基于UPLC的蒲黄指纹图谱和多成分含量测定方法研究

Study on UPLC Fingerprint and Multi-component Content Determination of Typhae Pollen

目的:采用UPLC法建立蒲黄指纹图谱和8个成分的含量测定方法。方法:采用Phenomenex Kinetex^(■)EVO C_(18)(100 mm×2.1 mm,1.7μm)色谱柱;流动相为乙腈-0.1%甲酸溶液,梯度洗脱;流速为0.2 mL/min;检测波长为254、280 nm;柱温为30℃;进样量为2μL。建立31批蒲黄UPLC指纹图谱,并对其中8个成分进行定量分析。结果:从蒲黄UPLC指纹图谱中共标定了15个共有峰,并指认了10个色谱峰。31批蒲黄指纹图谱的相似度均大于0.900。4-羟基苯甲酸、槲皮素-3-O-(2^(G)-α-L-鼠李糖基)-芸香糖苷、槲皮素-3-O-新橙皮糖苷、山柰酚-3-O-(2^(G)-α-L-鼠李糖基)-芸香糖苷、香蒲新苷、山柰酚-3-O-新橙皮糖苷、异鼠李素-3-O-新橙皮糖苷、柚皮素的含量分别为0.008%~0.018%、0.055%~0.135%、0.024%~0.078%、0.023%~0.067%、0.284%~0.533%、0.052%~0.116%、0.303%~0.498%、0.057%~0.156%。结论:采用UPLC建立的蒲黄指纹图谱和多成分含量测定方法简便、准确、重现性好,可为建立蒲黄的质量控制体系提供参考。

Objective:To establish the method of fingerprint and content determination of 8 components from Typhae Pollen by UPLC.Methods:Using Phenomenex Kinetex^(■)EVO C_(18)(100 mm×2.1 mm,1.7μm)column,the mobile phase was acetonitrile-0.1%formic acid solution with gradient elution,flow rate was 0.2 mL/min,detection wavelength was 254,280 nm,column temperature was 30℃,injection volume was 2μL.The UPLC fingerprint of 31 batches of Typhae Pollen was established and 8 components were quantitatively analyzed.Results:A total of 15 common peaks were calibrated and 10 chromatographic peaks were identified from UPLC fingerprint of Typhae Pollen.The similarity of fingerprints of 31 batches of Typhae Pollen was greater than 0.900.The contents of 4-hydroxybenzoic acid,quercetin-3-O-(2^(G)-α-L-rhamnol)-rutinoside,quercetin-3-O-neohesperidin,kaempferol-3-O-(2^(G)-α-L-rhamnol)-rutinoside,typhaneoside,kaempferol-3-O-neohesperidin,isorhamnetin-3-O-neohesperidin,naringin were 0.008%~0.018%,0.055%~0.135%,0.024%~0.078%,0.023%~0.067%,0.284%~0.533%,0.052%~0.116%,0.303%~0.498%,0.057%~0.156%,respectively.Conclusion:The fingerprint and multi-component determination method of Typhae Pollen established by UPLC is simple,accurate and reproducible,which can provide reference for the establishment of quality control system of Typhae Pollen.