CN108037197B

CN108037197B - Method for qualitatively and quantitatively analyzing chemical components of non-volatile secondary metabolites of houttuynia cordata wall-broken decoction pieces

Info

Publication number: CN108037197B
Application number: CN201711205106.5A
Authority: CN
Inventors: 成金乐; 姜志宏; 卢敬光; 邓雯; 张藏蔓; 王静蓉; 徐吉银; 王义娜
Original assignee: Zhongshan Zeus Medicine Group Co ltd
Current assignee: Zhongshan Zeus Medicine Group Co ltd
Priority date: 2017-11-27
Filing date: 2017-11-27
Publication date: 2020-08-07
Anticipated expiration: 2037-11-27
Also published as: CN108037197A

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

Method for qualitatively and quantitatively analyzing chemical components of non-volatile secondary metabolites of houttuynia cordata wall-broken decoction pieces

Technical Field

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

Background

The houttuynia cordata wall-broken decoction pieces are wall-broken decoction piece products prepared by using houttuynia cordata as a raw material and utilizing a traditional Chinese medicine wall-breaking technology. Herba Houttuyniae is dry aerial part of Houttuynia cordata (Houttuynia cordiata Thunb.) of Saururaceae, has effects of clearing heat and detoxicating, resolving carbuncle and expelling pus, inducing diuresis and treating stranguria, and is mainly used for treating lung carbuncle and empyema, phlegm heat and asthma cough, dysentery, heat stranguria, carbuncle and suppurative sore. The traditional Chinese medicine wall-breaking decoction pieces are prepared by processing plant traditional Chinese medicine decoction pieces which meet the requirements of legal standards and have cell structures into powder with the particle size D90 less than 45 mu m (more than 300 meshes) by a modern crushing technology, and preparing the uniformly-dried granular decoction pieces with the whole components of the original decoction pieces of 30-100 meshes by inducing the adhesiveness of substances per se. Compared with the traditional decoction pieces, the traditional Chinese medicine wall-broken decoction pieces have the advantages of uniform quality, high medicine utilization rate, good stability and convenient and quick application. However, since the traditional Chinese medicine wall-broken decoction pieces do not have the morphological characteristics of the traditional Chinese medicine decoction pieces, the change of chemical components such as effective components or index components can be brought after the wall breaking; therefore, the component analysis of the wall-broken decoction piece product has important significance.

The chemical components of the houttuynia cordata mainly comprise compounds such as volatile oil, phenolic acids, flavonoids, alkaloids and the like, the volatile oil components are mainly detected by GC-MS at present, and for the research on non-volatile secondary metabolites (compounds such as phenolic acids, flavonoids, alkaloids and the like), the prior art mainly focuses on the aspects of extraction, separation and identification, cannot comprehensively reflect the chemical substance basis of the houttuynia cordata, and cannot reflect the diversity difference and the characteristics of the effective components of the houttuynia cordata in different production places. Meanwhile, the research on the chemical components of the houttuynia cordata wall-broken decoction pieces in the prior art is not reported.

Disclosure of Invention

The invention aims to overcome the defects and defects of the analysis and detection technology of the components of the non-volatile secondary metabolites in the houttuynia cordata wall-broken decoction pieces in the prior art, and provides a method for qualitatively and quantitatively analyzing the chemical components of the non-volatile secondary metabolites in the houttuynia cordata wall-broken decoction pieces.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for qualitatively and quantitatively analyzing chemical components of a non-volatile secondary metabolite of a houttuynia cordata wall-broken decoction piece comprises the following steps:

s1: pretreating houttuynia cordata wall-broken decoction pieces to prepare a sample solution;

and S2, comparing the control product serving as a reference and the cracking fragment information with a related database by using a UHP L C-Q-TOF/MS technology, and qualitatively or quantitatively analyzing the chemical components of the non-volatile secondary metabolites in the sample, wherein the mobile phase A is a formic acid aqueous solution, and the mobile phase B is a formic acid acetonitrile solution.

