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ABSTRACT OBJECTIVE:To establish HPLC fingerprint of different products of suet oil-baked Epimedium brevicornum,and to
screen the optimal baking technology. METHODS:HPLC method was adopted. Using icariin as reference,HPLC fingerprints of 22
batches of samples were drawn. The similarity was evaluated by using Similarity Evaluation System of TCM Chromatographic
Fingerprint(2012 edition),and common peaks were confirmed. HCA,PCA and OPLS-DA analysis were performed by SIMCA
14.1 statistical software. Taking variable importance in the project >1 as criteria,biomarkers affecting the quality difference of suet
oil-baked E. brevicornum were screened;using mass marker as index,the baking technology was screened by baking technology.
RESULTS:There were 18 common peaks in 22 batches of samples. The similarities were between 0.831 and 0.991. Totally 7
common peaks were identified,i.e. epimedin A,epimedin B,epimedin C,icariin,sagittatoside A,sagittatoside B,baohuoside
Ⅰ. The 22 batches of samples were clustered into two categories,S19-S22 were clustered into category Ⅰ and S1-S18 were
clustered into category Ⅱ. The category Ⅱ was sub-clustered into category Ⅱa(S15-S18),category Ⅱb(S10-S14),category Ⅱc
(S1-S9);the result of PCA analysis was consistent with above results. OPLS-DA showed that the biomarkers affecting the quality
difference were icariin,sagittatoside B and baohuoside Ⅰ. The results of kinetic studies showed that the content of icariin when
baked at 180 ℃ for 25 min or 190 ℃ for 20 min,that of baohuoside Ⅰ when baked at 180 ℃ for 30 min or 190 ℃ for 15 min
and that of epimedin B when baked at 210 ℃ for 18 min were the highest;according to above results,the optimal baking
technology was baking at 180 ℃ for 25-30 min or 190 ℃ for 15-20 min. CONCLUSIONS:Established fingerprint is stable,
reliable and reproducible. The multivariate statistical analysis can be used for the changes of chemical components in E.
brevicornum under different baking condition and preliminary selection of baking technology.
KEYWORDS Epimedium brevicornum;Suet oil-based;HPLC;Fingerprint;Multivariate statistical analysis;Optimization of
baking technology
淫羊藿为小檗科植物淫羊藿(Epimedium brevicor- 应用于何首乌、姜炭、当归、栀子等饮片的炮制工艺筛
num Maxim.)、箭叶淫羊藿[Epimedium sagittatum(Sieb. 选 [13-16] 。基于此,本研究建立了不同淫羊藿羊脂油烘制
et Zucc.)Maxim.]、柔 毛 淫 羊 藿(Epimedium pubescens 品的高效液相色谱(HPLC)指纹图谱,同时借助聚类分
Maxim.)或朝鲜淫羊藿(Epimedium koreanum Nakai)的 析(HCA)、主成分分析(PCA)、正交偏最小二乘法-判别
干燥叶,具有补肾阳、强筋骨、祛风湿的功效,临床主要 分析(OPLS-DA)、烘制动力学等方法,筛选其最优烘制
用于治疗肾阳虚衰所致的阳痿遗精、筋骨痿软、风湿痹 工艺,旨在为淫羊藿羊脂油烘制工艺的规范化研究提供
[1]
痛、麻木拘挛,是临床常用中药 。淫羊藿含有黄酮类、 参考。
多糖类、木脂素类、生物碱类等化学成分,其主要成分淫 1 材料
羊藿总黄酮具有抗骨质疏松、促进生殖内分泌、抗肿瘤 1.1 仪器
[2]
等作用 。淫羊藿羊脂油烘制品为淫羊藿的炮制品(又 1260 型 HPLC 仪,包括自动进样器、四元泵洗脱系
称为炙淫羊藿),其传统炮制方法为每100 kg淫羊藿,加 统、柱温箱、二极管阵列检测器、OpenLab C.01.04[35]色
[1]
羊脂油(炼油)20 kg,经文火炒至均匀有光泽 。虽然, 谱工作站(美国Agilent公司);AB135-S型十万分之一电
目前已有相关研究对炙淫羊藿传统炮制工艺进行规范, 子天平、EP214C 型万分之一电子天平(瑞士 Mettler-To-
以淫羊藿苷、宝藿苷Ⅰ等黄酮类成分为评价指标,明确 ledo公司);KQ-250型超声波清洗器(昆山市超声仪器有
炒制温度、炒制时间等工艺参数 [3-5] 。但由于淫羊藿质地 限公司);101-E 型电热鼓风干燥箱(北京市永明医疗仪
轻浮、体积蓬松,在实际炮制中存在翻动、受热不均匀, 器厂)。
炒制火力难以控制,火候判断主观性强的问题,加之中 1.2 药品与试剂
药化学成分复杂,仅以单一或几种成分作为评价指标具 淫羊藿苷对照品(批号:110807-201306,纯度:≥
有局限性,可导致不同厂家、不同批次炙淫羊藿性状、外 98%)、宝藿苷Ⅰ对照品(批号:110807-201306,纯度:≥
观差别较大,质量参差不齐,使得其炮制工艺与现代化 98%)均购自中国食品药品检定研究院;朝藿定 A 对照
生产和质量控制要求存在较大差距。 品(批号:MUST-15010408,纯度:≥98%)、朝藿定 B 对
烘制可有效地保证温度均衡,且药材受热均匀、火 照品(批号:MUST-14062312,纯度:≥98%)、朝藿定 C
候易于控制 [6-7] 。有学者采用烘制法对蒲黄炭、金银花、 对照品(批号:MUST-14022312,纯度:≥98%)均购自成
荔枝核、杜仲、甘草、红芪等药材进行了炮制研究,获得 都曼斯特生物技术有限公司;箭藿苷 A 对照品(批号:
了各炮制品的具体烘制工艺参数,促进了炮制工艺的规 20140310,纯 度 :≥98%)、箭 藿 苷 B 对 照 品(批 号 :
范化 [6-11] 。中药指纹图谱能系统、全面地表征中药的内 201403012,纯度:≥98%)均购自宝鸡市辰光生物科技
[12]
在成分变化 ,其结合多元统计学分析方法可将信息充 公 司 ;羊 脂 油(河 北 龙 翔 油 脂 有 限 公 司 ,批 号 :
分整合,从整体角度筛选炮制工艺参数,目前该方法已 20190216);乙腈、磷酸为色谱纯,其余试剂均为分析纯,
中国药房 2020年第31卷第12期 China Pharmacy 2020 Vol. 31 No. 12 ·1481 ·