高效液相色谱法检测尿毒清颗粒(无糖型)中芍药苷、大车前苷和丹参酮ⅡA成分的含量

HPLC method for determination contents of paeoniflorin,plantamoside and tanshinone Ⅱ_(A) in Niaoduqing granules(sugar-free type)

目的建立检测尿毒清颗粒(无糖型)中芍药苷、大车前苷和丹参酮ⅡA成分含量的方法。方法取尿毒清颗粒(无糖型)适量,研细,取约1.0 g,精密称定,置50 mL棕色容量瓶中,加90%甲醇溶液超声处理30 min后,放冷定容,0.45µm滤膜滤过,Waters Spherisorb ODS1液相色谱柱分离。检测波长为芍药苷(230 nm)、大车前苷(330 nm)、丹参酮ⅡA(270 nm);流动相为乙腈(A)-0.2%磷酸溶液(B),梯度洗脱;流速为1.0 mL/min,柱温为25℃,进样量为10µL,二极管阵列检测器定量检测。结果芍药苷、大车前苷和丹参酮ⅡA线性范围分别为1.427~57.06µg/mL、2.255~90.2µg/mL和0.498~19.9µg/mL(R2为0.992~0.996);平均加样回收率分别为99.39%,97.99%和98.41%;重复性RSD分别为3.16%、3.38%和2.69%;样品稳定性良好。结论HPLC法重复性好、操作简单,弥补了单一指标成分含量测定作为质控手段的不足,为提高该产品的质量控制提供技术依据。

Objective To explore the method for determination contents of paeoniflorin,plantamoside and tanshinone ⅡA in Niaoduqing Granules(sugar-free type).Methods Appropriate amount of Niaoduqing granules(sugar-free type) was taken and grinded.1.0 g samples was weighed up precisely to 50 mL brown glass flask volumetric,90% methanol solution was added to the flask and ultrasound for 30 minutes.Cooling the samples and fixed to constant volume.After filtered by 0.45 μm membrane filter,samples was seperated on Waters Spherisorb ODS1 columm.Detection wavelength were as follows:paeoniflorin(230 nm),plantamoside(330 nm) and tanshinone ⅡA(270 nm).Mobile phase was acetonitrile(A)-0.2% phosphoric acid solution(B).Gradient elution,flow rate:1.0 mL/min,column temperature:25℃ and injection volumewas 10 μL.Diode array detector was for quantitative detection.Results Paeoniflorin,plantamoside and tanshinone ⅡA showed good linear relationships in following concentration range 1.427-57.06 μg/mL,2.255-90.2 μg/mL and 0.498-19.9 μg/mL,respectively.With correlation coefficients R2 all among 0.992-0.996.Average recoveries were 99.39%,97.99% and 98.41%,respectively.Relative standard deviation(RSD) of repeatability(n=6) was 3.16%,3.38% and 2.69%,respectively.Stability of samples was good.Conclusion The HPLC method has good repeatability and simple operation,which makes up for the deficiency of single index component content determination as a quality control method,and provides a technical basis for improving the quality control of the product.