CN108037197B - A method for qualitative and quantitative analysis of chemical constituents of non-volatile secondary metabolites in Houttuynia cordata decoction pieces - Google Patents

Abstract

The invention relates to a method for qualitatively and quantitatively analyzing chemical components of non-volatile secondary metabolites of houttuynia cordata wall-broken decoction pieces, which comprises the following steps of S1 preprocessing the houttuynia cordata wall-broken decoction pieces to prepare a sample solution, S2 comparing information of cracking fragments and related databases by using a UHP L C-Q-TOF/MS technology and using a reference substance as a reference, and qualitatively or quantitatively analyzing the chemical components of the non-volatile secondary metabolites in the sample, wherein a mobile phase A is a formic acid aqueous solution, and a mobile phase B is a formic acid acetonitrile solution.

Description

Qualitative and qualitative analysis of the chemical constituents of non-volatile secondary metabolites in a kind of Houttuynia cordata broken wall decoction pieces Methods of Quantitative Analysis

technical field

The invention relates to the technical field of detection of chemical components of traditional Chinese medicinal material preparations, in particular to a method for qualitative and quantitative analysis of chemical components of non-volatile secondary metabolites of Houttuynia cordata decoction pieces.

Background technique

Houttuynia cordata broken wall decoction piece is a broken wall decoction piece product made from Houttuynia cordata using traditional Chinese medicine wall breaking technology. Houttuynia cordata is the dry aerial part of Houttuynia cordata Thunb. , heat dysentery, hot stranguria, carbuncle sore and other diseases. Chinese herbal broken wall decoction pieces are plant Chinese medicine decoction pieces that meet the requirements of legal standards and have a cellular structure, and are processed to D90﹤45μm (above 300 mesh) powder by modern pulverization technology. ~100-mesh homogeneous dry granular decoction pieces with all the ingredients of the original decoction pieces. Compared with traditional decoction pieces, broken wall decoction pieces of traditional Chinese medicine have the advantages of uniform quality, high drug utilization rate, good stability and convenient and quick application. However, because the broken wall decoction pieces of traditional Chinese medicine do not have the morphological characteristics of traditional Chinese medicine decoction pieces, the broken wall will bring changes in chemical components such as active ingredients or index components; therefore, the composition analysis of broken wall decoction pieces is of great significance.

The chemical components of Houttuynia cordata mainly include volatile oils, phenolic acids, flavonoids and alkaloids. Compounds such as alkaloids and alkaloids), the existing technology also mainly focuses on extraction, separation and identification, which cannot fully reflect the chemical substance basis of Houttuynia cordata, and it is difficult to reflect the diversity of Houttuynia cordata active ingredients in different origins. Sexual differences and characteristics. At the same time, there is no report on the research on the chemical constituents of Houttuynia cordata broken wall decoction pieces in the prior art.

SUMMARY OF THE INVENTION

The purpose of the present invention is to overcome the defects and deficiencies of the non-volatile secondary metabolite component analysis and detection technology in the Houttuynia cordata wall-broken pieces in the prior art, and to provide a non-volatile secondary metabolite chemical composition of the Houttuynia cordata broken-wall pieces. Methods of qualitative and quantitative analysis. The method provided by the invention utilizes UHPLC-Q-TOF/MS technology for the first time, and realizes the qualitative and quantitative analysis of the chemical components of non-volatile secondary metabolites of Houttuynia cordata broken wall decoction pieces through the selection of a specific mobile phase. It has the advantages of strong specificity, low detection limit, strong qualitative and quantitative ability and so on.

For realizing the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

A method for qualitative and quantitative analysis of chemical components of non-volatile secondary metabolites of Houttuynia cordata decoction pieces, the detection method comprises the following steps:

S1: Pretreatment of the broken pieces of Houttuynia cordata to make a sample solution;

S2: Using UHPLC-Q-TOF/MS technology, using the reference substance as a control and comparing the fragmentation information with the relevant database, qualitative or quantitative analysis of the chemical components of non-volatile secondary metabolites in the sample is carried out. A is formic acid aqueous solution, and mobile phase B is formic acid acetonitrile solution.

The present invention utilizes UHPLC-Q-TOF/MS technology for the first time and realizes the qualitative and quantitative analysis of the chemical components of the non-volatile secondary metabolites of the broken pieces of Houttuynia cordata through the selection of a specific mobile phase, and a total of phenolic acids are detected. 36 kinds of non-volatile secondary metabolites, such as flavonoids and alkaloids, have the advantages of fast speed, strong specificity, low detection limit, strong qualitative and quantitative ability and so on.

