Page 77 - 2019年11月第30卷第21期

以离子液体为流动相添加剂的 HPLC 法分离并测定钩藤中钩藤

碱和异钩藤碱的含量 Δ

#
张媛媛 ，曾慧婷，陈 超，何小群，胡燕珍，陈 乐，虞金宝（江西省中医药研究院中药所，南昌 330046）
＊
中图分类号 R284.1 文献标志码 A 文章编号 1001-0408（2019）21-2952-06
DOI 10.6039/j.issn.1001-0408.2019.21.15

摘 要 目的：建立测定钩藤中钩藤碱和异钩藤碱含量的方法。方法：以离子液体1-丁基-3-甲基咪唑氯化盐（C4mimCl）为流动相
添加剂，与无添加剂的流动相以及加入传统添加剂三乙胺（对色谱柱有损伤）后对高效液相色谱（HPLC）法分离钩藤中钩藤碱和异
钩藤碱的分离度进行比较，筛选C4mimCl的最佳浓度，用新建立的方法测定江西4个产地钩藤中钩藤碱和异钩藤碱的含量。色谱
柱为Dikmatech Diamonsil Plus C18，流动相为乙腈-缓冲液（0.1％磷酸+3.0 mmol/L C4mimCl），梯度洗脱，紫外检测波长为245 nm，
流速为1 mL/min，进样量为10 μL。结果：当流动相中无添加剂时、加入3.0 mmol/L三乙胺或3.0 mmol/L C4mimCl作添加剂时，钩
藤碱与前峰分离度分别为1.02、1.23、1.72，与后峰分离度分别为1.06、6.00、4.25，对称因子分别为0.81、0.86、1.13；异钩藤碱与前峰
分离度分别为0.96、3.89、4.05，与后峰分离度分别为1.02、2.34、2.36，对称因子分别为0.88、0.81、0.96。钩藤碱、异钩藤碱检测质量
浓度线性范围分别为4.93～157.76（r＝0.999 9）、4.98～159.50 μg/mL（r＝1.000 0），定量限分别为0.486 4、0.793 6 μg/mL，精密度、
重复性、稳定性和耐用性试验中的RSD均小于5％（n＝6），回收率分别为102.9％～107.8％（RSD＝1.7％，n＝6）、95.4％～106.3％
（RSD＝3.9％，n＝6）。4个产地钩藤中钩藤碱和异钩藤碱的含量范围分别为0.758～1.343、1.511～1.823 mg/g。结论：C4mimCl加
入到流动相中能提高分离度，且以此建立的HPLC法快速、准确、重复性好，可用于钩藤中钩藤碱和异钩藤碱的含量测定。
关键词 离子液体；1-丁基-3-甲基咪唑氯化盐；流动相；添加剂；高效液相色谱法；钩藤碱；异钩藤碱；含量测定

Separation and Determination of Rhynchophylline and Isorhynchophylline in Uncaria rhynchophylla by
HPLC with Ionic Liquid as Mobile Phase Additives
ZHANG Yuanyuan，ZENG Huiting，CHEN Chao，HE Xiaoqun，HU Yanzhen，CHEN Le，YU Jinbao（Institute of
TCM Research，Jiangxi Academy of TCM Research，Nanchang 330046，China）

ABSTRACT OBJECTIVE：To establish a method to determine the contents of rhynchophylline and isorhynchophylline in
Uncaria rhynchophylla. METHODS：The separation degree of ionic liquid 1-butyl-3-methylimidazolium chloride （C4mimCl） as
mobile phase additive was compared with that of mobile phase without additives and with traditional additive triethylamine（which
damaged the chromatographic column）. The optimum concentration of C4mimCl was screened and the contents of rhynchophylline
and isorhynchophylline in U. rhynchophylla from 4 habitats in Jiangxi province were determined by the newly established method.
The determination was performed on Dikmatech Diamonsil Plus C18 column，the mobil phase was acetonitrile-buffer （0.1％
phosphoric acid+3.0 mmol/L C4mimCl），gradient elution. UV detection wavelength was set at 245 nm and the flow rate was 1
mL/min. Sample size was 10 μL. RESULTS：When mobile phase had no additives or 3.0 mmol/L triethylamine and 3.0 mmol/L
C4mimCl were added as additives，the separation of rhynchophylline from the front peak was 1.02，1.23 and 1.72，and the
separation from the back peak was 1.06，6.00 and 4.25，respectively. The symmetry factors were 0.81，0.86 and 1.13，respectively.
The separation of isorhynchophylline from the front peak was 0.96，3.89 and 4.05，and the separation from the back peak was
1.02， 2.34 and 2.36， respectively. The symmetry factors were 0.88， 0.81 and 0.96， respectively. The linear range of
rhynchophylline and isorhynchophylline were 4.93-157.76 （r＝0.999 9） and 4.98-159.50 μ g/mL （r＝1.000），respectively. The
quantitative limits were 0.486 4，0.793 6 μg/mL，respectively. RSDs of precision，repeatability，stability and durability tests were
all less than 5％（n＝6）. The recovery rates were 102.9％-107.8％（RSD＝1.7％，n＝6）and 95.4％-106.3％（RSD＝3.9％，n＝6），
respectively. The content of rhynchophylline and isorhynchophylline in U. rhynchophylla from 4 habitats were 0.758-1.343
and 1.511-1.823 mg/g， respectively. CONCLUSIONS：
Δ 基金项目：江西省卫生和计划生育委员会中医药科研课题 Addition of C4mimCl into mobile phase can enhance its
（No.2017A320）；2017 年中医药局公共卫生服务补助资金“全国中药
separation. Established HPLC method is rapid，accurate and
资源普查项目”（No.财社〔2017〕66号）；江西省自然科学基金资助项目
reproducible，which can be used for content determination of
（No.20161BAB205222）
rhynchophylline and isorhynchophylline in U. rhynchophylla.
＊研究实习员。研究方向：中药分析。 电话：0791-88501404 。
KEYWORDS Ionic liquid； 1-butyl-3-methylimidazolium
E-mail：1261115822@qq.com
# 通信作者：研究员。研究方向：中药分析和中药资源。 电话： chloride；Mobile phase；Additive；HPLC；Rhynchophylline；
0791-88501404 。E-mail：543349695@qq.com Isorhynchophylline；Content determination

·2952 · China Pharmacy 2019 Vol. 30 No. 21 中国药房 2019年第30卷第21期