目的 建立壁钱的HPLC指纹图谱和多指标成分的含量测定方法,并结合化学计量学方法评价其质量。方法 采用Waters Symmetry RP C18色谱柱,以甲醇(A)-体积分数0.1% 冰醋酸溶液(B)为流动相,梯度洗脱(0～15 min,0 A;15～18 min,0~10% A;18～30 min,10%~15% A),检测波长为262 nm,流速:0.5 mL·min-1,柱温:35 ℃。对壁钱药材、饮片和焙壁钱进行指纹图谱构建,并结合相似度分析和化学计量学分析,同时测定尿嘧啶、鸟嘌呤、次黄嘌呤、尿苷、肌苷和鸟苷的含量。结果 30批壁钱样品指纹图谱相似度均在0.96以上,共标定了17个共有峰,指认出6个色谱峰,分别是尿嘧啶、鸟嘌呤、次黄嘌呤、尿苷、肌苷和鸟苷。聚类分析将30批壁钱样品分为3类,不同炮制规格各自聚为一类。主成分分析表明,不同药材市场的壁钱样品间存在差异,不同炮制方式壁钱样品间差异较小。尿嘧啶、鸟嘌呤、次黄嘌呤、尿苷、肌苷及鸟苷的质量分数范围分别为0.20~0.94、0.56~0.94、1.43~3.30、0.13~0.58、0.87~1.36、0.53~1.57 mg·g-1,经方法学考察,各成分呈现良好的线性关系,平均回收率在95.75%~101.6%之间,RSD在0.33%~1.06%之间。结论 建立的HPLC指纹图谱和多指标含量测定方法稳定、可靠,可为壁钱药材、饮片和焙壁钱的质量评价提供依据。
Abstract
OBJECTIVE To establish the HPLC fingerprints and multi-components determination method of Uroctea compactilis, and evaluate its quality combined with chemometrics. METHODS Using Waters Symmetry RP-C18 column, methanol as mobile phase A and 0.1% glacial acetic acid solution as mobile phase B, gradient elution(0-15 min, 0 A; 15-18 min, 0-10% A; 18-30 min, 10%-15% A)was carried out. The detection wavelength was set at 262 nm, the flow rate was 0.5 mL·min-1 and the column temperature was maintained at 35 ℃. The fingerprints of 30 batches of Uroctea compactilis medicinal materials, decoction pieces of Uroctea compactilis and baked Uroctea compactilis were constructed. Combined with similarity analysis and chemometric analysis, the contents of uracil, guanine, hypoxanthine, uridine, inosine and guanosine were determined at the same time. RESULTS The similarity of fingerprints of 30 batches of Uroctea compactilis samples were more than 0.96. A total of 17 common peaks were calibrated and six chromatographic peaks were identified, namely uracil, guanine, hypoxanthine, uridine, inosine and guanosine. Cluster analysis divided 30 batches of Uroctea compactilis samples into three categories. Different processing specifications were grouped into one category. Principal component analysis showed that there were differences among Uroctea compactilis samples from different manufacturers, and there were little differences among Uroctea compactilis samples processed by different methods. The mass fractions of uracil, guanine, hypoxanthine, uridine, inosine, and guanosine were 0.20-0.94,0.56-0.94,1.43-3.30,0.13-0.58,0.87-1.36 mg·g-1, and 0.53-1.57 mg·g-1, respectively. Each component presented a good linear relationship, and the recovery was between 95.75%-101.6% and RSD was between 0.33%-1.06%. CONCLUSION The established HPLC fingerprint and multi index content determination method are stable and reliable, which can provide a basis for the quality evaluation of Uroctea compactilis, decoction pieces of Uroctea compactilis and baked Uroctea compactilis.
关键词
壁钱药材 /
壁钱饮片 /
焙壁钱 /
指纹图谱 /
聚类分析 /
主成分分析 /
含量测定
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Key words
Uroctea compactilis /
decoction pieces of Uroctea compactilis /
baked Uroctea compactilis /
finger-print /
cluster analysis /
principal component analysis /
content determination
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中图分类号：
R284
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参考文献
[1] Shandong provincial Chinese herbal medicine standard(2002)(山东省中药材标准).2002:274.
[2] Processing standard of traditional Chinese medicine in Anhui province(2019)(安徽省中药饮片炮制规范). 2019:343-344.
[3] Processing standard of traditional Chinese medicine in Fujian Province(2012)(福建省中药饮片炮制规范). 2012:245.
[4] Processing standard of traditional Chinese medicine in Shandong province(1990)(山东省中药炮制规范). 1990:286.
