WO2018076427A1

WO2018076427A1 - Method for detecting gallic acid in zhachong pills containing thirteen kinds of traditional chinese medicine ingredients

Info

Publication number: WO2018076427A1
Application number: PCT/CN2016/106660
Authority: WO
Inventors: 佘宜寰; 韩风雨; 王博一; 齐平; 于秀玲
Original assignee: 内蒙古天奇中蒙制药股份有限公司
Priority date: 2016-10-31
Filing date: 2016-11-21
Publication date: 2018-05-03
Also published as: CN106383111A

Abstract

A method for detecting gallic acid in Zhachong pills containing thirteen kinds of traditional Chinese medicine ingredients comprises the following steps: preparing a solution, screening experiment conditions, detecting the luminous intensity of a sample solution to be detected, drawing a standard curve, and finally substituting the related results into the standard curve to calculate the gallic acid content of the sample solution to be detected. The method for detecting gallic acid in Zhachong pills containing thirteen kinds of traditional Chinese medicine ingredients is combined with a chemiluminescence method, and provides high sensitivity; a good linear relationship may exist between the luminous intensity and the concentration of the detected matter in a range of 3-5 order of magnitudes, and the method has the advantages of high reproducibility and the like; and the method provides a theoretical basis for detection of active ingredients in the Zhachong pills containing thirteen kinds of traditional Chinese medicine ingredients and has important realistic significances in the aspects of medical research and environmental monitoring.

Description

Method for detecting gallic acid in Zhachong Xiewei Pill

This application claims the priority of the Chinese Patent Application filed on October 31, 2016, the Chinese Patent Office, the application number is 201610985206.3, and the invention is entitled "A method for detecting gallic acid in Zha Chong Xiewei Pill". This is incorporated herein by reference.

Technical field

The invention relates to a method for determining gallic acid, in particular to a method for determining the content of gallic acid in Zhachong Xiewei Pill.

Background technique

Zha Chong Xiewei Pill is one of the most commonly used traditional prescriptions in Mongolian medicine. It is also known as Yudi Thirteen Zha Chong Thirteen Pills. It is mainly made up of hazelnut, sarcophagus, woody, agarwood, and grass. It is composed of 13 kinds of traditional Chinese medicine ingredients. In clinical practice, the medicine is mainly used for hemiplegia, left and right sputum, sputum slanting, limb numbness, rheumatism and joint pain. Clinical studies have shown that Zhachong Xiewei Pill has obvious effects on promoting hematoma absorption, improving cerebral blood circulation, scavenging oxygen free radicals, inhibiting platelet aggregation, protecting myocardial ischemia and increasing cerebral blood flow. The scorpion is the prescription drug in the prescription, which has the effects of sputum diarrhea, stagnation of the lungs and cough, reducing the fire and pharynx. (Bao Naqin, Agula, Wang Lanying. The therapeutic effect of Zha Chong Thirteen Pills on cardiovascular and cerebrovascular diseases [ J]. Chinese Journal of Medicine and Pharmacy, 2005, 11(6): 9-10.).

Gallic acid is one of the main active ingredients in Zha Chong Xiewei Pills. Gallic acid (GA) is a natural phenolic compound widely found in fruits and Chinese herbal medicines. It has strong resistance. Oxidation, antibacterial, antiviral and antitumor effects (FU Yurong, ZHANG Wanming, CHEN Guimin. Determination of gallic acid and total phenolic acid in Jingtian Sanqi[J]. Chinese Patent Medicine, 2006, 28(7): 1016- 1018.).

At present, high-performance liquid chromatography, photometry, gas chromatography, thin-layer scanning, capillary electrophoresis, oscillating chemical reaction-pulse perturbation, and chemiluminescence are often used for the determination of gallic acid. Photometric and thin-layer scanning methods have poor sensitivity and narrow linear range; chromatography and capillary electrophoresis require more complicated and expensive instruments (Liu H, Ren J, Hao Y, et al. Determination of metoprolol tartrate in tablets and Human urine using flow-injection chemiluminescence method [J]. Journal of Pharmaceutical & Biomedical Analysis, 2006, 42(3): 384-388.). Chemiluminescence is a form of luminescence that is carried out by the chemical energy released by chemical reaction without any light, electricity or heat. The chemiluminescence method is generally considered to be highly sensitive, fast and simple. The advantage is a convenient and practical analysis method. Common luminescence systems mainly include luminol chemiluminescence system, luster and ruthenium chemiluminescence reaction system, peroxidic oxalic acid sulphate chemiluminescence system, nail complex chemiluminescence reaction, high Potassium manganate chemiluminescence reaction system.