Preferably, the gradient elution conditions are: 0.0-12.0min: mobile phase A: 90%→65%; 12.0-23.0min: mobile phase A: 65%→5%; 23.0-26.0min: mobile phase A: 5%; 26.0～27.0min: mobile phase A: 5%→90%.

Preferably, the volume fraction of formic acid in the mobile phases A and B is 0.1%.

Preferably, the mass concentration of the broken pieces of Houttuynia cordata in the sample solution of S1 is 10-30 mg/mL. More preferably, the mass concentration of Houttuynia cordata broken wall decoction pieces in the sample solution of S1 is 20 mg/mL.

The selected reference substance in the present invention is the reference substance solution commonly used in the prior art, which can be directly purchased.

Preferably, the reference substance is one or more mixed methanol solutions of chlorogenic acid, neochlorogenic acid, quercitrin, hypericin, cannabidiolamide or aristololactam I.

Preferably, the concentration of chlorogenic acid in the reference substance is at least 5 concentration values within the concentration range of 5.00-50.00 μg/ml. More preferably, the concentration of chlorogenic acid in the reference substance is 5.00 μg/ml, 10.00 μg/ml, 20.00 μg/ml, 30.00 μg/ml, 40.00 μg/ml and 50.00 μg/ml.

Preferably, the concentration of neochlorogenic acid in the reference substance is at least 5 concentration values within the concentration range of 6.00-48.00 μg/ml. More preferably, the concentration of neochlorogenic acid in the control substance is 6.00 μg/ml, 12.00 μg/ml, 24.00 μg/ml, 36.00 μg/ml and 48.00 μg/ml.

Preferably, the concentration of quercetin in the reference substance is at least 5 concentration values within the concentration range of 32.40-97.20 μg/ml. More preferably, the concentration of quercitrin in the control substance is 32.40 μg/ml, 48.60 μg/ml, 64.80 μg/ml, 81.00 μg/ml and 97.20 μg/ml.

Preferably, the concentration of hyperin in the reference substance is at least 5 concentration values within the concentration range of 4.00-40.00 μg/ml. More preferably, the concentration of chlorogenic acid in the reference substance is 4.00 μg/ml, 8.00 μg/ml, 16.00 μg/ml, 24.00 μg/ml, 32.00 μg/ml and 40.00 μg/ml.

Preferably, the concentration of aristololactam I in the reference substance is at least 5 concentration values within the concentration range of 1.84-9.20 μg/ml. More preferably, the concentration of quercetin in the control substance is 1.84 μg/ml, 3.68 μg/ml, 5.52 μg/ml, 7.36 μg/ml and 9.20 μg/ml.

Preferably, the concentration of cannabisin A in the reference substance is at least 5 concentration values within the concentration range of 0.80-16.08 μg/ml. More preferably, the concentration of cannabidiol in the control substance is 0.80 μg/ml, 1.61 μg/ml, 4.02 μg/ml, 8.04 μg/ml and 16.08 μg/ml.

Preferably, the chromatographic column is Agilent Eclips Plus C18, the flow rate is 0.3 mL/min, the column temperature is 25°C, and the injection volume is 2 μL.

Preferably, the mass spectrometry conditions are: gas temperature: 200°C; capillary voltage: 4500v(+), 3500v(-); end platform offset: 500; drying gas flow rate: 8L/min; spray gas pressure: 2.5Bar; acquisition rate : 2.0HZ; dwell time: 3.5sec; response absolute threshold: 213cts; scanning range: 50～1300m/z.

The invention also provides an efficient extraction method for preparing a sample solution containing broken wall decoction pieces of Houttuynia cordata.

Preferably, the sample solution in S1 is prepared by the following method: the broken pieces of Houttuynia cordata are crushed and dissolved in methanol solution for extraction, ultrasonication, centrifugation and filtration to take the supernatant to dilute to obtain the sample solution, the methanol solution The mass fraction of methanol is 50-100%.

Preferably, the mass fraction of methanol in the methanol solution is 70%.

Preferably, the number of times of extraction of the Houttuynia cordata broken wall decoction pieces is 2 times.

Preferably, the dosage of Houttuynia cordata broken wall decoction pieces and methanol solution is 50mg:1mL.

Preferably, the ultrasonic conditions are: ultrasonic frequency: 50-60 kHz, ultrasonic time: 30 minutes; the centrifugation conditions are: rotational speed: 4500 rpm, and centrifugation time: 5 minutes.

