摘要
目的:研究化学指纹图谱及DNA条形码分子鉴定技术在肉桂精准煮散饮片质量体系的应用。方法:应用psb Atrn H序列作为DNA条形码对肉桂进行鉴定;采用标准汤剂煎煮法,比较原饮片及不同规格煮散饮片的出膏率。建立肉桂HPLC指纹图谱,测定指标成分肉桂酸、肉桂醛的含量,同时标定指纹图谱共有峰,比较各样品共有峰相对峰面积及相似度评价。结果:psb A-trn H序列对肉桂药材可实现准确鉴定;精准煮散饮片平均出膏率及煎煮液中指标成分肉桂酸、肉桂醛含量略高于原饮片,且含量差异性明显减小,RSD从原饮片的5.19%、26.80%降低为0.36%、0.42%。10个共有峰相对峰面积均有所升高,均一性明显提高。结论:肉桂精准煮散饮片可提高原饮片的煎煮效率及均一性。
Objective: To study the application of precise powder decoction pieces( PPDP) of Cinnamonum cassia Presl( CCP)with chemical fingerprint chromatography and DNA molecular identification technology were used on quality system. Methods: Psb A-trn H sequence was used as DNA barcode to indentify CCP. Different specifications of PPDP were prepared,and their dry extract content rates were compared with that of original slices. HPLC fingerprint of CCP was established and the contents of cinnamic acid and cinnamic aldehyde were measured. Common peak of fingerprint was demarcated. The common peak relative peak areas and similarity evaluation of each sample were compared. Results: CCP could be accurately identified by psb A-trn H sequences.The extract rate of the concentration of cinnamic acid and cinnamic aldehyde of PPDP were slightly higher than that of the original pieces,and the content differences decreased significantly. RSD of inter-assay dissolution of cinnamic acid and cinnamic aldehyde of the original slices were 5. 19% and 26. 80%,respectively,which could be reduced to 0. 36% and 0. 42% after mixing and preparing into PPDP. The relative peak areas of the 10 common peaks were increased,and uniformity was significantly improved.Conclusion: The precise powder decoction pieces of CCP can improve the extraction efficiency and uniformity of original slices.
引文
[1]国家药典委员会.中华人民共和国药典[S].一部.北京:中国医药科技出版社,2015:136-137.
[2]韩亚明,蒋林,黄正恩,等.广西、云南产肉桂油化学成分及分子蒸馏技术纯化研究[J].中南药学,2005,3(4):215-218.
[3]黄亚非,黄际薇,陶玲,等.不同树龄肉桂挥发油的成分比较[J].中山大学学报:自然科学版,2005,44(1):82-85.
[4]Kanari M,Tomoda M,Gonda R,et al.A reticuloendothelial systemactivating arabinoxylan from the bark of Cinnamomum cassia[J].Chem Pharm Bull(Tokyo),1989,37(12):3191-3194.
[5]邱琴,崔兆杰,韦栋梁,等.肉桂挥发油化学成分的研究[J].上海中医药大学学报,2003,17(3):49-51.
[6]康廷国.中药鉴定学[M].北京:中国中医药出版社,2011:255.
[7]徐洋洋,王添敏,初正云,等.HPLC-DAD测定5种商品规格肉桂及两种伪品中桂皮醛的含量[J].中国实验方剂学杂志,2014,20(1):90-93.
[8]李睿,翟华强,田伟兰,等.中药煮散的历史源流及其与现代配方颗粒的对比性分析[J].中国中药杂志,2016,41(5):965-969.
[9]穆兰澄,曹京梅,李冀湘,等.中药煮散的历史沿革与现代研究概述[J].中国实验方剂学杂志,2008,14(7):74-75.
[10]Wu L,Sun W,Wang B,et al.An integrated system for identifying the hidden assassins in traditional medicines containing aristolochic acids[J].Sci Rep,2015,5:11318.
[11]Wang M,Zhao H X,Wang L,et al.Potential use of DNA barcoding for the identification of Salvia,based on cp DNA and nr DNA sequences[J].Gene,2013,528(2):206-215.
[12]Yuan QJ,Zhang B,Jiang D,et al.Identification of species and materia medica within Angelica L.(Umbelliferae)based on phylogeny inferred from DNA barcodes[J].Mol Ecol Resour,2015,15(2):358-371.
[13]Chen S,Pang X,Song J,et al.A renaissance in herbal medicine identification:from morphology to DNA[J].Biotechnol Adv,2014,32(7):1237-1244.
[14]陈士林,刘安,李琦,等.中药饮片标准汤剂研究策略[J].中国中药杂志,2016,41(8):1367-1375.