In the prior art, there is no report on the determination of gallic acid content in Zhachong Xiewei Pill by using luminol chemiluminescence system.

Summary of the invention

The object of the present invention is to overcome the deficiencies of the prior art and provide a method for detecting gallic acid in Zhachong Xiewei Pill, which provides a theoretical basis and basis for the detection of active ingredients in Zhachong Thirteen Flavor; for medical research and Environmental testing has important practical significance.

The invention provides a method for detecting gallic acid in Zhachong Xiewei Pill, comprising the following steps:

1S: preparation solution: preparing gallic acid solution, preparing luminol stock solution with alkaline solution, preparing potassium ferricyanide stock solution;

2S: Screening experimental conditions: screening the concentration conditions of the luminol solution, the NaOH solution, and the potassium ferricyanide solution;

3S: detecting luminous intensity: inserting the first flow tube into the luminol solution, inserting the second flow tube into the gallic acid standard solution or the sample solution to be tested, and inserting the third and fourth flow tubes into the potassium ferricyanide solution; The pump flushes the flow path of the whole system until a stable blank signal is obtained; the mixed solution of the luminol solution and the gallic acid standard solution or the sample solution to be tested is injected into the carrier through the injection valve, and then mixed with the potassium ferricyanide solution. After reaching the flow cell, it produces chemiluminescence and records the light signal;

4S: Develop a standard curve;

5S: Calculated content: According to the test method of step 3S, the sample solution to be tested is subjected to measurement of luminescence intensity, and is substituted into a standard curve for calculation to obtain a gallic acid content of the sample solution to be tested.

Wherein, the experimental condition of the step 2S is: the concentration of the luminol solution ranges from 1.5×10 ^-4 to 2.5×10 ^-4 mol/L; the concentration of the NaOH solution ranges from 0.5 mol/L to 0.7 mol/L; The concentration of the potassium ferricyanide solution ranges from 7.0 × 10 ^-5 to 6.0 × 10 ^-5 mol / L.

Wherein, the experimental condition of the step 2S is: the concentration of the luminol solution is 2.0×10 ^-4 mol/L; the concentration of the NaOH solution is 0.6 mol/L; and the concentration of the potassium ferricyanide solution is 6.0×10 ^-5 Mol/L.

Wherein, the step 4S is specifically: under the experimental conditions of the step 2S screening, respectively, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1000 μL of the gallic acid solution prepared in the step 1S are taken up, and the volume is set in a 100 mL volumetric flask; The luminous intensity was measured and a standard curve was prepared.

Wherein, the equation of the standard curve in the step 4S is LgΔI=0.92LgC+8.78, and the correlation coefficient is 0.986.

Wherein, after the step 4S, the interference test is carried out on a gallic acid standard solution having a concentration of 2.0×10 ^-7 g/mL; 1000 times methanol, ethanol, isopropanol, acetone, Ca ²⁺ , Na ⁺ , K ⁺ , NH ₄ ⁺ , 500 times benzoic acid, citric acid, 200 times ascorbic acid, 100 times tartaric acid, sulfosalicylic acid, oxalic acid, 50 times Zn ²⁺ , 10 times Pb ²⁺ , 5 The doubling of Co ²⁺ and 0.5 times of tannic acid did not interfere with the measurement.

The step 1S is specifically:

(1) Preparing a gallic acid solution having a concentration of 2.0×10 ^-4 g/mL: weigh 0.0100 g of gallic acid, dissolve it with methanol and dilute to 50 mL, and dilute to the desired concentration when used;

(2) Prepare a concentration of 0.01 mol/L luminol stock solution with NaOH solution: weigh 0.1772 g of luminol standard, dissolve it with 0.5 mol/L NaOH solution, transfer to 100 mL brown volumetric flask, use the same The concentration of NaOH solution is adjusted to the mark, shaken and stored in the refrigerator for use;

(3) Prepare a potassium ferricyanide stock solution with a concentration of 0.01 mol/L, store it in a brown volumetric flask, and set aside.

The invention also provides an application method for detecting gallic acid in Zha Chong Xiewei Pill in detecting the content of gallic acid in Zhachong Xiewei Pill.