The invention firstly utilizes UHP L C-Q-TOF/MS technology and realizes qualitative and quantitative analysis of chemical components of the non-volatile secondary metabolites of the houttuynia cordata wall-broken decoction pieces by selecting the specific mobile phase, and detects 36 non-volatile secondary metabolites such as phenolic acids, flavonoids and alkaloids together, and has the advantages of high speed, strong specificity, low detection limit, strong qualitative and quantitative capabilities and the like.

Preferably, the gradient elution conditions are: 0.0-12.0 min: mobile phase A: 90% → 65%; 12.0-23.0 min: mobile phase A: 65% → 5%; 23.0-26.0 min: mobile phase A: 5 percent; 26.0-27.0 min: 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 houttuynia cordata wall-broken decoction pieces in the sample solution of S1 is 10-30 mg/m L, and more preferably, the mass concentration of the houttuynia cordata wall-broken decoction pieces in the sample solution of S1 is 20mg/m L.

The reference substance selected by the invention is a reference substance solution commonly used in the prior art and can be directly purchased.

Preferably, the reference substance is one or more of chlorogenic acid, neochlorogenic acid, quercetin, hyperoside, cannabinamide or aristolactam I mixed methanol solution.

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 mug/ml. More preferably, the concentration of chlorogenic acid in the control is 5.00. mu.g/ml, 10.00. mu.g/ml, 20.00. mu.g/ml, 30.00. mu.g/ml, 40.00. mu.g/ml and 50.00. mu.g/ml.

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

Preferably, the concentration of quercetin in the control is at least 5 concentration values within the concentration range of 32.40-97.20 μ g/ml. More preferably, the concentration of quercetin in the control is 32.40. mu.g/ml, 48.60. mu.g/ml, 64.80. mu.g/ml, 81.00. mu.g/ml and 97.20. mu.g/ml.

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

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

Preferably, the concentration of cannabinamide (CannabisinA) in the control is at least 5 concentration values in the concentration range of 0.80-16.08 μ g/ml. More preferably, the concentration of cannabinamide in the control is 0.80. mu.g/ml, 1.61. mu.g/ml, 4.02. mu.g/ml, 8.04. mu.g/ml and 16.08. mu.g/ml.

Preferably, the chromatographic column is an Agilent Eclips Plus C18, the flow rate is 0.3m L/min, the column temperature is 25 ℃ and the sample size is 2 μ L.

Preferably, the mass spectrum conditions are that the gas temperature is 200 ℃, the capillary voltage is 4500v (+), 3500v (-), the end platform deflection is 500, the drying gas flow rate is 8L/min, the spraying gas pressure is 2.5Bar, the collection rate is 2.0HZ, the residence time is 3.5sec, the response absolute threshold is 213cts, and the scanning range is 50-1300 m/z.

The invention also provides an efficient extraction method for preparing the sample solution containing the houttuynia cordata wall-broken decoction pieces.

Preferably, the sample solution in S1 is prepared by the following method: crushing and dissolving the houttuynia cordata wall-broken decoction pieces into a methanol solution for extraction, performing ultrasonic treatment, centrifuging, filtering, and taking supernatant for dilution to obtain the sample solution, wherein the mass fraction of methanol in the methanol solution is 50-100%.

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

Preferably, the extraction times of the houttuynia cordata wall-broken decoction pieces are 2 times.

Preferably, the dosage of the houttuynia cordata wall-broken decoction pieces and the methanol solution is 50mg to 1m L.

Preferably, the ultrasound conditions are: ultrasonic frequency: 50-60kHz, ultrasonic time: 30 minutes; the centrifugation conditions were: rotating speed: 4500 rpm, centrifugation time: for 5 minutes.

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

the method for qualitatively and quantitatively analyzing the chemical components of the non-volatile secondary metabolites of the houttuynia cordata wall-broken decoction pieces can identify the types of the components of the non-volatile secondary metabolites of the houttuynia cordata and the houttuynia cordata wall-broken decoction pieces and perform quantitative analysis by applying the UHP L C-Q-TOF/MS technology, detects 36 non-volatile secondary metabolites such as phenolic acids, flavonoids and alkaloids together, has the advantages of high speed, strong specificity, low detection limit, strong qualitative and quantitative capabilities and the like, and can be applied to diversity difference and characteristic analysis of effective components of the houttuynia cordata in a production place.

