冬虫夏草及其发酵菌丝体多糖指纹特征
|
摘要
以冬虫夏草为试材,采用PMP衍生化结合HPLC分析建立冬虫夏草多糖指纹图谱分析方法,研究冬虫夏草与冬虫夏草菌丝体之间质量差异。以三氟乙酸水解冬虫夏草多糖,水解产物用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化结合HPLC分析测定冬虫夏草多糖指纹,色谱柱为Agilent Eclipse XDB C18(250mm×4.6mm,5μm),流动相为磷酸盐缓冲液(0.1mol·L-1,pH 6.7)-乙腈(80∶20,v/v),体积流量1mL·min-1,进样量20μL,柱温25℃。结果表明:不同产地12批次冬虫夏草中多糖指纹相似度均大于0.992,冬虫夏草与冬虫夏草发酵菌丝体多糖指纹存在较大差异,相似度在0.708～0.974。聚类分析与相似度分析结果基本一致,能够实现冬虫夏草与其发酵菌丝体的区分。建立的冬虫夏草多糖HPLC指纹图谱分析法操作简单、结果可靠,能够准确鉴别冬虫夏草与冬虫夏草菌丝体,可用于冬虫夏草质量的客观评价。
A pre-column derivation coupled HPLC analysis was applied to identify and characterize polysaccharides of Ophiocordyceps sinensis and relative cultivated mycelium from different origins.The polysaccharides extracted fromO.sinensis were hydrolyzed partially with trifluoroacetic acid and derivated by 1-phenyl-3-methyl-5-pyrazolone(PMP).The monosaccharide composition fingerprint was obtained by HPLC on a Agilent Eclipse XDB C18(250 mm×4.6 mm,5μm)column with phosphate buffered solution(0.1 mol·L-1,pH 6.7)and acetonitrile(80∶20,v/v)as mobile phase at the flow rate of 1 mL·min-1.The column temperature was at 25℃.The results showed that the similarity of12 batches of O.sinensis was more than 0.992.There were some differences between O.sinensis and cultivated mycelium,with the similarity value range from 0.708 to 0.974.The result of Cluster analysis(CA)largely agreed with the similarity evaluation,which distinguished O.sinensis from cultivated mycelium.The polysaccharides fingerprint by HPLC combined with CA were simple and reliable,which could precisely differentiate O.sinensis from cultivated mycelium and were useful for quality evaluation of O.sinensis.
A pre-column derivation coupled HPLC analysis was applied to identify and characterize polysaccharides of Ophiocordyceps sinensis and relative cultivated mycelium from different origins.The polysaccharides extracted fromO.sinensis were hydrolyzed partially with trifluoroacetic acid and derivated by 1-phenyl-3-methyl-5-pyrazolone(PMP).The monosaccharide composition fingerprint was obtained by HPLC on a Agilent Eclipse XDB C18(250 mm×4.6 mm,5μm)column with phosphate buffered solution(0.1 mol·L-1,pH 6.7)and acetonitrile(80∶20,v/v)as mobile phase at the flow rate of 1 mL·min-1.The column temperature was at 25℃.The results showed that the similarity of12 batches of O.sinensis was more than 0.992.There were some differences between O.sinensis and cultivated mycelium,with the similarity value range from 0.708 to 0.974.The result of Cluster analysis(CA)largely agreed with the similarity evaluation,which distinguished O.sinensis from cultivated mycelium.The polysaccharides fingerprint by HPLC combined with CA were simple and reliable,which could precisely differentiate O.sinensis from cultivated mycelium and were useful for quality evaluation of O.sinensis.
引文
[1]ZHU Z Y,YAO Q,LIU Y,et al.Highly efficient synthesis and antitumor activity of monosaccharide saponins mimicking components of Chinese folk medicine Cordycepssinensis[J].Journal of Asian Natural Products Research,2012,14(5):429-435.
[2]SHASHIDHAR M G,GIRIDHAR P,UDAYA SANKARK,et al.Bioactive principles from Cordyceps sinensis:A potent food supplement-a review[J].Journal of Functional Foods,2013(2):1013-1030.