The method for detecting gallic acid in Zhachong Xiewei Pill of the present invention mainly comprises the following steps:

1S: Preparation solution:

2S: screening experimental conditions; the concentration of luminol solution ranges from 1.5×10 ^-4 to 2.5×10 ^-4 mol/L; the concentration of NaOH solution ranges from 0.5 mol/L to 0.7 mol/L; potassium ferricyanide solution The concentration range is 7.0×10 ^-5 to 6.0×10 ^-5 mol/L. Further preferably, the concentration of the luminol solution is 2.0×10 ^-4 mol/L; the concentration of the NaOH solution is 0.6 mol/L; and the concentration of the potassium ferricyanide solution is 6.0×10 ^-5 mol/L.

The specific screening basis is as follows:

Screening of luminol solution concentration:

The following conditions are fixed: the inner diameter of the first to fourth flow tubes is 0.8 mm, the valve cell distance is 8 cm, the main pump speed is 100 rpm, the NaOH concentration is 0.6 mol/L, and the potassium ferricyanide concentration is 6.0×10 ^-5 mol. /L. When the concentration of luminol is in the range of 1.5×10 ^-4 to 2.5×10 ^-4 mol/L, the relative chemiluminescence intensity is large; further preferably, when the concentration of the luminol solution is 2.0×10 ^-4 mol/ When L is used, the instrument is relatively stable. When the concentration is exceeded, the signal stability is poor, the comprehensive reagent dosage, signal-to-noise ratio and other factors are determined, and the luminol concentration is finally determined to be 2.0×10 ^-4 mol/L.

Screening of NaOH solution concentration:

The following conditions are fixed: the inner diameter of the first to fourth flow tubes is 0.8 mm, the valve cell distance is 8 cm, the main pump speed is 100 rpm, the luminol concentration is 2.0×10 ^-4 mol/L, and the potassium ferricyanide concentration is 6.0 × 10 ^-5 mol / L. The chemiluminescence intensity increases with the increase of NaOH concentration, but decreases with the increase of concentration after reaching the peak. When the concentration of NaOH solution ranges from 0.5mol/L to 0.7mol/L, the relative chemiluminescence intensity is higher. Large; further preferably, when the concentration of the NaOH solution is 0.6 mol/L, the system has the best signal to noise ratio.

Screening of potassium ferricyanide solution concentration:

The following conditions were fixed: the inner diameter of the first to fourth flow tubes was 0.8 mm, the valve cell distance was 8 cm, the main pump speed was 100 rpm, the luminol concentration was 2.0×10 ^-4 mol/L, and the NaOH concentration was 0.6 mol/ L. The concentration of potassium ferricyanide solution in the range of 7.0×10 ^-5 to 6.0×10 ^-5 mol/L can obtain a large signal-to-noise ratio and a large relative chemiluminescence intensity; further preferably, when the potassium ferricyanide solution The concentration has a maximum signal-to-noise ratio of 6.0×10 ^-5 , and the concentration of potassium ferricyanide solution is 6.0×10 ^-5 mol/L.

3S: detecting luminous intensity: inserting the first flow tube into the luminol solution, inserting the second flow tube into the gallic acid standard solution or the sample solution to be tested, and inserting the third and fourth flow tubes into the potassium ferricyanide solution; The pump flushes the flow path of the whole system until a stable blank signal is obtained; the mixed solution of the luminol solution and the gallic acid standard or sample is injected into the carrier through the injection valve, and then mixed with the potassium ferricyanide solution to reach the flow cell. , generating chemiluminescence, recording optical signals; carrying current is Na ₂ SO ₃ solution.

4S: Standard curve was prepared; under the experimental conditions of step 2S screening, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1000 μL of gallic acid solution with a concentration of 2.0×10 ^-4 g/mL were taken up, and the volume was adjusted to a 100 mL volumetric flask. The luminescence intensity was measured and a standard curve was established. The linear regression equation is: LgΔI=0.92LgC+8.78, the correlation coefficient is 0.986, the detection limit is 9.6×10 ^-9 mol/L, and the concentration of 2.0×10 ^-7 mol/L gallic acid standard solution is adopted. Twelve parallel measurements were performed with a relative standard deviation of 2.4%.

Interference test was carried out on a standard solution of gallic acid at a concentration of 2.0×10 ^-7 g/mL, and found that 1000 times of methanol, ethanol, isopropanol, acetone, Ca ²⁺ , Na ⁺ , K ⁺ , NH ₄ ⁺ , 500 Benzoic acid, citric acid, 200 times ascorbic acid, 100 times tartaric acid, sulfosalicylic acid, oxalic acid, 50 times Zn ²⁺ , 10 times Pb ²⁺ , 5 times Co ²⁺ , 0.5 times single Ninic acid did not interfere with the assay.