Drawings

FIG. 1 is a study of the extraction efficiency of the wall-broken decoction pieces of houttuynia cordata in example 1 (50% methanol), example 2 (70% methanol) and example 3 (100% methanol);

FIG. 2 is a BPC spectrum in positive ion mode of a sample solution and a blank solution obtained by the detection method provided in example 2;

FIG. 3 is a BPC spectrum in negative ion mode of a sample solution and a blank solution obtained by the detection method provided in example 2;

FIG. 4 shows the absorption chart of 260-400 nm ultraviolet rays of decoction pieces of houttuynia cordata;

FIG. 5 is a positive ion mode MS/MS spectrum of chlorogenic acid components;

FIG. 6 is an MS/MS spectrum of compounds 1(5-CQA), 2(3-CQA), 3(4-CQA) in negative ion mode;

FIG. 7 is an MS/MS spectrum of compound 17Quercetin in negative ion mode;

FIG. 8 is MS/MS spectra of flavonoids Rutin, Vitexin and Afzelin in negative ion mode;

FIG. 9 is a comparison graph of the peak discharge time of the houttuynia cordata wall-broken decoction pieces and the reference Hyperoside in the negative ion mode;

FIG. 10 is a schematic diagram showing the classification of alkaloid components in houttuynia cordata;

FIG. 11 is a fragment diagram of secondary mass spectrometry in positive ion mode of Amides alkaloids;

FIG. 12 is a secondary mass spectrum of Aristolochia lactam compounds 20, 20', 23, and 25 in positive ion mode;

FIG. 13 is a MS/MS spectrum in positive ion mode for Compound 9;

FIG. 14 is an ESI-MS/MS spectrum of crotonine (Asimilabine) in positive ion mode in Massbankk and its structural formula.

Detailed Description

The invention is further illustrated by the following examples. These examples are intended to illustrate the invention and are not intended to limit the scope of the invention. Experimental procedures without specific conditions noted in the examples below, generally according to conditions conventional in the art or as suggested by the manufacturer; the raw materials, reagents and the like used are, unless otherwise specified, those commercially available from the conventional markets and the like. Any insubstantial changes and substitutions made by those skilled in the art based on the present invention are intended to be covered by the claims.

Example 1-3 qualitative and quantitative analysis method for chemical components of non-volatile secondary metabolites of houttuynia cordata wall-broken decoction pieces

1. A method for qualitatively and quantitatively analyzing chemical components of a non-volatile secondary metabolite of a houttuynia cordata wall-broken decoction piece comprises the following steps:

(1) pretreating houttuynia cordata wall-broken decoction pieces to prepare a sample solution;

the method comprises the steps of taking 3 batches of houttuynia cordata wall-broken decoction pieces in equal amount, uniformly mixing, crushing, sieving by a No. 2 sieve, precisely weighing 3 parts of 0.5g of powder, respectively adding 50% methanol, 70% methanol and 100% methanol each with the weight of 10m L, carrying out ultrasonic treatment (power: 220W and frequency: 50-60kHz) for 30 minutes, centrifuging at 4500 rpm for 5 minutes, taking supernate, extracting filter residues again for 1 time according to the same method, respectively combining the supernate, fixing the volume of the 3 parts of supernate to 25m L measuring bottles, shaking uniformly, filtering, and taking subsequent filtrate, namely the sample solution containing the houttuynia cordata wall-broken decoction pieces in the examples 1,2 and 3 respectively.

(2) Comparing the control product serving as a reference with the cracking fragment information and a related database by using UHP L C-Q-TOF/MS technology, and performing qualitative or quantitative analysis on the chemical components of the non-volatile secondary metabolites in the sample, wherein the mobile phase A is a formic acid aqueous solution, and the mobile phase B is a formic acid acetonitrile solution.

A proper amount of chlorogenic acid, neochlorogenic acid, quercetin, hyperoside and aristolocamide I reference substances (manufacturer: Chengdu Rui Fengsi Biotech limited, purity > 96%) and cannabinamide (self-made, purity > 96%) were weighed, and methanol was added to prepare a mixed reference substance solution with the concentration shown in Table 1.

