Quality control method of compound Gaoziba tablets

Abstract

The application discloses a quality control method of compound Gaozibo tablets, which comprises the following steps: 1) preparing a control solution; 2) preparing a test solution of the compound Gaozibo tablets; 3) diluting the control solution and injecting the test solution into a high performance liquid chromatograph for chromatographic condition detection; and 4) obtaining the content information of rutin, fumotoxin, zephyranthine and / or rosmarinic acid in the compound Gaozibo tablets according to the detection result. The method has excellent sensitivity, stability, repeatability and accuracy, and can be used for quality detection, quality evaluation or quality control of the compound Gaozibo tablets.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of medicines, in particular to a quality control method of compound Gaozi tablets. BACKGROUND

[0002] The quality evaluation mode of traditional Chinese medicine of "quantitative analysis of multi-components by single-marker" (QAMS) is a quantitative analysis method for simultaneously determining multiple components by using only one reference substance based on the relative correction factors among the components to be measured. The QAMS method is widely used in the quality control of traditional Chinese medicinal materials, extracts and prepared formulations, and is highly recognized in the traditional Chinese medicine industry. The 2020 edition of the Chinese Pharmacopoeia has applied the QAMS method to the content determination of many prepared formulations such as ginkgo leaf dripping pills / capsules and Keteling tablets / capsules. The research basis of ethnic medicines is very weak, and the reference substances of many main effective components are difficult to obtain, which brings great difficulties to the establishment and subsequent application of the quality control method. The establishment of the multi-component quantitative analysis method of ethnic medicines by the QAMS method not only makes up for the defects of some reference substances such as difficult preparation, high price, difficult batch supply and even no commercial products, but also provides a low-cost, high-efficiency, stable and reliable detection method for the quality evaluation and control of ethnic medicines.

[0003] The application takes the compound Gaozi tablets as an example to establish a simultaneous determination method (for example, the "quantitative analysis of multi-components by single-marker" method) of four components of rutin, fumotoxin, zephyranthine and rosmarinic acid, which can be used for the quality detection, quality evaluation or quality control of the compound Gaozi tablets.

[0004] At present, there is no literature report on the analysis and research method of the four chemical components of the above-mentioned compound Gaozi tablets. SUMMARY

[0005] Based on this, the present invention provides a quality control method for Compound Gaoziban Tablets, which includes the following steps: 1) Preparation of mixed reference solutions: preparing reference solutions of rutin, rosmarinic acid, naringin, and rosmarinic acid; 2) Preparation of Compound Gaoziban Tablets test solution: weighing an appropriate amount of Compound Gaoziban Tablets powder, adding alcohol for extraction, cooling, weighing, shaking, filtering, and collecting the filtrate to obtain the Compound Gaoziban Tablets test solution; 3) injecting the reference solutions and the test solution separately into a high-performance liquid chromatograph for chromatographic detection under the following conditions: using a column packed with octadecylsilane-bonded silica gel, and mobile phase A. The mobile phase B is an aqueous solution of acid, an aqueous solution of alkali, and / or an aqueous solution of buffer salt. The mobile phase B is selected from one or more of acetonitrile, methanol, and tetrahydrofuran. The gradient elution program is as follows: 0–10 min, 10% → 15% B; 10–40 min, 15% → 17% B; 40–50 min, 17% → 21% B; 50–60 min, 21% → 30% B. The flow rate is 0.4–1.5 ml / min, the column temperature is 20–50 °C, the detection wavelength is 200–350 nm, and the injection volume is 10–30 μl. 4) Based on the detection results, obtain the content information of rutin, rosmarinic acid, naringin, and / or rosmarinic acid in the compound Gaoziban tablets.

[0006] Furthermore, the method further includes calculating the relative retention value, relative correction factor and / or content of the target compound using rutin or narcissin as internal references, wherein the target compound is selected from one or more of the following: rutin, narcissin, narcissin and rosmarinic acid.

[0007] Further, the calculation of the relative retention value, relative correction factor, and / or content of the target compound using rutin or naringin as internal references includes: (a) determining standard curves for rutin, naringin, naringin, and rosmarinic acid based on the mass concentration of each reference standard in the mixed reference solution and the corresponding peak area in the chromatogram; (b) determining the internal reference, and according to the slope of the standard curve obtained in step (a), calculating the relative retention value, relative correction factor, and / or content of the target compound using formula f. s / x =K s / K x Calculate the relative correction factors for the target compounds, where f s / x K represents the relative correction factor of target compound x relative to internal reference s. s K represents the slope of the standard curve for the internal reference s. x (c) The slope of the standard curve for the target compound x; The chromatographic peaks of rutin, rosmarinic acid, naringin, and rosmarinic acid in the high performance liquid chromatogram of the compound Gaoziban tablets test solution were located using relative retention values, according to formula r x / s =t Rx / t Rs Calculate the relative retention value of the target compound, where rx / s represents the relative retention value of target compound x relative to internal reference s, t Rx is the retention time of target compound x, t Rs is the retention time of internal reference s; and (d) according to the mass concentration of the control solution of the target compound x in step (a) and the corresponding peak area, the relative correction factor in step (b) and the corresponding peak area of the chromatographic peak located in step (c), the formula C x = (A x / A s ) × C s × f s / x is used to calculate the content of rutin, farrerol, zephyranthine and rosmarinic acid in the test product solution of the compound Guaoshiba tablets, wherein C x represents the mass concentration of target compound x in the test product solution, A x represents the peak area of target compound x, A s represents the peak area of the control solution of internal reference s, C s represents the mass concentration of the control solution of internal reference s, f s / x represents the relative correction factor of target compound x.

[0008] Further, the standard curve of rutin is Y = 36085X, and the correlation coefficient (r) is 0.9999. Further, the standard curve of farrerol is Y = 29290X, and r = 0.9999. Further, the standard curve of zephyranthine is Y = 34361X, and r = 0.9999. Further, the standard curve of rosmarinic acid is Y = 19872X, and r = 0.9999. Further, when rutin is used as an internal reference, the relative correction factor of farrerol relative to rutin is about 1.38, and the relative retention value of farrerol relative to rutin is about 1.43; the relative correction factor of zephyranthine relative to rutin is about 1.02, and the relative retention value of zephyranthine relative to rutin is about 1.58; the relative correction factor of rosmarinic acid relative to rutin is about 1.70, and the relative retention value of rosmarinic acid relative to rutin is about 2.04. Further, when zephyranthine is used as an internal reference, the relative correction factor of rutin relative to zephyranthine is about 0.98, and the relative retention value of rutin relative to zephyranthine is about 0.63; the relative correction factor of farrerol relative to zephyranthine is about 1.35, and the relative retention value of farrerol relative to zephyranthine is about 0.91; the relative correction factor of rosmarinic acid relative to zephyranthine is about 1.67, and the relative retention value of rosmarinic acid relative to zephyranthine is about 1.29.

[0009] Further, the compound Gaozibo tablet is a compound Gaozibo tablet without coating. Further, the compound Gaozibo tablet powder is a sieved compound Gaozibo tablet powder. Further, the sieve is a No. 3 sieve. Further, the mass of the compound Gaozibo tablet powder is 0.1-10 g, for example, about 2.0 g. Further, the mass / volume (g / ml) ratio of the compound Gaozibo tablet powder to the alcohol is 0.01-0.2, for example, about 0.1. Further, the alcohol is methanol. Further, the mass percentage of the methanol is 10%-100%, for example, about 90%. Further, the extraction is reflux extraction, cold soak extraction, shaking extraction, and / or ultrasonic extraction, preferably, reflux extraction, for example, water bath reflux extraction. Further, the time of the water bath reflux extraction is 10-60 min, for example, about 30 min. Further, the cooling is to room temperature.

[0010] Further, the method for preparing the mixed reference solution comprises: weighing appropriate amounts of rutin, fumotoxin, zephyranthine, and rosmarinic acid reference substances, and adding methanol to prepare rutin reference solution, fumotoxin reference solution, zephyranthine reference solution, and rosmarinic acid reference solution, respectively; pipetting appropriate amounts of the rutin reference solution, the fumotoxin reference solution, the zephyranthine reference solution, and the rosmarinic acid reference solution into the same 10-ml volumetric flask to obtain No. 1 mixed reference solution; and diluting No. 1 mixed reference solution by 2, 5, 10, 25, and 50 to obtain No. 2-No. 6 mixed reference solutions, respectively. Further, the mass percentage of the methanol is 10%-100%, for example, about 90%. Further, the mass concentration of the rutin reference solution is 50-1000 μg / ml, for example, about 600 μg / ml. Further, the mass concentration of the fumotoxin reference solution is 50-1000 μg / ml, for example, about 642 μg / ml. Further, the mass concentration of the zephyranthine reference solution is 50-1000 μg / ml, for example, about 628 μg / ml. Further, the mass concentration of the rosmarinic acid reference solution is 50-1000 μg / ml, for example, about 496 μg / ml. Further, the volume of the rutin reference solution is 1-5 ml, for example, about 2 ml. Further, the volume of the fumotoxin reference solution is 1-5 ml, for example, about 2 ml. Further, the volume of the zephyranthine reference solution is 1-5 ml, for example, about 2 ml. Further, the volume of the rosmarinic acid reference solution is 1-5 ml, for example, about 4 ml. Further, the mass concentration of rutin in No. 1 mixed reference solution is about 120 μg / ml. Further, the mass concentration of fumotoxin in No. 1 mixed reference solution is about 128.4 μg / ml. Further, the mass concentration of zephyranthine in No. 1 mixed reference solution is about 125.6 μg / ml. Further, the mass concentration of rosmarinic acid in No. 1 mixed reference solution is about 198.4 μg / ml.

