CN111537653A - Method for measuring chemical component content in chrysanthemum - Google Patents

Abstract

The embodiment of the invention provides a method for measuring the content of chemical components in chrysanthemum, which adopts ultra-high performance liquid chromatography-mass spectrometry to simultaneously measure the content of 13 chemical components in chrysanthemum; the 13 chemical compositions comprise: isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin and cryptochlorogenic acid. By adopting the method, the content of 13 chemical components in the chrysanthemum can be simultaneously measured by reasonably selecting the chromatographic condition and the mass spectrum condition, and the method has the advantages of simplicity, convenience, high sensitivity, high analysis speed, strong specificity and the like, so that the method can be used for quality control of the chrysanthemum medicinal material.

Description

Method for measuring chemical component content in chrysanthemum

Technical Field

The invention relates to the technical field of traditional Chinese medicine component determination, in particular to a method for determining chemical component content in chrysanthemum.

Background

The flos Chrysanthemi, named as Chinese medicine, is the dried capitate of Chrysanthemum morifolium Ramat of Compositae, and has effects of dispelling pathogenic wind heat, suppressing liver yang, removing liver heat, improving eyesight, and clearing away heat and toxic materials. It can be used for treating wind-heat type common cold, early stage of epidemic febrile disease, vertigo due to liver-yang, excessive liver-wind, conjunctival congestion, blurred vision, sore, carbuncle, and toxic swelling. The chrysanthemum contains various chemical components such as flavonoids, volatile oils, organic acids, saccharides and amino acids, and the flavonoids and the organic acids are the main effective components. At present, the chrysanthemum medicinal materials in the market are various in types and different in production places, so that the quality difference of the chrysanthemum medicinal materials is large, and a quality control system is not complete.

Disclosure of Invention

The embodiment of the invention aims to provide a method for measuring the content of chemical components in chrysanthemum, which is used for simultaneously measuring the content of 13 chemical components of isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin and cryptochlorogenic acid in different chrysanthemum medicinal materials and can be used for quality control of the chrysanthemum medicinal materials. The specific technical scheme is as follows:

the application provides a method for measuring the content of chemical components in chrysanthemum, which adopts ultra-high performance liquid chromatography-mass spectrometry to simultaneously measure the content of 13 chemical components in chrysanthemum; the 13 chemical compositions comprise: isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin, cryptochlorogenic acid, the method comprising:

(1) establishing a standard curve of 13 chemical compositions

Preparing 5-10 mixed reference substance solutions containing 13 chemical components with different known concentrations by taking methanol with volume fraction of 60-100% as a solvent; wherein, the concentrations of hyperoside and isoquercitrin are 0.1-250ng/mL respectively, the concentration of quercetin is 0.1-1500ng/mL, the concentrations of hesperidin, acacetin and diosmetin are 0.1-2500ng/mL respectively, the concentration of luteolin is 0.1-5500ng/mL, the concentrations of luteolin-7-O-glucuronide, cosmosiin, apigenin and cryptochlorogenic acid are 0.1-15000ng/mL respectively, and the concentrations of chlorogenic acid and isochlorogenic acid are 0.5-25000ng/mL respectively;

under the same chromatographic and mass spectrometric conditions, volume V is subjected to₁Injecting each control mixed solution into an ultra-high performance liquid chromatograph, determining chemical components of each chromatographic peak through mass spectrum detection, and obtaining the chromatographic peak area of each chemical component;

wherein the chromatographic conditions comprise:

a chromatographic column: octadecylsilane chemically bonded silica chromatographic column;

mobile phase: the phase A is formic acid water solution with the volume fraction of 0.05-0.15%, and the phase B is acetonitrile; gradient elution is carried out by adopting 5-94% of phase A and 6-95% of phase B in volume fraction; flow rate: 0.2-0.4 mL/min; column temperature: 15-25 ℃; sample volume V₁：3-8μL；

Respectively establishing a standard curve of each chemical component by taking the peak area of each chemical component chromatographic peak as a vertical coordinate and the concentration of each chemical component as a horizontal coordinate;

