Method for measuring content of protocatechuic acid and chlorogenic acid in gynecological itching relieving tablet

Abstract

The invention relates to the technical field of medicine component detection, in particular to a method for measuring the content of protocatechuic acid and chlorogenic acid in gynecological itching relieving tablets, which comprises the following steps: preparing a control sample solution, weighing protocatechuic acid and chlorogenic acid, and preparing the control sample solution; preparing a test sample solution, respectively weighing quantitative patrinia, gynecological antipruritic tablet powder and negative control powder, and fixing the volume to obtain a first sample solution, a second sample solution and a third sample solution; detecting protocatechuic acid and chlorogenic acid of the control sample solution by using a dual-wavelength HPLC method; determining the content of protocatechuic acid and chlorogenic acid in the first sample solution, the second sample solution and the third sample solution by using a dual-wavelength HPLC (High Performance Liquid Chromatography) method; and obtaining the content of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablet powder. According to the method for measuring the content of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets, the content of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets is measured through a dual-wavelength method, sensitivity is high, and accuracy is high.

Description

Technical Field

[0001] The present invention relates to the technical field of drug component detection, and particularly relates to a method for determining the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets. Background Art

[0002] The main components of Fufang Zhiyang Tablets include Patrinia villosa Juss., Hedyotis diffusa Willd., Taraxacum mongolicum Hand.-Mazz., Memecylon lanceolatum Blanco, Rubia cordifolia L., Spatholobus suberectus Dunn, Angelica sinensis (Oliv.) Diels, and Corydalis yanhusuo W. T. Wang var. yanhusuo which is processed by vinegar roasting. Fufang Zhiyang Tablets have various effects such as clearing away heat and dampness, killing parasites and relieving itching, and are mainly used to treat diseases such as vaginitis caused by damp-heat. However, the relevant component standards of Fufang Zhiyang Tablets are not yet perfect. As one of the main raw materials in this prescription, Patrinia villosa Juss. is generally considered that the active ingredients in Fufang Zhiyang Tablets are in Patrinia villosa Juss. The organic acids in Patrinia villosa Juss. are mainly composed of protocatechuic acid and chlorogenic acid, and protocatechuic acid and chlorogenic acid play an important role in treating gynecological epigastric pain. Therefore, it is generally considered that Fufang Zhiyang Tablets rely on protocatechuic acid and chlorogenic acid in Patrinia villosa Juss. to provide medicinal effects. Therefore, determining the contents of protocatechuic acid and chlorogenic acid in Patrinia villosa Juss. and forming standards can provide a reference for formulating the standards of active ingredients in Fufang Zhiyang Tablets and improving the drug quality of Fufang Zhiyang Tablets.

[0003] In the prior art, the active ingredients of Patrinia villosa Juss. are often extracted by supercritical CO2 extraction method to obtain supercritical extracts, and then the contents of active ingredients in the extracts are determined by methods such as mass spectrometry or chromatography. However, the detection methods for the specific contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets are insufficient, and it is difficult to determine whether the protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets completely come from Patrinia villosa Juss. Therefore, it is difficult to quantify the contents of active ingredients in Fufang Zhiyang Tablets and there is a lack of corresponding standards.

[0004] Therefore, there is a need for a method that can determine the contents of protocatechuic acid and chlorogenic acid in Patrinia villosa Juss. in Fufang Zhiyang Tablets. Summary of the Invention

[0005] The main object of the present invention is to provide a method for determining the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets, aiming to solve the problem that the prior art lacks means to detect the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets.

[0006] To achieve the above object, the present invention provides a method for determining the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang Tablets, and the method includes the following steps:

[0007] Prepare a control sample solution, weigh 10 mg each of protocatechuic acid and chlorogenic acid, dissolve them in methanol and make up to 100 mL in a volumetric flask to obtain a 0.1 mg / mL control sample solution;

[0008] Prepare test sample solutions. Weigh a quantified amount of Patrinia scabiosaefolia, Fufang Zhiyang tablets powder, and negative control powder respectively, and make up to a constant volume to obtain the first sample solution, the second sample solution, and the third sample solution.