[0011] Further, the flow rate of the high performance liquid detection is 0.6-1.2 ml / min, for example, about 1.0 ml / min. Further, the column temperature is 30-40℃, for example, about 35℃. Further, the detection wavelength is 220-270 nm, for example, 254 nm. Further, the injection volume is 15-25 μl, for example, about 20 μl. Further, the resolution of the chromatographic peak corresponding to the target compound is greater than 1.5.

[0012] Further, the column is Kromasil C 18 Column, 4.6 x 250 mm, 5 μm, Phenomenex C 18 Column, 4.6 x 250 mm, 5 μm, MN C 18 Column, 4.6 x 250 mm, 5 μm, Agilent C 18 Column, 4.6 x 250 mm, 5 μm, or Thermo C 18 Column, 4.6 x 250 mm, 5 μm. Further, the column is Kromasil C 18 Column, 4.6 x 250 mm, 5 μm.

[0013] Further, the mobile phase B is acetonitrile. Further, the aqueous acid solution, aqueous alkali solution and / or aqueous buffer salt solution is selected from one or more of weak acids and their salts, weak alkalis and their salts at different concentrations. Further, the aqueous acid solution, aqueous alkali solution and / or aqueous buffer salt solution is selected from formic acid, glacial acetic acid, phosphoric acid, trifluoroacetic acid, formic acid and ammonium formate, acetic acid and sodium acetate, acetic acid and ammonium acetate, disodium hydrogen phosphate and sodium dihydrogen phosphate, disodium hydrogen phosphate and potassium dihydrogen phosphate, disodium hydrogen phosphate and citric acid, citric acid and sodium citrate, glycine and hydrochloric acid, or phthalic acid and hydrochloric acid at different concentrations. Further, the aqueous acid solution is 0.01%-2% aqueous acid solution. Further, the aqueous acid solution is 0.01%-2% formic acid aqueous solution. Further, the aqueous acid solution is about 0.2% formic acid aqueous solution. Further, the aqueous buffer salt solution is phosphate aqueous solution and / or acetate aqueous solution. Further, the pH value of the aqueous buffer salt solution is not greater than 7.0.

[0014] According to another aspect of the present application, there is provided a use of the above-mentioned quality control method in the quality detection and / or quality evaluation and / or quality control of Compound Gaozibo Tablets.

[0015] Advantages of the present application:

[0016] The method has excellent sensitivity, stability, repeatability and accuracy, can accurately and quickly determine the content of four components of rutin, fukurin, zephyranthine and rosmarinic acid in compound Gaozio tablet, and can be used for quality detection, quality evaluation or quality control of compound Gaozio tablet. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the embodiment description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without departing from the scope of the present application.

[0018] Figure 1 The HPLC chromatograms of the mixed control (A) and compound Gaozio tablet (B) of the present application. Wherein 1 represents rutin; 2 represents fukurin; 3 represents zephyranthine; 4 represents rosmarinic acid. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the present application will be described clearly and completely in the following with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of the present application.

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments or examples only and is not intended to be limiting of the application. As used herein, the term "and / or" is intended to cover any and all combinations of two or more of the associated listed items. The term "comprising" is intended to mean that the embodiments include at least the recited elements, but not excluding others.

[0021] In the present application, the technical features described in an open manner include both the closed technical solutions consisting of the listed features and the open technical solutions containing the listed features.

[0022] In the present application, the temperature parameters, if not particularly limited, allow both constant temperature treatment and treatment within a certain temperature range. The constant temperature treatment allows fluctuations within the accuracy range controlled by the instrument. The above applies to each parameter in the gradient elution program, flow rate, detection wavelength and injection volume.

[0023] In the present application, unless otherwise stated or there is a clear conflict in the context, the articles "a", "an" and "the" are intended to include "at least one" or "one or more". Therefore, these articles used in the present application refer to one or more than one (i.e. at least one) object.

[0024] As described in the background section, there is no literature report on the method of analyzing and researching the four chemical components of the above-mentioned compound Gaozibo tablets. In order to solve the above-mentioned problem, the present application provides a quality control method of compound Gaozibo tablets, which comprises the following steps: 1) preparation of mixed reference solution: prepare reference solutions of rutin, fukurin, zephyranthine and rosmarinic acid; 2) preparation of test solution of compound Gaozibo tablets: weigh an appropriate amount of compound Gaozibo tablet powder, add alcohol for extraction, then cool, supplement weight, shake well, filter, and take the filtrate to obtain the test solution of compound Gaozibo tablets; 3) dilute the reference solutions respectively and inject them into a high performance liquid chromatograph together with the test solution for chromatographic condition detection, the chromatographic conditions being: using a chromatographic column with octadecylsilane bonded silica gel as the filler, mobile phase A being an acid aqueous solution, an alkali aqueous solution and / or a buffer salt aqueous solution, mobile phase B being selected from one or more of acetonitrile, methanol and tetrahydrofuran, the gradient elution program being: 0-10 min, 10%→15% B; 10-40 min, 15%→17% B; 40-50 min, 17%→21% B; 50-60 min, 21%→30% B, the flow rate being 0.4-1.5 ml / min, the column temperature being 20-50°C, the detection wavelength being 200-350 nm, and the injection amount being 10-30 μl; and 4) obtaining the content information of rutin, fukurin, zephyranthine and / or rosmarinic acid in the compound Gaozibo tablets according to the detection results.

[0025] In the present application, unless otherwise stated or there is a clear conflict in the context, the articles "a", "an" and "the" are intended to include "at least one" or "one or more". Therefore, these articles used in the present application refer to one or more than one (i.e. at least one) object.

[0026] In a preferred embodiment, the method further comprises calculating the relative retention value, the relative correction factor and / or the content of the target compound using rutin or zephyranthine as the internal reference, wherein the target compound is selected from one or more of the following: rutin, fukurin, zephyranthine and rosmarinic acid.

[0027] In a preferred embodiment, the method uses rutin or narcissusin as the internal reference to calculate the relative retention value, the relative correction factor and / or the content of the target compound, respectively, comprising: (a) determining the standard curve of rutin, fumotoxin, narcissusin and rosmarinic acid according to the mass concentration of each reference in the mixed reference solution and the corresponding peak area in the chromatogram; (b) determining the internal reference, according to the slope of the standard curve obtained in step (a), calculating the relative correction factor of the target compound according to the formula f s / x = K s / K x , wherein f s / x represents the relative correction factor of the target compound x relative to the internal reference s, K s is the slope of the standard curve of the internal reference s, and K x is the slope of the standard curve of the target compound x; (c) using the relative retention value to locate the chromatographic peaks of rutin, fumotoxin, narcissusin and rosmarinic acid in the high performance liquid chromatogram of the compound Gaozibo tablet test solution, calculating the relative retention value of the target compound according to the formula r x / s = t Rx / t Rs , wherein r x / s represents the relative retention value of the target compound x relative to the internal reference s, t Rx is the retention time of the target compound x, and t Rs is the retention time of the internal reference s; and (d) according to the mass concentration and the corresponding peak area of the reference solution in step (a), the relative correction factor in step (b) and the corresponding peak area of the located chromatographic peak in step (c), calculating the content of rutin, fumotoxin, narcissusin and rosmarinic acid in the compound Gaozibo tablet test solution according to the formula C x = (A x / A s ) x C s x f s / x , wherein C x represents the mass concentration of the target compound x in the test solution, A x represents the peak area of the target compound x, A s represents the peak area of the reference solution of the internal reference s, C s represents the mass concentration of the reference solution of the internal reference s, and f s / x represents the relative correction factor of the target compound x.

[0028] In a preferred embodiment, the standard curve for rutin is Y = 36085X, with a correlation coefficient (r) = 0.9999. In a preferred embodiment, the standard curve for hyperoside is Y = 29290X, with r = 0.9999. In a preferred embodiment, the standard curve for zephyranthine is Y = 34361X, with r = 0.9999. In a preferred embodiment, the standard curve for rosmarinic acid is Y = 19872X, with r = 0.9999.