(2) obtaining the chromatographic peak area of a sample solution to be detected;

dissolving a sample to be detected with the mass of M into a volume V by using a methanol solution with the volume fraction of 60-80%₂Taking the supernatant fluid, filtering the supernatant fluid to be used as a sample solution to be detected, wherein M/V₂5-20 mg/mL;

taking the volume V under the same chromatographic and mass spectrometric conditions as in step (1)₁Injecting the sample solution to be detected into an ultra-high performance liquid chromatograph, determining chemical components of each chromatographic peak through mass spectrum detection, and obtaining the chromatographic peak area of each chemical component;

(3) determining the contents of 13 chemical components in a sample to be detected;

according to the established standard curve of each chemical component, the concentration C of each chemical component is respectively obtained from the chromatographic peak area of each chemical component in the sample solution to be detected₁Respectively calculating the contents C of 13 chemical components in the sample to be detected according to the following formula;

C＝C₁×V₂/M。

in some embodiments of the present application, the mixed control solution has hyperin and isoquercitrin concentrations of 1-200ng/mL, quercetin concentrations of 5-1000ng/mL, hesperidin, acacetin, diosmetin concentrations of 10-2000ng/mL, luteolin concentrations of 25-5000ng/mL, luteolin-7-O-glucuronide, cosmosiin, apigenin, cryptochlorogenic acid concentrations of 50-10000ng/mL, chlorogenic acid and isochlorogenic acid concentrations of 100-20000 ng/mL.

In the application, the methanol with the volume fraction of 60-100% refers to methanol or methanol aqueous solution with the volume fraction of more than or equal to 60%. In some embodiments of the present application, preferably, in step (1), the mixed control solution is prepared by using methanol as a solvent.

In some embodiments of the present application, in step (1), methanol with volume fraction of 60-100% is used as a solvent to prepare a mixed control stock solution containing 13 chemical components, wherein the concentrations of hyperin and isoquercitrin are 200-250ng/mL, the concentration of quercetin is 1000-1500ng/mL, the concentrations of hesperidin, farnesin and diosmetin are 2000-2500ng/mL, the concentration of luteolin is 5000-5500ng/mL, the concentrations of luteolin-7-O-glucuronide, cosmosiin, apigenin and cryptochlorogenic acid are 10000-15000ng/mL, and the concentrations of chlorogenic acid and isochlorogenic acid are 20000-25000 ng/mL;

and diluting the mixed reference substance stock solution by using methanol with the volume fraction of 60-100% to obtain 5-10 reference substance mixed solutions containing 13 chemical components with different known concentrations.

The solvent used for preparing the mixed reference stock solution and the solvent used for diluting the mixed reference stock solution can be the same or different, preferably, the solvent used for preparing the mixed reference stock solution and the solvent used for diluting the mixed reference stock solution are the same, and more preferably, the solvent used for preparing the mixed reference stock solution and the solvent used for diluting the mixed reference stock solution are methanol.

The inventors have found in their studies that better separation of the 13 chemical components can be achieved with the chromatographic conditions of the present application, and preferably, in some embodiments of the present application, the gradient elution is specifically: 0-2 minutes, 6% -25% B; 25% -26% of B in 2-4 minutes; 4-7 minutes, 26-75% of B, 7-8 minutes and 75-95% of B.

The type of mass spectrum is not limited in the present application as long as the object of the present invention can be achieved, and for example, triple quadrupole mass spectrometry can be used.