[0009] Use dual-wavelength HPLC to detect the control sample solution. Pass the control sample solution through the chromatographic column to determine protocatechuic acid and chlorogenic acid in it.

[0010] Use dual-wavelength HPLC to determine the first sample solution, the second sample solution, and the third sample solution, and determine the contents of protocatechuic acid and chlorogenic acid in the first sample solution, the second sample solution, and the third sample solution.

[0011] Compare the chromatogram of the control sample solution with the chromatograms of the first sample solution, the second sample solution, and the third sample solution to obtain the contents of protocatechuic acid and chlorogenic acid in the Fufang Zhiyang tablets powder.

[0012] Further, the step of using dual-wavelength HPLC to determine the contents of protocatechuic acid and chlorogenic acid in the first sample solution, the second sample solution, and the third sample solution includes:

[0013] Select two wavelengths at which protocatechuic acid and chlorogenic acid respectively have significant absorption in the ultraviolet-visible spectrum as the maximum absorption wavelength and the adjacent wavelength.

[0014] Measure the absorbance of the control sample solution at these two selected wavelengths and plot the standard curve.

[0015] Measure the absorbance of the first sample solution, the second sample solution, and the third sample solution at these two wavelengths and calculate through the standard curve to obtain the contents of protocatechuic acid and chlorogenic acid.

[0016] Further, when passing the control sample solution through the chromatographic column, the mobile phase is acetonitrile and / or phosphoric acid solution with a concentration of 0.1%.

[0017] Further, when passing the control sample solution through the chromatographic column, the column temperature is 40°C.

[0018] Further, when passing the control sample solution through the chromatographic column, the detection wavelengths are 260 nm and 325 nm.

[0019] The present invention provides a method for determining the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang tablets. This method determines the contents of protocatechuic acid and chlorogenic acid in Fufang Zhiyang tablets by the dual-wavelength method. The present invention has high sensitivity and a wide application range, and is used to provide data reference for formulating the quality standard of Patrinia scabiosaefolia in Fufang Zhiyang tablets, which is conducive to the development of new drugs and the improvement of the clinical application of Patrinia scabiosaefolia. Brief Description of the Drawings

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can also be obtained according to the processes shown in these drawings.

[0021] Figure 1 It is a schematic flowchart of an embodiment in the method for determining the contents of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets.

[0022] Figure 2 It is a chromatogram of a mixed reference substance at a wavelength of 260 nm in an embodiment of the present invention.

[0023] Figure 3 It is a chromatogram of a mixed reference substance at a wavelength of 325 nm in an embodiment of the present invention.

[0024] Figure 4 It is a chromatogram of protocatechuic acid in the gynecological antipruritic tablet sample of the present invention.

[0025] Figure 5 It is a chromatogram of chlorogenic acid in the gynecological antipruritic tablet sample of the present invention.

[0026] Figure 6 It is a chromatogram of a negative control sample at a wavelength of 260 nm in an embodiment of the present invention.

[0027] Figure 7 It is a chromatogram of a negative control sample at a wavelength of 325 nm in an embodiment of the present invention.

[0028] Figure 8 It is a chromatogram of protocatechuic acid in Patrinia scabiosaefolia in an embodiment of the present invention.

[0029] Figure 9 It is a chromatogram of chlorogenic acid in Patrinia scabiosaefolia in an embodiment of the present invention. Detailed implementation manners

[0030] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0031] It should be noted that all directional indications (such as up, down, left, right, front, back...) in the embodiments of the present invention are only used to explain the relative positional relationship, movement conditions, etc. between components in a certain specific posture (as shown in the attached drawings). If the specific posture changes, the directional indications will also change accordingly.

[0032] In addition, the descriptions involving "first", "second", etc. in the present invention are only for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include at least one such feature. In addition, the technical solutions between various embodiments can be combined with each other, but it must be based on the ability of those of ordinary skill in the art to implement. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such a combination of technical solutions does not exist and is not within the protection scope required by the present invention.

