目的 建立HPLC波长切换联合梯度洗脱法（HPLC-DVD法）同时测定茵栀黄口服液中14种活性成分新绿原酸、绿原酸、隐绿原酸、栀子苷、对羟基苯乙酮、野黄芩苷、异绿原酸B、异绿原酸A、异绿原酸C、千层纸素A苷、汉黄芩苷、黄芩素、汉黄芩素、千层纸素A及一种辅料成分苯甲酸的含量。方法 采用HPLC-DVD法，Welch Ultimate XB-C₁₈（250 mm×4.6 mm，5μm）色谱柱；乙腈-0.1%磷酸水溶液为流动相，进行梯度洗脱，体积流量前30 min为0.8 mL/min，后续为1.0 mL/min；新绿原酸、绿原酸、隐绿原酸检测波长为325 nm，栀子苷检测波长为238 nm，对羟基苯乙酮检测波长为275 nm，苯甲酸检测波长为228 nm，野黄芩苷、异绿原酸B、异绿原酸A、异绿原酸C检测波长为325 nm，千层纸素A苷、汉黄芩苷检测波长为203 nm，黄芩素、汉黄芩素、千层纸素A检测波长为274 nm；进样量为10 μL。结果 15种指标成分新绿原酸在74.46~1 574.42 ng（r=0.999 8）、绿原酸在34.58~741.10 ng（r=0.999 6）、隐绿原酸在34.65~742.62 ng（r=0.999 7）、栀子苷在234.42~5 024.45 ng（r=0.999 9）、对羟基苯乙酮在74.46~1 574.42 ng（r=0.999 9）、苯甲酸在321.79~6 897.10 ng（r=0.999 8）、野黄芩苷在44.70~958.08 ng（r=0.999 7）、异绿原酸B在32.53~697.22 ng（r=0.999 5）、异绿原酸A在8.60~184.38 ng（r=0.999 6）、异绿原酸C在31.93~684.30 ng（r=0.999 7）、千层纸素A苷在254.82~5 461.66 ng（r=0.999 5）、汉黄芩苷在10.18~218.12 ng（r=0.999 9）、黄芩素在92.81~1 989.33 ng（r=0.999 6）、汉黄芩素在31.17~668.00 ng（r=0.999 5）、千层纸素A在11.74~251.73 ng（r=0.999 8）与峰面积具有较好的线性关系；精密度良好，RSD ≤ 0.75%；重复性良好，RSD ≤ 1.95%；供试品溶液在室温条件下8 h内稳定，RSD ≤ 1.77%；平均加样回收率和相应的RSD分别为100.69%（0.55%）、101.99%（1.78%）、99.20%（0.72%）、100.13%（0.48%）、100.96%（1.74%）、100.02%（0.46%）、101.14%（0.27%）、99.87%（0.59%）、100.21%（1.56%）、102.18%（0.33%）、99.00%（1.02%）、98.82%（0.65%）、98.31%（0.58%）、96.01%（0.44%）、100.56（0.71%）。10批次供试品中新绿原酸、绿原酸、隐绿原酸、栀子苷、对羟基苯乙酮、苯甲酸、野黄芩苷、异绿原酸B、异绿原酸A、异绿原酸C、千层纸素A苷、汉黄芩苷、黄芩素、汉黄芩素、千层纸素A的含量分别为0.246~0.322、0.213~0.290、0.203~0.267、1.786~2.047、0.035~0.046、2.393~2.541、0.263~0.392、0.139~0.216、0.032~0.067、0.208~0.250、2.120~2.648、0.063~0.102、0.081~1.429、0.164~0.246、0.079~0.110 mg/mL。结论 建立的HPLC-DVD法同时测定茵栀黄口服液中的15种成分，方法简便、快速、准确，可为茵栀黄口服液质量控制提供科学依据。

Objective To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of 14 active components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, geniposide, 4-hydroxyacetophenone, scutellarin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, oroxylin A 7-O-glucuronide, wogonoside, baicalein, wogonin and oroxylin A and one auxiliary material (benzoic acid) in Yinzhihuang Oral Solution (YOS). Methods The chromatographic separation was achieved on Welch Ultimate XB-C₁₈ (250 mm×4.6 mm, 5 μm) column with acetonitrile-0.1% phosphoric acid solution as mobile phases for gradient elution, at the flow rate of 0.8 mL/min in the first 30 min and 1.0 mL/min in the follow-up; The detection wavelength was set at 325 nm for neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid, 238 nm for geniposide, 275 nm for 4-hydroxyacetophenone, 228 nm for benzoic acid, 325 nm for scutellarin, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C, 203 nm for oroxylin A 7-O-glucuronide and wogonoside, and 274 nm for baicalein, wogonin and oroxylin A. The volume of sample injection was 10 μL. Results The fifteen active components were well separated and showed good linearity, such as neochlorogenic acid 74.46-1 574.42 ng (r=0.999 8), chlorogenic acid 34.58-741.1 ng (r=0.999 6), cryptochlorogenic acid 34.65-742.62 ng (r=0.999 7), geniposide 234.42-5 024.45 ng (r=0.999 9), 4-hydroxyacetophenone 74.46-1 574.42 ng (r=0.999 9), benzoic acid 321.79-6 897.1 ng (r=0.999 8), scutellarin 44.7-958.08 ng (r=0.999 7), isochlorogenic acid B 32.53-697.22 ng (r=0.999 5), isochlorogenic acid A 8.6-184.38 ng (r=0.999 6), isochlorogenic acid C 31.93-684.3 ng (r=0.999 7), Oroxylin A 7-O-glucuronide 254.82-5 461.66 ng (r=0.999 5), wogonoside 10.18-218.12 ng (r=0.999 9), baicalein 92.81-1 989.33 ng (r=0.999 6), wogonin 31.17-668 ng (r=0.999 5), and oroxylin A 11.74-251.73 ng (r=0.999 8). The precision was good, and RSD was not more than 0.75%. The repeatability was good, and the RSD was not more than 1.95%. The stability was good in 8 h, and RSD was not more than 1.77%. The average recoveries and corresponding RSD values were 100.69% (0.55%), 101.99% (1.78%), 99.20% (0.72%), 100.13% (0.48%), 100.96% (1.74%), 100.02% (0.46%), 101.14% (0.27%), 99.87% (0.59%), 100.21% (1.56%), 102.18% (0.33%), 99.00% (1.02%), 98.82% (0.65%), 98.31% (0.58%), 96.01% (0.44%), and 100.56 (0.71%), respectively. The content of 10 batches of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, geniposide, 4-hydroxyacetophenone, benzoic acid, scutellarin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, oroxylin A 7-O-glucuronide, wogonoside, baicalein, wogonin and oroxylin A were 0.246-0.322, 0.213-0.290, 0.203-0.267, 1.786-2.047, 0.035-0.046, 2.393-2.541, 0.263-0.392, 0.139-0.216, 0.032-0.067, 0.208-0.250, 2.120-2.648, 0.063-0.102, 0.081-1.429, 0.164-0.246, and 0.079-0.110 mg/mL. Conclusion HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of 15 components in YOS. The method is simple, quick, accurate, and it can be used for content determination and quality control of YOS.

2019年国家药品评价性抽验中央补助地方经费项目（157）