Compared with the prior art, the present invention has the following beneficial effects:

The method for qualitative and quantitative analysis of the chemical components of non-volatile secondary metabolites in Houttuynia cordata broken wall decoction pieces provided by the present invention can identify non-volatile Houttuynia cordata and Houttuynia cordata broken wall decoction pieces by using UHPLC-Q-TOF/MS technology. The components of volatile secondary metabolites were quantitatively analyzed, and a total of 36 non-volatile secondary metabolites such as phenolic acids, flavonoids and alkaloids were detected. It has the advantages of strong quantitative ability, etc., and can be applied to the diversity difference and characteristic analysis of the active components of Houttuynia cordata in the producing area.

Description of drawings

Fig. 1 is the investigation of embodiment 1 (50% methanol), embodiment 2 (70% methanol) and embodiment 3 (100% methanol) on the extraction efficiency of Houttuynia cordata broken wall decoction pieces;

Fig. 2 is the BPC spectrum under the positive ion mode of the sample solution that the detection method provided by embodiment 2 obtains and blank solution;

Fig. 3 is the BPC spectrum under the negative ion mode of the sample solution that the detection method that embodiment 2 provides and blank solution obtain;

Figure 4 is the UV absorption diagram of Houttuynia cordata decoction pieces at 260-400 nm;

Figure 5 is the positive ion mode MS/MS spectrum of chlorogenic acids;

Fig. 6 is the MS/MS spectrum of compound 1 (5-CQA), 2 (3-CQA), 3 (4-CQA) under negative ion mode;

Figure 7 is the MS/MS spectrum of compound 17Quercetin in negative ion mode;

Figure 8 is the MS/MS spectrum of the flavonoids Rutin, Vitexin and Afzelin in negative ion mode;

Fig. 9 is a comparison chart of the peak time of Houttuynia cordata broken wall decoction pieces and the reference substance Hyperoside negative ion mode;

Figure 10 is a schematic diagram of the classification of alkaloids in Houttuynia cordata;

Figure 11. Fragmentation map of secondary mass spectrometry in positive ion mode of Amides alkaloids;

Figure 12 is the secondary mass spectrum of aristololactam compounds 20, 20', 23 and 25 in positive ion mode;

Figure 13 is the MS/MS spectrum of compound 9 in positive ion mode;

Figure 14 shows the ESI-MS/MS spectrum and structural formula of Asimilobine in Massbank in positive ion mode.

Detailed ways

The present invention is further described below in conjunction with the examples. These examples are only intended to illustrate the present invention and not to limit the scope of the present invention. The experimental methods that do not specify specific conditions in the following examples are usually in accordance with the conventional conditions in the field or the conditions suggested by the manufacturer; the raw materials, reagents, etc. used, unless otherwise specified, are available from commercial channels such as conventional markets. The obtained raw materials and reagents. Any insubstantial changes and substitutions made by those skilled in the art on the basis of the present invention fall within the scope of protection claimed by the present invention.

Embodiments 1-3 Methods for qualitative and quantitative analysis of the chemical constituents of non-volatile secondary metabolites in Houttuynia cordata decoction pieces

1. A method for qualitative and quantitative analysis of chemical components of non-volatile secondary metabolites of Houttuynia cordata decoction pieces, comprising the following steps:

(1) pretreatment of the broken wall decoction pieces of Houttuynia cordata is made into sample solution;

Take 3 batches of Houttuynia cordata broken wall decoction pieces, mix them evenly, smash through No. 2 sieve, accurately weigh 3 parts of 0.5g powder, add 50% methanol, 70% methanol, 100% methanol each 10mL, ultrasonic treatment (power : 220W, frequency: 50-60kHz) for 30 minutes, centrifuge at 4500 rpm for 5 minutes, and take the supernatant; then extract the filter residue once again in the same way, and combine the supernatants separately; Fill the volume to a 25mL volumetric flask, shake well, filter, and take the subsequent filtrate, which is the sample solution containing the broken-wall decoction pieces of Houttuynia cordata of Examples 1, 2 and 3, respectively.

(2) Using UHPLC-Q-TOF/MS technology, using the reference substance as a control and comparing the fragmentation information with the relevant database, qualitative or quantitative analysis of the chemical components of non-volatile secondary metabolites in the sample was carried out. Phase A is formic acid in water and mobile phase B is formic acid in acetonitrile.