[0029] In a preferred embodiment, the relative correction factor for hyperoside relative to rutin is about 1.38, and the relative retention value for hyperoside relative to rutin is about 1.43, using rutin as the internal standard. The relative correction factor for zephyranthine relative to rutin is about 1.02, and the relative retention value for zephyranthine relative to rutin is about 1.58. The relative correction factor for rosmarinic acid relative to rutin is about 1.70, and the relative retention value for rosmarinic acid relative to rutin is about 2.04.

[0030] In the present application, the term "about" or "approximately" with respect to a numerical value means ± 5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 1.38 means from 1.311 to 1.449, but also expressly includes exactly 1.38; "about" 1.43 means from 1.3585 to 1.5015, but also expressly includes exactly 1.43; "about" 1.02 means from 0.969 to 1.071, but also expressly includes exactly 1.02; "about" 1.58 means from 1.501 to 1.659, but also expressly includes exactly 1.58; "about" 1.70 means from 1.615 to 1.785, but also expressly includes exactly 1.70; "about" 2.04 means from 1.938 to 2.142, but also expressly includes exactly 2.04.

[0031] In a preferred embodiment, the relative correction factor for rutin relative to zephyranthine is about 0.98, and the relative retention value for rutin relative to zephyranthine is about 0.63, using zephyranthine as the internal standard. The relative correction factor for hyperoside relative to zephyranthine is about 1.35, and the relative retention value for hyperoside relative to zephyranthine is about 0.91. The relative correction factor for rosmarinic acid relative to zephyranthine is about 1.67, and the relative retention value for rosmarinic acid relative to zephyranthine is about 1.29.

[0032] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 0.98 means from 0.931 to 1.029, but also expressly includes exactly 0.98; "about" 0.63 means from 0.5985 to 0.6615, but also expressly includes exactly 0.63; "about" 1.35 means from 1.2825 to 1.4175, but also expressly includes exactly 1.35; "about" 0.91 means from 0.8645 to 0.9555, but also expressly includes exactly 0.91; "about" 1.67 means from 1.5865 to 1.7535, but also expressly includes exactly 1.67; "about" 1.29 means from 1.2255 to 1.3545, but also expressly includes exactly 1.29.

[0033] In a preferred embodiment, the compound Gaozhi tablet is a compound Gaozhi tablet without coating.

[0034] In a preferred embodiment, the compound Gaozhi tablet powder is a sieved compound Gaozhi tablet powder.

[0035] In a preferred embodiment, the sieve is a No. 3 sieve.

[0036] In a preferred embodiment, the mass of the compound Gaozhi tablet powder is 0.1-10 g, for example about 2.0 g.

[0037] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 2 means from 1.9 to 2.1, but also expressly includes exactly 2.

[0038] In a preferred embodiment, the mass / volume (g / ml) ratio of the compound Gaozhi tablet powder to the alcohol is 0.01-0.2, for example about 0.1.

[0039] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 0.1 means from 0.095 to 0.105, but also expressly includes exactly 0.1.

[0040] In a preferred embodiment, the alcohol is methanol.

[0041] In a preferred embodiment, the mass percentage of the methanol is 10%-100%, for example about 90%.

[0042] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 90 means from 85.5 to 94.5, but also expressly includes exactly 90.

[0043] In a preferred embodiment, the extraction is reflux extraction, cold soak extraction, shake extraction and / or ultrasonic extraction, preferably reflux extraction, such as water bath reflux extraction.

[0044] In a preferred embodiment, the water bath reflux extraction is for 10-60 min, such as about 30 min.

[0045] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 30 means from 28.5 to 31.5, but also expressly includes exactly 30.

[0046] In a preferred embodiment, the cooling is to room temperature.

[0047] In a preferred embodiment, the method for preparing the mixed control solution comprises: weighing appropriate amounts of rutin, hyperoside, zephyranthine and rosmarinic acid controls, and adding methanol to prepare rutin control stock solution, hyperoside control stock solution, zephyranthine control stock solution and rosmarinic acid control stock solution, respectively; pipetting appropriate amounts of the rutin control stock solution, the hyperoside control stock solution, the zephyranthine control stock solution and the rosmarinic acid control stock solution into the same 10 ml volumetric flask, respectively, to obtain No. 1 mixed control solution; and diluting No. 1 mixed control solution by 2, 5, 10, 25 and 50, respectively, to obtain No. 2 to No. 6 mixed control solutions.

[0048] In a preferred embodiment, the mass percentage of methanol is 10%-100%, such as about 90%.

[0049] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 90 means from 85.5 to 94.5, but also expressly includes exactly 90.

[0050] In a preferred embodiment, the mass concentration of the rutin stock solution is 50-1000 μg / ml, such as about 600 μg / ml.

[0051] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 600 means from 570 to 630, but also expressly includes exactly 600.

[0052] In a preferred embodiment, the mass concentration of the hyperoside stock solution is 50-1000 μg / ml, such as about 642 μg / ml.

[0053] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of that numerical value, but expressly includes the exact numerical value. For example, "about" 642 means from 609.9 to 674.1, but also expressly includes exactly 642.

[0054] In a preferred embodiment, the stock solution of rutin has a mass concentration of 50 to 1000 μg / ml, for example about 628 μg / ml.

[0055] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of that numerical value, but expressly includes the exact numerical value. For example, "about" 628 means from 596.6 to 659.4, but also expressly includes exactly 628.

[0056] In a preferred embodiment, the stock solution of rosmarinic acid has a mass concentration of 50 to 1000 μg / ml, for example about 496 μg / ml.

[0057] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of that numerical value, but expressly includes the exact numerical value. For example, "about" 496 means from 471.2 to 520.8, but also expressly includes exactly 496.

[0058] In a preferred embodiment, the stock solution of rutin has a mass concentration of 50 to 1000 μg / ml, for example about 628 μg / ml.

[0059] In a preferred embodiment, the stock solution of rutin has a mass concentration of 50 to 1000 μg / ml, for example about 628 μg / ml.

[0060] In a preferred embodiment, the stock solution of rutin has a mass concentration of 50 to 1000 μg / ml, for example about 628 μg / ml.

[0061] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of that numerical value, but expressly includes the exact numerical value. For example, "about" 2 means from 1.9 to 2.1, but also expressly includes exactly 2.

[0062] In a preferred embodiment, the stock solution of rosmarinic acid has a mass concentration of 50 to 1000 μg / ml, for example about 496 μg / ml.

[0063] In the present application, the term "about" or "approximately" in relation to a numerical value means ±5% of that numerical value, but expressly includes the exact numerical value. For example, "about" 4 means from 3.8 to 4.2, but also expressly includes exactly 4.

[0064] In a preferred embodiment, the stock solution of rutin has a mass concentration of 50 to 1000 μg / ml, for example about 628 μg / ml.

[0065] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 120 means from 114 to 126, but also expressly includes exactly 120.

[0066] In a preferred embodiment, the mass concentration of zephyranthine in the Mixed Reference Solution No. 1 is about 128.4 μg / ml.

[0067] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 128.4 means from 121.98 to 134.82, but also expressly includes exactly 128.4.

[0068] In a preferred embodiment, the mass concentration of zephyranthine in the Mixed Reference Solution No. 1 is about 128.4 μg / ml.

[0069] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 128.4 means from 121.98 to 134.82, but also expressly includes exactly 128.4.

[0070] In a preferred embodiment, the mass concentration of rosmarinic acid in the Mixed Reference Solution No. 1 is about 198.4 μg / ml.

[0071] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 198.4 means from 188.48 to 208.32, but also expressly includes exactly 198.4.

[0072] In a preferred embodiment, the flow rate for the HPLC detection is 0.6 to 1.2 ml / min, such as about 1.0 ml / min.

[0073] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 1 means from 0.95 to 1.05, but also expressly includes exactly 1.

[0074] In a preferred embodiment, the column temperature is 30 to 40 °C, such as about 35 °C.

[0075] In the present invention, the term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 35 means from 33.25 to 36.75, but also expressly includes exactly 35.

[0076] In a preferred embodiment, the detection wavelength is 220-270 nm, such as 254 nm.

[0077] In a preferred embodiment, the injection volume is 15-25 μl, such as about 20 μl.

[0078] In the present application, the term "about" or "approximately" in relation to a numerical value means ± 5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 20 means from 19 to 21, but also expressly includes exactly 20.

[0079] In a preferred embodiment, the separation of the chromatographic peaks corresponding to the target compounds is greater than 1.5.

[0080] In a preferred embodiment, the column is a Kromasil C 18 Column, 4.6 x 250 mm, 5 μm, Phenomenex C 18 Column, 4.6 x 250 mm, 5 μm, MN C 18 Column, 4.6 x 250 mm, 5 μm, Agilent C 18 Column, 4.6 x 250 mm, 5 μm, or Thermo C 18 Column, 4.6 x 250 mm, 5 μm. In a preferred embodiment, the column is a Kromasil C 18 Column, 4.6 x 250 mm, 5 μm.