Compared with the prior art, the present invention has the following advantages and benefits:

1. High sensitivity: This method is combined with chemiluminescence method and has high sensitivity. The detection of some metal ions can reach 10-11g/ml, which is 4~5 orders of magnitude lower than the photometric method.

2. Wide linear range: The luminous intensity and the concentration of the measured object can have a good linear relationship in the range of 3 to 5 orders of magnitude.

3. Good reproducibility, etc.: In general optical analysis, the reaction conditions are difficult to control and the reproducibility is poor. The sample and the reagent are highly reproducibly mixed in the flow, which significantly improves the reproducibility of the analysis results.

DRAWINGS

The accompanying drawings, which are incorporated in FIG The elements in the following description are some embodiments of the present invention, and not all of the embodiments. Those skilled in the art can obtain other drawings according to the drawings without any creative work. .

Figure 1 is a view showing the steps of a method for detecting gallic acid in Zhachong Xiewei Pill according to the present invention;

Figure 2 is a graph showing the relationship between luminol concentration and signal to noise ratio;

Figure 3 is a graph showing the relationship between NaOH concentration and signal to noise ratio;

Figure 4 is a graph showing the relationship between potassium ferricyanide concentration and signal to noise ratio;

Figure 5 is a plot of injection volume versus signal to noise ratio;

Figure 6 is a schematic view of a gallic acid detecting instrument; wherein: 1: first flow tube 2: second flow tube 3: third flow tube 4: fourth flow tube 5: injection valve 6: flow cell 7: negative high voltage power supply 8: Computer W: Waste liquid.

detailed description

The technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. It is a partial embodiment of the invention, and not all of the embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative efforts are within the scope of the present invention. It should be noted that, in the case of no conflict, the features in the embodiments and the embodiments in the present application may be arbitrarily combined with each other.

1S: Preparation solution: 2.0×10 ^-4 g/mL gallic acid solution configuration: Weigh 0.0100g of gallic acid, dissolve it with methanol and dilute to 50mL, dilute to the required concentration when used; 0.01mol/L luminol reserve Liquid configuration: Weigh 0.1772g luminol standard (Flkua, Bioehemika), dissolve it with a small amount of 0.5mol/L NaOH solution, transfer it to a 100mL brown volumetric flask, dilute to the mark with the same concentration of NaOH solution, shake After storage, store in the refrigerator for use; accurately arrange 0.01 mol / L of potassium ferricyanide stock solution, store in a brown volumetric flask, and set aside.

2S: screening experimental conditions: the concentration of luminol solution is 2.0×10 ^-4 mol/L; the concentration of NaOH solution is 0.6 mol/L; the concentration of potassium ferricyanide solution is 6.0×10 ^-5 mol/L, The sample volume is between 100 ul and 150 ul.

3S: insert the first flow tube into the luminol solution with a concentration of 2.0×10 ^-4 mol/L, insert the second flow tube into the standard solution of gallic acid or the sample solution to be tested, and insert the third and fourth flow tubes into the concentration. 6.0×10 ^-5 mol/L potassium ferricyanide solution; open the peristaltic pump to flush the flow path of the whole system until a stable blank signal is obtained; pass the mixed solution of 75ul luminol solution and gallic acid standard or sample solution The injection valve is injected into the carrier stream, and then mixed with the potassium ferricyanide solution to reach the flow cell to generate chemiluminescence and record the optical signal;

4S: Standard curve was prepared; under the experimental conditions of step 2S screening, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1000 μL of gallic acid solution with a concentration of 2.0×10 ^-4 g/mL were taken up, and the volume was adjusted to a 100 mL volumetric flask. The luminescence intensity was measured and a standard curve was established. The linear regression equation is: Lg ΔI = 0.92 LgC + 8.78, and the correlation coefficient is 0.986.

5S: Calculated content: According to the test method of step 3S, the luminescence intensity of the sample solution is measured, and the standard curve is substituted for calculation to obtain the gallic acid content of the sample solution to be tested.

Specific examples of the detection method of gallic acid in Zha Chong Xiewei Pill are given below:

Example

1. Equipment:

IFFM-D flow injection chemiluminescence analyzer; F-7000 fluorescence spectrometer; TU-1901 dual beam UV-visible spectrophotometer.

