目的: 通过制备15批一贯煎基准样品冻干粉,建立一贯煎基准样品HPLC指纹图谱,为该经典名方的质量控制及评价提供参考。方法: 采用Venusil C18色谱柱(250 mm×4.6 mm,5 μm),以甲醇(A)-0.1%磷酸水(B)为流动相梯度洗脱,建立15批一贯煎基准样品的HPLC指纹图谱,应用“中药色谱指纹图谱相似度评价系统”软件(2012版)进行相似度分析,并对共有峰进行归属。采用聚类分析(HCA),主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA)对指纹图谱数据进行评价。结果: 15批一贯煎基准样品与对照指纹图谱的相似度均≥0.771,共标定了31个共有峰,指认出了色谱峰11、20、23、25、26、30、31,分别为香草酸、阿魏酸、毛蕊花糖苷、异类叶升麻苷、补骨脂素、欧前胡素和蒿本内酯共7个成分;其中峰2、5~8、10、13、16~19、24、27归属于枸杞子,峰1、20、30、31归属于当归,峰21~23、25、28归属于生地黄,峰3归属于枸杞子和麦冬,峰4、29归属于枸杞子和生地黄,峰9归属于当归和沙参,峰11、12、14、15、26归属于枸杞子和当归,HCA、PCA及OPLS-DA可将15批基准样品大致分为2类。结论: 建立的一贯煎HPLC指纹图谱方法专属性强,分离度好,灵敏度高,除川楝子药味以外均有表征,可为该方剂复方制剂的质量控制与评价提供参考。
Objective: To establish the Yiguanjian substance benchmark HPLC fingerprint by preparing 15 batches of Yiguanjian material benchmark freeze-dried powder, and provide a reference for the quality control and evaluation of this classic recipe. Methods: Using a Venusil C18 column (250 mm×4.6 mm, 5 μm) and methanol (A)-0.1%phosphoric acid water (B) as mobile phase gradient elution, 15 batches of Yiguan decoction material benchmark HPLC fingerprints were established. And the "Chinese Medicine" chromatographic fingerprint similarity evaluation system" software(2012 version) was performed to analyze similarity and assign common peaks. Cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to evaluate the fingerprint data. Results: The similarity between the 15 batches of Yiguanjian material benchmark and the control fingerprint was ≥0.771. A total of 31 common peaks were calibrated, and the chromatographic peaks 11, 20, 23, 25, 26, 30, and 31 were identified, which were vanilla. Acid, ferulic acid, verbascoside, xenocimicifugaoside, psoralen, imperatorin, and artemisinol, including 7 components. Peaks 2, 5-8, 10, 13, 16-19, 24 and 27 were attributed to Lycium barbarum L., peaks 1, 20, 30, and 31 were attributable to Angelica sinensis (Oliv.) Diels, peaks 21-23, 25, and 28 belonged to Rehmannia glutinosa Libosch., peak 3 was attributable to Lycium barbarum L. and Ophiopogon japonicus (L.f) Ker-Gawl., peaks 4 and 29 were attributable to Lycium barbarum L. and Rehmannia glutinosa Libosch., peak 9 was attributed to Angelica sinensis (Oliv.) Diels and Glehnia littoralis Fr. Schmindt ex Miq., and peaks 11, 12, 14, 15, and 26 were attributed to Lycium barbarum L. and Angelica sinensis (Oliv.) Diels. HCA, PCA and OPLS-DA can roughly divide the 15 batches of substance standards into two categories. Conclusion: The established HPLC fingerprint method of Yiguanjian has strong specificity, good resolution, and high sensitivity. It has characteristics except for the medicinal taste of Melia toosendan Sieb. et Zucc., which can provide a reference for the quality control and evaluation of the compound preparation of this formula.
