed solution into a mixed standard intermediate solution with the concentration of 1 mu g/mL;

preparation of internal standard solution: prepared at 10. mu.g/mL¹⁸Marking the perchlorate intermediate solution with O, and then adding¹⁸The O-marked perchlorate intermediate solution is prepared into 50ng/mL¹⁸Marking perchlorate working solution with O;

standard series working solutions: mixing standard intermediate solution and¹⁸marking perchlorate working solution with O, preparing standard series working solution with the concentration of 0.4, 1.0, 2.0, 5.0, 10.0, 15.0, 30.0 and 60.0ng/mL by pure water, wherein,¹⁸the concentration of the O-labeled perchlorate solution is 1-4 ng/mL.

Preferably, the control solution is prepared as follows:

preparing an external standard solution: precisely sucking 100 mu L of each of the perchlorate standard solution and the chlorate standard solution into a 10mL volumetric flask, adding water to a constant volume to a scale, preparing a 10 mu g/mL mixed solution, and storing at 4 ℃; precisely sucking 1.0mL of the mixed solution into a 10mL volumetric flask, adding water to a constant volume to a scale, preparing a mixed standard intermediate solution with the volume of 1 mu g/mL, and storing at 4 ℃;

preparing an internal standard solution: precise suction¹⁸Marking 100 mu L of perchlorate solution in a 10mL volumetric flask, adding water to constant volume to scale, and preparing into 1 mu g/mL¹⁸Marking the perchlorate intermediate solution with O, and storing at 4 ℃; then precisely absorb¹⁸Marking 500 mu L of perchlorate intermediate solution in a 10mL volumetric flask, adding water to constant volume to scale, and preparing into 50ng/mL¹⁸Marking perchlorate working solution with O, and storing at 4 ℃;

standard series working solutions: sucking a certain amount of mixed standard intermediate liquid¹⁸Marking perchlorate working solution with O, preparing standard series working solution with concentration of 0.4, 1.0, 2.0, 5.0, 10.0, 15.0, 30.0 and 60.0ng/mL by pure water, wherein¹⁸The concentration of the O-marked perchlorate solution is 1-4 ng/mL, the solution is used as a preparation, preferably,¹⁸the concentration of the O-labeled perchlorate solution is 2 ng/mL.

As a preferred embodiment of the method of the present invention, in the step (2), the fast extraction conditions are: the extraction temperature is 80-200 ℃, the volume of the flushing solvent is 30-60%, the heat balance time is 6min, the static extraction time is 3-10 min, the extraction pressure is 1500psi, the cyclic extraction is carried out for 1-2 times, and the nitrogen purging time is 100-200 s; the filter membrane used for filtration was a 0.22 μm polyethersulfone filter membrane.

Preferably, in the step (2), the fast extraction conditions are as follows: the extraction temperature is 120 ℃, the volume of the flushing solvent is 30%, the heat balance time is 6min, the static extraction time is 5min, the extraction pressure is 1500psi, the cyclic extraction is carried out for 2 times, and the nitrogen purging time is 150 s.

As a preferred embodiment of the method of the present invention, in the step (2), 0.5 to 5.0g of notoginseng powder is placed in a 34mL extraction cell, and 0.5 to 2.0g of basic alumina, silica, and silica are added,0.2-1.0 g of graphite carbon black powder, 50-200 mu L of graphite carbon black powder with the concentration of 1 mu g/mL¹⁸Marking the perchlorate intermediate solution by O, and using diatomite as the rest in the extraction tank.

Preferably, in the step (2), dried pseudo-ginseng is taken and crushed, and is screened by a No. 4 screen for standby; precisely weighing 2g (to 0.01g) of Notoginseng radix powder sample, placing in 34mL stainless steel extraction tank (one piece of glass fiber filter membrane is placed in the extraction tank in advance), sequentially placing 1g of alkaline alumina and 0.4g of graphite carbon black powder, and adding 100 μ L¹⁸Marking the perchlorate intermediate solution (1 mu g/mL) by O, and filling the rest part with diatomite; and (3) placing the extraction pool on an extraction disc of a rapid solvent extraction instrument after the extraction pool is assembled, performing rapid extraction and online purification, transferring all liquid into a 50mL volumetric flask after extraction and purification are finished, performing constant volume with water, shaking up, filtering through a 0.22 mu m polyether sulfone filter membrane, and determining by a liquid chromatography-tandem mass spectrometer.

Preferably, the blank matrix solution: taking a negative pseudo-ginseng sample, and obtaining a solution which is a matrix solution according to the operation; blank test: the preparation steps are carried out according to the preparation steps of the test sample except that no sample is added.

As a preferred embodiment of the method of the present invention, in the step (3), the liquid chromatography-tandem mass spectrometer has the following liquid chromatography conditions:

a chromatographic column: a C18 chromatography column;

mobile phase: taking 0.1-1.0% formic acid water solution as mobile phase A and 0.1-0.5% acetic acid methanol solution as mobile phase B to carry out gradient elution; the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (100% -50%) in the mobile phase and B (0-50%) in the mobile phase are in a liquid phase; 1-5.5 min, mobile phase A (100-50% → 60% -0), and mobile phase B (0-50% → 40% -100%); 5.5-9.5 min, wherein A (60-0) in the mobile phase and B (40-100) in the mobile phase are mixed; 9.5-10.0 min, mobile phase A (60% -0 → 20% -0), mobile phase B (40% -100% → 80% -100%); 10.0-11.0 min, wherein A (20-0) in the mobile phase and B (80-100) in the mobile phase are mixed; 11.0-11.1 min, mobile phase A (20% -0 → 100% -50%), and mobile phase B (80% -100% → 0-50%); 11.1-14.0 min, wherein A (100-50%) in the mobile phase and B (0-50%) in the mobile phase are mixed;

flow rate: 0.4-0.6 mL/min;

sample introduction amount: 10 mu L of the solution;

column temperature: 35 ℃ is carried out.

