用高效液相色谱指纹图谱定量评价4种不同剂型六味地黄丸质量和工艺差异

目的建立4种剂型六味地黄丸（Liuweidihuang Pill，LWDHP）HPLC指纹图谱，以系统指纹定量法（SQFM）综合鉴定其质量和考察不同剂型的工艺情况。方法采用RP—HPLC法以Century SIL C18 BDS柱（250mm×4．6mm，5μ），流动相为1％HAc溶液和1％HAc甲醇溶液梯度洗脱，紫外检测波长265nm，柱温（30．00±20．15）℃，进样量10μL。以29批传统剂型LWDHPs建立代表传统剂型LWDHPs药效物质基础特点的对照指纹图谱（RFP），以其评价39批LWDHPs质量。结果以j羟甲基糠醛为参照峰，确定33个共有指纹峰，建立了HPLC指纹图谱。以宏定性相似度Sm和校正宏定量相似度为指标按SQFM鉴定4种不同剂型LWDHPs整体质量。结果LWDHP胶囊的整体化学成分数量和分布比例以及化学成分含量显著低于传统剂型，而浓缩丸与RFP相当；以SQFM分别评价4类LWDHPs批间质量差异的结果表明浓缩丸剂型的工艺稳定性较好，样品化学成分数量和分布比例及含量的差别显著低于其他3个剂型。水蜜丸工艺稳定性最差，胶囊的工艺稳定较好但化学物质基础与传统剂型差异太大。结论采用SQFM可迅速、准确地评价不同剂型和不同批次的中成药质量。该法可实现控制和解决我国中成药药效物质基础极不稳定的状况。

Quantitative identification of the quality of 4 dosage forms of Liuweidihuang pills and evaluation of the formulation difference by HPLC fingerprint

Objective To set up the HPLC fingerprint of 4 dosage forms of Liuweidihuang pills （LWDHP）, in which the overall quality of LWDHPs was evaluated and the formulation difference were investigated by systematic quantified fingerprint method （SQFM）. Methods The chromatographic fingerprints （CFPs） were determined by RP-HPLC and injecting 10 μL of the sample solution each time on a Century SIL C18 BDS column （250 mm×4. 6 mm, 5μm） with the gradient elution solvent system composed of 1 % acetate acid water and 1%0 acetate acid methanol. The flow rate was 1 mL · min- 1, the column temperature was （30.00 ±0.15）C , and the detective wavelength was 265 nm. The referential fingerprint （RFP） which represented characteristics of traditional formulation material bases for efficacy was produced from 29 batches of traditional LWDHPs in terms of average model. Taking the RFP as a norm, 39 batches I.WDHPs was assessed by SQFM. Results The LWDHP HPLOfingerprint containing 33 co-possessing peaks was obtained when choosing 5-hydroxymethyl-2-furfural （5 HMF） peak as the referential peak. The 4 dosage forms of LWDHPs were identified by SQFM using maero qualitative similarity （Sm） and the corrected macro quantitative similarity （Pm） as the evaluating parameters. Among the 4 different dosage forms of LWDHPs, the number of chemical fingerprints, distribution ratio and content of chemical components in the capsule samples had lower similarity than the traditional formulation while the concentrated pill samples had little difference with RFP. Quality differences among batches of the 4 dosage forms LWDHPs were assessed by SQFM in dosage form separately. The formulation stability of concentrated pills was well, and the difference in chemical composition volume, distribution ratio and the content of chemical components among different batches of concentrated pills was less than the other 3 dosage forms, while the formulation stability of water-honeyed pills was poor. The difference in chemical substance bases between the capsule and the 3 traditional formulations were significant although the capsule stability was good. Conclusion SOFM can quickly and accurately evaluate the entire quality of traditional Chinese medicine of different formulations and different batches, which can control and settle the instability of chemical substance bases of traditional Chinese medicine.