[1] 魏玉璜. 续名医类案卷十八[M].北京:人民卫生出版社,1957:1770
WEI YH. Renewed Cases of Famous Doctors Eighteenth[M].Beijing: People’s Medical Publishing House, 1957: 1770
[2] 宗全和. 中医方剂通释卷三[M].石家庄: 河北科学技术出版社, 1995: 2
ZONG QH. General Explanation of Chinese Medicine Formulas Volume 3[M].Shijiazhuang: Hebei Science & Technology Press, 1995: 2
[3] 孙福慧, 车念聪, 唐佐青, 等. 基于Notch通路研究一贯煎逆转肝纤维化的作用机制[J].环球中医药, 2019, 12(9): 1308
SUN FH, CHE NC, TANG ZQ, et al. Research on the mechanism of Yiguan Decoction reversing liver fibrosis based on Notch pathway[J].Global J Tradit Chin Med, 2019, 12(9): 1308
[4] 乔天阳. 基于FGF2-DLK1信号通路研究一贯煎促进骨髓MSCs逆转肝纤维化的作用机制[D].北京: 首都医科大学, 2017
QIAO TY. Study Themechanism of Yiguanjian Promoting Bone Marrow MSCs on Reversing Liver Fibrosis Based on FGF2-DLK1 Signal Pathway[D].Beijing: Capital Medical University, 2017
[5] 张鹏飞, 丛琳, 杨瑜婷, 等. 叶建州教授治疗带状疱疹及其后遗神经痛经验[J].云南中医中药杂志, 2016, 37(8): 5
ZHANG PF, CONG L, YANG YT, et al. Professor Ye Jianzhou’s experience in treating herpes zoster and its postherpetic neuralgia[J].Yunnan J Tradit Chin Med Mater Med, 2016, 37(8): 5
[6] 徐加艳. 芳香疗法并一贯煎治疗经断前后诸证肾虚肝郁型临床研究[D].昆明: 云南中医药大学, 2020
XU JY. A Clinical Study on the Treatment of Kidney Deficiency and Liver Depression before and after Jingduan by Aromatherapy Combined with YiguanJian[D].Kunming: Yunnan University of Chinese Medicine, 2020
[7] 余功, 李璐, 胡桥, 等. 一贯煎中川楝子佐制配伍对荷H22肝癌小鼠胃肠消化吸收功能的影响[J].江西中医药大学学报, 2020, 32(4): 91
YU G, LI L, HU Q, et al. Effect of compatibility of fructus Meliae toosendan in Yiguan decoction on tumor proliferation in H22 mice[J].J Jiangxi Univ Tradit Chin Med, 2020, 32(4): 91
[8] 林存博. 一贯煎合失笑散加味治疗慢性萎缩性胃炎(阴虚血瘀证)的临床观察[D].长春:长春中医药大学, 2020
LIN CB. Clinical Observation on Treating Chronic Atrophic Gastritis (Yin Deficiency and Blood Stasis Syndrome) by Yiguanjian Combined with Shixiao Prescription[D].Changchun: Changchun University of Chinese Medicine, 2020
[9] 宋维海, 李琴, 茅建春. 一贯煎对干燥综合征模型大鼠及颌下腺PI3K/Akt/eNOS通路的影响[J].中药材, 2020, 43(7): 1721
SONG WH, LI Q, MAO JC. Effects of Yiguanjian decoction on Sjögren’s syndrome model rats and submandibular gland PI3K/Akt/eNOS pathway[J].J Chin Med Mater, 2020, 43(7): 1721
[10] 祝明, 陈碧莲, 石上梅. 中药指纹图谱技术在中国药典2015年版一部中的应用[J].中国现代应用药学, 2016, 33(5): 611
ZHU M, CHEN BL, SHI SM. Application of fingerprint technology on traditional Chinese medicine in Chinese pharmacopoeia(2015 edition) volume Ⅰ[J].Chin J Mod Appl Pharm, 2016, 33(5): 611
[11] 孙立丽, 王萌, 任晓亮. 化学模式识别方法在中药质量控制研究中的应用进展[J].中草药, 2017, 48(20): 4339
SUN LL, WANG M, REN XL. Application progress on chemical pattern recognition in quality control of Chinese materia medica[J].Chin Tradit Herb Drugs, 2017, 48(20): 4339
[12] 唐维维, 梁献葵, 马驰虹, 等. 不同采收季节怀牛膝指纹图谱研究[J].中药材, 2019, 42(9): 2079
TANG WW, LIANG XK, MA CH, et al. Study on fingerprint of Achyranthes Bidentata in different harvesting seasons[J].J Chin Med Mater, 2019, 42(9): 2079
[13] 刘颖, 王青, 王放, 等. 苦碟子注射液HPLC指纹图谱与化学模式识别分析[J].中国药学杂志, 2013, 48(24): 2097
LIU Y, WANG Q, WANG F, et al. HPLC fingerprint and chemical pattern recognization of Kudiezi injection[J].Chin Pharm J, 2013, 48(24): 2097
[14] 刘慧, 肖金超, 张庆捷, 等. 金骨莲胶囊HPLC指纹图谱及化学模式识别研究[J].中草药, 2021, 52(14): 4185
LIU H, XIAO JC, ZHANG QJ, et al. HPLC fingerprint and chemical pattern recognition of Jingulian capsules[J].Chin Tradit Herb Drugs, 2021, 52(14): 4185
[15] 管咏梅, 万鑫浩, 吴文婷, 等. 经典名方桂枝加葛根汤标准汤剂HPLC指纹图谱研究[J].中草药, 2021, 52(18): 5538
GUAN YM, WAN XH, WU WT, et al. HPLC fingerprint of classic prescription Guizhi Jia Gegen Decoction[J].Chin Tradit Herb Drugs, 2021, 52(18): 5538