As a preferred embodiment of the method of the present invention, in the step (3), the mass spectrometry conditions of the liquid chromatography-tandem mass spectrometer are as follows:

ionization mode: electrospray ion source, scanning mode: negative ion mode, detection mode: multiple reactive ion monitoring, ionization voltage: -4.0 to-5.0 kV, air curtain gas: 69-276 kPa, collision gas: 34-62 kPa, ionization temperature: 250-450 ℃, auxiliary heating gas 1: 275 to 482kPa, auxiliary heating gas 2: 275 to 482 kPa.

As a preferred embodiment of the method of the present invention, in the liquid chromatography conditions, the flow rate is 0.5mL/min, and the chromatographic column is Phenomenex Synergi MAX-RP 80A (4.6 mm. times.250 mm, 4 μm); in the mobile phase of the liquid chromatogram, 0.5% formic acid aqueous solution is taken as a mobile phase A, and 0.2% acetic acid methanol solution is taken as a mobile phase B for gradient elution; the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (85%) in the mobile phase and B (15%) in the mobile phase are in a liquid phase; 1-5.5 min, wherein A (85% → 40%) in the mobile phase and B (15% → 60%) in the mobile phase; 5.5-9.5 min, wherein A (40%) in the mobile phase and B (60%) in the mobile phase are obtained; 9.5-10.0 min, wherein A (40% → 10%) in the mobile phase and B (60% → 90%) in the mobile phase; 10.0-11.0 min, wherein A (10%) in the mobile phase and B (90%) in the mobile phase are mixed; 11.0-11.1 min, wherein A (10% → 85%) in the mobile phase and B (90% → 15%) in the mobile phase; 11.1-14.0 min, A (85%) in the mobile phase and B (15%) in the mobile phase.

As a preferred embodiment of the method of the present invention, in the mass spectrometry conditions, the ionization voltage: 4.5kV, gas curtain gas: 137kPa, collision gas: 48kPa, ionization temperature: 450 ℃, auxiliary heating gas 1: 413kPa, auxiliary heating gas 2: 482 kPa.

As a preferred embodiment of the method of the present invention, the mass spectrometric conditions wherein the internal standard quantitation of ion pairs comprises¹⁸O-labeled perchlorate quantitative ion pair, and internal standard qualitative ion pair comprising¹⁸Marking a perchlorate qualitative ion pair by O, wherein the target quantitative ion pair comprises the perchlorate quantitative ion pair and the chlorate quantitative ion pair, and the target qualitative ion pair comprises the perchlorate qualitative ion pair and the chlorate qualitative ion pair;

the mass-to-charge ratio conditions of the multi-reaction monitoring ion scanning MRM of the internal standard quantitative ion pair and the internal standard qualitative ion pair are as follows:¹⁸the mass-to-charge ratio of the parent ion of the O-labeled perchlorate quantitative ion pair is 106.9, and the mass-to-charge ratio of the corresponding daughter ion is 88.9;¹⁸the mass-to-charge ratio of the parent ion of the O-labeled perchlorate qualitative ion pair is 108.9, and the mass-to-charge ratio of the corresponding daughter ion is 90.9;

the mass-to-charge ratio conditions of the multi-reaction monitoring ion scanning MRM of the target quantitative ions and the target qualitative ions are as follows: the mass-to-charge ratio of parent ions of the perchlorate quantitative ion pairs is 98.8, and the mass-to-charge ratio of corresponding daughter ions is 83.0; the mass-to-charge ratio of parent ions of the chlorate quantitative ion pairs is 82.9, and the mass-to-charge ratio of corresponding daughter ions is 66.9; the mass-to-charge ratio of parent ions of the perchlorate qualitative ion pairs is 100.8, and the mass-to-charge ratio of corresponding daughter ions is 84.9; the mass-to-charge ratio of the parent ion of the chlorate qualitative ion is 84.9, and the mass-to-charge ratio of the corresponding daughter ion is 68.9.

The mass spectrum parameters of the compounds are shown in table 1.

TABLE 1

Quantitative ions

As a preferred embodiment of the method of the present invention, in the step (3), the qualitative detection method comprises: measuring a test solution and a reference solution, and judging the existence of perchlorate and chlorate by taking the retention time and the relative abundance of perchlorate and chlorate as qualitative basis; the quantitative detection method comprises the following steps: according to perchlorates and chlorates and¹⁸o-labelled perchloroAnd (4) calculating the content of the perchlorate and the chlorate in the sample according to the peak area ratio of the acid salt on the internal standard curve.

The qualitative determination of the invention is as follows: and (3) measuring the test sample and the standard series working solution according to the chromatographic mass spectrum conditions, and if the retention time of the detected mass spectrum peak is consistent with that of the standard sample, the relative abundance of each qualitative ion is consistent with that of the standard working solution with equivalent concentration, and the allowable deviation of the relative abundance does not exceed the range specified in the table 2, judging that the corresponding measured object exists in the sample.

TABLE 2

Relative ion abundance	>50	20-50	10-20	<10
					Allowable relative deviation%	±20	±25	±30	±50

The quantitative determination of the invention is: the response values of the tested object in the standard series working solution and the test sample solution are both in the linear range of the instrument, the equal volume of the standard series working solution and the test sample solution is added into the sample for measurement, the internal standard method is used for quantification,¹⁸o-labeled perchlorate as internal perchlorate and chlorate standardAnd (4) if the content of the measured substance in the test sample exceeds the linearity of the working curve, diluting the substance as required and then measuring the substance. Blank values need to be deducted from the calculation result. The liquid chromatogram tandem mass spectrogram of the standard solution is shown in figures 1-6,¹⁸the MRM map of the O-labeled perchlorate quantitative ion pair is shown in figure 1,¹⁸the MRM spectrum of the O-labeled perchlorate qualitative ion pair is shown in fig. 2, the MRM spectrum of the perchlorate quantitative ion pair is shown in fig. 3, the MRM spectrum of the perchlorate qualitative ion pair is shown in fig. 4, the MRM spectrum of the chlorate quantitative ion pair is shown in fig. 5, and the MRM spectrum of the chlorate qualitative ion pair is shown in fig. 6.

