基于多指标成分含量的冬虫夏草冻干和晒干法比较
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摘要
该文建立HPLC同时测定冬虫夏草中主要核苷及碱基成分,考察冻干和晒干2种加工方法对其含量的影响。以次黄嘌呤、黄嘌呤、尿苷、肌苷、鸟苷、腺苷等6种主要核苷及碱基成分为对照品,采用GL Inertsustain AQ-C_(18)色谱柱(4. 6 mm×250mm,5μm);水(A)-乙腈(B)为流动相,梯度洗脱(0～10 min,0～1%B; 10～65 min,1%～3%B),流速1. 0 mL·min~(-1),检测波长260 nm,柱温30℃,进样量5μL。次黄嘌呤、黄嘌呤、尿苷、肌苷、鸟苷和腺苷分别在1. 025～20. 50(r=0. 999 8),0. 545～10. 90(r=0. 999 9),4. 051～81. 02(r=0. 999 8),4. 044～80. 88(r=0. 999 9),2. 075～41. 50(r=0. 999 9),4. 032～80. 64(r=0. 999 9)mg·L~(-1)呈良好的线性关系,平均回收率分别为102. 3%(RSD 2. 1%),101. 1%(RSD 2. 4%),97. 80%(RSD 1. 7%),101. 8%(RSD 1. 8%),98. 90%(RSD 2. 0%),98. 10%(RSD 1. 4%)。每批野生冬虫夏草样品采用冻干和晒干2种干燥方式干燥后用于比较研究。测定结果表明,不同干燥方式对冬虫夏草中的6种核苷及碱基类成分的含量存在一定影响,冻干法中核苷类成分的总含量高于晒干法。该研究为冬虫夏草干燥方式及质量评价提供了科学依据。
The aim of this paper was to compare the influence of freeze-drying and sun-drying on six main nucleosides and nucleobases of Cordyceps sinensis by HPLC. Hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were reference substances. HPLC analysis was performed on a GL Inertsustain AQ-C_(18) column( 4. 6 mm×250 mm,5 μm),with mobile phase consisting of water( A)-acetonitrile( B) at the flow rate of 1. 0 mL·min~(-1)( 0-10 min,0-1% B; 10-65 min,1%-3% B). The detection wavelength was 260 nm,the column temperature was controlled at 30 ℃,and the injection volume was 5 μL. The linear ranges of hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were 1. 025-20. 50( r = 0. 999 8),0. 545-10. 90( r = 0. 999 9),4. 051-81. 02( r = 0. 999 8),4. 044-80. 88( r= 0. 999 9),2. 075-41. 50( r= 0. 999 9),4. 032-80. 64( r = 0. 999 9) mg·L~(-1),respectively. The average recoveries of them( n = 6)were as follows: 102. 3%( RSD 2. 1%),101. 1%( RSD 2. 4%),97. 80%( RSD 1. 7%),101. 8%( RSD 1. 8%),98. 90%( RSD2. 0%) and 98. 10%( RSD 1. 4%),respectively. Each sample was processed by freeze-drying and sun-drying so as to compare the difference between the two drying methods. The contents of six index ingredients were significantly different between freeze-drying and sun-drying sample of C. sinensis. The total contents of six index ingredients in sun-drying sample were higher than that in the corresponding freeze-drying sample. This research results provide the scientific basis for the drying methods and quality evaluation of C. sinensis.
The aim of this paper was to compare the influence of freeze-drying and sun-drying on six main nucleosides and nucleobases of Cordyceps sinensis by HPLC. Hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were reference substances. HPLC analysis was performed on a GL Inertsustain AQ-C_(18) column( 4. 6 mm×250 mm,5 μm),with mobile phase consisting of water( A)-acetonitrile( B) at the flow rate of 1. 0 mL·min~(-1)( 0-10 min,0-1% B; 10-65 min,1%-3% B). The detection wavelength was 260 nm,the column temperature was controlled at 30 ℃,and the injection volume was 5 μL. The linear ranges of hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were 1. 025-20. 50( r = 0. 999 8),0. 545-10. 90( r = 0. 999 9),4. 051-81. 02( r = 0. 999 8),4. 044-80. 88( r= 0. 999 9),2. 075-41. 50( r= 0. 999 9),4. 032-80. 64( r = 0. 999 9) mg·L~(-1),respectively. The average recoveries of them( n = 6)were as follows: 102. 3%( RSD 2. 1%),101. 1%( RSD 2. 4%),97. 80%( RSD 1. 7%),101. 8%( RSD 1. 8%),98. 90%( RSD2. 0%) and 98. 10%( RSD 1. 4%),respectively. Each sample was processed by freeze-drying and sun-drying so as to compare the difference between the two drying methods. The contents of six index ingredients were significantly different between freeze-drying and sun-drying sample of C. sinensis. The total contents of six index ingredients in sun-drying sample were higher than that in the corresponding freeze-drying sample. This research results provide the scientific basis for the drying methods and quality evaluation of C. sinensis.
