知母HPLC指纹图谱的4种相似度评估

HPLC fingerprints of Rhizoma anemarrhenae based on 4 kinds of similarities

目的:用双定性双定量相似度法评价知母HPLC指纹图谱。方法:采用反相高效液相色谱法,使用Century SIL C18BDS柱(20 cm×4.6 mm,5μm),以1%醋酸水-1%醋酸乙腈为流动相,梯度洗脱,检测波长265 nm,柱温(30.0±0.15)℃,进样量5μL,测定10批知母HPLC指纹图谱。以双定性相似度SF和SF′、双定量相似度C%和P%评价知母HPLC指纹图谱。分别考察在大峰缺失和小峰缺失两种情况下,4个相似度指标变化特征。结果:以芒果苷为参照物峰,确定21个共有峰,建立了知母HPLC指纹图谱。SF能反映化学成分分布比例,受大峰影响严重,无法反映小峰丢失;SF′对所有指纹峰等权,反映小峰缺失灵敏。C%能反映样品总体含量,但受大峰影响严重,无法反映小峰缺失;P%对所有峰积分值等权,能较好反映小峰变动。结论:双定性相似度SF与SF′、双定量相似度C%与P%能准确反映大峰和小峰的含量变动与缺失。所建立的HPLC指纹图谱可用于知母质量控制。

AIM： The method of the double qualitative similarities and the double quantitative similarities was carried out for the evaluation of the chromatographic fingerprints and applied to HPLC fingerprints of Rhizoma anemarrhenae. METHODS ： The chromatographic fingerprints were obtained by injecting 5 μL of the sample solution each time on a CenturySIL C18 BDS column （20 cm ×4.6 mm, 5 μm） with the gradient elution solvent system composed of 1% acetic acid-water and 1% acetic acid-acetonitrile. The column temperature was maintained at （30 ± 0.15） ℃ and the detection wavelength was set at 265 nm. The chromatographic fingerprints were evaluated by the normalized similarity SF and normalizing similarity S′F and the projecting similarity C% and quautitative similarity P%, changes in characteristics were investigated in the cases of big peaks-free and small peaks-free respectively. RESULTS： 21 co-possessing peaks were selected as the fingerprint peaks of Rhizoma anernarrhenae by choosing rutin peak as the reference peak. SF could clearly reflect the chemical constituent distribution, and C% could reflect the total contents of the sample, but both of them were seriously influenced by the big peaks and could not reflect the missing of small peaks. S′F and P% were equivalent weight to all fingerprint peaks, and they could reflect sensitively the small peaks drop-out. CONCLUSION： The double qualitative similarities and the double quantitative similarities are composed of SF, S′F, C% and P%, so the changes or lacks of both big peaks and small peaks can be punctually simultaneously monitored to be a novel method for evaluating fingerprints. The HPLC fingerprints can be useful in the quality control of Rhizoma anemarrhenae.