2. Materials and reagents:

Methanol, sodium hydroxide, potassium ferricyanide, acetone, produced by Sinopharm Chemical Reagent Co., Ltd.

2.1 Solution preparation: 2.0×10 ^-4 g/mL gallic acid solution configuration: Weigh 0.0100g of gallic acid, dissolve it with methanol and dilute to 50mL, dilute to the required concentration when used; 0.01mol/L luminol stock solution Configuration: Weigh 0.1772g luminol standard (Flkua, Bioehemika), dissolve it with a small amount of 0.5mol / L NaOH solution, transfer it to a 100mL brown volumetric flask, dilute to the mark with the same concentration of NaOH solution, shake up Store in the refrigerator for later use; accurately configure 0.01 mol / L of potassium ferricyanide stock solution, stored in a brown volumetric flask, and set aside.

2.2 Preparation of the sample solution to be tested: Take a different batch of Zha Chong Xiewei Pills, grind it, place it in a Soxhlet extractor, add 50mL of acetone and heat it on the water bath for 8h, and dilute the extract to 50mL.

3. Experimental conditions: the concentration of luminol solution is 2.0×10 ^-4 mol/L; the concentration of NaOH solution is 0.6 mol/L; the concentration of potassium ferricyanide solution is 6.0×10 ^-5 mol/L; The inner diameter of the fourth flow tube is 0.8 mm, the valve cell distance is 8 cm, and the main pump speed is 100 rpm.

4. Detection:

4.1 Insert the first flow tube into the luminol solution with a concentration of 2.0×10 ^-4 mol/L, and insert the second flow tube into the 2.0×10 ^-7 g/mL gallic acid standard or the sample solution to be tested, third and fourth. The flow tube is inserted into a potassium ferricyanide solution at a concentration of 6.0×10 ^-5 mol/L; the peristaltic pump is turned on to flush the flow path of the whole system until a stable blank signal is obtained; 75 ul of luminol solution and 100 ul to 150 ul of gallops are used. The acid standard or the mixed solution of the sample solution to be tested is injected into the carrier stream through the injection valve, and then mixed with the potassium ferricyanide solution to reach the flow cell to generate chemiluminescence, and the optical signal is recorded;

4.2 Develop a standard curve; under the experimental conditions of step 2S screening, absorb 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1000 μL of gallic acid solution at a concentration of 2.0×10 ^-4 g/mL, and dilute to a 100 mL volumetric flask; The luminescence intensity was measured separately, and a standard curve was prepared. The linear regression equation is: Lg ΔI = 0.92 LgC + 8.78, and the correlation coefficient is 0.986.

4.3 Interference experiment: Interference test was carried out on a standard solution of gallic acid with a concentration of 2.0×10 ^-7 g/mL, and found that 1000 times of methanol, ethanol, isopropanol, acetone, Ca ²⁺ , Na ⁺ , K ⁺ , NH ₄ ⁺ , 500 times benzoic acid, citric acid, 200 times ascorbic acid, 100 times tartaric acid, sulfosalicylic acid, oxalic acid, 50 times Zn ²⁺ , 10 times Pb ²⁺ , 5 times Co ²⁺ , 0.5 times the tannic acid did not interfere with the assay.

4.4 Calculating the content: According to the test method of step 3S, the sample solution to be tested is measured for the luminescence intensity, and is substituted into a standard curve for calculation to obtain the gallic acid content of the sample solution to be tested.

4.5 spike recovery rate determination: the results are shown in Table 1.

The results of determination of gallic acid in the sample of Table 1 (n=3)

In summary, the detection method of gallic acid in Zhachong Xiewei Pill according to the present invention has the advantages of high sensitivity, wide linear range, good reproducibility, and the like, and significantly improves the reproducibility of the analysis result; The determination and detection of active ingredients in scent provides a theoretical basis and has important practical significance for medical research and environmental testing.

It should be noted that the term "comprises", "comprises" or any other variants thereof is intended to encompass a non-exclusive inclusion, such that a process, method, article, or device comprising a series of elements includes those elements. And also includes other elements not explicitly listed, or elements that are inherent to such a process, method, item, or device. An element that is defined by the phrase "comprising a ..." does not exclude the presence of additional elements in the process, method, article, or device that comprises the element.

The above embodiments are only used to illustrate the technical solutions of the present invention, and are not limited thereto. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or equivalently replace some of the technical features. Modifications or substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the invention.