[0033] The Gynecological Itch-Relieving Tablets have various effects such as clearing heat and drying dampness, killing insects and relieving itching, etc. It is mainly used to treat diseases such as vaginitis caused by damp-heat. Patrinia scabiosaefolia is one of the main raw materials that make up the Gynecological Itch-Relieving Tablets. Detecting whether the active ingredients in the Gynecological Itch-Relieving Tablets completely come from Patrinia scabiosaefolia and obtaining the specific content of the active ingredients can provide a reference for the formulation of standards for the Gynecological Itch-Relieving Tablets.

[0034] Patrinia scabiosaefolia is composed of the dried whole herbs of Patrinia scabiosaefolia Fisch. ex Link or Patrinia villosa Juss. of the family Valerianaceae. It was initially recorded in "Shennong's Classic of Materia Medica" and was listed as a medium-grade herb in this book. In ancient medical practices, Patrinia scabiosaefolia was often used as a drug for treating gynecological-related diseases and could be used to treat various diseases such as gynecological epigastric pain, postpartum stasis, postpartum hemorrhage, furuncles and carbuncles, and eczema.

[0035] Dual-wavelength HPLC (High-Performance Liquid Chromatography) method is an analytical technique that uses two detectors with different wavelengths to simultaneously detect multiple components in a sample. This method has the advantages of being able to simultaneously detect two different wavelengths, improving selectivity and sensitivity.

[0036] In view of this, since the response values of protocatechuic acid and chlorogenic acid in the active ingredients are different at the same wavelength, the present invention selects the dual-wavelength HPLC method to determine the contents of protocatechuic acid and chlorogenic acid in the Gynecological Itch-Relieving Tablets and Patrinia scabiosaefolia, and the present invention proposes a method for determining the contents of protocatechuic acid and chlorogenic acid in the Gynecological Itch-Relieving Tablets.

[0037] Refer to Figures 1-9 , the embodiments of the present application provide a method for determining the contents of protocatechuic acid and chlorogenic acid in the Gynecological Itch-Relieving Tablets. The main instruments and experimental reagents used in the present invention are as follows:

[0038] Table 1 Main experimental instruments

[0039]

[0040]

[0041] Table 2 Main experimental reagents

[0042]

[0043] The experimental steps disclosed by the present invention are specifically as follows:

[0044] Step S1: Prepare a control sample solution. Weigh 10 mg each of protocatechuic acid and chlorogenic acid, mix the 10 mg of protocatechuic acid and 10 mg of chlorogenic acid, and make up the mixed powder to 100 mL in a volumetric flask with methanol to obtain a 0.1 mg / mL control sample solution;

[0045] Step S2: Weigh 1 g of Patrinia scabiosaefolia, add 25 mL of methanol, place it in an ultrasonic cleaner for ultrasonic extraction for 30 min. After extraction, filter the solution to obtain the total extract of Patrinia scabiosaefolia. Concentrate the total extract of Patrinia scabiosaefolia to 10 mL and then filter it through a microporous membrane with a pore size of 0.45 μm to obtain the first sample solution;

[0046] Weigh 6 g of Fufang Zhiyang tablets powder, add 25 mL of methanol, place it in an ultrasonic cleaner for ultrasonic extraction for 30 min. After extraction, filter the solution. After filtration, take 1 mL of the Fufang Zhiyang tablets solution and place it in a 10 mL volumetric flask for volume fixing. After volume fixing, filter the Fufang Zhiyang tablets solution through a microporous membrane with a pore size of 0.45 μm to obtain the second sample solution, and refrigerate the second sample solution for later use;

[0047] Weigh 5 g of negative control powder, put it into a beaker containing 400 mL of pure water, heat it to 80 - 100 °C and keep it for 2 h, then filter. Dissolve the filter residue with 100 mL of pure water, heat it to 80 - 100 °C again and maintain it for 2 h, then filter. Combine the two filtrates and concentrate them to dehydration. Add the dehydrated solid to 3 mL of methanol for dissolution, then take 1 mL of the solution, transfer it to a 10 mL volumetric flask for volume fixing. After volume fixing, filter it through a microporous membrane with a pore size of 0.45 μm to obtain the third sample solution, and refrigerate the third sample solution for later use;