Compared with the prior art, the invention has the beneficial effects that: the invention takes precious Chinese medicinal material pseudo-ginseng as a research object, establishes an HPLC-MS/MS detection method for detecting perchlorate and chlorate in Chinese medicinal materials such as pseudo-ginseng and the like, fills the research blank of perchlorate which is an environmental pollutant in the field of Chinese medicinal materials, provides technical support for the research of perchlorate in Chinese medicinal materials, protects the medication safety of consumers, and promotes and develops great economic and social benefits of Chinese medicinal materials. The detection method of the invention has strong practicability, simple and convenient operation, accurate result and good reproducibility.

Drawings

FIG. 1 is a drawing of¹⁸And (3) marking the MRM map of the perchlorate quantitative ion pair by O.

FIG. 2 is¹⁸And (3) marking the MRM map of the perchlorate qualitative ion pair.

FIG. 3 is an MRM map of perchlorate quantitative ion pairs.

FIG. 4 is an MRM map of perchlorate qualitative ion pairs.

FIG. 5 is an MRM profile of chlorate quantitative ion pairs.

FIG. 6 is an MRM map of chlorate qualitative ion pairs.

FIG. 7 is a total ion flow graph of negative samples.

FIG. 8 is a total ion flow graph of perchlorate and chloric acid added to a negative sample.

FIG. 9 is a statistical chart of the recovery rates obtained for different extraction solvents.

FIG. 10 is a bar graph of recovery for different extraction temperatures.

FIG. 11 is a statistical plot of the recovery results of the first purge optimization test.

FIG. 12 is a statistical plot of the recovery results of the second purge optimization test.

Figure 13 is a total ion flow graph of perchlorate and chlorate as in example 4.

Figure 14 is a total ion flow graph of perchlorate and chlorate as in example 5.

Figure 15 is a total ion flow graph of perchlorate and chlorate as in example 6.

Figure 16 is a total ion flow graph of perchlorate and chlorate as in example 7.

Figure 17 is a total ion flow graph of perchlorate and chlorate as in example 8.

Figure 18 is a total ion flow graph of perchlorate and chlorate as in example 9.

Figure 19 is a total ion flow graph of perchlorate and chlorate as in example 10.

Figure 20 is a total ion flow graph of perchlorate and chlorate as in example 11.

Figure 21 is a total ion flow graph of perchlorate and chlorate as in example 12.

Figure 22 is a total ion flow graph of perchlorate and chlorate as in example 13.

Figure 23 is a total ion flow graph of perchlorate and chlorate as in example 14.

Figure 24 is a total ion flow graph of perchlorate and chlorate as in example 15.

Figure 25 is a total ion flow graph of perchlorate and chlorate as in example 16.

Figure 26 is a total ion flow graph of perchlorate and chlorate as in example 17.

Detailed Description

To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.

The instruments and reagents used in the examples of the present invention were as follows: triple Quad5500 (AB, USA) and ASE350 type rapid solvent extractionTaking an instrument (Samera Feishell science and technology, USA), a sartorius CP225D electronic balance, a perchlorate standard solution (0.999g/ml, MC-CLOX663048, Inorgnic vents, USA), a chlorate standard solution (0.999g/ml, M2-CLOX660055, Inorgnic vents, USA),¹⁸o-labeled perchlorate solution (100. mu.g/ml, SDFF-012, Cambridge Isotope Laboratories, USA), formic acid (chromatographically pure, Macklin), acetic acid (chromatographically pure, Codow), methanol (chromatically pure, Honeywell), acetonitrile (chromatographically pure, Honeywell), alumina (Shanghai PentaIV Chemicals Co., Ltd.), diatomaceous earth (Thermo Scientific Co., Ltd.), graphite black (Shanghai Ann spectral Co., Ltd.), and experimental water were each Drech drinking water.

Example 1

A method for detecting perchlorate and chlorate in pseudo-ginseng, the method comprising the steps of:

(1) preparation of control solutions:

standard series working solutions: sucking a certain amount of mixed standard intermediate liquid¹⁸Marking perchlorate working solution with O, preparing standard series working solution with concentration of 0.4, 1.0, 2.0, 5.0, 10.0, 15.0, 30.0 and 60.0ng/mL by pure water, wherein¹⁸The concentration of the O-marked perchlorate solution is 2ng/mL, and the solution is prepared for use;

(2) preparation of a test solution:

pulverizing dried Notoginseng radix, and sieving with No. 4 sieve; precisely weighing 2g (to 0.01g) of Notoginseng radix powder sample, placing in 34mL stainless steel extraction tank (one piece of glass fiber filter membrane is placed in the extraction tank in advance), sequentially placing 1g of alkaline alumina and 0.4g of graphite carbon black powder, and adding 100 μ L¹⁸Marking the perchlorate intermediate solution (1 mu g/mL) by O, and filling the rest part with diatomite; the extraction pond is placed on an extraction disc of a rapid solvent extraction instrument after being well assembled, rapid extraction and online purification are carried out, and the rapid extraction conditions are as follows: the extraction temperature is 120 ℃, the volume of the flushing solvent is 30%, the heat balance time is 6min, the static extraction time is 5min, the extraction pressure is 1500psi, the cyclic extraction is carried out for 2 times, and the nitrogen purging time is 150 s; after extraction and purification are finished, transferring all liquid into a 50mL volumetric flask, performing constant volume with water, shaking up, filtering through a 0.22 mu m polyethersulfone filter membrane, and determining by a liquid chromatography-tandem mass spectrometer;

blank matrix solution: taking a negative pseudo-ginseng sample, and obtaining a solution which is a matrix solution according to the operation; blank test: except that no sample is added, the preparation steps are carried out according to the preparation steps of the test sample;

