ã€Abstract】Objective To establish a method for the determination of glycyrrhizic acid in Sanyue Mahuang Granule by reversed-phase high performance liquid chromatography (RP- HPLC) . The method of determination using RP-HPLC Ephedra Mitsukoshimae particles glycyrrhizic acid, KromasilODS 1C18 column (5μm, 4.6mm × 250mm), mobile phase of acetonitrile 0.05M potassium dihydrogen phosphate (pH5 with triethylamine adjusted .75 ) ( 26.5 : 73.5 ), detection wavelength is 250 , column temperature is 40 °C , flow rate is 1.0ml/min . Results The linear range of glycyrrhizic acid in Sanyue Mahuang Granules was 3.75 ~ 154.5μg / ml (r = 0.9998) , the recovery rate was 100.2% , RSD = 2.5% . Conclusion The determination of glycyrrhizic acid by RP-HPLC method is simple, accurate and reproducible, and it can provide a basis for the quality control of Sanyue Mahuang Granule. Key words: high performance liquid chromatography, glycyrrhizic acid, Sanyue ephedra Sanyue Mahuang Granules are made from 10 herbs ( ephedra, gypsum, bitter almond, radix radicans, licorice, tangerine peel, Bupleurum, Shegan, Houttuynia, Fritillaria ) . The licorice in the middle is used as a medicine. Glycyrrhizic acid is one of the main indicators in the medicinal herbs. It is proposed that the content of the component can reflect and control the intrinsic quality of the product. The HPLC method was established to determine the content of glycyrrhizic acid in this product, in order to achieve the purpose of accurate quantification.   1 , instruments and reagents    1.1 Instrument Waters2690 high performance liquid chromatography, Waters2487 UV detector, KromasilODS 1C18 column ( 5μm, 4.6mm × 250mm ) and guard column, electronic analytical balance ( BS110S , Beijing Sartorius Co., Ltd.).    1.2 reagent acetonitrile is chromatographically pure (grade 1), water is re-distilled water, other reagents are of analytical grade.    1.3 The control product monoammonium glycyrrhizinate ( Aldrich Chem Co. , lot No. 23,224 -6 ) was dried in a desiccator to a constant weight before use.   2 , method    2.1 Chromatographic conditions The mobile phase was acetonitrile  0.05 M potassium dihydrogen phosphate ( pH 5.55 adjusted with triethylamine ) ( 26.5 : 73.5 ); detection wavelength 250 nm ; column temperature 40 ° C ; flow rate 1.0 ml / min .    2.2 Solution preparation    3 , the results    3.1 System suitability experiment takes "    3.2 Determination of linear range Accurately take 10μl of the solution of the monoammonium glycyrrhizinate reference substance solution , record the chromatogram, and determine the peak area. The results are shown in Table 1 . The concentration ( C ) is regressed with the peak area value ( A ) to obtain the standard curve equation: A = 5834.7C-8388.1, r = 0.9998 . The above results show that the peak area value ( A ) of glycyrrhizic acid has a good linear relationship with the concentration ( C ) in the range of 3.75 to 154.5 μg/ml .    3.3 Precision Experiment The above reference solution was accurately aspirated and the injection was repeated 5 times. The RSD of the glycyrrhizic acid peak area value was 0.28 %. The results are shown in Table 2 .    3.4 Stability test The test solution (batch number 20000729 ) was taken at 0 , 2 , 4 , 6 , 8 and 10 h , respectively. The results are shown in Table 3 . The experimental results show that the test solution has good stability within 10 hours .    3.5 Repetitive experiments According to the proposed content determination method, 6 samples were prepared for the same batch of test materials (20020915) , and the peak area was measured and the glycyrrhizic acid content was calculated.    3.6 Recovery rate experiment Take 9 parts of the known content (batch number 20020825 ), add 0.7725mg/ml glycyrrhizic acid reference solution 0.46 , 0.58 , 0.7ml , according to the above preparation method and chromatographic conditions, prepare The test solution was sampled and injected into a liquid chromatograph to calculate the recovery.    3.7 For the determination of the test sample, accurately draw the appropriate amount of the reference solution and the test solution, and determine according to the above chromatographic conditions. The results are shown in Table 6 .   4 , discussion The pre-experiment was carried out by HPLC conditions under the licorice in the Chinese Pharmacopoeia . The results showed that the effective separation of glycyrrhizic acid in the compound preparation could not be achieved. It was found that it was possible to achieve the expected chromatographic peak separation when pre-tested using the literature conditions. The mobile phase system is further optimized based on the basis of the mobile phase system. The results show that the introduction of a certain amount of triethylamine while increasing the acidity of the mobile phase can not only improve the separation effect, but also protect the column, and finally determine the mobile phase composition. 2% methanol -0.1% phosphoric acid and acetonitrile and 0.1% triethylamine (25: 75), and in that chromatographic conditions as the basis for a methodological study. In this experiment, a high performance liquid chromatography method for the determination of glycyrrhizic acid in Sanyue Mahuang Granules was established. According to the methodological investigation, the method meets the requirements of the establishment of relevant quality standards in the Chinese Pharmacopoeia. It has the advantages of rapid sensitivity, accuracy, reliability, simplicity, and repeatability, and can be used to control the quality of Sanyue ephedra granules. Commercial Waffle Maker,Tartlets Machine,Egg Tart Shell Machine,Egg Tart Machine Guangzhou New Power Catering Equipment Manufacturing Co.,Ltd , https://www.gznewpower.com