[0048] Step S3: Use the dual-wavelength HPLC method to detect the control sample solution. Pass the control sample solution into the chromatographic column. The mobile phase is acetonitrile and a 0.1% phosphoric acid solution. At a flow rate of 1.0 mL / min, with a column temperature of 40 °C, an injection volume of 10 μL, and wavelengths of 260 nm and 325 nm, detect to obtain the contents of protocatechuic acid and chlorogenic acid and obtain the standard curve;

[0049] Step S4: Introduce the first sample solution, the second sample solution, and the third sample solution into the chromatographic column. Set the mobile phase as acetonitrile and a phosphoric acid solution with a concentration of 0.1%. At a flow rate of 1.0 mL / min, with a column temperature of 40 °C, an injection volume of 10 μL, and detect at wavelengths of 260 nm and 325 nm. Measure the absorbance of the first sample solution, the second sample solution, and the third sample solution at these two wavelengths, and calculate through the standard curve to obtain the contents of protocatechuic acid and chlorogenic acid;

[0050] Step S5: Compare the chromatogram of the control sample solution with the chromatograms of the first sample solution, the second sample solution, and the third sample solution to obtain the contents of protocatechuic acid and chlorogenic acid in the powder of the gynecological antipruritic tablets.

[0051] Among them, the negative control powder disclosed in the present invention is the powder of gynecological antipruritic tablets that do not contain Patrinia scabiosaefolia in the prescription.

[0052] Specifically, the mobile phase gradient elution conditions in Step S3 are shown in Table 3 as follows:

[0053] Table 3 Mobile phase gradient elution conditions

[0054]

[0055] As Figures 2-3 shown, the mixed reference substance is a solution obtained by mixing the control sample solutions prepared from protocatechuic acid and chlorogenic acid. In the control sample solution at a wavelength of 260 nm, the response of protocatechuic acid is higher, while the response of chlorogenic acid is lower; at a wavelength of 325 nm, the response of chlorogenic acid is higher and the response of protocatechuic acid is lower. Therefore, in the present invention, the dual-wavelength HPLC method is used to simultaneously determine the contents of protocatechuic acid and chlorogenic acid in the sample.

[0056] Specifically, the present invention also discloses Examples 1-10 of specific applications. Examples 1-10 are respectively ten different batches of gynecological antipruritic tablets, as follows:

[0057] Table 4 Sample batch numbers of the gynecological antipruritic tablets in Examples 1-10

[0058]

[0059] From Figure 4 、 Figure 5It can be obtained that 5.831 min is the retention time of protocatechuic acid, and 10.211 min is the retention time of chlorogenic acid. Their contents are calculated based on the peak areas. The content of protocatechuic acid is between 0.8674 mg / g and 1.4463 mg / g, with an average content of 1.1360 mg / g; the content of chlorogenic acid is between 0.2640 mg / g and 0.6070 mg / g, with an average content of 0.4263 mg / g.

[0060] Depend on Figure 6 , Figure 7 It can be seen that by comparing the control chromatogram of the negative control powder with the chromatogram of the gynecological antipruritic tablet sample, at the same retention time, the negative control powder also contains protocatechuic acid and chlorogenic acid standards, indicating that the protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablet are not only derived from the Patrinia lappa herb, but also from other medicinal materials in the gynecological antipruritic tablet.

[0061] The step S5 disclosed in the present invention further includes the following steps:

[0062] S51, conduct a linear relationship investigation, take the control sample solution and prepare test solutions with concentrations of 0.01 mg / mL, 0.02 mg / mL, 0.03 mg / mL, 0.05 mg / mL, 0.09 mg / mL, and 0.10 mg / mL, and perform chromatographic inspection on the test solution of each concentration by injecting 10 μL according to the chromatographic conditions and measure the peak area of ​​the test solution of each concentration, set the peak area Y (A) as the ordinate, and the mass concentration X (mg / mL) as the abscissa, and establish a linear regression equation;

[0063] In detail, the linear regression equation is shown in Table 5, and from the experimental results, it can be seen that the peak area and mass concentration of the two reference substances have a good linear relationship.