(3) and (3) detection: precisely measuring the test solution and the reference solution, injecting into a liquid chromatogram-tandem mass spectrometer, and performing qualitative detection or quantitative detection;

the liquid chromatography-tandem mass spectrometer has the following liquid chromatography conditions:

a chromatographic column: phenomenex Synergi MAX-RP 80A (4.6 mm. times.250 mm, 4 μm);

mobile phase: performing gradient elution by using 0.5% formic acid water solution as a mobile phase A and 0.2% acetic acid methanol solution as a mobile phase B; the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (85%) in the mobile phase and B (15%) in the mobile phase are in a liquid phase; 1-5.5 min, wherein A (85% → 40%) in the mobile phase and B (15% → 60%) in the mobile phase; 5.5-9.5 min, wherein A (40%) in the mobile phase and B (60%) in the mobile phase are obtained; 9.5-10.0 min, wherein A (40% → 10%) in the mobile phase and B (60% → 90%) in the mobile phase; 10.0-11.0 min, wherein A (10%) in the mobile phase and B (90%) in the mobile phase are mixed; 11.0-11.1 min, wherein A (10% → 85%) in the mobile phase and B (90% → 15%) in the mobile phase; 11.1-14.0 min, wherein A (85%) in the mobile phase and B (15%) in the mobile phase are mixed;

flow rate: 0.5 mL/min;

sample introduction amount: 10 mu L of the solution;

column temperature: 35 ℃;

the mass spectrum conditions of the liquid chromatography-tandem mass spectrometer are as follows:

ionization mode: electrospray ion source, scanning mode: negative ion mode, detection mode: multiple reactive ion monitoring, ionization voltage: 4.5kV, gas curtain gas: 137kPa, collision gas: 48kPa, ionization temperature: 450 ℃, auxiliary heating gas 1: 413kPa, auxiliary heating gas 2: 482 kPa;

under the mass spectrum condition, the internal standard quantitative ion pair comprises¹⁸O-labeled perchlorate quantitative ion pair, and internal standard qualitative ion pair comprising¹⁸Marking a perchlorate qualitative ion pair by O, wherein the target quantitative ion pair comprises the perchlorate quantitative ion pair and the chlorate quantitative ion pair, and the target qualitative ion pair comprises the perchlorate qualitative ion pair and the chlorate qualitative ion pair;

the mass-to-charge ratio conditions of the multi-reaction monitoring ion scanning MRM of the target quantitative ions and the target qualitative ions are as follows: the mass-to-charge ratio of parent ions of the perchlorate quantitative ion pairs is 98.8, and the mass-to-charge ratio of corresponding daughter ions is 83.0; the mass-to-charge ratio of parent ions of the chlorate quantitative ion pairs is 82.9, and the mass-to-charge ratio of corresponding daughter ions is 66.9; the mass-to-charge ratio of parent ions of the perchlorate qualitative ion pairs is 100.8, and the mass-to-charge ratio of corresponding daughter ions is 84.9; the mass-to-charge ratio of the parent ion of the chlorate qualitative ion is 84.9, and the mass-to-charge ratio of the corresponding daughter ion is 68.9;

the qualitative determination is as follows: determining the sample and the standard series working solution according to the above chromatographic mass spectrum conditions, if the retention time of the detected mass spectrum peak is consistent with that of the standard sample, the relative abundance of each qualitative ion is consistent with that of the standard working solution with equivalent concentration, and the allowable deviation of the relative abundance does not exceed the range specified in table 2, determining that the corresponding object exists in the sample,

the quantitative determination was: the response values of the tested object in the standard series working solution and the test sample solution are both in the linear range of the instrument, the equal volume of the standard series working solution and the test sample solution is added into the sample for measurement, the internal standard method is used for quantification,¹⁸marking perchlorate as an internal standard substance of perchlorate and chlorate by O, and if the content of the measured substance in the sample exceeds the linearity of the working curve, diluting the sample as required and then measuring the sample. Blank values need to be deducted from the calculation result. The liquid chromatogram tandem mass spectrum of the standard solution is shown in figures 1-6.

Example 2 methodological validation

The following methodology was set up and tested on the basis of example 1.

(1) Linear relation

Precisely measuring a proper amount of mixed standard intermediate solution (1 mu g/mL) and¹⁸the O-labeled perchlorate working solution (50ng/mL) is subjected to constant volume by pure water, standard series working solutions with the concentrations of 0.4 (detection limit), 1, 2, 5, 10, 15, 30 and 60ng/mL (the concentrations of internal standard substances are all 2ng/mL) are respectively prepared, 10 mu L of working solution is absorbed, and the solution is determined by adopting a liquid chromatogram-tandem mass spectrometry method. And (3) preparing a standard curve by taking the peak area ratio of each characteristic ion mass chromatographic peak to the corresponding isotope internal standard as a vertical coordinate and the concentration of the reference solution as a horizontal coordinate.

The experimental results are as follows: linear regression equations obtained after background subtraction of perchlorate and chlorate are respectively 0.75904x +0.22321 and 0.15480x +0.02758, and correlation coefficient R²0.9988 and 0.9988, respectively. Therefore, the perchlorate and chlorate have good linear relation in the concentration range of 1.0-60 ng/mL, and the correlation coefficient (R)²) Are all larger than 0.998. According to the characteristic ion mass chromatographic peak signal-to-noise ratio S/N more than or equal to 3 as a detection limit, referring to relevant data at home and abroad, the final detection limit is 10 mug/kg, and the quantitative limit is 25 mug/kg.

(2) Matrix effect

And (3) evaluating the matrix effect of the established pretreatment method of the HPLC-MS/MS method by adopting a common post-extraction and standard-adding method. By comparing response values of the perchlorate and chlorate in the standard sample matrix and the standard blank solution, not only qualitative analysis can be carried out, but also the influence value (SSE) of matrix effect can be quantitatively analyzed. The procedure of example 1 was followed using the pseudo-ginseng powder reference as a sample to obtain a matrix solution. The working solution is diluted into a working solution with the concentration same as the linear relation of the above investigation by using pure water and a sample matrix solution respectively, namely, the addition concentration gradients of perchlorate and chlorate are 1, 2, 5, 10, 15, 30 and 60ng/mL (the internal standard solution is 2 ng/mL). And establishing a standard curve by an internal standard method, wherein the ratio of the slope of the standard curve of the sample matrix solution to the slope of the standard curve of the blank solvent is the matrix effect of the sample. The results are shown in Table 3.

TABLE 3

Compound (I)	Regression equation for matrix solution	Regression equation for blank solution	SSE value%
				Perchlorate salt	y＝0.74828x+0.66997	y＝0.75904x+0.22321	98.6％
Chlorate salt	y＝0.16098x+0.00288	y＝0.15480x+0.02758	103.9％

When the SSE value is between 80% and 120%, the matrix effect is negligible. As can be seen from table 3, the SSE values were substantially in the satisfied range when the perchlorate and chlorate were simultaneously quantitatively detected using the internal standard method.