Weigh chlorogenic acid, neochlorogenic acid, quercitrin, hypericin, aristololactam I reference substance (manufacturer: Chengdu Refensi Biotechnology Co., Ltd., purity> 96%), cannabidiol (self-made, Purity>96%) appropriate amount, add methanol to make the mixed reference solution with the concentration shown in Table 1.

Table 1 Concentration of reference substance series

(3) Analysis conditions

Selected instrument: Agilent 1290series UHPLC, Bruker Q-TOF MS, chromatographic column: AgilentEclips Plus C18 (2.1×150mm, 1.8μm), control flow rate: 0.3ml/min, column temperature: 25°C, injection volume: 2μl; Chromatographic conditions shown in Table 2 and mass spectrometry conditions shown in Table 3.

Table 2 Gradient elution conditions

Table 3 Mass Spectrometry Conditions

2. Measurement results

As shown in Figure 1, it is the investigation of the extraction efficiency of Houttuynia cordata broken wall decoction pieces using different extraction methods in Examples 1-3. It can be seen from the figure that 50-100% methanol solution can extract the non-volatile secondary metabolites in the broken pieces of Houttuynia cordata.

2.1 Component identification of nonvolatile secondary metabolites

The detection method provided in Example 2 (that is, the concentration of methanol solution was 70%, extracted twice) was selected for detection. As shown in Figure 2, it is the BPC (Base Peak Chromatogram) spectrum of the sample solution and blank sample solution in positive ion mode, Figure 3 is the BPC spectrum in negative ion mode, and Figure 4 is the UV absorption map of Houttuynia cordata decoction pieces at 260-400 nm. As can be seen from the figure, a total of 36 non-volatile secondary metabolites were detected by this method.

Further identification and content determination of each component are carried out below.

(1) Identification of phenolic acids

Using this detection method, a total of 3 phenolic acid components were identified, namely compound 1: 5-CQA (neochlorogenic acid), compound 2: 3-CQA (chlorogenic acid) and compound 3: 4-CQA (crypto green ortho acid). The UV absorption peaks of compounds 1, 2, and 3 are at 210 nm and 325 nm; [M+H] ⁺ are all m/z 355.10, and there is no difference in MS/MS in positive ion mode, as shown in Figure 5. In the negative ion mode [MH] ^- both m/z 353.08 (presumably the molecular formula is C ₁₆ H ₁₈ O ₉ ), as shown in Figure 6, in the MS/MS spectrum, the base peak ions of compounds 1 and 2 are m/z 191 It is a quinic acid fragment, among which the fragment of the higher abundance caffeic acid group [MH-174] ^- m/z 179 can be seen in compound 1, which is identified as 5-CQA (neochlorogenic acid); the base peak ion of compound 3 [ MH-162-18] ^- m/z 173 was produced by the loss of H ₂ O from quinic acid and was identified as 4-CQA (cryptochlorogenic acid). The retention time and MS/MS fragments of compound 2 and the reference substance chlorogenic acid were consistent, so compound 2 was 3-CQA (chlorogenic acid).

(2) Identification of flavonoids

The molecular ion peak [MH] ^- of compound 17 in negative ion mode is m/z 301.0353 (presumably the molecular formula is C ₁₅ H ₁₀ O ₇ ), and fragment 151.0035 can be obtained by RDA fragmentation in its MS spectrum. One molecule of CO is lost to form the m/z 273.0411 ion. There is also a prominent fragment m/z 178.9982 (C ₈ H ₃ O ₅ ^- ) in the MS/MS spectrum, which was formed after the loss of the fragment (CO) with a molecular weight of 27.9949 Da. In addition, the classical fragment of quercetin in negative ion mode is shown in Figure 7, and compound 17 was identified as quercetin. The cleavage rule of quercetin is as follows:

The molecular ion peak [MH] ^of compound 10 in negative ion mode is m/z 609.1461 (C ₂₇ H ₃₀ O ₁₆ ), in its MS spectrum, a molecule of rutino group is lost by the fragmentation of the parent ion to generate aglycone free radical Ion [MH-309] ^– m/z 300 and aglycone ion [MH-308] ^– m/z 301. It is obvious that in the secondary fragmentation spectrum, the relative abundance of the aglycon radical ion m/z 300 is higher than that of the aglycon ion m/z 301, as shown in Figure 8, indicating the structure of compound 10. The rutose group in quercetin is connected to the C-3 position of quercetin, so it is identified as Rutin.