[0081] In a preferred embodiment, the mobile phase B is acetonitrile. In a preferred embodiment, the aqueous acid, aqueous base and / or aqueous buffer salt solution is selected from one or more of weak acids and their salts, weak bases and their salts, at different concentrations. In a preferred embodiment, the aqueous acid, aqueous base and / or aqueous buffer salt solution is selected from formic acid, glacial acetic acid, phosphoric acid, trifluoroacetic acid, formic acid and ammonium formate, acetic acid and sodium acetate, acetic acid and ammonium acetate, disodium hydrogen phosphate and sodium dihydrogen phosphate, disodium hydrogen phosphate and potassium dihydrogen phosphate, disodium hydrogen phosphate and citric acid, citric acid and sodium citrate, glycine and hydrochloric acid, or phthalic acid and hydrochloric acid, at different concentrations. In a preferred embodiment, the aqueous acid is an aqueous acid at 0.01-2%. In a preferred embodiment, the aqueous acid is an aqueous formic acid at 0.01-2%. In a preferred embodiment, the aqueous acid is an aqueous formic acid at about 0.2%.

[0082] In the present application, the term "about" or "approximately" in relation to a numerical value means ± 5% of the numerical value, but expressly includes the exact numerical value. For example, "about" 0.2 means from 0.19 to 0.21, but also expressly includes exactly 0.2.

[0083] In a preferred embodiment, the buffered saline solution is a phosphate buffered saline solution and / or an acetate buffered saline solution. In a preferred embodiment, the buffered saline solution has a pH of no more than 7.0.

[0084] According to another aspect of the present application, there is provided use of the above-mentioned quality control method in quality detection and / or quality evaluation and / or quality control of Compound Gaozibo Tablets.

[0085] The present application is further described in conjunction with the following examples. It should be understood that these examples are only used to illustrate the present application and not intended to limit the scope of the present application. The experimental methods in the following examples, if not otherwise specified, are generally carried out according to the conventional conditions or the conditions suggested by the manufacturers.

[0086] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. In addition, any method and material similar or equivalent to those described herein can be used in the practice of the present application. The preferred methods and materials described herein are described only as examples.

[0087] The above-mentioned features mentioned in the present application, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in the present patent specification can be used in any combination. Each feature disclosed in the present specification can be replaced by any alternative feature that provides the same, equivalent or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equivalent or similar features.

[0088] Examples

[0089] 1 Materials

[0090] Shimadzu LC-20AT high performance liquid chromatograph (Japan Shimadzu Corporation); Waters alliance e2695 high performance liquid chromatograph (WATERS Corporation, USA); Agilent 1200 high performance liquid chromatograph (Agilent Corporation, USA); FW100 type high-speed universal pulverizer (Tianjin Test Instrument Co., Ltd.); KQ-250DB type digital ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); ME104 Mettler one-hundredth electronic balance (Mettler-Toledo Instruments (Shanghai) Co., Ltd.); MS105DU Mettler one-thousandth electronic balance (Mettler-Toledo Instruments (Shanghai) Co., Ltd.); XMTD-6000 type electric heating constant temperature water bath (Shanghai Shulai Instrument & Meter Co., Ltd.).

[0091] Kromasil C 18 Chromatographic column (4.6 x 250 mm, 5 μm), Phenomenex C18 Column (4.6 x 250 mm, 5 μm), MN C 18 Column (4.6 x 250 mm, 5 μm), Agilent Eclipse XDB-C 18 Column (4.6 x 250 mm, 5 μm), Thermo C 18 Column (4.6 x 250 mm, 5 μm).

[0092] Rutin (Batch No. 100080-202012, content 91.6%), Narcissoside (Batch No. 111997-202302, content 93.1%) and Rosmarinic acid (Batch No. 111871-202007, content 98.1%) were purchased from China Institute for Drug Control; Nicotiflorin (Batch No. CHB201218, purity ≥98%) was purchased from Chengdu Keluoma Biological Technology Co., Ltd.; Compound Gaoziba Tablets (Batch Nos. 2201135, 2303134 and 230564) were produced by Xinjiang Uygur Pharmaceutical Co., Ltd.

[0093] 2 Method and Results

[0094] 2.1 Preparation of mixed reference solution

[0095] An appropriate amount of Rutin, Nicotiflorin, Narcissoside and Rosmarinic acid reference substances were precisely weighed and added to 90% methanol to prepare reference stock solutions with mass concentrations of 600, 642, 628 and 496 ug·mL -1 , respectively. 2 mL of each of the above Rutin, Nicotiflorin and Narcissoside stock solutions and 4 mL of the Rosmarinic acid stock solution were precisely measured into the same 10 mL volumetric flask to obtain No. 1 mixed reference solution with mass concentrations of 120, 128.4, 125.6 and 198.4 ug·mL -1 , respectively. No. 2 to No. 6 mixed reference solutions were prepared by diluting No. 1 mixed reference solution by 2, 5, 10, 25 and 50 times, respectively.

[0096] 2.2 Preparation of test solution

[0097] Take the powder (pass through No. 3 sieve) of compound Gaozibo tablets (batch number 2201135, remove the coating) about 2.00 g, accurately weighed, placed in a conical flask with a stopper, accurately added 90% methanol 20 mL, weighed, respectively, using water bath reflux for 60 min, ultrasonic for 60 min (500 W, 53 kHz), cold soak overnight (18 h) for sample extraction, after treatment, placed at room temperature, weighed, added 90% methanol to make up the lost mass, shake, filter, take the filtrate as the test sample solution, sample analysis. The content of 4 kinds of ingredients under different extraction methods is shown in Table 1. Compared with ultrasonic and cold soak, water bath reflux has higher extraction efficiency for 4 kinds of ingredients, so the extraction method is selected as water bath reflux.

[0098] Table 1 Content of 4 kinds of ingredients in compound Gaozibo tablets under different extraction methods (n = 2)

[0099]

[0100] Take the sample (batch number 2201135) powder about 2.00 g, respectively, 40% methanol, 65% methanol and 90% methanol 20 mL as extraction solvent, water bath reflux for 60 min for sample extraction, preparation of test sample solution, sample analysis, the content of 4 kinds of ingredients under different extraction solvents is shown in Table 2. Compared with 40% methanol and 65% methanol, 90% methanol has higher extraction efficiency for 4 kinds of ingredients, so 90% methanol is selected as the extraction solvent.

[0101] Table 2 Content of 4 kinds of ingredients in compound Gaozibo tablets under different extraction solvents (n = 2)

[0102]

[0103] Take the sample (batch number 2201135) powder about 2.00 g, with 90% methanol 20 mL as extraction solvent, respectively, water bath reflux for 30, 60, 90 min, preparation of test sample solution, sample analysis. The content of 4 kinds of ingredients under different extraction times is shown in Table 3. There is no obvious difference in extraction efficiency of 4 kinds of ingredients with extraction time, from the perspective of reducing energy consumption and improving work efficiency, the extraction time is selected as 30 min.

[0104] Table 3 Content of 4 kinds of ingredients in compound Gaozibo tablets under different extraction times (n = 2)

[0105]

[0106] Sample (batch number 2201135) powder about 2.00g, using 90% methanol as the extraction solvent, solvent dosage was 75 times (15mL), 100 (20mL), 125 (25mL), 150 times (30mL), water bath reflux 30min, preparation of test solution, sample analysis, the content of four components under different solvent dosage as shown in Table 4. Under different solvent dosage, the extraction efficiency of four components has no obvious difference, from the perspective of convenient experimental operation and reduce energy consumption, the solvent dosage is 100 times.

[0107] Table 4 content of four components in compound Gaozibo tablets under different solvent dosage (n = 2)

[0108]

[0109] In summary, the final test solution preparation method is as follows: compound Gaozibo tablets (remove the coating) powder (through three sieve) about 2.00g, accurately weighed, placed in a conical flask with plug, accurately added 90% methanol 20mL, weighed, water bath reflux 30min, placed at room temperature, weighed again, supplemented with the loss of quality with 90% methanol, shake, filter, take the filtrate, get.

[0110] 2.3 HPLC chromatographic conditions

[0111] The separation effect of elution system such as pure water-acetonitrile, pure water-methanol, acid aqueous solution (0.2% formic acid or 0.5% phosphoric acid or 1% acetic acid)-methanol and acid aqueous solution (0.2% formic acid or 0.5% phosphoric acid or 1% acetic acid)-acetonitrile under different gradient elution was investigated, for example:

[0112] Elution condition ①: pure water (A)-acetonitrile (B) gradient elution, 0-10min, 12%→15% B; 10-40min, 15→20% B; 40-50min, 20→25% B; 50-60min, 25→30% B.

[0113] Elution condition ②: pure water (A)-methanol (B) gradient elution, 0-10min, 15%→20% B; 10-40min, 20→23% B; 40-50min, 23→28% B; 50-60min, 28→40% B.

[0114] Elution condition ③: 0.2% formic acid aqueous solution (A)-methanol (B) gradient elution, 0-10min, 15%→20% B; 10-40min, 20→23% B; 40-50min, 23→28% B; 50-60min, 28→40% B.