(3) Accuracy of

After sample pretreatment and optimization of mass spectrum parameter conditions, a standard addition method is adopted, and low-medium (25, 50 and 250 mu g/kg) and high-medium (3 kinds of mixed liquor of perchlorate and chlorate with different final concentrations (the concentration of the internal standard is 2ng/mL) are added into a sample for a recovery rate test, each concentration is subjected to a parallel test for 6 times, and the results are shown in Table 4.

And (3) low concentration: a sample solution of low concentration for accuracy test was prepared by precisely aspirating 50. mu.L of a mixed standard intermediate solution (1. mu.g/mL) and mixing the resulting solution with 2g of a sample, and the sample recovery rate was calculated by the measurement according to the method in example 1.

Medium concentration: a sample solution of low concentration for accuracy evaluation was prepared by precisely aspirating 100. mu.L of a mixed standard intermediate solution (1. mu.g/mL) and mixing the resulting solution with 2g of a sample, and the sample recovery rate was calculated by the following method.

High concentration: a500. mu.L portion of the mixed standard intermediate solution (1. mu.g/mL) was precisely aspirated, mixed with 2g of the sample, and subjected to the procedure of example 1 to prepare a low-concentration sample solution for accuracy evaluation.

TABLE 4

(4) Precision degree

And adding the mixed solution of perchlorate and chlorate into the blank extracting solution, wherein the detection limit of perchlorate and chlorate is 0.01mg/kg and the quantification limit is 0.025mg/kg according to the characteristic ion mass chromatographic peak signal-to-noise ratio S/N which is more than or equal to 3 and the quantification limit of S/N which is more than or equal to 10. Under the conditions of sample pretreatment and mass spectrum parameter optimization, the ratio of perchlorate to chlorate is measured for multiple times by selecting the intermediate concentration of 0.25mg/kg, and the result is shown in table 5.

TABLE 5

Composition (I)	Ratio 1	Ratio 2	Ratio 3	Ratio 4	Ratio 5	Ratio 6	Mean value of	RSD(％)
									Perchlorate salt	5.385	5.37	5.311	5.337	5.311	5.316	5.34	0.6
Chlorate salt	1.907	1.889	1.851	1.878	1.893	1.888	1.88	1.0

(5) Method specificity

The total ion flow graph of the negative sample and the total ion flow graph of the negative sample with perchlorate and chlorate added are respectively shown in fig. 7 and fig. 8. As can be seen from FIGS. 7 and 8, the negative sample did not interfere with the measurement, and the specificity of the method was good.

Example 3 chromatographic Condition optimization

The following conditions were selected and tested on the basis of example 1.

(1) Selection of chromatography columns

This experiment compares IC-Pak^TMAnion HR column (4.6 mm. times.75 mm, 6 μm) and Synergi^TMMAX-RP 80A chromatography column (4.6 mm. times.250 mm, 4 μm), mobile phase was 0.5% formic acid water-0.2% acetic acid methanol. The chromatographic behavior of a mixed solution of perchlorate and chlorate at a concentration of 10ng/ml was compared on different chromatographic columns. As a result, it was found that perchlorate is present in IC-Pak^TMThe retention time on the Anion HR chromatographic column is about 13min, the sensitivity is low, the peak shape is wide and asymmetric, and the separation degree of the perchlorate peak and the chlorate peak is low. In Synergi^TMMAX-RP 80A chromatographic columnIn addition, the perchlorate and chlorate are well separated, the peak shape is symmetrical, and the response value is higher. Therefore Synergi was used in this experiment^TMMAX-RP 80A chromatographic column.

(2) Selection of mobile phase

In the experiment, a 20mmol/L ammonium formate-acetonitrile system, a water-0.05% ammonia water-acetonitrile system and a 0.1% formic acid-methanol system are selected as comparison, and the result shows that when the 20mmol/L ammonium formate-acetonitrile system is used as a mobile phase, the retention time of perchlorate and chlorate is longer, although the peak shape of perchlorate is symmetrical, the peak shape response value of chlorate is smaller, and the separation degree of perchlorate and chlorate is smaller. When a water-0.05% ammonia water acetonitrile system is used as a mobile phase, the peak shapes of perchlorate and chlorate are found to be symmetrical, the retention time is 3.5min, the response values are high, but the separation degrees of the perchlorate and the chlorate are poor, after multiple tests, the mobile phase system is found to have large fluctuation relative to the peak shapes, the response value is high and low, and the retention time is easy to drift. In a 0.1% formic acid-methanol system, the retention time of perchlorate is 11min and the retention time of chlorate is 8min, the peak shapes are symmetrical, the response value is stable and the retention time cannot drift although the response value is not high in a water-0.05% ammonia acetonitrile system. The 0.1% formic acid-methanol system was selected as the mobile phase for comprehensive consideration.

The peak shape can be modified by adding acid in the mobile phase, and in order to make the peak shape more symmetrical, a proper amount of acetic acid can be added in methanol. After determining the formic acid water-methanol system as the mobile phase, different concentrations of formic acid and acetic acid methanol were examined. Experiments compared a 0.2% aqueous formic acid-0.2% acetic acid methanol system, a 0.2% aqueous formic acid-0.5% acetic acid methanol system, and a 0.5% aqueous formic acid-0.2% acetic acid methanol system. The result shows that the retention time of perchlorate and chlorate in a 0.2 percent formic acid water-0.2 percent acetic acid methanol system is respectively 10.4min and 9.3min, the peak shape is symmetrical, and the response value is higher. In the 0.2% aqueous formic acid-0.5% acetic acid methanol system, the peak shape is symmetrical, the retention time is basically the same as that of the 0.2% aqueous formic acid-0.2% acetic acid methanol system, but the response value of the hetero peak is increased. The retention time of perchlorate and chlorate in a 0.5 percent formic acid water-0.2 percent acetic acid methanol system is respectively 8.4min and 7.6min, which is shorter than that of a 0.2 percent formic acid water-0.2 percent acetic acid methanol system, and the system has symmetrical peak shape and higher sensitivity. The 0.5% aqueous formic acid-0.2% methanolic acetic acid system was selected as the mobile phase for this experiment.