The molecular ion peaks [MH] ^of compounds 11 and 15 in negative ion mode are both m/z 431.0987 (C ₂₁ H ₂₀ O ₁₀ ), and in the MS/MS spectrum of compound 15, [Y] that has lost rhamnosyl (146Da) can be seen ⁰ -H] ^- m/z 284, the relative abundance is higher than that of the aglycone ion [Y ⁰ ] ^- , as shown in Figure 8, the compound 15 is identified as Afzelin. In the MS/MS spectrum of compound 11, the base peak [MH-120] ^- m/z 311, the fragment peak [MH-90] ^- m/z 341, [MH-120-CO] ^- m/z 283, this cleavage pathway Consistent with the cleavage pathway of Vitexin/Isovitexin, as shown in Figure 8, compound 11 was identified as Vitexin/Isovitexin.

The molecular ion peak [MH] ^of compound 13 in negative ion mode is m/z 447.0940 (C ₂₁ H ₂₀ O ₁₁ ), and the MS/MS spectrum shows that the relative abundance of m/z 301, which loses rhamnosyl group (146Da), is higher than m/z 300, indicating that the compound is a 7-O-flavonoid glycoside, presumably Quercitrin.

Compounds 12 and 12' in negative ion mode have molecular ion peaks [MH] ^- both at m/z 463.0886 (C ₂₁ H ₂₀ O ₁₂ ) both produce aglycon radical ions [MH-163] ^- m/z 300 and aglycone Ion [MH-162] ^– m/z 301, and the relative abundance of aglycon radical ion m/z 300 is higher than that of aglycon ion m/z 301, indicating that compounds 12 and 12' are both 3- O-flavonoid glycosides. By comparing the peak time of the reference substance Hyperoside (as shown in Figure 9 ), it was determined that the compound 12 was Hyperoside, and the other compound 12' was Isoquercitrin.

(3) Identification of alkaloids

The detection method provided in Example 2 identifies aporphines in the broken wall decoction pieces of Houttuynia cordata, which is the first time found in this medicinal material.

The alkaloids in Houttuynia cordata are divided into 3 categories:

(1) Amides, such as compounds 16 (Moupinamide) and 24 (Asperglaucide);

(2) Aristolactams, mainly including compounds 23 (Cepharanone B), 19 (Caldensin), 20 (AristolactamAII), 20' (PiperolactamA);

(3) Aporphine category, which can be divided into 3 subcategories:

①Oxoaporphines, mainly including compounds 21(Splendidine), 18(Lysicamine), 14(4-hydroxy-1,2,3-trimethoxy-7H-dibenzo-quinolin-7-one);

②5,4-Dioxoaporphines, mainly including compounds 19 (Noraritolodione), 21' (Cepharadione B), 22 (Norcepharadione B), 14' (Ouregidione);

③Aporphines, mainly include compounds 4 (4a, 4a', 4b, 4b'), 5, 5', 6, 6', 7 (7a, 7b, 7c) and compound 9 (Asimilobine), as shown in Figure 10.

(3.1) Amides class

The reference substance Moupinamide is [M+H] ⁺ m/z 314.1408 in positive ion mode, and the MS/MS spectra of compounds 16 and 24 in the reference substance and Houttuynia cordata decoction pieces are shown in Figure 11.

(3.2) Aristolactams class

According to the literature, the aristololactam alkaloids in Houttuynia cordata include AristolactamA I, PiperolactamA, Cepharanone B and Caldensin.

(3.3) Aporphine class

According to the reference, the cleavage rule of ESI-MS ⁿ of apophenanthine alkaloids is that RNH ₂ is lost first, and NH ₃ (17Da) is lost when the 6-position N is unsubstituted to produce [M+H-17] ⁺ fragments, and when the 6-position N is unsubstituted, the [M+H-17] + fragment is generated. Loss of _{CH3NH2 (} 31 Da) with methyl substitution yields the [M+H-31] ⁺ fragment. Subsequently, CH ₃ OH is lost in alkaloids containing vicinal hydroxyl and methoxy groups, followed by loss of CO. The structural formulas and cracking rules of apophthene alkaloids are as follows:

The [M+H] ⁺ m/z of compound 9 in positive ion mode is 268.1333, and the MS/MS spectrum is shown in Figure 13 .

It is speculated that its fragmentation fragments and pathways are as follows

Consistent with the fragmentation rule of aporphine compounds in positive ion mode, and comparing the ESI-MS ₂ spectrum of Asimilobine in the Massbank website (as shown in Figure 14), it is reasonable to speculate that the compound is Asimilobine.

