目的: 建立五子衍宗丸指纹图谱并进行化学模式识别分析,测定五子衍宗丸中金丝桃苷、毛蕊花糖苷、山柰酚、五味子醇甲、五味子甲素和五味子乙素的含量,为该制剂的质量控制提供参考。方法: 采用高效液相色谱法,使用Agilent SB-C18色谱柱(250 mm×4.6 mm,5 μm),以乙腈-0.05%磷酸水溶液为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长254 nm,柱温30 ℃,进样量20 μL。以五味子醇甲为参照,使用《中药色谱指纹图谱相似度评价系统软件(2012A版)》建立30批五子衍宗丸的指纹图谱,确定共有峰并进行相似度评价,通过与对照品溶液比对指认色谱峰。采用Simca-P 14.1软件对共有峰进行评价。采用上述HPLC方法测定30批样品中指认化学成分的含量。结果: 建立五子衍宗丸指纹图谱,共标定17个共有峰,相似度不低于0.90;共指认化学成分6个,分别为金丝桃苷、毛蕊花糖苷、山柰酚、五味子醇甲、五味子甲素、五味子乙素。经化学模式分析,30批样品聚成3类,筛选出8个差异性标志物。30批样品中上述6个成分的含量分别为1.158~3.695、0.089~1.965、1.158~3.695、0.089~1.965、1.158~3.695、0.089~1.965 mg·g-1。结论: 所建立的五子衍宗丸HPLC分析方法简便、稳定、可行,同时指纹图谱结合化学模式识别分析可为五子衍宗丸质量控制标准的制定提供参考。
Objective: To establish fingerprint of Wuziyanzong pills, analyze its chemical pattern recogniton, and determine the contents of hyperoside, verbascoside, kaempferol, schisandrin, deoxyschizandrin and γ-schisandrin in Wuziyanzong pills, so as to provide reference for the quality control of the preparation. Methods: HPLC method was adopted. The determination was performed on an Agilent SB-C18 column(250 mm×4.6 mm, 5 μm) with acetonitrile-0.05% phosphoric acid water as the mobile phase(gradient elution) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 254 nm, and the column temperature was 30 ℃. The sample volume was 20 μL. With schisandrin as the reference, the HPLC fingerprints of 30 batches of Wuziyanzong pills were established with Similarty Evalutation System for TCM Chromatographic Fingerprint (2012A edition); common peaks were determined and the similarity was evaluated. The chromatographic peaks were identified by comparing with the reference. Simca-P 14.1 software was used to evaluate the common peaks. The contents of the identified components in 30 batches of samples was determined by the abvoe HPLC method. Results: The fingerprint of Wuziyanzong pills was established, and 17 common peaks were calibrated, and the similarity was no less than 0.90. There were 6 chemical components identified,which were hyperoside, verbascoside, kaempferol, schisandrin, deoxyschizandrin and γ-schisandrin. 30 batches of samples were classified into three groups and eight difference markers were selected by chemical pattern recognition. The contents of above 6 components in 30 samples were 1.158-3.695 mg·g-1, 0.089-1.965 mg·g-1, 1.158-3.695 mg·g-1, 0.089-1.965 mg·g-1, 1.158-3.695 mg·g-1, 0.089-1.965 mg·g-1, respectively. Coclusion: The established HPLC analysis method of Wuziyanzong pills is sample,stable and feasible, and the fingerprint combined with chemical pattern recognition can provide reference for the establishment of the quality control standard of the Wuziyanzong pills.
[1] 中华人民共和国药典2020年版. 一部[S].2020: 611
ChP 2020. Vol Ⅰ[S].2020: 611
[2] 李艳荣, 樊慧杰, 柴智, 等. 五子衍宗丸对帕金森病小鼠的神经保护及抗炎作用[J].中华中医药杂志, 2020, 35(7):3623
LI YR, FAN HJ, CHAI Z, et al. Neuroprotective and anti-inflammatory effects of Wuzi Yanzong pill on Parkinson's disease mice[J].China J Tradit Chin Med Pharm, 2020, 35(7):3623
[3] 王雪宁, 丛慧芳, 陈思宇. 五子衍宗丸加减治疗早发性卵巢功能不全的研究进展[J].中西医结合心血管病电子杂志, 2020, 8(17):152
WANG XN, CONG HF, CHEN SY. Research progress of Wuzi Yanzong pill in the treatment of early-onset ovarian insufficiency[J].Cardiovasc Dis Electronic J Integrated Tradit Chin West Med, 2020, 8(17):152
[4] 王坤, 王娟, 刘小金, 等. 五子衍宗丸加减对肾虚精亏证黄体功能不全所致不孕患者内分泌及代谢的影响[J].中药材, 2021, 44(3):722
WANG K, WANG J, LIU XJ, et al. Effect of Wuziyanzong pill on endocrine and metabolism in infertile patients with luteal insufficiency of kidney deficiency and essence deficiency syndrome[J].J Chin Med Mater, 2021, 44(3):722
[5] 刘广楠. 五子衍宗丸治疗化疗后肝损伤的临床疗效分析[D].哈尔滨: 黑龙江中医药大学, 2018