(3) Selection of extraction solvent

Perchlorate and chlorate are generally easy to dissolve in water, and according to the property, 4 extraction solvents of water, 0.2 percent acetic acid water, acetonitrile-water (1:1) and acetonitrile-0.2 percent acetic acid water (1:1) are selected for comparison, and the recovery rate results obtained by extracting samples by the 4 extraction solvents are shown in figure 9, and as can be seen from figure 9, when water is taken as the extraction solvent, the recovery rates of perchlorate and chlorate are optimal, and the method selects water as the extraction solvent.

(4) Selection of temperature for rapid extraction apparatus

Taking the extraction solvent as pure water, the volume of the washing solvent is 30%, the heat balance time is 6min, the static extraction time is 5min, the extraction pressure is 1500psi, the circulation extraction is carried out for 2 times, the fixed conditions are adopted, respectively sampling is carried out, and 1 mu g/mL 500 mu L of mixed standard intermediate solution and¹⁸o-labeled perchlorate intermediate solution (1. mu.g/Ml, 100. mu.l) was mixed with 100. mu.l of perchlorate (final perchlorate/chlorate concentration: 250. mu.g/kg), extracted at 80, 100, 120 and 150 ℃ respectively in accordance with the procedure of example 1, and the recovery was calculated by measuring the contents of perchlorate and chlorate in the extract, and the results are shown in Table 6 and FIG. 10.

TABLE 6

Composition (I)	Temperature (. degree.C.)	Recovery (%)
			Perchlorate salt	80	68
Chlorate salt	80	117
			Perchlorate salt	100	82
Chlorate salt	100	92
			Perchlorate salt	120	94
Chlorate salt	120	91
			Perchlorate salt	120	115
Chlorate salt	120	53

As can be seen from the table, the recovery rate of perchlorate and chlorate at a certain temperature is increased along with the increase of the extraction temperature, but the recovery rate of chlorate at 150 ℃ is reduced, so that the extraction temperature of 120 ℃ is considered to be the extraction temperature of the experiment.

(5) Selection of purification materials

The method considers four purifying materials of aluminum oxide, C18, PSA and GCB and a combination mode of the purifying materials, 1g of each purifying material is added, the operation is carried out according to 1.4 steps, the result shows that a large amount of white bubbles are generated in an extraction collecting bottle added with C18, and the on-machine result shows that the test solution chlorate added with C18 has a double-peak phenomenon; both PSA and GCB are affected by adsorbed pigments, and the liquid to which GCB is added is relatively clear in appearance of the extract, while the liquid to which PSA is added is relatively cloudy, and the recovery results are shown in FIG. 11. Then we have performed a second purification material optimization test, selecting two materials, namely alumina and GCB, and examining the amounts of the purification materials, wherein the alumina is added in 1g, 2g and 3g, respectively, and the GCB is added in 0.4g, 0.8g and 1.2g, and the combination of these amounts is clear in appearance, the recovery result is shown in fig. 12, and we finally select the purification mode of alumina 1g and GCB0.4 g.

Example 4

Example 4 differs from example 1 in that: in the step (2), the amount of the notoginseng powder was 0.5g, the amount of the basic alumina was 0.5g, and the amount of the graphite carbon black powder was 0.2g, and the rest of the methods were the same as in example 1. The total ion flow of perchlorate and chlorate in this example is shown in figure 13 and the retention times are shown in table 7.

TABLE 7

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.76(8.08)	1.023e+04
GLS 1	98.8/83.0	7.77(8.10)	1.238e+06
				GLS 2	100.8/84.9	7.77(8.10)	4.461e+05
LS 1	82.9/66.9	7.15(7.44)	3.087e+05
				LS 2	84.9/68.9	7.15(7.44)	1.048e+05

Example 5

Example 5 differs from example 1 in that: in the step (2), the amount of the notoginseng powder is 5g, the amount of the basic alumina is 2g, and the amount of the graphite carbon black powder is 1g, and the rest of the method is the same as that of the example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 14, and the retention time is shown in table 8.

TABLE 8

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.83(8.08)	1.141e+04
GLS 1	98.8/83.0	7.83(8.10)	7.373e+05
				GLS 2	100.8/84.9	7.82(8.10)	2.767e+05
LS 1	82.9/66.9	7.19(7.44)	2.033e+05
				LS 2	84.9/68.9	7.20(7.44)	6.812e+04

Example 6

Example 6 differs from example 1 in that: in the step (2), the rapid extraction conditions are as follows: the extraction temperature was 80 ℃, the volume of the rinsing solvent was 30%, the heat equilibration time was 6min, the static extraction time was 3min, the extraction pressure was 1500psi, the cyclic extraction was performed 2 times, the nitrogen purge time was 100s, and the rest of the procedure was the same as in example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 15, and the retention time is shown in table 9.

TABLE 9

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.91(8.08)	9.147e+03
GLS 1	98.8/83.0	7.91(8.10)	8.341e+05
				GLS 2	100.8/84.9	7.91(8.10)	3.042e+05
LS 1	82.9/66.9	7.27(7.44)	2.150e+05
				LS 2	84.9/68.9	7.27(7.44)	7.462e+04

Example 7

Example 7 differs from example 1 in that: in the step (2), the rapid extraction conditions are as follows: the extraction temperature was 200 ℃, the volume of the rinsing solvent was 60%, the heat equilibration time was 6min, the static extraction time was 10min, the extraction pressure was 1500psi, the cyclic extraction was performed 1 time, the nitrogen purge time was 200s, and the rest of the methods were the same as in example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 16, and the retention time is shown in table 10.

Watch 10

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.88(8.08)	1.253e+04
GLS 1	98.8/83.0	7.88(8.10)	9.657e+05
				GLS 2	100.8/84.9	7.88(8.10)	3.370e+05
LS 1	82.9/66.9	7.26(7.44)	7.682e+04
				LS 2	84.9/68.9	7.26(7.44)	2.925e+04

Example 8

Example 8 differs from example 1 in that: in the step (3), gradient elution was performed using a 0.1% formic acid aqueous solution as a mobile phase a and a 0.1% acetic acid methanol solution as a mobile phase B, and the rest of the procedure was the same as in example 1. The total ion flow of perchlorate and chlorate in this example is shown in figure 17 and the retention times are shown in table 11.

