在线梯度提取-液相色谱法分析冬虫夏草中的化学成分
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摘要
该研究建立了在线梯度提取-液相色谱法同时分析冬虫夏草中大极性和小极性类化学成分。将冬虫夏草样品与硅藻土均匀混合,置于提取罐中,并安装于高效液相系统中,于70℃下用流动相进行在线梯度提取。分析柱为Zorbax SB-AQ(4. 6 mm×150 mm,5μm),流动相为0. 1%甲酸水-甲醇,梯度洗脱,流速1 mL·min~(-1),柱温30℃,在260 nm波长下检测。结果发现,该方法可以同时提取和分离冬虫夏草中大极性和小极性类成分,并通过与对照品色谱峰进行比对的方法,鉴定了其中6个大极性成分(尿嘧啶、尿苷、胸腺嘧啶、肌苷、鸟苷、腺苷)和1个小极性成分(麦角甾醇)。结果表明,所建立的分析方法快速、稳定、环保,有助于提升冬虫夏草的综合质量评价水平。
Online gradient extraction-high performance liquid chromatography( HPLC) method was developed for simultaneous determination of high and low polar components in Cordyceps. The sample powder of Cordyceps was uniformly mixed with diatomaceous earth,packed into extraction tank,and installed into the HPLC system. Online gradient extraction was conducted with mobile phase at 70 ℃. The separation was performed on Zorbax SB-AQ( 4. 6 mm×150 mm,5 μm) column with 0. 1% formic acid solution-methanol as the mobile phase for gradient elution at 1. 0 mL·min~(-1). The column temperature was 30 ℃,and detection wavelength was set at 260 nm. The results showed that the high and low polar components in Cordyceps could be simultaneously extracted and separated by the developed method. Meanwhile,six high polar compounds( uracil,uridine,thymine,inosine,guanosine and adenosine) and one low polar compound( ergosterol) were identified by comparison with the reference peaks. The established method is rapid,stable and environment friendly,which is helpful to improve the quality evaluation level for Cordyceps.
Online gradient extraction-high performance liquid chromatography( HPLC) method was developed for simultaneous determination of high and low polar components in Cordyceps. The sample powder of Cordyceps was uniformly mixed with diatomaceous earth,packed into extraction tank,and installed into the HPLC system. Online gradient extraction was conducted with mobile phase at 70 ℃. The separation was performed on Zorbax SB-AQ( 4. 6 mm×150 mm,5 μm) column with 0. 1% formic acid solution-methanol as the mobile phase for gradient elution at 1. 0 mL·min~(-1). The column temperature was 30 ℃,and detection wavelength was set at 260 nm. The results showed that the high and low polar components in Cordyceps could be simultaneously extracted and separated by the developed method. Meanwhile,six high polar compounds( uracil,uridine,thymine,inosine,guanosine and adenosine) and one low polar compound( ergosterol) were identified by comparison with the reference peaks. The established method is rapid,stable and environment friendly,which is helpful to improve the quality evaluation level for Cordyceps.
引文
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[13]过立农,张美,刘杰,等.冬虫夏草人工繁育品与野生品基于甾醇特征图谱的比较研究[J].中国药事,2017,31(8):951.
[2] Liu Y,Wang J H,Wang W,et al. The chemical constituents and pharmacological actions of Cordyceps sinensis[J]. Evid Based Complement Alternat Med,2015,doi:10. 1155/2015/575063.
[3]钱正明,李文庆,王传喜,等.高效液相色谱定量分析比较4种虫草药材的核苷类成分[J].中国中药杂志,2016,41(13):2493.
[4]白雪莲,杨淑贤,单宇,等.冬虫夏草人工繁育品对人白血病K562细胞增殖和凋亡的影响[J].中国中药杂志,2018,43(10):2134.
[5]昝珂,黄莉莉,过立农,等.基于特征图谱及多指标成分含量的冬虫夏草野生与人工繁育品比较研究[J].中国中药杂志,2017,42(20):3957.
[6]诸葛定娟,陈爱瑛,程敏,等.虫草多糖调节小鼠T淋巴细胞及其亚群数量的机制研究[J].中国现代应用药学,2017,34(2):171.
[7]钱正明,孙敏甜,李华,等.冬虫夏草水提液和醇提液HPLC指纹图谱分析[J].世界中医药,2017,12(12):3133.
[8] Song Y L,Song Q Q,Li J,et al. Chromatographic analysis of Polygalae Radix by online hyphenating pressurized liquid extraction[J]. Sci Rep,2016,6:27303.
[9] Song Q Q,Li J,Liu X,et al. Home-made online hyphenation of pressurized liquid extraction,turbulent flow chromatography,and high performance liquid chromatography,Cistanche deserticola as a case study[J]. J Chromatogr A,2016,1438:189.
[10]秦伟瀚,花雷,郭延垒,等.UPLC-Q-TOF-MS结合代谢组学分析冬虫夏草不同部位的差异性[J].中国实验方剂学杂志,2018,24(21):69.
[11] Tong C Y,Peng M J,Tong R N,et al. Use of an online extraction liquid chromatography quadrupole time-of-flight tandem mass spectrometry method for the characterization of polyphenols in Citrus paradisi cv. Changshanhuyu peel[J]. J Chromatogr A,2018,1533:87.
[12]钱正明,李春红,李文庆,等.在线提取HPLC法测定两种虫草中腺苷和虫草素的含量[J].今日药学,2018,28(6):387.
[13]过立农,张美,刘杰,等.冬虫夏草人工繁育品与野生品基于甾醇特征图谱的比较研究[J].中国药事,2017,31(8):951.