[0115] Elution condition IV: 0.4% phosphoric acid (A) - acetonitrile (B) gradient elution, 0-15 min, 10%→15% B; 15-30 min, 15→17% B; 30-50 min, 17→21% B; 50-60 min, 21→30% B.

[0116] Elution condition V: 1% acetic acid (A) - acetonitrile (B) gradient elution, 0-20 min, 10%→15% B; 20-30 min, 15→17% B; 30-50 min, 17→21% B; 50-60 min, 21→30% B.

[0117] Elution condition VI: 0.2% formic acid (A) - acetonitrile (B) gradient elution, 0-10 min, 10%→15% B; 10-40 min, 15%→17% B; 40-50 min, 17%→21% B; 50-60 min, 21%→30% B.

[0118] Under elution conditions I-III, when using pure water-acetonitrile (methanol) elution system, the chromatographic peak of rosmarinic acid has obvious tailing phenomenon, and when using acid aqueous solution-methanol elution system, the chromatographic peak of the target compound has impurity interference and baseline drift.

[0119] Under elution conditions IV-VI, 4 components of rutin, yanhuaoside, narcissus glycoside and rosmarinic acid are overlapped with impurity peaks under conditions IV and V, while under condition VI, the 4 components of rutin, yanhuaoside, narcissus glycoside and rosmarinic acid have excellent separation effect with impurity peaks.

[0120] The optimal HPLC conditions finally determined in the test are as follows: Kromasil C 18 column (4.6×250 mm, 5 μm); mobile phase: 0.2% formic acid (A) - acetonitrile (B) gradient elution, 0-10 min, 10%→15% B; 10-40 min, 15%→17% B; 40-50 min, 17%→21% B; 50-60 min, 21%→30% B. Column temperature 35°C, flow rate 1.0 mL·min -1 , detection wavelength 254 nm, injection volume: 20 μL. Under the current chromatographic conditions, the HPLC chart of the mixed reference substance and compound Gaotiao tablet is shown in Figure 1 , the separation degree R of each chromatographic peak is >1.5, the tailing factor T is 0.95-1.05, and the chromatographic separation effect is good.

[0121] 2.4 Methodology investigation

[0122] 2.4.1 Linear range and detection limit, quantification limit

[0123] Take 20 μL of each of the six concentrations of the mixed control solution under 2.1, inject and analyze, and determine the peak areas of rutin, rhodaxanthin, zephyranthine, and rosmarinic acid. Linear regression processing (passing through the origin) was performed on the peak area Y against the mass concentration X (μg·mL -1 ) to obtain the regression equation, correlation coefficient (r), and linear range for each component; at the same time, the No. 6 mixed control was diluted step by step, injected and analyzed, and the peak areas were determined. The mass concentration at a signal-to-noise ratio (S / N) of 3 was taken as the detection limit, and the mass concentration at an S / N of 10 was taken as the quantification limit, and the results are shown in Table 5. The four components had good linear relationships in a wide mass concentration range (50 times) and high sensitivity.

[0124] Table 5 Regression equation, linear range, detection limit, and quantification limit of the four components

[0125]

[0126] 2.4.2 Precision

[0127] Take the No. 3 mixed control solution, inject continuously for six times, determine the peak areas, and calculate the RSD. The RSD (n = 6) of the peak areas of rutin, rhodaxanthin, zephyranthine, and rosmarinic acid was 0.27%, 0.39%, 0.34%, and 0.35%, respectively. The above results show that the precision of the instrument is good.

[0128] 2.4.3 Stability

[0129] Stability of the control solution: take the No. 3 mixed control solution, inject and analyze at 0, 6, 12, 24, 36, and 48 h after preparation, determine the peak areas, and calculate the RSD. The RSD (n = 6) of the peak areas of rutin, rhodaxanthin, zephyranthine, and rosmarinic acid was 0.26%, 0.39%, 0.29%, and 0.71%, respectively. This shows that the control solution has good stability within 48 h.

[0130] Stability of the test solution: prepare the test solution according to the method under item 2.2, inject and analyze at 0, 6, 12, 17, 36, and 48 h after preparation, determine the peak areas, and calculate the RSD. The RSD (n = 6) of the peak areas of rutin, rhodaxanthin, zephyranthine, and rosmarinic acid was 0.86%, 1.78%, 0.52%, and 1.36%, respectively. This shows that the test solution has good stability within 48 h.

[0131] 2.4.4 Repeatability

[0132] Take the same batch (batch number 2201135) of compound Gaoziban tablets (remove the coating) powder about 2.00 g, precision weighing, 6 parallel, according to the method of 2.2 to prepare the sample solution, sample analysis, determination of peak area, calculate the content and RSD. The contents of rutin, fumotoxin, zephyranthine and rosmarinic acid were 0.473, 0.481, 0.286 and 1.24 mg·g -1 , respectively, and the RSDs were 0.96%, 1.32%, 0.84% and 0.68%, respectively, indicating that the repeatability of the method was good.

[0133] 2.4.5 Sample recovery rate

[0134] Take the compound Gaoziban tablets (batch number 2201135) powder with known ingredient content about 1.00 g, 6 parallel, precision weighing, respectively, according to the sample amount and the amount of reference substance about 1:1 to add reference substance solution, according to the method of 2.2 to prepare the sample solution, sample analysis, determine and calculate the sample recovery rate of each component and RSD. The average recovery rates of rutin, fumotoxin, zephyranthine and rosmarinic acid were 102.2%, 98.06%, 97.62% and 99.25%, respectively; RSD (n=6) were 2.26%, 1.72%, 1.32% and 2.47%, respectively, indicating that the accuracy of the method was good.

[0135] 2.5 Establishment of relative correction factor (with rutin as internal reference)

[0136] 2.5.1 Calculation of relative correction factor

[0137] Prepare mixed reference solution 1-6 according to the method of 2.1, and determine the peak area of rutin, fumotoxin, zephyranthine and rosmarinic acid according to the chromatographic conditions of 2.3. Linear regression (passing through the origin) was performed on the peak area Y with the mass concentration X (mg·L -1 ) to obtain the regression equation, slope and correlation coefficient (r) of each component.

[0138] Using the slope method, with rutin as the internal reference (s), the relative correction factor (RCF) of fumotoxin, zephyranthine and rosmarinic acid was calculated according to the formula f s / x = K s / K x , where f s / x is the RCF of component x relative to internal reference s, K s is the slope of the standard curve of internal reference s, and K xLet x be the slope of the standard curve for component x. The results are shown in Table 6. When using the slope method to calculate RCFs, the origin is included in the standard curve. The larger the correlation coefficient, the more accurate the RCFs calculated using the slope method. The correlation coefficient r of the regression equations (passing through the origin) for the four components is ≥0.9999, indicating a good linear relationship and demonstrating the feasibility of using the slope method to calculate the RCFs of each component.

[0139] Table 6 shows the results of the relative correction factors calculated using the slope method with rutin as an internal reference.

[0140]

[0141] 2.5.2 Repeatability of relative correction factors

[0142] Mixed reference solutions 1–6 were prepared according to the method described in section 2.1, in six parallel replicates. The RCFs, mean values, and RSDs of rutin, rosmarinic acid, naringin, and rosmarinic acid were calculated according to the method described in section 2.5.1. The results are shown in Table 7. 芦丁 / 烟花苷 f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸 The mean values ​​were 1.34, 1.01 and 1.67, respectively, and the RSDs were 1.12%, 0.61% and 1.23%, respectively, indicating that the repeatability of the relative correction factors was good.

[0143] Table 7. Repeatability test results of relative correction factors (with rutin as an internal reference) (n=6)

[0144]

[0145] 2.5.3 Robustness test of relative correction factor

[0146] 2.5.3.1 Effect of different HPLC instruments on relative correction factor

[0147] Following the method described in section 2.5.1, Kromasil C was used. 18 Chromatographic columns were used on three HPLC systems: a Shimadzu LC-20A, an Agilent 1200, and a Waters e2695, respectively, to detect f. 芦丁 / 烟花苷 f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸 The measurements were performed, and the results are shown in Table 8. 芦丁 / 烟花苷 f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸 The mean values ​​on the three HPLC instruments were 1.39, 1.03 and 1.72, respectively, with RSDs of 3.41%, 0.26% and 3.38%, respectively, all <4.00%, indicating that the relative correction factor has good robustness on different HPLC instruments.

[0148] Table 8 Influence of different HPLC instruments on relative correction factors (using rutin as internal standard) (n = 3)

[0149]

[0150]

[0151] 2.5.3.2 Influence of different chromatographic columns on relative correction factors

[0152] According to the method under item 2.5.1, using Shimadzu LC-20A HPLC instrument and Phenomenex C 18 , Thermo C 18 , Agilent C 18 , MN C 18 and Kromasil C 18 chromatographic columns, f 芦丁 / 烟花苷 , f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸 were determined respectively, and the results are shown in Table 9. The average values of f 芦丁 / 烟花苷 , f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸 on the five chromatographic columns were 1.32, 1.01 and 1.64 respectively, and the RSDs were 0.51%, 0.57% and 0.56% respectively, all < 1.00%, indicating that the relative correction factors had good robustness to different chromatographic columns.

