Page 74 - 《中国药房》2022年10期

银杏叶片中5种银杏双黄酮类成分同时测定方法的建立

刘丽娜 ，李海亮 ，李耀磊 ，金红宇 ，昝 珂 （1.中国食品药品检定研究院，北京 100050；2.北京中医药
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大学中药学院，北京 100029）
中图分类号 R917 文献标志码 A 文章编号 1001-0408（2022）10-1220-05
DOI 10.6039/j.issn.1001-0408.2022.10.11

摘 要 目的 建立同时测定银杏叶片中穗花杉双黄酮、白果黄素、银杏双黄酮、异银杏双黄酮、金松双黄酮含量的方法。方法 银
杏叶片经甲醇超声提取后，采用超高效液相色谱法测定。以Waters Acquity UPLC HSS T3为色谱柱，以乙腈-0.4％磷酸溶液为流
动相进行梯度洗脱，流速为0.4 mL/min，柱温为35 ℃，检测波长为340 nm，进样量为1 μL（对照品）、10 μL（供试品）。采用一测多
评法，以穗花杉双黄酮为参照物，计算白果黄素、银杏双黄酮、异银杏双黄酮、金松双黄酮的相对校正因子，以相对保留时间法定位
色谱峰，再计算上述成分的含量并与外标法测定结果（穗花双黄酮除外）进行比较。结果 穗花杉双黄酮、白果黄素、银杏双黄酮、
异银杏双黄酮、金松双黄酮检测进样量的线性范围分别为0.10～8.21、0.24～19.34、0.16～12.98、0.22～17.66、0.06～4.86 ng（r均大
于0.999），定量限分别为0.10、0.24、0.16、0.22、0.06 ng；精密度、重复性、稳定性（36 h）试验的RSD均小于3.00％，平均加样回收率
为99.77％～102.85％，RSD为1.90％～4.40％（n＝6）。白果黄素、银杏双黄酮、异银杏双黄酮、金松双黄酮的平均相对校正因子分
别为 0.91、0.93、0.96、0.95，平均相对保留时间分别为 1.08、1.18、1.19、1.30；一测多评法计算结果与外标法测得结果的相对偏差
在±3.00％以内。结论 所建方法准确、稳定，可用于银杏叶片中银杏双黄酮类成分的测定及质量控制。
关键词 超高效液相色谱法；一测多评法；银杏叶片；银杏双黄酮；含量测定

Establishment of determination method for five Ginkgo biflavones in Ginkgo biloba leaves tablets
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LIU Lina ，LI Hailiang ，LI Yaolei ，JIN Hongyu ，ZAN Ke（1. National Institutes for Food and Drug Control，
Beijing 100050，China；2. College of TCM，Beijing University of Chinese Medicine，Beijing 100029，China）
ABSTRACT OBJECTIVE To establish a method for the determination of amentoflavone，bilobetin，ginkgetin，isoginkgetin and
sciadopitysin in Ginkgo biloba leaves tablets. METHODS After extracted with methanol，ultra-performance liquid chromatography
（UPLC）was adopted to determine G. biloba leaves tablets. The determination was performed on Waters Acquity UPLC HSS T3
column with acetonitrile-0.4％ phosphoric acid as mobile phase（gradient elution）at the flow rate of 0.4 mL/min. The column
temperature was set at 35 ℃，and the detection wavelength was 340 nm. The sample size were 1 μL（substance control）and 10 μL
（test sample）. The relative correction factors（RCFs）of bilobetin，ginkgetin，isoginkgetin and sciadopitysin were calculated by
quantitative analysis of multicomponents by single marker（QAMS）using amentoflavone as control. The chromatographic peak was
located with the relative retention time method. Then the contents of the above components were calculated，and the results were
compared with those of external standard method （ESM）（except for amentoflavone）. RESULTS The linear ranges of
amentoflavone，bilobetin，ginkgetin，isoginkgetin and sciadopitysin were 0.10-8.21，0.24-19.34，0.16-12.98，0.22-17.66，0.06-4.86
ng，respectively（all r＞0.999）. The quantitation limits were 0.10，0.24，0.16，0.22，0.06 ng，respectively. RSDs of precision，
repeatability and stability tests（36 h）were all lower than 3.00％. The average recoveries were 99.77％-102.85％，and RSDs were
1.90％-4.40％（n＝6）. The average RCFs of bilobetin，ginkgetin，isoginkgetin and sciadopitysin were 0.91，0.93，0.96 and 0.95，
respectively. The average relative retention times were 1.08，1.18，1.19 and 1.30，respectively. The relative deviation between the
calculation result of QAMS and ESM was within ±3.00％. CONCLUSIONS The established method is accurate and stable，and
can be applied to the determination of Ginkgo biflavones in G. biloba leaves tablets and control the quality.
KEYWORDS ultra-performance liquid chromatography；quantitative analysis of multicomponents by single marker；Ginkgo
biloba leaves tablets；Ginkgo biflavones；content determination

银杏叶片是以银杏叶提取物为原料，加适宜辅料制 的药品。银杏叶提取物中不仅含有黄酮、萜类内酯、有
备而成的中药制剂，是临床重要的治疗心脑血管疾病 机酸、烷基酚、酚酸、甾体、聚戊烯等30余种成分，还含有
＊副研究员，硕士研究生。研究方向：中药、天然药物质量控制及 双黄酮类成分 [1－2] 。银杏双黄酮是由两分子黄酮母核通
有害残留物分析。电话：010-53852466。E-mail：kllln@163.com 过 C—C 键聚合而成的一类化合物，现已从银杏叶中分
#a 通信作者：主任药师。研究方向：中药、民族药质量控制与评
离鉴定出8种双黄酮类成分，其中苷元6种，分别为穗花
价。电话：010-53852076。E-mail：jhyu@nifdc.org.cn
杉双黄酮、金松双黄酮、银杏双黄酮、异银杏双黄酮、白
#b通信作者：副研究员，博士。研究方向：中药、民族药质量控制
与评价。电话：010-53852092。E-mail：6206310@qq.com 果黄素、5′-甲氧基白果黄素；双黄酮苷2种，分别为银杏

·1220 · China Pharmacy 2022 Vol. 33 No. 10 中国药房 2022年第33卷第10期