Page 71 - 2019年8月第30卷第16期

博落回提取物中血根碱的离子交换树脂分离纯化工艺研究 Δ

2，3
2 #
1＊
邓霖芳 ，袁 帅 ，刘江云 （1.浙江中医药大学附属第三医院药剂科，杭州 310005；2.苏州大学药学院，江
苏 苏州 215123；3.无锡市中医医院药剂科，江苏 无锡 214071）
中图分类号 R284.2 文献标志码 A 文章编号 1001-0408（2019）16-2226-06
DOI 10.6039/j.issn.1001-0408.2019.16.13

摘 要 目的：建立博落回提取物中血根碱的离子交换树脂分离纯化工艺。方法：采用高效液相色谱法（HPLC）测定博落回提取
物中血根碱含量，色谱柱为Cosmosil C18-R-Ⅱ（250 mm×4.6 mm，5 μm），流动相为乙腈-0.2％醋酸溶液（25∶75，V/V），流速为1 mL/min，
检测波长为270 nm，柱温为30 ℃，进样量为20 μL。通过静态吸附与解吸附试验，对比8种离子交换树脂对血根碱的吸附和解吸
附性能；采用优选离子交换树脂，考察最佳上样液浓度、上样pH值以及上样体积；采用APPS 10D液相制备系统，考察动态洗脱条
件，获得博落回精提物溶液；采用反相C18色谱柱对博落回精提物溶液脱盐、洗脱并干燥，获得精制纯化物。采用HPLC法测定所
得精制纯化物的纯度，并采用HPLC法、紫外分光光度法、质谱法、核磁共振法分别对其进行结构确证。结果：筛选获得CM-FF型
树脂用于博落回提取物中血根碱的分离纯化，以含20％甲醇、0.25 mol/L氯化钠的20 mmol/L醋酸铵溶液100 mL进行洗脱。优化
的动态吸附条件为上样浓度6.0 mg/mL、上样pH值5.5、上样体积25 mL；洗脱精提物经脱盐和精制后，最终获得纯度为97％的精
制纯化产物（纯化收率为71％），并经结构确证其为血根碱。结论：优选的离子交换树脂分离纯化工艺绿色环保、安全高效、易于
操作，可用于博落回提取物中血根碱单体的分离纯化，适合工业化生产。
关键词 博落回；血根碱；离子交换树脂；分离；纯化

Study on Separation and Purification Technology of Sanguinarine from the Extract of Macleaya cordata by
Ion Exchange Resin
DENG Linfang ，YUAN Shuai ，LIU Jiangyun（1. Dept. of Pharmacy，the Third Affiliated Hospital of Zhejiang
1
2，3
2
Chinese Medical University，Hangzhou 310005，China；2. School of Pharmacy，Soochow University，Jiangsu
Suzhou 215123，China；3. Dept. of Pharmacy，Wuxi Hospital of TCM，Jiangsu Wuxi 214071，China）
ABSTRACT OBJECTIVE：To establish the separation and purification technology of sanguinarine from the extract of Macleaya
cordata with ion exchang resin. METHODS：The content of sanguinarine from the extract of M. cordata was determined by HPLC，
with Cosmosil C18-R-Ⅱ column（250 mm×4.6 mm，5 μm），mobile phase of acetonitrile-0.2％ acetic acid solution（25 ∶ 75，V/V），
the flow rate of 1 mL/min，detection wavelength of 270 nm，column temperature of 30 ℃，and sample size of 20 μL. Static
adsorption and desorption tests were carried out to compare the adsorption and desorption properties of 8 ion exchange resins for
sanguinarine. The optimum concentration of sample solution，pH value and volume of sample were investigated by optimum ion
exchange resin. APPS 10D liquid phase preparation system was used to investigate the dynamic elution conditions and obtain M.
cordata refined extract solution. The refined purified product of M. cordata was obtained by desalination， elution on a
reversed-phase（RP）C 18 column and drying. The purity of the purified product was analyzed by HPLC. The structure of the purified
product was confirmed by HPLC，UV spectrophotometry，MS and NMR. RESULTS：CM-FF resin was screened for the separation
and purification of sanguinarine from M. cordata extract. It was eluted with 20 mmol/L ammonium acetate solution 100 mL
containing 20％ methanol and 0.25 mol/L sodium chloride. The optimal dynamic absorption condition included that the
concentration of sample was 6.0 mg/mL at pH 5.0，and the loading amount was 25 mL；after desalination and refinement，for the
eluted refined extract，the purified product with 97％ purity （purified yield of 71％） was obtained，and its structure was
confirmed to be sanguinarine. CONCLUSIONS：The optimal separation and purification technology by ion exchange resin is green，
safe，efficient and easy to operate，which can be used for the
Δ 基金项目：江苏省重点研发专项资金项目（No.BE2018322）；江
苏省第十四批“六大人才高峰”高层次人才资助项目（No.SWYY-167）； separation and purification of sanguinarine from M. cordata
浙江中医药大学附属第三医院院级医药卫生科技计划项目（No. extract and is suitable for industrial production.
ZS17YA01） KEYWORDS Macleaya cordata；Sanguinarine；Ion exchange
＊药师，硕士。研究方向：天然药物化学。电话：0571-88393553。 resin；Separation；Purification
E-mail：850588794@qq.com
# 通信作者：副教授，硕士生导师，博士。研究方向：天然药物化
学。电话：0512-65884301。E-mail：liujiangyun@suda.edu.cn

·2226 · China Pharmacy 2019 Vol. 30 No. 16 中国药房 2019年第30卷第16期