[0153] 1.32, 1.01 and 1.64, and the RSDs were 0.51%, 0.57% and 0.56% respectively, all < 1.00%, indicating that the relative correction factors had good robustness to different chromatographic columns.

[0154] Table 9 Influence of different chromatographic columns on relative correction factors (using rutin as internal standard) (n = 5)

[0155]

[0156] 2.5.3.3 Influence of different flow rates on relative correction factors

[0157] According to the method under item 2.5.1, using Shimadzu LC-20A HPLC instrument and Kromasil C 18 chromatographic column, f -1 , f 芦丁 / 烟花苷 and f 芦丁 / 水仙苷 were determined respectively under different flow rates (0.90, 1.00, 1.10 mL·min 芦丁 / 迷迭香酸 ), and the results are shown in Table 10. The average values of f 芦丁 / 烟花苷 , f 芦丁 / 水仙苷 and f 芦丁 / 迷迭香酸The mean values of f Rutin, f Rutin and f Rutin at different flow rates were 1.35, 1.02 and 1.66, and the RSDs were 0.09%, 0.30% and 0.34%, respectively, indicating that the relative correction factors had good robustness to different flow rates.

[0158] Table 10 Influence of different flow rates on relative correction factors (using rutin as internal reference) (n = 3)

[0159]

[0160]

[0161] 2.5.3.4 Influence of different column temperatures on relative correction factors

[0162] According to the method under item 2.5.1, using Shimadzu LC-20A HPLC instrument and Kromasil C 18 chromatographic column, f Rutin, f Rutin and f Rutin were determined at different column temperatures (30, 35 and 40 °C), and the results are shown in Table 11. The mean values of f Rutin, f Rutin and f Rutin at different column temperatures were 1.35, 1.02 and 1.67, and the RSDs were 0.15%, 0.37% and 0.33%, respectively, indicating that the relative correction factors had good robustness to different column temperatures. 芦丁 / 烟花苷 芦丁 / 水仙苷 芦丁 / 迷迭香酸 芦丁 / 烟花苷 芦丁 / 水仙苷 芦丁 / 迷迭香酸

[0163] Table 11 Influence of different column temperatures on relative correction factors (using rutin as internal reference) (n = 3)

[0164]

[0165] 2.5.4 Investigation of reproducibility of relative correction factors

[0166] f Rutin, f Rutin and f Rutin were determined by different test personnel on different working days on HPLC instruments in three different laboratories, and the results are shown in Table 12. The mean values of f Rutin, f Rutin and f Rutin determined in different scenarios were 1.38, 1.02 and 1.70, and the RSDs were 4.31%, 0.84% and 3.97%, respectively, indicating that the relative correction factors had good reproducibility, and the RCFs were finally determined as follows: f Rutin = 1.38, f Rutin = 1.02 and f Rutin = 1.70. 芦丁 / 烟花苷 芦丁 / 水仙苷 芦丁 / 迷迭香酸 芦丁 / 烟花苷 芦丁 / 水仙苷 芦丁 / 迷迭香酸 芦丁 / 烟花苷 芦丁 / 水仙苷 芦丁 / 迷迭香酸 ​​​​​​​​​​​​​​​

[0167] Table 12 Repeatability of relative correction factors (using rutin as the internal standard) (n = 3)

[0168]

[0169]

[0170] 2.6 Establishment of relative correction factors (using tinamareine as the internal standard)

[0171] 2.6.1 Calculation of relative correction factors

[0172] Prepare the mixed reference solution No. 1-6 according to the method under item 2.1, and determine the peak areas of rutin, chrysorooside, tinamareine and rosmarinic acid according to the chromatographic conditions under item 2.3. Linearly regress (passing through the origin) the peak area Y against the mass concentration X (mg-L -1 ) of each component to obtain the regression equation, slope and correlation coefficient (r) of each component.

[0173] Using the slope method, calculate the RCFs of rutin, chrysorooside and rosmarinic acid using tinamareine as the internal standard (s), and the results are shown in Table 13. The correlation coefficients r of the regression equations (passing through the origin) of the four components are all > 0.9999, indicating that the linear relationship is good, and that it is feasible to calculate the RCFs of each component using the slope method.

[0174] Table 13 Relative correction factors calculated using the slope method with tinamareine as the internal standard

[0175]

[0176] 2.6.2 Repeatability of relative correction factors

[0177] Prepare the mixed reference solution No. 1-6 according to the method under item 2.1, in six replicates, and calculate the RCFs, mean values and RSDs of rutin, chrysorooside, tinamareine and rosmarinic acid according to the method under item 2.5.1, and the results are shown in Table 14.f 水仙苷 / 芦丁 , and the mean values of f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 are 0.99,

[0178] 1.32 and 1.65, respectively, and the RSDs are 0.62%, 0.57% and 0.95%, respectively, indicating that the repeatability of the relative correction factors is good.

[0179] Table 14 Repeatability of relative correction factors (using tinamareine as the internal standard) (n = 6)

[0180]

[0181]

[0182] 2.6.3 Robustness test of relative correction factor

[0183] 2.6.3.1 Effect of different HPLC instruments on relative correction factor

[0184] Following the method described in section 2.6.1, Kromasil C was used. 18 Chromatographic columns were used on three HPLC systems: a Shimadzu LC-20A, an Agilent 1200, and a Waters e2695, respectively, to detect f. 水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The measurements were performed, and the results are shown in Table 15. 水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The mean values ​​on the three HPLC instruments were 0.98, 1.35 and 1.67, respectively, with RSDs of 1.00%, 4.14% and 3.38%, respectively, indicating that the relative correction factor has good robustness on different HPLC instruments.

[0185] Table 15 Effect of different HPLC instruments on relative correction factors (with narcissin as internal reference) (n=3)

[0186]

[0187] 2.6.3.2 Effect of different chromatographic columns on relative correction factors

[0188] Following the method described in section 2.6.1, a Shimadzu LC-20A HPLC system and a Phenomenex C2000 HPLC system were used. 18 ThermoC 18 Agilent C 18 MN C 18 and Kromasil C 18 Five different chromatographic columns were used to treat f. 水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The measurements were performed, and the results are shown in Table 16. 水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The mean values ​​on the five columns were 0.99, 1.31, and 1.62, respectively, with RSDs of 0.58%, 0.29%, and 0.80%, respectively, indicating that the relative correction factor was robust to different columns.

[0189] Table 16. Effects of different chromatographic columns on relative correction factors (using narcissin as an internal reference) (n=5)

[0190]

[0191] 2.6.3.3 Effect of different flow rates on relative correction factors

[0192] The Shimadzu LC-20A HPLC instrument and Kromasil C 18 chromatographic column were used to determine f -1 , f 水仙苷 / 芦丁 and f 水仙苷 / 烟花苷 at different flow rates (0.90, 1.00, 1.10 mL·min 水仙苷 / 迷迭香酸 ) respectively, and the results are shown in Table 17. The average values of f 水仙苷 / 芦丁 , f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 at different flow rates were 0.98, 1.32 and 1.63 respectively, and the RSDs were 0.31%, 0.23% and 0.07% respectively, indicating that the relative correction factors had good robustness to different flow rates.

[0193] 1.63, RSD respectively were 0.31%, 0.23% and 0.07%, indicating that the relative correction factors had good robustness to different flow rates.

[0194] Table 17 Effect of different flow rates on relative correction factors (using tulipin as the internal reference) (n = 3)

[0195]

[0196] 2.6.3.4 Effect of different column temperatures on relative correction factors

[0197] The Shimadzu LC-20A HPLC instrument and Kromasil C 18 chromatographic column were used to determine f 水仙苷 / 芦丁 , f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 at different column temperatures (30, 35, 40 °C) respectively, and the results are shown in Table 18. The average values of f 水仙苷 / 芦丁 , f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 at different column temperatures were 0.98, 1.32 and 1.64 respectively, and the RSDs were 0.39%, 0.30% and 0.46% respectively, indicating that the relative correction factors had good robustness to different column temperatures.

[0198] Table 18 Effect of different column temperatures on relative correction factors (using tulipin as the internal reference) (n = 3)

[0199]

[0200] 2.6.4 Investigation of the reproducibility of relative correction factors

[0201] The Shimadzu LC-20A HPLC instrument and Kromasil C水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The measurements were performed, and the results are shown in Table 19. f was measured under different scenarios. 水仙苷 / 芦丁 f 水仙苷 / 烟花苷 and f 水仙苷 / 迷迭香酸 The mean values ​​were 0.98, 1.35, and 1.67, respectively, with RSDs of 1.00%, 4.14%, and 3.38%, indicating good reproducibility of the relative correction factors. The RCFs were ultimately determined to be f. 水仙苷 / 芦丁 =0.98, f 水仙苷 / 烟花苷 =1.35 and f 水仙苷 / 迷迭香酸 =1.67.