TABLE 11

Target analysisArticle (A)	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.65(8.08)	1.056e+02
GLS 1	98.8/83.0	8.43(8.10)	1.670e+03
				GLS 2	100.8/84.9	10.58(8.10)	1.475e+07
LS 1	82.9/66.9	9.37(7.44)	2.897e+06
				LS 2	84.9/68.9	9.37(7.44)	1.005e+06

Example 9

Example 9 differs from example 1 in that: in step (3), gradient elution was performed using a 1.0% formic acid aqueous solution as mobile phase a and a 0.5% acetic acid methanol solution as mobile phase B, and the rest of the procedure was the same as in example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 18, and the retention time is shown in table 12.

TABLE 12

Example 10

Example 10 differs from example 1 in that: in step (3), the flow rate under liquid chromatography conditions was 0.4mL/min, and the rest of the procedure was the same as in example 1. The total ion flow graph of the perchlorate and chlorate in this example is shown in fig. 19, and the retention time is shown in table 13.

Watch 13

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	8.35(8.08)	3.300e+01
GLS 1	98.8/83.0	9.95(8.10)	1.536e+06
				GLS 2	100.8/84.9	9.95(8.10)	5.647e+05
LS 1	82.9/66.9	9.23(7.44)	3.202e+05
				LS 2	84.9/68.9	9.23(7.44)	1.057e+05

Example 11

Example 11 differs from example 1 in that: in step (3), the flow rate under liquid chromatography conditions was 0.6mL/min, and the rest of the procedure was the same as in example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 20, with retention times as shown in table 14.

TABLE 14

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.91(8.08)	3.302e+01
GLS 1	98.8/83.0	6.72(8.10)	1.007e+06
				GLS 2	100.8/84.9	6.72(8.10)	3.715e+05
LS 1	82.9/66.9	6.14(7.44)	2.009e+05
				LS 2	84.9/68.9	6.14(7.44)	6.920e+04

Example 12

Example 12 differs from example 1 in that: in the step (1), the step (c),¹⁸the concentration of the O-labeled perchlorate solution was 1ng/mL, and the rest of the method was the same as in example 1. The total ion flow of perchlorate and chlorate in this example is shown in figure 21, and the retention times are shown in table 15.

Watch 15

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.86(8.08)	4.377e+03
GLS 1	98.8/83.0	7.86(8.10)	9.119e+05
				GLS 2	100.8/84.9	7.86(8.10)	3.300e+05
LS 1	82.9/66.9	7.23(7.44)	2.580e+05
				LS 2	84.9/68.9	7.22(7.44)	8.679e+04

Example 13

Example 13 differs from example 1 in that: in the step (1), the step (c),¹⁸the concentration of the O-labeled perchlorate solution was 4ng/mL, and the rest of the method was the same as in example 1. The total ion flow of the perchlorate and chlorate in this example is shown in fig. 22, and the retention time is shown in table 16.

TABLE 16

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	7.88(8.08)	2.236e+04
GLS 1	98.8/83.0	7.88(8.10)	8.333e+05
				GLS 2	100.8/84.9	7.88(8.10)	2.946e+05
LS 1	82.9/66.9	7.25(7.44)	2.447e+05
				LS 2	84.9/68.9	7.24(7.44)	8.299e+04

Example 14

Example 14 differs from example 1 in that: in the step (3), the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (100%) in the mobile phase and B (0) in the mobile phase are obtained; 1-5.5 min, wherein A (100% → 60%) in the mobile phase and B (0 → 40%) in the mobile phase; 5.5-9.5 min, wherein A (60%) in the mobile phase and B (40%) in the mobile phase are obtained; 9.5-10.0 min, wherein A (60% → 20%) in the mobile phase and B (40% → 80%) in the mobile phase; 10.0-11.0 min, wherein A (20%) in the mobile phase and B (80%) in the mobile phase are mixed; 11.0-11.1 min, mobile phase A (20% → 100%), mobile phase B (80% → 0); the rest of the procedures are the same as in example 1 except that the procedures are 11.1-14.0 min, A (100%) in the mobile phase and B (0) in the mobile phase. The total ion flow graph of the perchlorate and chlorate in this example is shown in fig. 23, and the retention time is shown in table 17.

TABLE 17

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	8.41(8.08)	2.690e+05
GLS 1	98.8/83.0	8.41(8.10)	1.971e+07
				GLS 2	100.8/84.9	8.41(8.10)	7.236e+06
LS 1	82.9/66.9	7.94(7.44)	1.536e+06
				LS 2	84.9/68.9	7.94(7.44)	5.168e+05

Example 15

Example 15 differs from example 1 in that: in the step (3), the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (50%) in the mobile phase and B (50%) in the mobile phase are in a liquid phase; 1-5.5 min, mobile phase A (50% → 0), mobile phase B (50% → 100%); 5.5-9.5 min, wherein A (0) in the mobile phase and B (100%) in the mobile phase are obtained; 9.5-10.0 min, wherein A (0 → 20%) in the mobile phase and B (100% → 80%) in the mobile phase; 10.0-11.0 min, wherein A (20%) in the mobile phase and B (80%) in the mobile phase are mixed; 11.0-11.1 min, wherein A (20% → 50%) in the mobile phase and B (80% → 50%) in the mobile phase; the rest of the procedures are the same as example 1 except that A (50%) in the mobile phase and B (50%) in the mobile phase are carried out for 11.1-14.0 min. The total ion flow of the perchlorate and chlorate in this example is shown in figure 24 and the retention times are shown in table 18.

Watch 18

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	8.38(8.08)	1.360e+03
GLS 1	98.8/83.0	6.21(8.10)	7.114e+06
				GLS 2	100.8/84.9	6.21(8.10)	2.530e+06
LS 1	82.9/66.9	6.20(7.44)	1.018e+06
				LS 2	84.9/68.9	6.20(7.44)	3.469e+05

Example 16

Example 16 differs from example 1 in that: in the step (3), the mass spectrum conditions are as follows: ionization mode: electrospray ion source, scanning mode: negative ion mode, detection mode: multiple reactive ion monitoring, ionization voltage: 4.0kV, air curtain gas: 69kPa, collision gas: 34kPa, ionization temperature: 250 ℃, auxiliary heating gas 1: 275kPa, auxiliary heating gas 2: 275kPa, the rest of the process was the same as in example 1. The total ion flow of perchlorate and chlorate in this example is shown in figure 25 with retention times in table 19.