[0064] Table 5 Standard curve equations of protocatechuic acid and chlorogenic acid in control sample solutions (n=6)

[0065]

[0066] S52, perform a precision experiment, inject 10 μL of the mixed reference solution into the chromatograph according to the chromatographic conditions and repeat the injection 6 times, and calculate the RSD of each component based on the peak area data of the chromatogram;

[0067] In detail, the precision test results of the mixed reference solution are shown in Table 6.

[0068] Table 6 Precision test results of mixed reference solution (n=6)

[0069]

[0070] S53. Conduct a stability test. Take the mixed reference solution and inject 10 μL according to the chromatographic conditions. Multiple injections are carried out at 0 h, 4 h, 8 h, 12 h, 18 h, and 24 h. After injection, record the chromatogram and calculate the RSD based on the peak area.

[0071] Specifically, the stability test results of the mixed reference solution are shown in Table 7. After measurement and calculation, the RSD values of the stability of protocatechuic acid and chlorogenic acid are both < 3%, indicating that the stability of protocatechuic acid and chlorogenic acid is good.

[0072] Table 7 Stability test results (n = 6)

[0073]

[0074] S54. Conduct a repeatability test. Weigh 6 portions of the powder of Fufang Zhiyang Tablets, each portion being 6 g. Inject according to the chromatographic conditions and calculate the RSD of the repeatability of the powder of Fufang Zhiyang Tablets based on the peak area data of the chromatogram.

[0075] Table 8 Repeatability test results (n = 6)

[0076]

[0077] Specifically, as shown in Table 8, the RSD of the repeatability of protocatechuic acid and chlorogenic acid in the powder of Fufang Zhiyang Tablets is within 3%, indicating that the repeatability of protocatechuic acid and chlorogenic acid in the powder of Fufang Zhiyang Tablets is good.

[0078] S55. Conduct a spiking recovery test. Weigh 6 portions of the powder of Fufang Zhiyang Tablets, each portion being 3 g. Add equal amounts of protocatechuic acid and chlorogenic acid reference substances in equal proportion and prepare them into solutions. Inject 10 μL according to the chromatographic conditions, record the chromatographic results and calculate the recoveries of protocatechuic acid and chlorogenic acid.

[0079] Table 9 Determination results of the recovery test (n = 6)

[0080]

[0081] Specifically, as shown in Table 9, the recoveries of protocatechuic acid and chlorogenic acid are between 95% and 105%, and the RSD values are both within 3%, indicating that the recovery test scheme is feasible.

[0082] S56. Determine the contents of protocatechuic acid and chlorogenic acid in Patrinia scabiosaefolia Fisch. Take 10 μL of the first sample solution (Patrinia scabiosaefolia Fisch. solution) and conduct chromatographic detection under dual-wavelength HPLC.

[0083] Specifically, the chromatograms of protocatechuic acid and chlorogenic acid in Patrinia scabiosaefolia Fisch. are as shown in Figure 8 . Figure 9As shown, the retention times in Patrinia scabiosaefolia and the contents of protocatechuic acid and chlorogenic acid in Patrinia scabiosaefolia are shown in Table 10.

[0084] Table 10 Determination Results of the Contents of Protocatechuic Acid and Chlorogenic Acid in Patrinia scabiosaefolia

[0085]

[0086] S57, The contents of protocatechuic acid and chlorogenic acid in ten batches of different production batch numbers 23081801 (Sample 1), 23081802 (Sample 2), 23081803 (Sample 3), 23081804 (Sample 4), 23081805 (Sample 5), 23121105 (Sample 6), 23121101 (Sample 7), 23121104 (Sample 8), 23121102 (Sample 9), 23121103 (Sample 10) of the Gynecological Itching-Relieving Tablets were determined. Ten batches of different production batch numbers of Gynecological Itching-Relieving Tablets were configured into solutions, and then injected with 10 μL according to the chromatographic conditions. After injection, the chromatograms were recorded and the contents of protocatechuic acid and chlorogenic acid were calculated;

[0087] Table 11 Contents of Protocatechuic Acid and Chlorogenic Acid in the Gynecological Itching-Relieving Tablets of Ten Different Batches (n = 10)

[0088]

[0089] Experimental conclusion: By using the dual-wavelength high-performance HPLC method to determine the contents of protocatechuic acid and chlorogenic acid in Patrinia scabiosaefolia and the Gynecological Itching-Relieving Tablets, it is concluded that the protocatechuic acid and chlorogenic acid in the Gynecological Itching-Relieving Tablets not only come from Patrinia scabiosaefolia, but also from other medicinal materials in the Gynecological Itching-Relieving Tablets.