[0202] Table 19. Reproducibility study results of the relative correction factor (using narcisin as an internal reference) (n=3)

[0203]

[0204] 2.7 Localization of chromatographic peaks

[0205] Kromasil C 18 MN C18, Agilent C 18 The retention times (t) of the four components were determined using three HPLC instruments: a Shimadzu LC-20A, an Agilent 1200, and a Waters e2695. R .

[0206] Using rutin as an internal reference (s), according to formula r x / s =t Rx / t Rs and r x-s =t Rx -t Rs Calculate the relative retention values ​​and retention time differences of rosmarinic acid, narcissin, and rosmarinic acid, where r is the relative retention value and retention time difference. x / s r x-s Let t represent the relative retention value and retention time difference of component x relative to internal reference s. Rx Let t be the retention time of component x. Rs The retention times of the internal reference s are shown in Tables 20 and 21. The RSDs of the relative retention values ​​of the three analytes ranged from 1.61% to 3.38%, and the RSDs of the retention time differences ranged from 8.60% to 10.6%. These results indicate that the relative retention value method is more accurate in chromatographic peak localization compared to the retention time difference method. Therefore, when rutin was used as the internal reference, the relative retention values ​​of rosmarinic acid, naringin, and rosmarinic acid were finally determined to be 1.43, 1.58, and 2.04, respectively.

[0207] Table 20 Relative retention values of the test components when rutin was used as the internal standard (n = 9)

[0208]

[0209] Table 21 Retention time differences of the test components when rutin was used as the internal standard (n = 9)

[0210]

[0211]

[0212] According to the same method described above, the relative retention values and retention time differences of rutin, narciclasine and rosmarinic acid were calculated when narciclasine was used as the internal standard, and the results are shown in Table 22 and Table 23. The RSDs of the relative retention values of the three test components were 0.63% to 1.29%, and the RSDs of the retention time differences were -9.12% to 11.4%. The above results show that, compared with the retention time difference method, the relative retention value method can achieve more accurate positioning of the chromatographic peaks. Therefore, when narciclasine was used as the internal standard, the relative retention values of rutin, narciclasine and rosmarinic acid were finally determined to be 0.63, 0.91 and 1.29, respectively.

[0213] Table 22 Relative retention values of the test components when narciclasine was used as the internal standard (n = 9)

[0214]

[0215]

[0216] Table 23 Retention time differences of the test components when narciclasine was used as the internal standard (n = 9)

[0217]

[0218] 2.8 Determination of sample content and verification of QAMS method

[0219] The contents of rutin, narciclasine, narciclasine and rosmarinic acid in compound Gaoziba tablets were determined by the external standard method (ESM) and the QAMS method, respectively. When the QAMS method was used, the calculation was performed according to the formula C x = (A x / A s ) x C s x f s / x , where C x represents the mass concentration of the test component x in the test solution, A x represents the peak area of the test component, A s represents the peak area of the control solution of the internal standard, C sThe mass concentration of the control solution representing the internal reference substance, f s / x The relative correction factor of the component x to be measured.

[0220] 2.8.1 Verification of QAMS method (with rutin as internal reference substance)

[0221] The contents of the above-mentioned four components in Compound Gaozhi Tablets were calculated by the external standard method and the QAMS method with rutin as internal reference substance, respectively, and the relative deviation (RD) between them was compared, and the results are shown in Table 24. The results show that there is no significant difference between the results obtained by the two methods, and the relative deviation is less than 2.0%, indicating that the QAMS method with rutin as internal reference substance has good accuracy for the determination of the contents of the above-mentioned four components in Compound Gaozhi Tablets.

[0222] Table 24 Comparison of the contents of four components in Compound Gaozhi Tablets by ESM method and QAMS method (with rutin as internal reference substance)

[0223] (mg·g -1 , n = 2)

[0224]

[0225] 2.8.2 Verification of QAMS method (with zephyranthine as internal reference substance)

[0226] The contents of the above-mentioned four components in Compound Gaozhi Tablets were calculated by the external standard method and the QAMS method with zephyranthine as internal reference substance, respectively, and the relative deviation (RD) between them was compared, and the results are shown in Table 25. The results show that there is no significant difference between the results obtained by the two methods, and the relative deviation is less than 2.0%, indicating that the QAMS method with zephyranthine as internal reference substance has good accuracy for the determination of the contents of the above-mentioned four components in Compound Gaozhi Tablets.

[0227] Table 25 Comparison of the contents of four components in Compound Gaozhi Tablets by ESM method and QAMS method (with zephyranthine as internal reference substance) -1 (mg·g

[0228]

[0229] The above has introduced the embodiments of the present application in detail, and the principles and implementation manners of the present application have been described by applying specific examples. The above description of the embodiments is only for helping to understand the method of the present application and its core idea. Meanwhile, the changes or deformations made by the person skilled in the art according to the idea of the present application, based on the specific implementation manners and application scope of the present application, all belong to the scope of protection of the present application. In summary, the content of the present description should not be understood as a limitation of the present application.

Claims

1. A quality control method for compound Gao Zi Ban tablets, characterized in that, The quality control method includes the following steps: 1) Preparation of mixed reference solutions: Prepare reference solutions of rutin, rosmarinic acid, narcissin, and rosmarinic acid; 2) Preparation of Compound Gaoziban Tablets Test Solution: Weigh an appropriate amount of Compound Gaoziban Tablets powder, add methanol solution for extraction, cool, weigh, shake, filter, and take the filtrate to obtain the Compound Gaoziban Tablets Test Solution. 3) The reference solution was diluted separately and injected into a high-performance liquid chromatograph (HPLC) along with the test solution for chromatographic detection. The chromatographic conditions were as follows: a column packed with octadecylsilane-bonded silica gel was used; mobile phase A was an acid-water solution; mobile phase B was acetonitrile; the gradient elution program was: 0–10 min, 10% → 15% B; 10–40 min, 15% → 17% B; 40–50 min, 17% → 21% B; 50–60 min, 21% → 30% B; the flow rate was 0.4–1.5 ml / min; the column temperature was 20–50 °C; the detection wavelength was 200–350 nm; and the injection volume was 10–30 μl. 4) Based on the test results, obtain the content information of rutin, rosmarinic acid, naringin and / or rosmarinic acid in the compound Gaoziban tablets.

2. The quality control method according to claim 1, characterized in that, The method further includes calculating the relative retention value, relative correction factor and / or content of the target compound using rutin or narcissin as internal references, wherein the target compound is selected from one or more of the following: rutin, narcissin, narcissin and rosmarinic acid.

3. The quality control method according to claim 2, characterized in that, The calculation of the relative retention value, relative correction factor, and / or content of the target compound using rutin or narcissin as internal references includes: (a) Based on the mass concentration of each reference standard in the mixed reference solution and the corresponding peak area in the chromatogram, determine the standard curves for rutin, rosmarinic acid, narcissin, and rosmarinic acid, respectively. (b) Determine the internal reference material, based on the slope of the standard curve obtained in step (a), according to formula f. s / x =K s / K x Calculate the relative correction factor for each target compound. Among them, f s / x K represents the relative correction factor of target compound x relative to internal reference s. s K represents the slope of the standard curve for the internal reference s. x The slope of the standard curve for target compound x; (c) The chromatographic peaks of rutin, rosmarinic acid, naringin, and rosmarinic acid in the high performance liquid chromatogram of the compound Gaoziban tablets test solution were located using relative retention values, and the peaks were determined according to formula r. x / s =t Rx / t Rs Calculate the relative retention value of the target compound. Where, r x / s t represents the relative retention value of target compound x relative to internal reference s. Rx t represents the retention time of the target compound x. Rs The retention time of the internal reference substance s; and (d) Based on the mass concentration and corresponding peak area of ​​the reference solution described in step (a), the relative correction factor described in step (b), and the corresponding peak area of ​​the chromatographic peak located in step (c), according to formula C x =(A x / A s )×C s ×f s / x Calculate the contents of rutin, rosmarinic acid, naringin, and rosmarinic acid in the test solution of the compound Gaoziban tablets. Among them, C x A represents the mass concentration of target compound x in the test solution. x A represents the peak area of ​​target compound x. s C represents the peak area of ​​the reference solution of internal reference s. s f represents the mass concentration of the reference solution of internal reference substance s. s / x This represents the relative correction factor for the target compound x.

4. The quality control method according to claim 2, characterized in that, The standard curve for rutin is Y = 36085X, and the correlation coefficient (r) is 0.9999.

5. The quality control method according to claim 2, characterized in that, The standard curve for the pyrolysin is Y = 29290X, r = 0.9999.

6. The quality control method according to claim 2, characterized in that, The standard curve for narcisin is Y = 34361X, r = 0.9999.