引文
[1]中国药典.一部[S]. 2015:115.
[2]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.
[3]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.
[4]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.
[5]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.
[6]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.
[7]杨平,赵晓霞,张永文,等.发酵虫草菌类产品的标准比较与评价[J].中国中药杂志,2018,43(3):463.
[8] Zhao J,Xie J,Wang L Y,et al. Advanced development in chemical analysis of Cordyceps[J]. J Pharm Biomed Anal,2014,87(1434):271.
[9] Zhao H Q,Wang X,Li H M,et al. Characterization of nucleosides and nucleobases in natural Cordyceps by HILIC-ESI/TOF/MS and HILIC-ESI/MS[J]. Molecules,2013,18(8):9755.
[10] Yang F Q,Li D Q,Feng K,et al. Determination of nucleotides,nucleosides and their transformation products in Cordyceps by ionpairing reversed-phase liquid chromatography-mass spectrometry[J]. J Chromatogr A,2010,1217(34):5501.
[11]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.
[12]金士元.中药材传统鉴别经验[M].北京:中国中医药出版社,2010:296.
[13]任迪峰,毛志怀.我国中草药干燥的现状及发展趋势[J].农业工程学报,2001,17(2):5.
[14]桑迎迎,周国燕,王爱民,等.中药材干燥技术研究进展[J].中成药,2010,32(12):2140.
[15]刘苗苗,叶利春,陈立军,等.真空冷冻干燥技术在中药研究中的应用[J].中药材,2014,37(5):909.
[16]詹娟娟,伍振峰,王雅琪,等.中药材及制剂干燥工艺与装备现状及问题分析[J].中国中药杂志,2015,40(23):4715.
[17]马茹,龚燚婷,刘璘,等.低真空度下冷冻干燥技术在中药鲜药保存方面的应用分析[J].中国实验方剂学杂志,2017,23(21):30.
[18]钱正明,甄达明,李文庆,等.冬虫夏草水提取过程中腺苷转化途径研究[J].世界中医药,2016,11(5):758.
[2]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.
[3]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.
[4]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.
[5]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.
[6]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.
[7]杨平,赵晓霞,张永文,等.发酵虫草菌类产品的标准比较与评价[J].中国中药杂志,2018,43(3):463.
[8] Zhao J,Xie J,Wang L Y,et al. Advanced development in chemical analysis of Cordyceps[J]. J Pharm Biomed Anal,2014,87(1434):271.
[9] Zhao H Q,Wang X,Li H M,et al. Characterization of nucleosides and nucleobases in natural Cordyceps by HILIC-ESI/TOF/MS and HILIC-ESI/MS[J]. Molecules,2013,18(8):9755.
[10] Yang F Q,Li D Q,Feng K,et al. Determination of nucleotides,nucleosides and their transformation products in Cordyceps by ionpairing reversed-phase liquid chromatography-mass spectrometry[J]. J Chromatogr A,2010,1217(34):5501.
[11]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.
[12]金士元.中药材传统鉴别经验[M].北京:中国中医药出版社,2010:296.
[13]任迪峰,毛志怀.我国中草药干燥的现状及发展趋势[J].农业工程学报,2001,17(2):5.
[14]桑迎迎,周国燕,王爱民,等.中药材干燥技术研究进展[J].中成药,2010,32(12):2140.
[15]刘苗苗,叶利春,陈立军,等.真空冷冻干燥技术在中药研究中的应用[J].中药材,2014,37(5):909.
[16]詹娟娟,伍振峰,王雅琪,等.中药材及制剂干燥工艺与装备现状及问题分析[J].中国中药杂志,2015,40(23):4715.
[17]马茹,龚燚婷,刘璘,等.低真空度下冷冻干燥技术在中药鲜药保存方面的应用分析[J].中国实验方剂学杂志,2017,23(21):30.
[18]钱正明,甄达明,李文庆,等.冬虫夏草水提取过程中腺苷转化途径研究[J].世界中医药,2016,11(5):758.