[3]CHENG Y,CHENG S,TENG Y,et al.Cordyceps sinensis prevents apoptosis in mouse liver with d-galactosamine/lipopolysaccharide-induced fulminant hepatic failure[J].American Journal of Chinese Medicine,2014,42(2):427-441.
[4]BENSKY D,GAMBLE A,CLAVEY S,et al.Materia medica:Chinese herbal medicine[M].3ed.Seattle,Washington:Eastland Press,2004.
[5]胡贤达,周菲,黄雪,等.冬虫夏草中虫草多糖的药理研究进展[J].中国实验方剂学杂志,2016,22(6):224-229.
[6]ZHAO J,XIE J,WANG L Y,et al.Advanced development in chemical analysis of Cordyceps[J].Journal of Pharmaceutical and Biomedical Analysis,2014,87(SI):271-289.
[7]朱振元,原静,李盛峰,等.古尼虫草多糖硫酸酯化修饰及其抗氧化活性[J].现代食品科技,2011,27(1):1-5.
[8]NIE S,CUI S W,XIE M,et al.Bioactive polysaccharides fromCordyceps sinensis:Isolation,structure features and bioactivities[J].Bioactive Carbohydrates and Dietary Fibre,2013,1(1):38-52.
[9]余晓斌,罗长才,缪静.冬虫夏草多糖提取工艺的优化[J].中草药,2002,33(12):33-34.
[10]许峰,吴玲芳,林善,等.发酵冬虫夏草菌丝体中虫草多糖含量的检测及结构鉴定[J].基因组学与应用生物学,2014,33(6):1294-1302.
[11]张安强,王小臣,王星丽,等.冬虫夏草多糖抗氧化活性及其单糖组成的研究[J].食药用菌,2015,23(4):237-242.
[12]GUAN J,ZHAO J,FENG K,et al.Comparison and characterization of polysaccharides from natural and cultured Cordyceps using saccharide mapping[J].Analytical and Bioanalytical Chemistry,2011,399(10):3465-3474.
[2]SHASHIDHAR M G,GIRIDHAR P,UDAYA SANKARK,et al.Bioactive principles from Cordyceps sinensis:A potent food supplement-a review[J].Journal of Functional Foods,2013(2):1013-1030.
[3]CHENG Y,CHENG S,TENG Y,et al.Cordyceps sinensis prevents apoptosis in mouse liver with d-galactosamine/lipopolysaccharide-induced fulminant hepatic failure[J].American Journal of Chinese Medicine,2014,42(2):427-441.
[4]BENSKY D,GAMBLE A,CLAVEY S,et al.Materia medica:Chinese herbal medicine[M].3ed.Seattle,Washington:Eastland Press,2004.
[5]胡贤达,周菲,黄雪,等.冬虫夏草中虫草多糖的药理研究进展[J].中国实验方剂学杂志,2016,22(6):224-229.
[6]ZHAO J,XIE J,WANG L Y,et al.Advanced development in chemical analysis of Cordyceps[J].Journal of Pharmaceutical and Biomedical Analysis,2014,87(SI):271-289.
[7]朱振元,原静,李盛峰,等.古尼虫草多糖硫酸酯化修饰及其抗氧化活性[J].现代食品科技,2011,27(1):1-5.
[8]NIE S,CUI S W,XIE M,et al.Bioactive polysaccharides fromCordyceps sinensis:Isolation,structure features and bioactivities[J].Bioactive Carbohydrates and Dietary Fibre,2013,1(1):38-52.
[9]余晓斌,罗长才,缪静.冬虫夏草多糖提取工艺的优化[J].中草药,2002,33(12):33-34.
[10]许峰,吴玲芳,林善,等.发酵冬虫夏草菌丝体中虫草多糖含量的检测及结构鉴定[J].基因组学与应用生物学,2014,33(6):1294-1302.
[11]张安强,王小臣,王星丽,等.冬虫夏草多糖抗氧化活性及其单糖组成的研究[J].食药用菌,2015,23(4):237-242.
[12]GUAN J,ZHAO J,FENG K,et al.Comparison and characterization of polysaccharides from natural and cultured Cordyceps using saccharide mapping[J].Analytical and Bioanalytical Chemistry,2011,399(10):3465-3474.