LIU GN. The Chinical Effect of Wuzi Yanzong Pill in the Treatment of Liver Injury after Chemotherapy[D].Haerbin: Heilongjiang University of Chinese Medicine, 2018
[6] 周卫东, 卢汉祺, 陈泽伟, 等. 五子衍宗丸对化疗性肠黏膜炎小鼠的保护作用及其机制研究[J].光明中医, 2021, 36(5):729
ZHOU WD, HU HQ, CHEN ZW, et al. Research on the protection and mechanism of Wuzi Yanzong pill on chemotherapy-induced intestinal inflammation in mice[J].Guangming J Chin Med, 2021, 36(5):729
[7] 杨婵娟, 樊慧杰, 柴智, 等. 五子衍宗丸预防神经管畸形的机制研究[J].中国中医基础医学杂志, 2021, 27(12):1887
YANG CJ, FAN HJ, CHAI Z, et al. Mechanism of Wuzi Yanzong pill in preventing neural tube defects[J].J Basic Chin Med, 2021, 27(12):1887
[8] 孙梦瀛, 樊慧杰, 柴智, 等. 五子衍宗丸对脱髓鞘模型小鼠脑组织星形胶质细胞神经营养因子分泌的影响[J].中医杂志, 2021, 62(8):712
SUN MY, FAN HJ, CHAI Z, et al. Effect of Wuzi Yanzong pills on the secretion of neurotrophic factors by astrocytes in brain tissues of demyelination medol mice[J].J Tradit Chin Med, 2021, 62(8):712
[9] 亓翠玲, 郭平, 于爱莲. 五子衍宗丸对糖尿病周围血管病变患者体内NO含量的探讨[J].实用糖尿病杂志, 2015, 11(4):12
QI CL, GUO P, YU AL. Study on the content of NO in patients with diabetic peripheral vascular disease by Wuzi Yanzong pills[J].J Pract Diabetol, 2015, 11(4):12
[10] 刘移兰, 蔡卫华, 窦志华, 等. 一测多评法同时测定五味子中12个木脂素类成分的含量[J].药物分析杂志, 2019, 39(7):1207
LIU YL, CAI WH, DOU ZH, et al. Simultaneous determination of 12 lignans in Schisandra chinensis by QAMS[J].Chin J Pharm Anal, 2019, 39(7):1207
[11] 董秋菊, 周园, 冯薇, 等. 指纹图谱结合一测多评法在五子衍宗丸质量评价中的应用[J].中药材, 2019, 42(3):579
DONG QJ, ZHOU Y, FENG W, et al. Application of fingerprint combined with QAMS in quality evaluation of Wuzi Yanzong pills[J].J Chin Med Mater, 2019, 42(3):579
[12] 廖宇娇, 敖明月, 彭颖, 等. 基于指纹图谱及化学模式识别研究盐制对五子衍宗丸化学成分的影响[J].中草药, 2021, 52(11):3224
LIAO YJ, AO MY, PENG Y, et al. Study on the effect of salt preparation on the chemical composition of Wuzi Yanzong pills based on fingerprint and chemical pattern recognition[J].Chin Tradit Herb Drugs, 2021, 52(11):3224
[13] 刘伟, 周建良, 陈碧莲, 等. 五子衍宗丸高效液相色谱指纹图谱研究[J].医药导报, 2014, 33(10):1360
LIU W, ZHOU JL, CHEN BL, et al. HPLC fingerprint analysis of Wuzi Yanzong pills[J].Her Med, 2014, 33(10):1360
[14] 刘伟, 周建良, 陈碧莲, 等. HPLC同时测定五子衍宗丸中5种活性成分的含量[J].中国实验方剂学杂志, 2014, 20(3):74
LIU W, ZHOU JL, CHEN BL, et al. Simultaneous determination of five ingredients in Wuzi Yanzong pill by HPLC[J].Chin J Exp Tradit Med Form, 2014, 20(3):74
[15] 郭蕊蕊. 菟丝子中黄酮类成分对人精子细胞活力的影响[D].兰州: 兰州大学, 2018
GUO RR. Effects of Flavonoids from Cuscuta chinensis on Human Sperm Viability[D].Lanzhou: Lanzhou University, 2018
[16] 白晓蕾, 宋琳, 郑远武, 等. 五味子醇甲药理研究进展[J].实用中医内科杂志, 2021, 35(2):18
BAI XL, SONG L, ZHENG YW, et al. Research progress on pharmacology of schisandrin A[J].J Pract Tradit Chin Intern Med, 2021, 35(2):18
[17] 李博怿. 五味子甲素通过SCF/c-kit信号通路调控少、弱精子症大鼠模型精子凋亡作用机制研究[D].北京: 北京中医药大学, 2018
LI BY. Study on the Mechanism of Schisandrin A Regulating Sperm Apoptosis in Oligozoospermia and Asthenospermia Rat Model Through SCF/c-kit Signal Pathway[D].Beijing: Beijing University of Chinese Medicine, 2018
[18] 陈振羽. 五味子乙素通过Nrf2/ARE信号通路对肾病综合征大鼠的保护作用[J].西部中医药, 2020, 33(1):45
CHEN ZY. Protective effects of schisandrin B on rat model with nephrotic syndrome through Nrf2/ARE signaling pathway[J].West J Tradit Chin Med, 2020, 33(1):45
[19] 谢莹, 林鑫, 袁静, 等. 毛蕊花糖苷对糖尿病肾病小鼠肾组织中高迁移率族蛋白1及核因子κB表达的影响[J].中华肾脏病杂志, 2021, 37(10):817
XIE Y, LIN X, YUAN J, et al. Effects of acteoside on the expressions of high mobility group box 1 and nuclear factor-κB in the renal tissues of diabetic nephropathy mice[J].Chin J Nephrol, 2021, 37(10):817
[20] 邵静媛. 质量源于设计理念在若干中药分析方法中的应用研究[D].杭州: 浙江大学, 2019
SHAO JY. Researches on Several Analytical Methods for Traditional Chinese Medicines Based on QbD Concept[D].Hangzhou: Zhejiang University, 2019