[0090] Combining all the above technical solutions, a method for determining the contents of protocatechuic acid and chlorogenic acid in the Gynecological Itching-Relieving Tablets provided by the present invention, by using the dual-wavelength high-performance HPLC method to determine the contents of protocatechuic acid and chlorogenic acid in Patrinia scabiosaefolia and the Gynecological Itching-Relieving Tablets, proves that the protocatechuic acid and chlorogenic acid in the Gynecological Itching-Relieving Tablets not only come from Patrinia scabiosaefolia, but also from other medicinal materials. This method has high sensitivity and a wide application range, provides data reference for the standard formulation of the Gynecological Itching-Relieving Tablets, is conducive to the development of new drugs and the improvement of the clinical application of Patrinia scabiosaefolia.

[0091] The above are only the preferred embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. Any equivalent structural transformation made under the inventive concept of the present invention by using the content of the specification and drawings of the present invention, or directly / indirectly applied in other related technical fields, is included in the patent protection scope of the present invention.

Claims

1. A method for determining the contents of protocatechuic acid and chlorogenic acid in Fuke Zhiyang tablets, characterized in that, It includes the following steps: Prepare a control sample solution. Weigh 10 mg each of protocatechuic acid and chlorogenic acid, dissolve them in methanol and make up the volume to 100 mL in a volumetric flask to obtain a 0.1 mg / mL control sample solution; Prepare test sample solutions. Weigh a certain amount of Patrinia scabiosaefolia, Fufang Zhiyang Tablets powder and negative control powder respectively, and make up the volume to obtain the first sample solution, the second sample solution and the third sample solution; Use dual-wavelength HPLC to detect the control sample solution. Inject the control sample solution into the chromatographic column to determine the protocatechuic acid and chlorogenic acid in it; Use dual-wavelength HPLC to determine the first sample solution, the second sample solution and the third sample solution, and determine the contents of protocatechuic acid and chlorogenic acid in the first sample solution, the second sample solution and the third sample solution; Compare the chromatograms of the control sample solution with those of the first sample solution, the second sample solution and the third sample solution to obtain the contents of protocatechuic acid and chlorogenic acid in the Fufang Zhiyang Tablets powder.

2. The method for determining the contents of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets as described in claim 1, characterized in that, The step of using dual-wavelength HPLC to determine the contents of protocatechuic acid and chlorogenic acid in the first sample solution, the second sample solution and the third sample solution includes: Select two wavelengths at which protocatechuic acid and chlorogenic acid respectively have significant absorption in the ultraviolet-visible spectrum as the maximum absorption wavelength and the adjacent wavelength; Measure the absorbance of the control sample solution at these two selected wavelengths and plot a standard curve; Measure the absorbance of the first sample solution, the second sample solution and the third sample solution at these two wavelengths and calculate through the standard curve to obtain the contents of protocatechuic acid and chlorogenic acid.

3. The method for determining the contents of protocatechuic acid and chlorogenic acid in the Gynecological Itch-relieving Tablets according to claim 1, wherein, When injecting the control sample solution into the chromatographic column, the mobile phase is acetonitrile and / or 0.1% phosphoric acid solution.

4. The method for determining the contents of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets as claimed in claim 1, characterized in that, When injecting the control sample solution into the chromatographic column, the column temperature is 40 °C.

5. The method for determining the contents of protocatechuic acid and chlorogenic acid in the gynecological antipruritic tablets according to claim 1, characterized in that, When injecting the control sample solution into the chromatographic column, the detection wavelengths are 260 nm and 325 nm.

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