In order to effectively obtain molecular ion peaks and characteristic fragments of the chromatographically separated chemical components so as to obtain more accurate identification results of the chemical components, in some embodiments of the present application, the mass spectrometry conditions include: an electrospray ion source is adopted, multiple reaction ion monitoring is used as a detection mode, and a negative ion full scanning mode is adopted; capillary voltage: 3500 ℃ and 4500V; the temperature of the capillary tube is set to be 250 ℃ and 350 ℃, and the drying airflow speed is 5-10L/min; atomizer pressure 30-40 psi;

the characteristic ion peaks of the 13 chemical components in the mass spectrum detection comprise:

the method for determining the chemical component content in chrysanthemum provided by the embodiment of the invention adopts an ultra-high performance liquid chromatography-mass spectrometry combined technology (UPLC-MS/MS), can simultaneously determine the content of 13 chemical components in chrysanthemum by reasonably selecting chromatographic conditions and mass spectrometry conditions, and has the advantages of simplicity, high sensitivity, high analysis speed, strong specificity and the like, thereby being applicable to the quality control of chrysanthemum medicinal materials.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1, panel a, is a multiple reactive ion monitoring profile (MRM profile) of a control of 13 chemical compositions;

FIG. 1, panel b, is a multiple reactive ion monitoring spectrum (MRM chart) of 13 chemical components in chrysanthemum (lot 201810);

in fig. 1, each numeral symbol represents: 1. hesperidin; 2. isochlorogenic acid C; 3. isoquercitrin; 4. hyperin; 5. luteolin-7-O-glucuronide; 6. cosmosiin; 7. chlorogenic acid; 8. cryptochlorogenic acid; 9. quercetin; 10. diosmetin; 11. luteolin; 12. farnesin; 13. apigenin.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Instrument for measuring the position of a moving object

Agilent model 1290 high performance liquid chromatograph (Agilent, USA); agilent 6470 triple quadrupole tandem mass spectrometer (Agilent, USA); agilent Mass Hunter analysis software (Agilent, USA); a one hundred thousand balance of type AX205 (Mettler Toledo, Switzerland); SB-1000YDTD ultrasonic cleaning tank (Ningbo Xinzhi Biotechnology Co., Ltd.); model 3K15 high speed centrifuge (Sigma, usa); XW-80A vortex mixer (Shanghai province of analytical instruments); Milli-Q ultrapure water preparation apparatus (Millipore Corp.).

Reagent

The control product of isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin, cryptochlorogenic acid from Doudster Biotech limited; methanol and acetonitrile were chromatographically pure and purchased from Fisher corporation; formic acid is chromatographically pure and purchased from ROE corporation; ultrapure water was prepared by a Milli-Q ultrapure water meter.

Material

The chrysanthemum is produced in Anhui, Hubei, Zhejiang, Henan, Jiangxi and Hebei, respectively.

Example 1

1. Chromatographic condition determination

A chromatographic column: an ACQUITY UPLC BEH C18 column (2.1X 100mm,1.7 μm); mobile phase: phase A is 0.1% formic acid water, phase B is acetonitrile; gradient elution, the elution procedure is: 0-2min, 6% -25% of B; 2-4min, 25% -26% of B; 4-7min, 26-75% of B, 7-8min and 75-95% of B. Flow rate: 0.3 mL/min; column temperature: 20 ℃; sample introduction amount: 5 μ L.

2. Mass spectrometry condition determination

An electrospray ionization source (ESI) is adopted, multiple reactive ion monitoring (MRM) is used as a detection mode, and a negative ion full-scanning mode is adopted; capillary voltage: 4000V; the temperature of the capillary tube is set to be 300 ℃, and the flow rate of the drying gas is 7L/min; atomizer pressure 35 psi. The ion pairs and mass spectral parameters of the 13 analytes are shown in Table 1.

TABLE 1 ion Pair and Mass Spectrometry parameter information for 13 Compounds in Chrysanthemum

3. Preparation of stock solution of mixed reference substance

Accurately weighing 5.00mg of each of isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin and cryptochlorogenic acid as reference substances, placing the reference substances in a 5mL measuring flask, dissolving the reference substances in methanol, fixing the volume, preparing a reference substance stock solution with the concentration of 1mg/mL, and placing the reference substance stock solution in a refrigerator at 4 ℃ for cold storage for later use.

