目的 通过指纹图谱分析黄花菜Hemerocallis citrina叶的特征成分，进一步对其中指标成分绿原酸、芦丁、槲皮素-3-O-葡萄糖苷、槲皮苷、槲皮素进行含量测定，以期为黄花菜叶利用及其质量评价提供依据。方法 采用乙醇提取法制备黄花菜叶提取物，选用色谱柱DIKMA spursil C₁₈柱（250 mm×4.6 mm，5 mm），流动相为乙腈-0.5%乙酸水，进样量10 mL，检测波长为190～400 nm，柱温30 ℃，体积流量1 mL/min，采集时间80 min；基于最大吸收波长原则，分别在时间段0～5.5 min和23～48 min选择258 nm，5.5～23 min选择300 nm，48～80 min选择370 nm作为检测波长，构建了3波长切换指纹图谱。采用一测多评法（quantitative analysis of multi-components by single-marker，QAMS）测定黄花菜叶乙醇提取物中绿原酸、芦丁、槲皮素-3-O-葡萄糖苷、槲皮苷、槲皮素5种酚类成分的含量。结果 10批黄花菜叶的指纹图谱相似度在0.90以上，所构建的3波长切换指纹图谱共包括42个共有峰（占总峰面积90%以上），其中峰11、25、26、32、37被鉴定为绿原酸、芦丁、槲皮素-3-O-葡萄糖苷、槲皮苷、槲皮素。以槲皮素（S）为内参物，QAMS法获得绿原酸、芦丁、槲皮素-3-O-葡萄糖苷、槲皮苷的相对校正因子分别为1.402、1.896、1.268、1.221。10批黄花菜叶中上述5种成分的平均含量为0.079、0.086、0.374、0.325、0.037 mg/g。外标法与QAMS法的含量测定结果无显著性差异。结论 3波长切换指纹图谱较任何单一波长指纹图谱所包含的色谱峰数均有显著提升，所构建的多波长切换指纹图谱可用于黄花菜叶化学成分表征，进一步构建的QAMS法可以一次性实现绿原酸、芦丁、槲皮素-3-O-葡萄糖苷、槲皮苷、槲皮素5种成分的含量测定，为全面评价黄花菜叶质量提供了方法学依据和参考。

Objective Huanghuacai (Hemerocallis citrina) is a traditional vegetable with anti-inflammatory and anti-depressant effects. Its leaves are widely used in Taiwan and other regions to promote sleep, however, there are few reports on its chemical composition. This article aims to analyze the characteristic components of H. citrina leaves by fingerprint spectrum and to further determine the content of the index components, including chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin, providing a basis for better utilization and quality evaluation of H. citrina leaves. Methods Firstly, the ethanol extraction method was used to prepare H. citrina leaves extract. Chromatographic separation was performed by DIKMA spursil C₁₈ column (250 mm × 4.6 mm, 5 mm) maintained at 30 ℃ and the mobile phase was acetonitrile-0.5% acetic acid water. The injection volume was 10 mL, the detection wavelength was 190—400 nm and the flow rate was 1 mL/min. Based on the principle of maximum absorption wavelength, the wavelengths of 258 nm was selected in two periods of 0—5.5 and 23—48 min, the wavelengths of 300 and 370 nm were selected in two periods of 5.5—23 and 48—80 min respectively, and a three-wavelength switching fingerprint was constructed. Secondly,a quantitative analysis of multi-components by single marker (QAMS) was used to determine the contents of chlorogenic acid, rutin, quercetin-3-O-glucoside, quercetin, and quercetin in ethanol extract of H. citrina leaves. Results The similarity of fingerprint spectrums of 10 batches of H. citrina leaves was above 0.90, and the three-wavelength switching fingerprint spectrums included 42 common peaks (accounting for more than 90% of the total peak area), of which 11, 25, and 26, 32 and 37 were identified as chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin. With quercetin (S) as the internal reference, the relative correction factors of chlorogenic acid, rutin, quercetin-3-O-glucoside, and quercitrin were obtained by the QAMS as 1.402, 1.896, 1.268, 1.221. The average contents of the above five components in 10 batches of H. citrina leaves are 0.079, 0.086, 0.374, 0.325, 0.037 mg/g. There was no significant difference between the external standard method and QAMS. Conclusion The three-wavelength switching fingerprint spectrum constructed in this study has significantly increased the number of chromatographic peaks contained in any single wavelength fingerprint spectrum and can be used to characterize the chemical composition of H. citrina leaves. The QAMS further constructed can be used to determine the content of chlorogenic acid, rutin, quercetin-3-O-glucoside, quercitrin, and quercetin at one time. This study provides a methodological basis and reference for the comprehensive evaluation of H. citrina leaves quality.

R282.6

国家自然科学基金资助项目（81972018）；国家重点研发计划项目（2018YFC1706300，2018YFC17063003）；中央高校基本业务费重点项目和自由探索项目（lzujbky-2020-sp13，lzujbky-2020-sp25）；甘肃省重点研发计划项目（20YF2FA019）；甘肃省技术创新引导计划-民生专项（20CX4FK014）；兰州市人才创新创业项目（2017-RC115，2020-RC-41）