7. The quality control method according to claim 2, characterized in that, The standard curve for rosmarinic acid is Y = 19872X, r = 0.9999.

8. The quality control method according to claim 2, characterized in that, When rutin was used as an internal control, the relative correction factor of rosmarinic acid relative to rutin was approximately 1.38, and the relative retention value of rosmarinic acid relative to rutin was approximately 1.43; the relative correction factor of narcissin relative to rutin was approximately 1.02, and the relative retention value of narcissin relative to rutin was approximately 1.58; the relative correction factor of rosmarinic acid relative to rutin was approximately 1.70, and the relative retention value of rosmarinic acid relative to rutin was approximately 2.

04.

9. The quality control method according to claim 2, characterized in that, When narcissin was used as an internal control, the relative correction factor of rutin to narcissin was approximately 0.98, and the relative retention value of rutin to narcissin was approximately 0.63; the relative correction factor of rosmarinic acid to narcissin was approximately 1.35, and the relative retention value of rosmarinic acid to narcissin was approximately 0.91; the relative correction factor of rosmarinic acid to narcissin was approximately 1.67, and the relative retention value of rosmarinic acid to narcissin was approximately 1.

29.

10. The quality control method according to claim 1, characterized in that, The compound Gaoziban tablets are compound Gaoziban tablets with the coating removed.

11. The quality control method according to claim 1, characterized in that, The compound Gaoziban tablet powder is the sieved compound Gaoziban tablet powder.

12. The quality control method according to claim 11, characterized in that, The sieve in question is a No. 3 sieve.

13. The quality control method according to claim 1, characterized in that, The mass of the compound Gaoziban tablet powder is 0.1-10g.

14. The quality control method according to claim 13, characterized in that, The mass of the compound Gaoziban tablet powder is approximately 2.0g.

15. The quality control method according to claim 1, characterized in that, The mass / volume ratio of the compound Gaoziban tablet powder to the methanol solution is 0.01 to 0.2, with units of g / ml.

16. The quality control method according to claim 15, characterized in that, The mass / volume ratio of the compound Gaoziban tablet powder to the methanol solution is approximately 0.1, in g / ml.

17. The quality control method according to claim 1, characterized in that, The methanol solution has a mass-volume percentage of 10% to 100%.

18. The quality control method according to claim 17, characterized in that, The methanol solution has a mass-volume percentage of approximately 90%.

19. The quality control method according to claim 1, characterized in that, The extraction is performed by reflux extraction, cold soaking extraction, shaking extraction, and / or ultrasonic extraction.

20. The quality control method according to claim 19, characterized in that, The extraction method is reflux extraction.

21. The quality control method according to claim 20, characterized in that, The reflux extraction is a water bath reflux extraction.

22. The quality control method according to claim 21, characterized in that, The water bath reflux extraction time is 10–60 min.

23. The quality control method according to claim 22, characterized in that, The water bath reflux extraction time is approximately 30 minutes.

24. The quality control method according to claim 1, characterized in that, The cooling process refers to cooling the material to room temperature.

25. The quality control method according to claim 1, characterized in that, The method for preparing the mixed reference solution includes: weighing appropriate amounts of rutin, safflowerin, narcissin, and rosmarinic acid reference standards, and adding methanol solution to prepare rutin reference standard stock solutions, safflowerin reference standard stock solutions, narcissin reference standard stock solutions, and rosmarinic acid reference standard stock solutions, respectively; separately pipetting appropriate amounts of the rutin reference standard stock solutions, safflowerin reference standard stock solutions, narcissin reference standard stock solutions, and rosmarinic acid reference standard stock solutions into the same 10 ml volumetric flask to obtain mixed reference solution No. 1; and diluting mixed reference solution No. 1 by 2 times, 5 times, 10 times, 25 times, and 50 times, respectively, to prepare mixed reference solutions No. 2 to No.

6.

26. The quality control method according to claim 25, characterized in that, The methanol solution has a mass-volume percentage of 10% to 100%.

27. The quality control method according to claim 26, characterized in that, The methanol solution has a mass-volume percentage of approximately 90%.

28. The quality control method according to claim 25, characterized in that, The mass concentration of the rutin stock solution is 50–1000 μg / ml.

29. The quality control method according to claim 28, characterized in that, The mass concentration of the rutin stock solution is approximately 600 μg / ml.

30. The quality control method according to claim 25, characterized in that, The mass concentration of the pyrolysin stock solution is 50–1000 μg / ml.

31. The quality control method according to claim 30, characterized in that, The mass concentration of the pyrolysin stock solution is approximately 642 μg / ml.

32. The quality control method according to claim 25, characterized in that, The mass concentration of the narcissin stock solution is 50–1000 μg / ml.

33. The quality control method according to claim 32, characterized in that, The mass concentration of the narcisin stock solution is approximately 628 μg / ml.

34. The quality control method according to claim 25, characterized in that, The mass concentration of the rosmarinic acid stock solution is 50–1000 μg / ml.

35. The quality control method according to claim 34, characterized in that, The mass concentration of the rosmarinic acid stock solution is approximately 496 μg / ml.

36. The quality control method according to claim 25, characterized in that, The volume of the rutin stock solution is 1-5 ml.

37. The quality control method according to claim 36, characterized in that, The volume of the rutin stock solution is approximately 2 ml.

38. The quality control method according to claim 25, characterized in that, The volume of the pyrolysis glycoside stock solution is 1-5 ml.

39. The quality control method according to claim 38, characterized in that, The volume of the pyrolysis glycoside stock solution is approximately 2 ml.

40. The quality control method according to claim 25, characterized in that, The volume of the narcissin stock solution is 1–5 ml.

41. The quality control method according to claim 40, characterized in that, The volume of the narcissin stock solution is approximately 2 ml.

42. The quality control method according to claim 25, characterized in that, The volume of the rosmarinic acid stock solution is 1–5 ml.

43. The quality control method according to claim 42, characterized in that, The volume of the rosmarinic acid stock solution is approximately 4 ml.

44. The quality control method according to claim 25, characterized in that, The mass concentration of rutin in the No. 1 mixed reference solution is approximately 120 μg / ml.

45. The quality control method according to claim 25, characterized in that, The mass concentration of pyroside in the No. 1 mixed reference solution was approximately 128.4 μg / ml.

46. ​​The quality control method according to claim 25, characterized in that, The mass concentration of narcisin in the No. 1 mixed reference solution was approximately 125.6 μg / ml.

47. The quality control method according to claim 25, characterized in that, The mass concentration of rosmarinic acid in the No. 1 mixed reference solution was approximately 198.4 μg / ml.

48. The quality control method according to claim 1, characterized in that, The flow rate for the high-performance liquid chromatography (HPLC) detection is 0.6–1.2 ml / min.

49. The quality control method according to claim 48, characterized in that, The flow rate for the high-performance liquid chromatography detection is approximately 1.0 ml / min.

50. The quality control method according to claim 1, characterized in that, The column temperature is 30–40°C.

51. The quality control method according to claim 50, characterized in that, The column temperature is approximately 35°C.

52. The quality control method according to claim 1, characterized in that, The detection wavelength is 220–270 nm.

53. The quality control method according to claim 52, characterized in that, The detection wavelength is 254 nm.

54. The quality control method according to claim 1, characterized in that, The injection volume is 15–25 μl.

55. The quality control method according to claim 54, characterized in that, The injection volume is approximately 20 μl.

56. The quality control method according to claim 2, characterized in that, The resolution of the chromatographic peak corresponding to the target compound is greater than 1.

5.

57. The quality control method according to claim 1, characterized in that, The chromatographic column was Kromasil C10. 18 Chromatographic column, 4.6 × 250 mm, 5 μm, Phenomenex C100000 ppm. 18 Chromatographic column, 4.6 × 250 mm, 5 μm, MN C 18 Chromatographic column, 4.6 × 250 mm, 5 μm, Agilent C10. 18 Chromatographic column, 4.6 × 250 mm, 5 μm, or Thermo C 18 Chromatographic column, 4.6×250mm, 5μm.

58. The quality control method according to claim 57, characterized in that, The chromatographic column was Kromasil C10. 18 Chromatographic column, 4.6×250mm, 5μm.

59. The quality control method according to claim 1, characterized in that, The acidic aqueous solution is selected from formic acid, glacial acetic acid, phosphoric acid, trifluoroacetic acid, or a mixture of phthalic acid and hydrochloric acid of different concentrations.

60. The quality control method according to claim 1, characterized in that, The acidic aqueous solution is a 0.01% to 2% acidic aqueous solution.

61. The quality control method according to claim 60, characterized in that, The acidic aqueous solution is a 0.01% to 2% formic acid aqueous solution.

62. The quality control method according to claim 61, characterized in that, The acidic aqueous solution is an aqueous solution of approximately 0.2% formic acid.

63. Use of the quality control method according to any one of claims 1 to 62 in the quality testing and / or quality evaluation and / or quality control of Compound Gao Zi Ban Tablets.

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