TABLE 1 control line row concentrations

(3) Conditions of analysis

The chromatographic condition shown in the table 2 and the mass spectrum condition shown in the table 3 are selected from an instrument of Agilent 1290series UHP L C and Bruker Q-TOF MS, a chromatographic column of Agilent eclipss Plus C18(2.1 × 150mm,1.8 mu m), a controlled flow rate of 0.3ml/min, a sample amount of 2 mu l at 25 ℃ and the like.

TABLE 2 gradient elution conditions

TABLE 3 Mass Spectrometry conditions

2. Measurement results

As shown in fig. 1, the method for examining the extraction efficiency of the houttuynia cordata wall-broken decoction pieces in examples 1 to 3 by adopting different extraction methods. As can be seen from the figure, 50-100% of methanol solution can extract the non-volatile secondary metabolites from the houttuynia cordata wall-broken decoction pieces.

2.1 identification of the composition of non-volatile secondary metabolites

The detection method provided in example 2 (i.e., the concentration of the methanol solution was 70%, and the extraction was performed twice) was selected for detection. As shown in fig. 2, the BPC (base peak chromatogram) spectra of the sample solution and the blank sample solution in the positive ion mode, fig. 3 is the BPC spectrum in the negative ion mode, and fig. 4 is the ultraviolet absorption spectrum of 260-400 nm of the houttuynia cordata decoction pieces. As can be seen from the figure, the method has 36 kinds of detection of nonvolatile secondary metabolites.

Further identification and content measurement of each component were carried out as follows.

(1) Identification of phenolic acid component

The detection method is selected, and 3 phenolic acid components are identified in total, namely a compound 1: 5-CQA (neochlorogenic acid), compound 2: 3-CQA (chlorogenic acid) and compound 3: 4-CQA (cryptochlorogenic acid). The absorption peaks of ultraviolet absorption diagrams of the

compounds

1,2 and 3 are 210nm and 325 nm; [ M + H ]]⁺Both are m/z 355.10, there is no difference in MS/MS in positive ion mode, as shown in FIG. 5. In negative ion mode [ M-H]^-Are all m/z 353.08 (the molecular formula is presumed to be C₁₆H₁₈O₉) As shown in FIG. 6, in the MS/MS spectrum, the peak ion M/z 191 of the base peaks of the compounds 1 and 2 is a quinic acid fragment, wherein caffeoyl [ M-H-174 ] with higher abundance is visible in the compound 1]^-A fragment of m/z 179 identified as 5-CQA (neochlorogenic acid); peak ion [ M-H-162-18 ] of Compound 3]^-m/z 173 loss of H for quinic acid₂O was produced and identified as 4-CQA (cryptochlorogenic acid). The retention time and MS/MS fragment of chlorogenic acid of compound 2 and the control are consistent, so that compound 2 is 3-CQA (chlorogenic acid).

(2) Identification of flavonoid components

Compound 17 molecular ion peak [ 2 ] in the anion modeM-H]^-Is m/z 301.0353 (presumed to be of the formula C)₁₅H₁₀O₇) Cleavage of the RDA to give fragment 151.0035 was observed in the secondary mass spectrum. One molecule of CO is lost, forming m/z273.0411 ions. There is also a significant fragment m/z 178.9982 (C) in the MS/MS spectrum₈H₃O₅ ^-) This fragment is formed after loss of a fragment (CO) having a molecular weight of 27.9949 Da. In addition, the classical fragmentation of quercetin in the negative ion mode is shown in fig. 7, identifying compound 17 as quercetin. The cracking rule of quercetin is as follows:

molecular ion peak [ M-H ] of compound 10 in anion mode]^-Is m/z 609.1461 (C)₂₇H₃₀O₁₆) In its secondary mass spectrum, cleavage of a molecule of rutinosyl group by the parent ion produces the aglycone radical ion [ M-H-309]^–M/z300 and aglycone ion [ M-H-308 ]]^–m/z 301. It is clear that in the secondary cleavage map, the relative abundance of the aglycone radical ion m/z300 is higher than that of the aglycone ion m/z301, as shown in FIG. 8, indicating that the rutinose group in the structure of Compound 10 is linked to the C-3 position of quercetin, and thus is identified as Rutin.

