双色谱系统定量指纹图谱评价石斛夜光丸质量

Evaluation of Shihu Yeguang pills by quantity fingerprints monitored by dual chromatographic elution systems

目的研究在双色谱系统条件下用系统指纹定量法(SQFM)对石斛夜光丸(SHYGP)进行评价所产生的变动情况,同时考察SHYGP同一75%乙醇提取液经5℃放置70 d后指纹图谱变化。方法 SQFM的3个指标宏定性相似度(Sm)、宏定量相似度(Pm)和变动系数(α)间存在密切相关特性,在洗脱程序EP1条件下用高效液相色谱测定12批SHYGPs在203、286和326 nm波长处的指纹图谱,分别以均值法整合三波长下定量指纹图谱的鉴定结果;经70 d后重新提取样品在显著变动的洗脱程序EP2条件下同法试验和评价指纹图谱,对2次获得指纹图谱群的评价结果进行误差对比,并讨论质量鉴定等级和宏定量相似度变化。同时对75%乙醇提取液5℃存放70 d后采用EP2方法试验,同法考察变化。结果以柚皮苷(NG)为参照物峰,EP1确定38(203 nm),34(286 nm)和30(326 nm)个共有指纹峰,EP2确定32(203 nm)、37(286 nm)和26(326 nm)个共有指纹峰。经SQFM评价后的三波长指纹图谱结果按照均值法整合,对S2、S3、S4和S12等4批样品鉴定结果一致,除S6、S11外,余下6批样品鉴定结果波动仅1级,不考虑S1、S6和S11,Sm平均波动<0.04,Pm误差基本<10%(S9、S10除外)。对EP1和EP2三波长HPLC-FPs鉴定数据取平均,最终鉴定S1、S2、S3、S5和S7质量极好(1级),S4、S8和S9质量很好(2级),S10和S12质量好(3级),S11质量较好(4级),S6质量中等(5级)。结论在有效期内SQFM对同一总体样品以定量指纹图谱进行评价时,系统梯度洗脱条件的显著变化基本不影响鉴定结果。

Objective To evaluate the change in Shihu Yeguang pills（SHYGP） by systematically quantified fingerprint method（SQFM） under dual chromatographic elution system. Meanwhile, the fingerprint of the same solution extracted by 75% ethanol stored under 5 ℃ for 70 d was investigated. Methods The 3 indicators of SQFM, namely Sm, Pm and α, had inherent ties among each other. The 203, 286 and 326 nm HPLC fingerprints of 12 batches of SHYGPs under elution program 1（EP1） were established, respectively, in which the 3 quantitative fingerprint identification results were integrated by the average method. After 70 d, the same procedures were repeated in the same samples under significantly changed elution program 2（EP2） to determine the changes. For the two fingerprints, the changes in quality grade and macro quantitative similarity within them were discussed. Results While naringin（NG） was selected as the reference peak, 38（203 nm）, 34（286 nm） and 30（326 nm） common peaks were identified under EP1, also 32（203 nm）, 37（286 nm） and 26（326 nm） common peaks under EP2. After evaluation by SQFM through averaging the 3 wavelengths fingerprints, S2, S3, S4 and S12 were identified to be consistent for both EP conditions, and the remaining 6 batches of samples fluctuated by only 1 grade（except for S6 and S11）. The micro qualitative similarity of each sample under EP2 changed within 0.04, and the micro quantitative similarities errors were all within ±10% excluding S9 and S10. Finally 3-wavelength HPLC-FPs identification data under EP1 and EP2 were averaged to identify S1, S2, S3, S5 and S7 as the best quality（grade 1）, and S4, S8 and S9 as better quality（grade 2）, S10 and S12 as good quality（grade 3）, S11 as fine quality（grade 4）, and S6 as moderate qualiyt（grade 5）. Conclusion In the period of validity, when the same overall samples are evaluated by SQFM, significant change of gradient elution conditions do not affect the identification.