Study on the Quality Standard of Tetrastigma hemsleyanum Stem and Leaves Dispensing Granules and Content Determination of 5 Active Ingredients / 中国药房

ABSTRACT

OBJECTIVE：To estab lish the quality standard of Tetrastigma hemsleyanum stem and leaves dispensing granules ， and to determine the contents of 5 active ingredients simultaneously. METHODS ： Chlorogenic acid ， isoorientin and isoorientin-2′-O-rhamnoside in T. hemsleyanum stem and leaves dispensing granules were identified by TLC. The total flavonoids of the 3 batches granules were determined by UV spectrophotometry （by isoorientin ）. The granule sizes ，moisture contents ， dissolvability，and content uniformity were determined. Using isoorientin as internal reference ，relative correction factors of other components were established. QAMS method was adopted to determine the contents of neochlorogenic acid ，chlorogenic acid ， orientin，isoorientin and isoorientin- 2′-O-rhamnoside，which were compared with the results of ESM method. RESULTS ：TLC spots of chlorogenic acid ，isoorientin and isoorientin- 2′-O-rhamnoside were clear and well-separated ，without interference from negative control. The linear range of isoorientin were 7.73-61.82 μ g/mL（r＝0.999 9）. RSDs of precision ，stability and reproducibility tests were all lower than 2％. The recoveries were 93.75％-97.85％（RSD＝1.41％，n＝6）. The average percentages that the 3 batches granules could not pass through sieve No. 1 - moisture contents were 4.63％，5.18％ and 4.03％（n＝3）. were dissolved within 5 min ，and content uniformity were 4.8％-5.0％. Which were all in line with the relevant provi-sions of granules in 2015 edition of Chinese Pharmacopoeia . The linear range of 5 ingredients were 2.325-93 μg/mL（r＝0.999 9），5.125-205 μg/mL（r＝0.999 9），1.150- 46 μg/mL（r＝0.999 3），2.625-105 μg/mL（r＝0.999 9），4.725-189 μg/mL（r＝0.999 9），respectively. RSD of precision ，stability and reproducibility tests were all lower than 3％. The recoveries were 99.78％-106.13％（RSD＝2.33％，n＝6），95.07％-103.32％ （RSD＝2.72％，n＝6），97.17％-105.43％（RSD＝2.98％，n＝6），95.52％-101.33％（RSD＝2.46％，n＝6），99.42％-105.56％ （RSD＝2.34％，n＝6）. Using isoorientin as internal reference ，relative correction factors of neochlorogenic acid ，chlorogenic acid ， isoorientin and isoorientin- 2′-O-rhamnoside were 0.731，0.805，0.821，0.590，respectively. The contents were 0.828-1.123， 2.379-3.118，0.281-0.880，1.039-1.393，2.121-3.209 mg/g by QAMS method ，while the contents were 0.803-1.099，2.345-3.085， 0.269-0.872，1.309-1.393，2.113-3.201 mg/g by ESM method ，there was no significant difference in the content determination results between QAMS and ESM method （P＞0.05）. CONCLUSIONS ：Established quality standard is simple and rapid ，and can be used for quality control of T. hemsleyanum stem and leaves dispensing granules. Established QAMS method is accurate and efficient，and it can be used for simultaneous determination of 5 active ingredients of T. hemsleyanum stem and levels dispensing granules.