基于Box-Behnken设计-响应面法与质量综合评价优化葛根芩连汤煎煮工艺  

Decocting process of Gegen Qinlian Decoction was optimized based on Box-Behnken design-response surface methodology and comprehensive quality evaluation

作　　者：徐蓓蕾[1,2,3] 韩晓宇 刘晶晶 张文君[1,2,3] 孙志伟[1,2,3] 胡扬[1,2,3] 綦峥 杨雪晶[1,2,3] 孙文斌[1] 杨娜娜 李文兰[1,2,3] XU Bei-lei;HAN Xiao-yu;LIU Jing-jing;ZHANG Wen-jun;SUN Zhi-wei;HU Yang;QI Zheng;YANG Xue-jing;SUN Wen-bin;YANG Na-na;LI Wen-lan(School of Pharmacy,Harbin University of Commerce,Harbin 150076,China;Engineering Research Center of Natural Anti-cancer Drugs,Ministry of Education,Harbin 150076,China;Heilongjiang Key Laboratory of Preventive and Therapeutic Drug Research of Senile Diseases,Harbin 150076,China;National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]哈尔滨商业大学药学院,黑龙江哈尔滨150076 [2]抗肿瘤天然药物教育部工程研究中心,黑龙江哈尔滨150076 [3]黑龙江省预防与治疗老年性疾病药物研究重点实验室,黑龙江哈尔滨150076 [4]中国食品药品检定研究院,北京100050

出　　处：《中草药》2022年第22期7070-7081,共12页Chinese Traditional and Herbal Drugs

基　　金：黑龙江省自然科学基金联合指导计划(LH2020H069);哈尔滨商业大学青年学术骨干支持计划(2020CX12)。

摘　　要：目的优化和科学评价葛根芩连汤(Gegen Qinlian Decoction,GQD)的煎煮工艺,为其经典方剂中药复方制剂的研究与开发奠定基础以及提供参考。方法以指纹图谱概貌结合多指标定量为综合评价,采用Box-Behnken设计-响应面法对加水量、浸泡时间、煎煮时间、煎煮次数进行优化。指纹图谱研究采用Waters Acquity UPLC BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),流动相为乙腈-0.1%甲酸水溶液(梯度洗脱),体积流量为0.3 mL/min,柱温为35℃,进样量为1μL;采用电喷雾离子源,以正、负离子模式检测,在质荷比(m/z)50~1200进行扫描,建立29批GQD指纹图谱并对共有峰进行指认。对图谱信息进行主成分分析,建立UPLC多波长检测方法测定木兰花碱、葛根素、药根碱、巴马汀、黄芩苷、大豆苷元、黄芩素、甘草苷8个成分含量,计算各样品综合得分,优化煎煮工艺。结果建立了29个试验号GQD样品的UPLC-Q-TOF-MS正、负离子模式下指纹图谱。正、负离子模式下分别有共有峰23、19个,并对其进行指认,指认出葛根素、大豆苷元、黄芩素、黄芩苷、巴马汀、药根碱、木兰花碱、甘草苷、甘草次酸、大豆苷、异甘草苷、汉黄芩苷、汉黄芩素、甘草酸、glyasperins D、甘草醇、绿原酸、韧黄芩素Ⅱ。对共有峰中8个指标成分进行含量测定,以计算综合得分,综合得分结果显示,26号工艺为最佳工艺。最终确定GQD最佳煎煮工艺为加10倍量水,浸泡0.5 h,煎煮3次,每次0.5 h。结论所得出的GQD最佳煎煮工艺主要成分溶出率高,稳定可行,可为经典方剂GQD复方制剂的进一步研究和开发提供一定的依据。Objective To optimize and scientifically evaluate the decoction process of Gegen Qinlian Decoction(GQD,葛根芩连汤)based on Box-Behnken design-response surface methodology in order to lay the foundation and provide reference for the research and development of traditional Chinese medicine compound preparations.Methods Taking fingerprint profile combined with multi-index quantification as the comprehensive evaluation,the amount of water,soaking time,decocting time and decocting times were optimized by Box-Behnken design response surface method.The fingerprint analysis was performed on a Waters Acquity UPLC BEH C_(18) column(100 mm×2.1 mm,1.7μm)with mobile phase consisted of acetonitrile-0.1%formic acid aqueous solution(gradient elution)at the volume flow of 0.3 mL/min.The column temperature was 35℃,and the injection volume was 1μL.The fingerprints of 29 batches of GQD were established by scanning in the range of mass charge ratio(m/z)of 50-1200 with ESI source and detected in positive and negative ion modes,and common peaks were identified.The principal components were analyzed,and contents of 8 main components,including magnoflorine,puerarin,jatrorrhizine,palmatine,baicalin,daidzein,baicalein and liquiritin were determined by UPLC multi-wavelength detection method.And the comprehensive score of each sample was calculated to optimize the decoction process..Results The UPLC-Q-TOF-MS fingerprints of 29 test number of GQD samples were established in positive and negative ion mode.There were 23 and 19 common peaks in positive and negative ion mode respectively,and 18 components of them were identified,including puerarin,daidzein,baicalein,baicalin,palmatine,jatrorrhizine,magnoflorine,liquiritin,glycyrrhetinic acid,daidzin,isoliquiritin,wogonoside,wogonin,glycyrrhizic acid,glyasperins D,glycyrol,chlorogenic acid and tenaxin II.The content of eight index components of the common peaks was determined to calculate the comprehensive score,and the comprehensive score results showed that test number 26 was the optim

关 键 词：葛根芩连汤 指纹图谱 Box-Behnken设计-响应面法 煎煮工艺 UPLC-Q-TOF-MS 葛根素 大豆苷元 黄芩素 黄芩苷 巴马汀 药根碱 木兰花碱 甘草苷 甘草次酸 大豆苷 异甘草苷 汉黄芩苷 汉黄芩素 甘草酸 glyasperins D 甘草醇 绿原酸 韧黄芩素Ⅱ 

分 类 号：R283.6[医药卫生—中药学]