Molecular ion peaks [ M-H ] in anion mode of compounds 11 and 15]^-Are all m/z 431.0987 (C)₂₁H₂₀O₁₀) In the MS/MS spectrum of Compound 15, [ Y ] with loss of rhamnosyl group (146Da) was observed⁰-H]^-m/z 284, the relative abundance is higher than that of the aglycone ion [ Y⁰]^- Compound 15 was identified as Afzelin, as shown in figure 8. The MS/MS spectrum of the compound 11 obtains a base peak [ M-H-120]^-M/z 311, debris peak [ M-H-90]^-m/z341，[M-H-120-CO]^-m/z 283, which cleavage pathway is consistent with that of Vitexin/Isovitexin, as shown in FIG. 8, identifying Compound 11 as Vitexin/Isovitexin.

Molecular ion peak [ M-H ] of compound 13 in anion mode]^-Is m/z 447.0940 (C)₂₁H₂₀O₁₁) The relative abundance of m/z301 lost rhamnosyl (146Da) in the MS/MS map is higher than that of m/z300, which indicates that the compound is 7-O-flavonoid glycoside and is presumed to be Quercitin.

Molecular ion peaks [ M-H ] of Compounds 12 and 12' in anion mode]^-Are all m/z 463.0886 (C)₂₁H₂₀O₁₂) All generate aglycone free radical ion [ M-H-163]^–M/z300 and aglycone ion [ M-H-162 ]]^–m/z301, and the relative abundance of the aglycone radical ion m/z300 is higher than that of the aglycone ion m/z301, indicating that both compounds 12 and 12' are 3-O-flavonoid glycosides. Compound 12 was identified as Hyperoside and Compound 12' as Isoquercitrin by comparing the time to peak of the control Hyperoside (see FIG. 9).

(3) Identification of alkaloid components

The detection method provided by the embodiment 2 identifies Aporphines compounds in the houttuynia cordata wall-broken decoction pieces, and the Aporphines compounds are found in the medicinal materials for the first time.

The alkaloid in the houttuynia cordata is divided into 3 types:

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

(2) aristoloctams, which mainly includes compounds 23(Cepharanone B), 19(Caldensin), 20 (AristoloctamAII), 20' (Piperoctacta A);

(3) aporphine, which can be divided into 3 subclasses:

① Oxoamorphines, mainly including compounds 21(Splendidine), 18 (L physicamine), 14(4-hydroxy-1,2, 3-trimethyl-7H-dibenzo-quinolin-7-one);

② 5,4-Dioxoaporphines, mainly comprising compounds 19 (Noraritolodene), 21 '(Cepharadiene B), 22 (Norcephaladine B), 14' (Ouregenine);

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

(3.1) Amides

Reference product Moupinamide [ M + H ] in positive ion mode]⁺m/z 314.1408, control and decoction pieces of herba HouttuyniaeThe MS/MS spectra of

compounds

16, 24 are shown in FIG. 11.

(3.2) Aristoloctams

According to the documents Aristolochialactam alkaloids in houttuynia cordata, such as Aristoloctam A I, Piperoctam A, Cepharanone B and Caldensin, the secondary mass spectrum cleavage pattern of 4 components is shown in FIG. 12.

(3.3) Aporphines

According to the reference, the aporphine alkaloid ESI-MSⁿThe lysis law is that RNH is lost first₂Loss of NH when N at position 6 is unsubstituted₃(17Da) to produce [ M + H-17]⁺Fragment, loss of CH when N in position 6 is substituted by methyl₃NH₂(31Da) to produce [ M + H-31]⁺And (4) fragmenting. Subsequent loss of CH in ortho-hydroxyl and methoxy containing alkaloids₃OH, followed by loss of CO. The structural formula and the cracking rule of the aporphine alkaloid are shown as follows:

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

The cleavage fragments and pathways thereof are presumed to be shown below

Accords with the cracking rule of Aporphine compounds in a positive ion mode, and compares with ESI-MS of Asimilobine in a Massbank website₂Map (as in fig. 14), it is reasonable to assume that the compound is Asimilobine.