[5] LV A J, WU H. Research progress of nucleosides in traditional Chinese Medicine. Inf Tradit Chin Med(中国中医药信息杂志),2006,(7): 94-97.
[6] ZHOU H Y, CHEN Y, XU L L. Progress in determination of nucleosides of Cordyceps. Chin Tradit Pat Med(中成药), 2011, 33(11):1955-1960.
[7] WANG B, LI N, DENG T X, et al. Determination of Nucleosides and HPLC fingerprints of cordyceps. J Chin Med Mater(中药材), 2015, 38(5): 952-956.
[8] JI Y B, WANG Y, LIN W H. Study on nucleotidoids from n-butyl alcohol part of sponge Cinachyrellaaustraliensis . Chin Tradit Herb Drugs(中草药),2004,35(11):15-17.
[9] LIAO X J, XU S H, HUANG Q C, et al. Studies on chemical constituents from Callyspongia Fibrosa. Chin J Spectrosc Lab(光谱实验室), 2005, 22(2):281-283.
[10] SUN J, TIAN F, MAO R Q, et al. Study on characteristic chromatogram and quantitation method of five nucleoside components of Guang Dilong decoction pieces. Chin J Pharm Anal(药物分析杂志), 2019, 39(11):2010-2019.
[11] GUAN S Q, ZHOU G L, DENG J J, et al. Simultaneous Determination of five nucleosides in Pheretima aspergillum(E.Perrier)by HPLC. J Guangxi Norm Univ Nat Sci Ed(广西师范大学学报:自然科学版), 2020, 38(3):85-91.
[12] LIU Y X, LIU Y, FENG L Y, et al. Research progress on the snake venom of Agkistrodonacutus. Biol Chem Eng(生物化工), 2018, 4(6):137-141.
[13] BI X L, LIU M Y, CHEN W T, et al. Study on quality control methods of 3 nucleosides in Opisthoplatia Orientalis Burm . J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2018, 20(4):15-18.
[14] ZHANG Q, ZHU Y H, WEN H M, et al. Simultaneous Determination of Eight Nucleosides in Scolopendra of Different Varieties and Origins by HPLC-PDA. Chin J Exp Tradit Med Form(中国实验方剂学杂志), 2017, 23(14):57-61.
[15] YANG R R, JI L, LI E W, et al. HPLC fingerprint analysis and pattern recognition of Spatholobus suberectus. Chin Tradit Herb Drugs(中草药), 2017, 48(21):4530-4536.
[16] LI Y, HE W Y, WANG K Y, et al. Quality assessment of Asarum heterotropoides var. mandshuricum based on HPLC multi-component determination and multiple pattern recognition method of fingerprint . Chin Tradit Herb drugs(中草药), 2022, 53(1):238-243.
[17] CHEN F, WEI C J, DING Q, et al. Quality evaluation of Chrysanthemum morifolium based on UPLC fingerprint and multi index content determination. J Chin Med Mater(中药材), 2021(10):2377-2381.
[18] LI Y, CHEN J, ZHANG Y, et al. Determination of multi-components of Paeoniae Alba Radix based on fingerprints and chemical pattern recognition. Chin Tradit Herb drugs(中草药), 2022, 53(1):231-237.
[19] CAI H Y, WANG X L, WANG G H, et al. Quality evaluation of Trollius chinensis Bge. based on fingerprint combined with chemometrics methods and content determination . Chin J Hosp Pharm(中国医院药学杂志),2022, 42(4):341-346.
[20] CHEN J, YANG R, ZHANG Q, et al. Specific chromatograms of Glycyrrhiza uralensis Fisch. Flavonoids in different growth years by HPLC coupled with chemometric analysis. Chin Pharm J(中国药学杂志), 2020, 55(17):1415-1420.
[21] ZHANG L L, ZHANG W T, TANG D F, et al. Quality evaluation of Ningxinbao capsules based on combinative method of HPLC fingerprint,quantitative analysis of multi-components and Chemometrics. Chin Pharm J(中国药学杂志), 2019, 54(17):1425-1431.
[22] Nanjing University of Chinese Medicine.The Dictionary of Chinese Herbal Medicine(中药大辞典 下册). Vol 2. Shanghai: Shanghai Scientific & Technical Publishers,2006:3763-3764.
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脚注
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基金
山东省重点研发计划(软科学项目)资助(2020RKB24001);山东省人文社会科学课题资助(2021-YYGL-44);山东省中药材及饮片标准研究资助(2020-065,2020-066);中药配方颗粒质量识别与控制关键技术研究及产业化应用资助(2021CXGC010511)
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