Arborphine alkaloids containing one or more methoxy groups that are not adjacent to the hydroxyl group often have a dipole loss of 15Da(CH3*) or 31Da(OCH3*) in the secondary mass spectrometry. This regular fragmentation was also observed in apophine alkaloids containing vicinal hydroxyl and methoxy groups, but in very low abundance. Compound 7c lost 17Da first, indicating that N was not substituted, and there was no loss of 32Da(CH ₃ OH) fragment. It was speculated that there was no hydroxyl group and methylene group at the 1-position and 11-position, so compound 7C was speculated to be Laurotetanine. According to this rule, a series of similar components such as 4a, 4b, 5, 5', 6, 6', 7a, 7b and other compounds are deduced.

(3.4) Megastigmane Glycosides

The molecular ion peak [MH] ^of compound 8 in negative ion mode is m/z 385.1866 (C ₁₉ H ₃₀ O ₈ ), resulting in the fragment m/z 223 that loses the glucosyl group (162Da). Combined with references, it is speculated that compound 8 is

(E)-4-Hydroxy-4-[3'-(β-D-glucopyranosyloxy)butylidene]-3,5,5-trimethyl-2-cyclophexen-1-one.

2.2 Identification and content determination results of non-volatile secondary metabolites

The non-volatile secondary metabolites in Houttuynia cordata decoction pieces were identified according to the above-mentioned chemical composition mass spectrometry fragmentation rules and the accurate molecular weight provided by MS. The detection results of each chemical composition and its content are shown in Table 4.

Table 4 Identification results of chemical components of Houttuynia cordata decoction pieces

Table 56 batches of Houttuynia cordata decoction pieces content determination results

Note: The absolute content measured according to the reference substance is marked with *, and the relative content is not marked; the content of aristololactam alkaloids marked with aristolochia lactam I is calculated as the reference substance, and the content of Aporphine marked with b The content of alkaloids was calculated with Cannabisin A as the reference substance.

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, The simplification should be equivalent replacement manners, which are all included in the protection scope of the present invention.

Claims (8)

1. a method for qualitative and quantitative analysis of the non-volatile secondary metabolite chemical composition of Houttuynia cordata broken wall decoction pieces, is characterized in that, comprises the steps: S1: Pretreatment of the broken pieces of Houttuynia cordata to make a sample solution; S2: Using UHPLC-Q-TOF/MS technology, using the reference substance as a control, the fragmentation information is compared with the relevant database, and the chemical components of non-volatile secondary metabolites in the sample are qualitatively and quantitatively analyzed. Among them, the mobile phase A is formic acid aqueous solution, mobile phase B is formic acid acetonitrile solution; gradient elution conditions are: 0.0～12.0min: mobile phase A: 90%→65%; 12.0～23.0min: mobile phase A: 65%→5%; 23.0 ~26.0min: mobile phase A: 5%; 26.0~27.0min: mobile phase A: 5%→90%; the chromatographic column is Agilent Eclips Plus C18; The mass spectrometry conditions are: gas temperature: 200°C; capillary voltage: 4500v+, 3500v-; end platform offset: 500; drying gas flow rate: 8L/min; spray gas pressure: 2.5Bar; acquisition rate: 2.0HZ; dwell time: 3.5sec; response absolute threshold: 213cts; scanning range: 50～1300m/z. 2 . The method according to claim 1 , wherein the mass concentration of Houttuynia cordata broken wall decoction pieces in the sample solution of S1 is 10-30 mg/mL. 3 . 3. The method according to claim 2, wherein the mass concentration of Houttuynia cordata broken wall decoction pieces in the sample solution of S1 is 20 mg/mL. 4. method according to claim 1, is characterized in that, described reference substance is a kind of in chlorogenic acid, neochlorogenic acid, quercitrin, hypericin, aristololactam I or cannabamide Or several mixed methanol solutions. 5 . The method according to claim 1 , wherein the flow rate of the chromatographic column is 0.3 mL/min, the column temperature is 25° C., and the injection volume is 2 μL. 6 . 6. The method according to claim 1, wherein the sample solution in S1 is prepared by the following method: the broken wall decoction pieces of Houttuynia cordata are pulverized and dissolved in methanol solution for extraction, and supernatant is obtained by ultrasonication, centrifugation and filtration. The sample solution is obtained by diluting, and the mass fraction of methanol in the methanol solution is 50-100%. 7. The method according to claim 6, wherein the mass fraction of methanol in the methanol solution is 70%. 8. method according to claim 7, is characterized in that, the consumption ratio of described Houttuynia cordata broken wall decoction pieces and methanol solution is 50mg: 1mL.

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