Watch 19

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	8.05(8.08)	5.985e+04
GLS 1	98.8/83.0	8.04(8.10)	5.182e+06
				GLS 2	100.8/84.9	8.05(8.10)	1.855e+06
LS 1	82.9/66.9	7.41(7.44)	6.794e+05
				LS 2	84.9/68.9	7.40(7.44)	2.279e+05

Example 17

Example 17 differs from example 1 in that: in the step (3), the mass spectrum conditions are as follows: ionization mode: electrospray ion source, scanning mode: negative ion mode, detection mode: multiple reactive ion monitoring, ionization voltage: 5.0kV, gas curtain gas: 276kPa, collision gas: 62kPa, ionization temperature: 450 ℃, auxiliary heating gas 1: 482kPa, auxiliary heating gas 2: 482kPa and the rest of the process was the same as in example 1. The total ion flow of perchlorate and chlorate in this example is shown in fig. 26, with retention times as shown in table 20.

Watch 20

Target analyte	Mass range	Retention time (min)	Area (cps)
				GLS IS 2	108.9/90.9	8.04(8.08)	5.429e+05
GLS 1	98.8/83.0	8.04(8.10)	3.582e+07
				GLS 2	100.8/84.9	8.04(8.10)	1.392e+07
LS 1	82.9/66.9	7.40(7.44)	3.247e+06
				LS 2	84.9/68.9	7.40(7.44)	1.155e+06

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for detecting perchlorate and chlorate in pseudo-ginseng is characterized by comprising the following steps:

(1) preparation of control solutions:

(2) preparation of a test solution:

2. The method of claim 1, wherein in step (1), the control solution is prepared by:

standard series working solutions: mixing standard intermediate solution and¹⁸marking the perchlorate working solution by O,preparing standard series working solution with concentration of 0.4, 1.0, 2.0, 5.0, 10.0, 15.0, 30.0, 60.0ng/mL with pure water, wherein,¹⁸the concentration of the O-labeled perchlorate solution is 1-4 ng/mL.

3. The method of claim 1, wherein in step (2), the fast extraction conditions are: the extraction temperature is 80-200 ℃, the volume of the flushing solvent is 30-60%, the heat balance time is 6min, the static extraction time is 3-10 min, the extraction pressure is 1500psi, the cyclic extraction is carried out for 1-2 times, and the nitrogen purging time is 100-200 s; the filter membrane used for filtration was a 0.22 μm polyethersulfone filter membrane.

4. The method according to claim 1, wherein in the step (2), 0.5-5.0 g of notoginseng powder is placed in a 34mL extraction cell, 0.5-2.0 g of basic alumina, 0.2-1.0 g of graphite carbon black powder and 50-200 μ L of 1 μ g/mL of the powder¹⁸Marking the perchlorate intermediate solution by O, and using diatomite as the rest in the extraction tank.

5. The method according to claim 1, wherein in the step (3), the liquid chromatography-tandem mass spectrometer has the following liquid chromatography conditions:

a chromatographic column: a C18 chromatography column;

flow rate: 0.4-0.6 mL/min;

sample introduction amount: 10 mu L of the solution;

column temperature: 35 ℃ is carried out.

6. The method of claim 1, wherein in step (3), the mass spectrometry conditions of the liquid chromatography-tandem mass spectrometer are as follows:

7. The method of claim 5, wherein the liquid chromatography conditions are conditions of 0.5mL/min flow rate, a column of Phenomenex synergy MAX-RP 80A (4.6mm x 250mm, 4 μ ι η); in the mobile phase of the liquid chromatogram, 0.5% formic acid aqueous solution is taken as a mobile phase A, and 0.2% acetic acid methanol solution is taken as a mobile phase B for gradient elution; the conditions of the mobile phase gradient elution are as follows: 0-1 min, wherein A (85%) in the mobile phase and B (15%) in the mobile phase are in a liquid phase; 1-5.5 min, wherein A (85% → 40%) in the mobile phase and B (15% → 60%) in the mobile phase; 5.5-9.5 min, wherein A (40%) in the mobile phase and B (60%) in the mobile phase are obtained; 9.5-10.0 min, wherein A (40% → 10%) in the mobile phase and B (60% → 90%) in the mobile phase; 10.0-11.0 min, wherein A (10%) in the mobile phase and B (90%) in the mobile phase are mixed; 11.0-11.1 min, wherein A (10% → 85%) in the mobile phase and B (90% → 15%) in the mobile phase; 11.1-14.0 min, A (85%) in the mobile phase and B (15%) in the mobile phase.

8. The method of claim 6, wherein, in the mass spectrometry condition, the ionization voltage: 4.5kV, gas curtain gas: 137kPa, collision gas: 48kPa, ionization temperature: 450 ℃, auxiliary heating gas 1: 413kPa, auxiliary heating gas 2: 482 kPa.

9. The method of claim 6, wherein quantifying ion pairs within the mass spectrometry conditions comprises quantifying ion pairs using an internal standard¹⁸O-labeled perchlorate quantitative ion pair, and internal standard qualitative ion pair comprising¹⁸Marking a perchlorate qualitative ion pair by O, wherein the target quantitative ion pair comprises the perchlorate quantitative ion pair and the chlorate quantitative ion pair, and the target qualitative ion pair comprises the perchlorate qualitative ion pair and the chlorate qualitative ion pair;

10. The method of claim 1, wherein in the step (3), the qualitative detection method comprises: measuring a test solution and a reference solution, and judging the existence of perchlorate and chlorate by taking the retention time and the relative abundance of perchlorate and chlorate as qualitative basis; the quantitative detection method comprises the following steps: according to perchlorates and chlorates and¹⁸and marking the peak area ratio of the perchlorate on the internal standard curve by O, and calculating the content of the perchlorate and the chlorate in the sample.