Taking each control stock solution with different volumes, and taking methanol as a solvent to prepare a mixed control stock solution with the concentrations of hyperin and isoquercitrin of 200ng/mL, quercetin of 1 mu g/mL, hesperidin, acacetin, diosmetin of 2 mu g/mL, luteolin of 5 mu g/mL, luteolin-7-O-glucuronide, cosmosiin, apigenin, cryptochlorogenic acid of 10 mu g/mL, and chlorogenic acid and isochlorogenic acid of 20 mu g/mL.

4. Preparation of sample solution to be tested

Crushing 10 kinds of chrysanthemum (sieving by a first sieve), precisely weighing 250mg of chrysanthemum coarse powder into a 25mL volumetric flask, fixing the volume to scale by 70% methanol, carrying out ultrasonic treatment (power 250W and frequency 50KHz) for 40min, taking out, standing to room temperature, adding 70% methanol to complement weight loss, filtering the taken supernatant by an organic microporous filter membrane of 0.45 mu m, and storing the stock solution in a refrigerator at 4 ℃ for later use. The sources of the 10 varieties of chrysanthemum are shown in table 2.

Table 210 sources of Chrysanthemum

Example 2 creation of a Standard Curve

The method comprises the steps of accurately measuring a proper amount of a mixed reference substance stock solution prepared in example 1, taking methanol as a solvent, carrying out gradient dilution on the mixed reference substance stock solution sequentially by 2 times, 2.5 times, 2 times and 2 times to obtain mixed reference substance solutions with 8 concentrations, respectively taking the mixed reference substance solutions with 8 concentrations, carrying out sample injection by 5 mu L to analyze, and obtaining multiple reaction ion monitoring maps of each reference substance under different concentrations, wherein the multiple reaction ion monitoring maps of the mixed reference substance solution after the second dilution are shown in a picture in figure 1. The concentration (X) of the analyte was taken as the abscissa and the peak area (Y) of the analyte was taken as the ordinate, regression calculation was performed by the weighted least squares method with the weight coefficient set to 1/X, the regression equation and the correlation coefficient of each compound were calculated, the concentration of each control at S/N (Signal-to-noise ratio) 10 was taken as the lowest limit of quantitation (LLOQ), and the results were recorded in Table 3.

TABLE 3 regression equation of standard curve for 13 chrysanthemum components

Example 3 precision test

1. Precision in the day: a mixed reference stock solution is prepared according to the method of example 1, the peak areas of the compounds are obtained by continuous 6 times of sample injection according to the chromatographic condition and the mass spectrum condition of example 1, the peak areas of 13 chemical components are recorded, and the RSD value is calculated, so that the method has good precision in day.

Table 4 in-day precision experimental data table of 13 chrysanthemum components (n ═ 6)

2. Precision in the daytime: a mixed control stock solution was prepared according to the method of example 1, and the injection was repeated 2 times for three consecutive days according to the chromatographic and mass spectrometric conditions of example 1, and the RSD value was calculated by recording the peak areas of 13 chemical components. The results are reported in table 5 and show that the method is good in daytime accuracy.

TABLE 5 day precision experimental data table for 13 ingredients of chrysanthemum (n ═ 6)

Example 4 repeatability test

Precisely weighing 6 parts of sample 1 (batch 201810) chrysanthemum powder (sieved by a No. one sieve), weighing 250mg of each part, preparing 6 parts of sample solution to be detected according to the method of example 1, performing sample injection on the sample solution to be detected in parts by 5 mu L to obtain peak areas of 13 components, calculating RSD values, and recording the result in 6, wherein the obtained result shows that the method has better repeatability.

Table 6 chrysanthemum 13 components repeatability test data table (n ═ 6)

Example 5 stability test

Sample 1 (lot 201810) was precisely weighed and 6 parts of chrysanthemum powder (sieved through a sieve) each weighing 250mg, 6 parts of a sample solution to be measured was prepared according to the method of example 1, and 5. mu.L of each sample solution was injected at time points 0, 2, 4, 8, 12, and 24 hours to obtain peak areas of 13 components to examine the stability of the samples in the sample injector and calculate RSD values, and the results are recorded in Table 7, which indicates that the stability of the 13 components in the samples was good after the samples were left in the sample injector for 24 hours.

