血塞通滴丸皂苷类成分含量测定及批次一致性评价研究

目的建立血塞通滴丸超高压液相色谱(UPLC)的指纹图谱方法并测定其主要成分含量,并结合相关化学计量学方法对产品质量批次一致性进行系统、全面和科学的评价。方法采用Agilent 1290 UPLC,Acquity UPLC BEH C18色谱柱,以乙腈-水为流动相进行梯度洗脱,对5种皂苷类成分三七皂苷R_1(R_1)及人参皂苷Rg1、Re、Rb1、Rd(Rg1、Re、Rb1、Rd)进行含量测定,并建立了血塞通滴丸的指纹图谱,通过对指纹图谱的相似度分析、主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)对血塞通滴丸的批次一致性进行全面评价,同时采用Hotelling’s T2和DModX 2种统计量,为不同批次产品的质量设定控制范围。结果 14批样品中,5种皂苷含量较为稳定,设定的质控范围分别为R_1 24.69～30.03 mg/g、Rg_1 102.60～122.44 mg/g、Re 12.93～15.45 mg/g、Rb_1 99.74～112.12 mg/g和Rd 23.35～31.75 mg/g。建立的指纹图谱共标定15个共有峰,经对照品化学指认共鉴定出R_1、Rg_1、Re、Rb_1、Rd 5个色谱峰。14批样品的相似度均大于0.998。PCA分析中14批样品无异常批次,表明血塞通滴丸的批次一致性较高,但存放时间差异在指纹图谱上有所体现;OPLS-DA分析得出,在造成14批样品有渐变趋势的变量中,已定量的Rb1和Rd影响较大;Hotelling’s T~2和DModX的控制上限分别为31.17和1.82。结论所建方法科学准确,简便可靠,不但可以用于血塞通滴丸的质量控制,而且可以对其批次质量一致性进行全面评价,可为解决中药质量批次一致性的共性问题提供借鉴。

Determination of saponins in Xuesaitong Drop Pills and study on batch consistency evaluation techniques

Objective To establish the fingerprint of Xuesaitong Dropping Pills (XDP) using ultra-high pressure liquid chromatography (UPLC), determine the content of its main components and propose a chemometrics method for the systemic, comprehensive and scientific quality evaluation. Methods The Agilent 1290 UPLC and Acquity UPLC BEH C₁₈ column were used for the establishment of the UPLC fingerprints of 14 batches of XDP with acetonitrile and water as mobile phase for gradient elution, and the content of five kinds of notoginsenosides wsa determined. Then, the further quality assessment of XDP was carried out with similarity evaluation, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Meanwhile, the Hotelling''s T² and DModX control ranges were set for the different batches of samples. Results Among the 14 batches, the content of five kinds of notoginsenosides was relatively stable, and the quality control ranges for notoginsenosides R, ginsenoside Rg₁, Re, Rb₁, Rd were set at 24.69-30.03 mg/g, 102.60-122.44 mg/g, 12.93-15.45 mg/g, 99.74-112.12 mg/g, and 23.35-31.75 mg/g, respectively. In this research, 15 chromatographic peaks were recognized as common peaks in the fingerprints, five peaks were identified with standard references compounds, which were notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Re, ginsenoside Rb₁ and ginsenoside Rd. The similarity values of the drugs were all above 0.998. In the PCA analysis, 14 batches of samples had no abnormal batch, indicating that the quality was relatively stable, however, the difference of the storage time was reflected in the fingerprint. According to the OPLS-DA result, the quantitative ginsenoside Rb₁ and ginsenoside Rd had greater influence on the variables causing the gradual trend in 14 batches of samples. The upper limit of control for Hotelling''s T² and DModX was 31.17 and 1.82, respectively. Conclusion The established method was accurate, reliable, and simple. It can be used not only for the quality control of XDP, but also for the comprehensive evaluation of batch consistency. The study also provides a reference for solving the common problem of the consistency evaluation of Chinese materia medica.