HPLC指纹图谱控制复方丹参片质量

目的采用双定性双定量相似度作为评价指标,建立复方丹参片(CSMT)HPLC指纹图谱的全质量控制方法。方法采用RP-HPLC法以Century SIL C18BDS柱(250 mm×4.6 mm,5μm);以色谱指纹图谱指数F为目标函数优化选择指纹图谱检测条件,确定流动相组成是质量分数为1%的醋酸水溶液-质量分数为1%的醋酸甲醇溶液低压梯度洗脱,紫外检测波长290 nm,柱温(30.00±0.15)℃,进样量5μL。以双定性相似度均值和双定量相似度均值为参量通过对20批复方丹参片进行系统聚类分析确定用其中10批生成复方丹参片对照指纹图谱(RFP),以此RFP为标准对总计20批复方丹参片采用双定性双定量相似度法进行质量评价。结果以丹酚酸B(SAB)为参照物峰,确定40个指纹峰,建立了复方丹参片HPLC指纹图谱。用双定性相似度评价15批样品的化学成分数量和分布比例都合格,但其中仅8批双定量相似度合格,为质量完全合格品。其它7批含量明显偏低,生产时应根据双定量相似度偏低程度提高投料量或者适当增大服用剂量。其余5批都存在化学成分和分布比例不合格特征,有2批含量相似度合格和3批含量相似度明显偏低。结论在OTC中随机选择的20批复方丹参片,无明显的劣质品出现,若按指纹图谱技术控制则出现明显的质量差异,因此所建立的具有较好精密度和重现性的复方丹参片的HPLC指纹图谱,完全适用于复方丹参片的生产质量控制。试验证明了双定性双定量相似度法是宏观定性定量评价中药复方制剂质量的最理想和最客观的指纹图谱评价技术。

Overall quality control of compound salvia miltrorrhiza tablets by HPLC fingerprints

Objective To establish an overall quality control method for compound salvia miltrorrhiza tablets（CSMTs）by HPLC fingerprints using the dual qualitative and dual quantitative similarities (DQDQS) as the assessing parameters. Method Applying the chromatographic fingerprint (CFP) index (F) to optimize the CFP conditions, the CFPs were obtained by injecting 5 μL of the sample solution each time in a CenturySIL C18 BDS column (250 mm×4. 6 mm, 5 μm) with the gradient elution solvent composed of 1% acetic acid water and 1% acetic acid methanol. The flow rate was 1 mL/min. The column temperature was maintained at (30.00 ±0.15)℃, and the detection wavelength was set at 290 nm. 20 batches of CSMTs were classified by hierarchical clustering analysis which used both the mean of dual qualitative similarities and the mean of dual qualitative similarities as the variables. Total 20 batches of CSMTs were evaluated by DQDQS based on the referential fingerprint (RFP), which was developed from 10 batches of CSMTs according to the results of classification. Results All of 40 co-possessing peaks were selected as fingerprint peaks while the salvianolic acid B peak was chosen as the referential peak. Evaluated by DQDQS, 15 batches of CSMTs were qualified both in quantity and distribution proportion of the chemical compositions. Only 8 batches of them were completely qualified while the other 7 ones were not when evaluated by dual quantitative similarities. The other 5 batches of them were all characterized as disqualification in chemical compositions and their distribution proportion, in which the contents of 2 batches were qualified while those of the other 3 batches were obviously lower. Conclusions There were evident quality differences when evaluated by fingerprint indexes, although no obviously poor quality appeared among these 20 batches of CSMTs selected randomly in OTCs. This novel HPLC method with good precision and reproducibility can be perfectly applied in the manufacturing quality control of CSMTs. is the test demonstrated that the DQDQS method is the most reasonable and objective technique to qualitatively and quantitatively assess the quality of Chinese traditional medicine compound preparation.