Table 7 chrysanthemum 13 ingredients stability at room temperature for 24h (n ═ 3)

EXAMPLE 6 sample recovery test

Precisely weighing 6 parts of sample 1 (batch No. 201810) chrysanthemum powder (sieved by a No. one sieve), weighing 125mg of each part, adding a proper amount of mixed reference substance stock solution, fixing the volume to 25mL by methanol, subsequently carrying out ultrasonic treatment according to the method of example 1, preparing a sample solution to be detected in parallel, injecting 5 muL of sample volume, calculating the sample recovery rate and the corresponding RSD value, and recording the results in a table 8, wherein the sample recovery rate of the 13 components is 97.5-110.1%, and the RSD value is 0.4-3.6%.

Table 8 chrysanthemum 13 ingredients sample application recovery rate experimental data table (n ═ 6)

Example 7 sample determination

Take 1 in Table 20 batches of different chrysanthemum powder (screened by a No. one sieve) in 6 parts, weighing 250mg of each part, preparing a sample solution to be detected according to the method of example 1, and measuring according to the chromatographic condition and the mass spectrum condition of example 1 to obtain a multi-reaction ion chromatogram of each chemical component in the chrysanthemum sample, as shown in a b diagram in figure 1; determining the concentration C corresponding to the peak area of each chemical component in the chrysanthemum sample from the standard curve of each chemical component in Table 3₁Respectively calculating the contents of 13 chemical components in the sample to be detected according to the following formula; the results are reported in Table 9-1 and Table 9-2.

The content of each chemical component

TABLE 9-1 content of 13 ingredients (μ g/g) in different chrysanthemum species (n ═ 6)

Table 9-2 content of 13 ingredients (μ g/g) in different chrysanthemum (n ═ 6)

The method establishes a UPLC-MS/MS analysis method, and simultaneously determines the contents of 13 components, namely isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin and cryptochlorogenic acid in chrysanthemum.

All the embodiments in the present specification are described in a related manner, and the same and similar parts among the embodiments may be referred to each other, and each embodiment focuses on the differences from the other embodiments. In particular, for the system embodiment, since it is substantially similar to the method embodiment, the description is simple, and for the relevant points, reference may be made to the partial description of the method embodiment.

The above description is only for the preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.

Claims (6)

1. A method for measuring the chemical component content in chrysanthemum is characterized in that the ultra-high performance liquid chromatography-mass spectrometry is adopted, and the content of 13 chemical components in chrysanthemum is measured simultaneously; the 13 chemical compositions comprise: isochlorogenic acid C, hesperidin, cosmosiin, quercetin, acacetin, hyperoside, chlorogenic acid, luteolin-7-O-glucuronide, luteolin, apigenin, diosmetin, isoquercitrin, cryptochlorogenic acid, the method comprising:

(1) establishing a standard curve of 13 chemical compositions

Preparing 5-10 mixed reference substance solutions containing 13 chemical components with different known concentrations by taking methanol with volume fraction of 60-100% as a solvent; wherein, the concentrations of hyperoside and isoquercitrin are 0.1-250ng/mL respectively, the concentration of quercetin is 0.1-1500ng/mL, the concentrations of hesperidin, acacetin and diosmetin are 0.1-2500ng/mL respectively, the concentration of luteolin is 0.1-5500ng/mL, the concentrations of luteolin-7-O-glucuronide, cosmosiin, apigenin and cryptochlorogenic acid are 0.1-15000ng/mL respectively, and the concentrations of chlorogenic acid and isochlorogenic acid are 0.5-25000ng/mL respectively;

under the same chromatographic and mass spectrometric conditions, volume V is subjected to₁Injecting each control mixed solution into an ultra-high performance liquid chromatograph, determining chemical components of each chromatographic peak through mass spectrum detection, and obtaining the chromatographic peak area of each chemical component;

wherein the chromatographic conditions comprise:

a chromatographic column: octadecylsilane chemically bonded silica chromatographic column;

mobile phase: phase A is by volumeFormic acid water solution with the number of 0.05-0.15%, and acetonitrile as a phase B; gradient elution is carried out by adopting 5-94% of phase A and 6-95% of phase B in volume fraction; flow rate: 0.2-0.4 mL/min; column temperature: 15-25 ℃; sample volume V₁：3-8μL；

