高效液相色谱定量分析比较4种虫草药材的核苷类成分
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摘要
为比较4种常见虫草类药材(冬虫夏草、蛹虫草、虫草菌丝体和蝉花)中核苷类成分的差异,该研究以0.5%磷酸溶液提取样本,色谱柱为Zorbax SB-Aq(4.6 mm×150 mm,5μm),流动相为0.04 mol·L-1磷酸二氢钾溶液(A)和乙腈(B),梯度洗脱,流速为0.8 m L·min-1,检测波长260 nm,柱温30℃,对4类药材产品进行分析。已知核苷以外标一点法进行计算,未知成分以腺苷为对照进行计算,并对测定结果进行聚类分析,同时利用指纹图谱相似度软件对不同样本进行了比较。结果显示,冬虫夏草、蛹虫草、虫草菌丝体、蝉花4种药材中的核苷类成分差异较大,聚类分析和指纹图谱分析均可有效区分冬虫夏草和其他3种虫草药材。该研究建立的核苷测定方法准确可靠,可用于虫草类产品的品质评价。
To compare the main nucleosides in Cordyceps genus herbs( C. sinensis,C. millitaris,Hirsutella sinensis and C.sobolifera),an HPLC method for simultaneous determination of uridine,inosine,guanosine,adenosine and cordycepine in Cordyceps genus herbs was developed. The sample was extracted with 0. 5 % phosphoric acid solution to prepare test solution. The separation was performed on a Zorbax SB-Aq( 4. 6 mm × 150 mm,5 μm) column with gradient elution by 0. 04 mol· L- 1potassium dihydrogen phosphate solution and acetonitrile,column temperature 30 ℃,flow rate 0. 8 m L · min- 1,and detection wavelength 260 nm. The content of nucleosides in four Cordyceps genus herbs was evaluated by fingerprint analysis and hierarchical cluster analysis( HCA). The calibration curves of five nucleosides showed good linear regression( r > 0. 99) and the average recoveries were between 95. 0 % and 105. 0 %. The contents of the five nucleosides in the four Cordyceps genus herbs were different and could be obviously distinguished by HCA. The fingerprint analysis result showed that the similarity between C. sinensis and the others was less than 0. 9. The method was accurate and reliable,which can be used for quality control of Cordyceps genus herbs.
To compare the main nucleosides in Cordyceps genus herbs( C. sinensis,C. millitaris,Hirsutella sinensis and C.sobolifera),an HPLC method for simultaneous determination of uridine,inosine,guanosine,adenosine and cordycepine in Cordyceps genus herbs was developed. The sample was extracted with 0. 5 % phosphoric acid solution to prepare test solution. The separation was performed on a Zorbax SB-Aq( 4. 6 mm × 150 mm,5 μm) column with gradient elution by 0. 04 mol· L- 1potassium dihydrogen phosphate solution and acetonitrile,column temperature 30 ℃,flow rate 0. 8 m L · min- 1,and detection wavelength 260 nm. The content of nucleosides in four Cordyceps genus herbs was evaluated by fingerprint analysis and hierarchical cluster analysis( HCA). The calibration curves of five nucleosides showed good linear regression( r > 0. 99) and the average recoveries were between 95. 0 % and 105. 0 %. The contents of the five nucleosides in the four Cordyceps genus herbs were different and could be obviously distinguished by HCA. The fingerprint analysis result showed that the similarity between C. sinensis and the others was less than 0. 9. The method was accurate and reliable,which can be used for quality control of Cordyceps genus herbs.
引文
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[2]胡敏,皮惠敏,郑元梅.冬虫夏草的化学成分及药理作用[J].时珍国医国药,2008,19(11):2804.
[3]郭海平,杨智敏.冬虫夏草药理作用研究进展[J].中草药,1999,30(3):231.
[4]董彩虹,李文佳,李增智,等.我国虫草产业发展现状、问题及展望——虫草产业发展金湖宣言[J].菌物学报,2016,35(1):1.
[5]吴雪晶,马慧君,刘宁.北虫草与冬虫夏草的药用价值比较[J].浙江食用菌,2009,17(5):12.
[6]杨钊,迟少云,张春辉,等.冬虫夏草及其代用品中腺苷和虫草素的LC-MS-MS定量分析研究[J].中国中药杂志,2007,32(19):2018.
[7]梅雪艳,李媛媛,沈于兰.冬虫夏草、虫草菌丝体及蛹虫草中核苷类成分的测定[J].药学与临床研究,2015,23(1):27.
[8]中国药典.一部[S].2015:115.
[9]谢丽娜.虫草花与冬虫夏草的区别[J].海峡药学,2010,22(7):54.
[10]中国药典.一部[S].2015:832.
[11]杨明静.蝉花发酵生产免疫活性组分ISP的研究[D].重庆:重庆大学,2008.
[12]钱正明,孙培培,李文庆,等.冬虫夏草高效液相特征图谱分析[J].世界科学技术——中医药现代化,2014,16(2):279.
[13]钱正明,周妙霞,孙敏甜,等.冬虫夏草不同部位核苷类成分比较分析[J].世界科学技术——中医药现代化,2014,16(11):2390.