目的建立中药复方总苷类成分的定量测定方法——差式蒽酮-糖脎比色法。方法用蒽酮比色法测定中药总糖苷(包括糖类)的量,然后采用糖脎硫酸显色法测定还原性糖类的量,两者相减,即得中药总苷类成分的量。结果蒽酮比色法的反应在6 h内完成;测定波长为620 nm;线性范围在80~180μg/mL,R2=0.994;回收率为99.2%,RSD为1.35%。糖脎硫酸显色法的反应在4 h内完成;测定波长为412 nm;线性范围在2~12 mg/mL,R2=0.991;回收率为99.5%,RSD为1.98%。采用本法测得补阳还五汤、六味地黄丸、穿心莲片、黄芪注射剂、茯苓多糖口服液5种中成药总苷的量依次为15.1、26.0、6.68 mg/g及21.3、16.9 mg/mL。结论首次创立了中药复方中总苷类成分定量测定方法,通过蒽酮比色法与糖脎硫酸显色法差值测算建立起以糖苷键为特征的成分群分析方法。方法学证明本法有较好的精确度,能用于中药总苷成分群的定量测定。
Objective To establish a method for the determination of total glycosides in Chinese materia medica (CMM)--differential anthrone-sugar hydrazone colorimetry. Methods The content of total glycosides (including sugar) in CMM was determined by anthrone colorimetry, and then sugar hydrazone-sulfuric acid colorimetric method was used to determine the content of reducing sugars. The content of total glycosides came from the two subtraction. Results Anthrone colorimetric reaction was completed within 6 h, detection wavelength was 620 nm, linear range was 80--180 p,g/mL, correlation coefficient R2 = 0.994, and recovery was 99.2%, RSD was 1.35%. Sugar hydrazone-sulfuric acid color reaction method was completed within 4 h, detection wavelength was 412 nm,linear range was 2--12 mg/mL, correlation coefficient R2 = 0.991, and recovery was 99.5%, RSD was 1.98%. The total glycosides contents in five Chinese patent medicines such as Buyang Huanwu Decoction, Liuwei Dihuang Pill, Andrographis Tablet, Astragalus Injection, and Pachymaran Oral Liquid were 15.1, 26.0, 6.68 mg/g, and 21.3, 16.9 mg/mL. Conclusion This method is firstly established for the quantitative determination of total glycosides in CMM, and the ingredient group analysis method is established by anthrone colorimetry and sugar hydrazone-sulfuric acid colorimetric method. The methods have better accuracy, and can be used for the determination of total glycosides ingredients group of CMM.