Respectively establishing a standard curve of each chemical component by taking the peak area of each chemical component chromatographic peak as a vertical coordinate and the concentration of each chemical component as a horizontal coordinate;

(2) obtaining the chromatographic peak area of a sample solution to be detected;

dissolving a sample to be detected with the mass of M into a volume V by using a methanol aqueous solution with the volume fraction of 60-80%₂Taking the supernatant fluid, filtering the supernatant fluid to be used as a sample solution to be detected, wherein M/V₂5-20 mg/mL;

taking the volume V under the same chromatographic and mass spectrometric conditions as in step (1)₁Injecting the sample solution to be detected into an ultra-high performance liquid chromatograph, determining chemical components of each chromatographic peak through mass spectrum detection, and obtaining the chromatographic peak area of each chemical component;

(3) determining the contents of 13 chemical components in a sample to be detected;

according to the established standard curve of each chemical component, the concentration C of each chemical component is respectively obtained from the chromatographic peak area of each chemical component in the sample solution to be detected₁Respectively calculating the contents C of 13 chemical components in the sample to be detected according to the following formula;

C＝C₁×V₂/M。

2. the method as claimed in claim 1, wherein the concentration of hyperin and isoquercitrin in the mixed control solution is 1-200ng/mL, the concentration of quercetin is 5-1000ng/mL, the concentration of hesperidin, acacetin and diosmetin is 10-2000ng/mL, the concentration of luteolin is 25-5000ng/mL, the concentration of luteolin-7-O-glucuronide, cosmosiin, apigenin and cryptochlorogenic acid is 50-10000ng/mL, and the concentration of chlorogenic acid and isochlorogenic acid is 100-20000 ng/mL.

3. The method according to claim 1, wherein the mixed control solution is prepared in step (1) using methanol as a solvent.

4. The method as claimed in claim 1 or 2, wherein in step (1), methanol with volume fraction of 60-100% is used as solvent to prepare a mixed control stock solution containing 13 chemical components, wherein the concentrations of hyperin and isoquercitrin are respectively 200-250ng/mL, the concentration of quercetin is 1000-1500ng/mL, the concentrations of hesperidin, farnesin and diosmetin are respectively 2000-2500ng/mL, the concentration of luteolin is 5000-5500ng/mL, the concentrations of luteolin-7-O-glucuronide, cosmosiin, apigenin and cryptochlorogenic acid are respectively 10000-15000ng/mL, and the concentrations of chlorogenic acid and isochlorogenic acid are 20000-25000 ng/mL;

diluting the mixed reference substance stock solution with methanol with volume fraction of 60-100% to obtain 5-10 mixed reference substance solutions containing 13 chemical components with different known concentrations.

5. The method according to claim 1, characterized in that the gradient elution is in particular: 0-2 minutes, 6% -25% B; 25% -26% of B in 2-4 minutes; 4-7 minutes, 26-75% of B, 7-8 minutes and 75-95% of B.

6. The method of claim 1, wherein the mass spectrometry is triple quadrupole tandem mass spectrometry, and wherein the mass spectrometry conditions comprise: an electrospray ion source is adopted, multiple reaction ion monitoring is used as a detection mode, and a negative ion full scanning mode is adopted; capillary voltage: 3500 ℃ and 4500V; the temperature of the capillary tube is set to be 250 ℃ and 350 ℃, and the drying airflow speed is 5-10L/min; atomizer pressure 30-40 psi;

the characteristic ion peaks of the 13 chemical components in the mass spectrum detection comprise:

Images