Industrial applicability

The detection method of gallic acid in Zhachong Xiewei Pill of the invention has the advantages of high sensitivity, wide linear range and good reproducibility, and the reproducibility of the analysis result is remarkably improved; Measurement and detection provide a theoretical basis and have important practical significance for medical research and environmental testing.

Claims

A method for detecting gallic acid in Zhachong Xiewei Pill, characterized in that it comprises the following steps:

1S: preparation solution: preparing gallic acid solution, preparing luminol stock solution with alkaline solution, preparing potassium ferricyanide stock solution;

2S: Screening experimental conditions: screening the concentration conditions of the luminol solution, the NaOH solution, and the potassium ferricyanide solution;

3S: detecting luminous intensity: inserting the first flow tube into the luminol solution, inserting the second flow tube into the gallic acid standard solution or the sample solution to be tested, and inserting the third and fourth flow tubes into the potassium ferricyanide solution; The pump flushes the flow path of the whole system until a stable blank signal is obtained; the mixed solution of the luminol solution and the gallic acid standard solution or the sample solution to be tested is injected into the carrier through the injection valve, and then mixed with the potassium ferricyanide solution. After reaching the flow cell, it produces chemiluminescence and records the light signal;

4S: Develop a standard curve;

5S: Calculated content: According to the test method of step 3S, the sample solution to be tested is subjected to measurement of luminescence intensity, and is substituted into a standard curve for calculation to obtain a gallic acid content of the sample solution to be tested.
The method for detecting gallic acid in Zhachong Xiewei Pill according to claim 1, wherein the experimental conditions of the step 2S are:

The concentration of the luminol solution ranges from 1.5×10 -4 to 2.5×10 -4 mol/L; the concentration of the NaOH solution ranges from 0.5 mol/L to 0.7 mol/L; and the concentration of the potassium ferricyanide solution ranges from 7.0× 10 -5 to 6.0 × 10 -5 mol/L.
The method for detecting gallic acid in Zhachong Xiewei Pill according to claim 2, wherein the experimental conditions of the step 2S are:

The concentration of the luminol solution is 2.0×10 -4 mol/L; the concentration of the NaOH solution is 0.6 mol/L; and the concentration of the potassium ferricyanide solution is 6.0×10 -5 mol/L.
The method for detecting gallic acid in the Zha Chong Xiewei Pill according to any one of claims 1 to 3, wherein the step 4S is specifically: in the experimental condition of the step 2S screening, respectively, the suction step 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, and 1000 μL of the gallic acid solution prepared in 1S were adjusted to a 100 mL volumetric flask; the luminescence intensity was measured and a standard curve was prepared.
The method for detecting gallic acid in Zhachong Xiewei Pill according to claim 4, wherein the equation of the standard curve in the step 4S is Lg ΔI = 0.92 LgC + 8.78, and the correlation coefficient is 0.986.
The method for detecting gallic acid in Zhachong Xiewei Pill according to claim 4, further comprising, after said step 4S, a standard solution of gallic acid having a concentration of 2.0×10 -7 g/mL. Perform interference test; 1000 times methanol, ethanol, isopropanol, acetone, Ca 2+ , Na + , K + , NH 4 + , 500 times benzoic acid, citric acid, 200 times ascorbic acid, 100 times tartaric acid, sulphur Salicylic acid, oxalic acid, 50 times Zn 2+ , 10 times Pb 2+ , 5 times Co 2+ , 0.5 times tannic acid did not interfere with the determination.
The method for detecting gallic acid in Zhachong Xiewei Pill according to claim 1, wherein the step 1S is specifically:

(1) Preparing a gallic acid solution having a concentration of 2.0×10 -4 g/mL: weigh 0.0100 g of gallic acid, dissolve it with methanol and dilute to 50 mL, and dilute to the desired concentration when used;

(2) Prepare a concentration of 0.01 mol/L luminol stock solution with NaOH solution: weigh 0.1772 g of luminol standard, dissolve it with 0.5 mol/L NaOH solution, transfer to 100 mL brown volumetric flask, use the same The concentration of NaOH solution is adjusted to the mark, shaken and stored in the refrigerator for use;

(3) Prepare a potassium ferricyanide stock solution with a concentration of 0.01 mol/L, store it in a brown volumetric flask, and set aside.
The method for detecting gallic acid in Zhachong Xiewei Pill according to any one of claims 1 to 7 for detecting the content of gallic acid in Zhachong Xiewei Pill.