An aphrphine containing one or more methoxy-substituted groups not adjacent to the hydroxy groupThe base often had a 15Da (CH 3) or 31Da (OCH 3) dipole loss in the secondary mass spectrum. This regular cleavage fragment is also observed in aporphine alkaloids containing vicinal hydroxyl and methoxy groups, but in very low abundance. The first 17Da loss for compound 7c indicates no substitution for N, and no 32Da (CH) loss₃OH) fragment, assuming that both hydroxyl and methylene groups are not present at positions 1 and 11, compound 7C is assumed to be L aurotenine.

(3.4) Megastigmane Glycosines class

Molecular ion peak [ M-H ] of compound 8 in negative ion mode]^-Is m/z 385.1866 (C)₁₉H₃₀O₈) Generating a fragment m/z 223 with a missing glucosyl group (162Da), the compound is a component of the Megastigmane glycoids class, and the compound 8 is presumed to be

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

2.2 identification of non-volatile secondary metabolites and measurement of the content thereof

Identifying the non-volatile secondary metabolites in the houttuynia cordata decoction pieces according to the chemical component mass spectrum cracking rule and the accurate molecular weight provided by MS, wherein the detection results of the chemical components and the content thereof are shown in Table 4.

TABLE 4 identification result of each chemical component of houttuynia cordata wall-broken decoction pieces

TABLE 56 measurement results of the contents of the ingredients in houttuynia cordata decoction pieces

Note: the absolute content is measured according to a reference substance, and the relative content is not marked; the content of aristololactam alkaloid in the label a is calculated by taking aristololactam I as a reference substance, and the content of Aporphine alkaloid in the label b is calculated by taking cannabiamide (CannabisinA) as a reference substance.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A method for qualitatively and quantitatively analyzing chemical components of a non-volatile secondary metabolite of a houttuynia cordata wall-broken decoction piece is characterized by comprising the following steps of:

s2, comparing the cracking fragment information with a related database by using a UHP L C-Q-TOF/MS technology and taking a reference substance as a reference, and qualitatively and quantitatively analyzing the chemical components of the non-volatile secondary metabolites in the sample, wherein the mobile phase A is a formic acid aqueous solution, the mobile phase B is a formic acid acetonitrile solution, and the gradient elution conditions are 0.0-12.0 min, 90% → 65% of the mobile phase A, 12.0-23.0 min, 65% → 5% of the mobile phase A, 23.0-26.0 min, 5% of the mobile phase A, 26.0-27.0 min, 5% → 90% of the mobile phase A and Agilent eclipss Plus C18;

the mass spectrum conditions comprise gas temperature of 200 ℃, capillary voltage of 4500v + and 3500v-, end platform deflection of 500, drying gas flow rate of 8L/min, spray gas pressure of 2.5Bar, acquisition rate of 2.0HZ, residence time of 3.5sec, response absolute threshold of 213cts and scanning range of 50-1300 m/z.

2. The method as claimed in claim 1, wherein the mass concentration of the houttuynia cordata wall-broken decoction pieces in the sample solution of S1 is 10-30 mg/m L.

3. The method as claimed in claim 2, wherein the mass concentration of the houttuynia cordata wall-broken decoction pieces in the sample solution of S1 is 20mg/m L.

4. The method of claim 1, wherein the control is a mixed methanol solution of one or more of chlorogenic acid, neochlorogenic acid, quercetin, hyperoside, aristolactam I, or cannabinamide.

5. The method of claim 1, wherein the flow rate of the chromatographic column is 0.3m L/min, the column temperature is 25 ℃, and the sample size is 2 μ L.

6. The method of claim 1, wherein the sample solution in S1 is prepared by: crushing and dissolving the houttuynia cordata wall-broken decoction pieces into a methanol solution for extraction, performing ultrasonic treatment, centrifuging, filtering, and taking supernatant for dilution to obtain the sample solution, wherein 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. The method as claimed in claim 7, wherein the ratio of the herba houttuyniae wall-broken decoction pieces to the methanol solution is 50mg:1m L.