HPLC-DAD法对不同生长年限瓦布贝母10种核苷和碱基含量的测定

doi:10.5246/jcps.2017.05.036

中国药学(英文版) ›› 2017, Vol. 26 ›› Issue (5): 346-354.DOI: 10.5246/jcps.2017.05.036

HPLC-DAD法对不同生长年限瓦布贝母10种核苷和碱基含量的测定

潘峰¹, 吴卫^1*, 董品利², 胡博¹, 官玲亮³, 舒玲¹

1. 四川农业大学农学院, 四川成都 611130
2. 四川省茂县农业局, 四川茂县 623200
3. 中国热带农业科学院热带作物品种资源研究所, 海南儋州 571737

收稿日期:2017-03-13 修回日期:2017-04-10 出版日期:2017-05-26 发布日期:2017-04-30
通讯作者: Tel./Fax: +86-028-86290870, E-mail: ewuwei@sicau.edu.com
基金资助:
The Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant No. 20115103110009) and “211” Project Double-Support Plan of Sichuan Agricultural University (Grant No. 03570313) and Modernization of Chinese Traditional Medicines in Hainan Province (Grant No. ZY201410).

Simultaneous determination of 10 nucleosides and nucleobases from different cultivation years of Fritillaria unibracteata var. wabuensis by HPLC-DAD

Feng Pan¹, Wei Wu^1*, Pinli Dong², Bo Hu¹, Lingliang Guan³, Ling Shu¹

1. Agronomy College, Sichuan Agricultural University, Chengdu 611130, Sichuan, China
2. Agriculture Bureau of Maoxian County, Maoxian 623200, Sichuan, China
3. Chinese Academy of Tropical Agricultural Sciences, Tropical Crops Genetic Resources Institute, Danzhou 571737, Hainan, China

Received:2017-03-13 Revised:2017-04-10 Online:2017-05-26 Published:2017-04-30
Contact: Tel./Fax: +86-028-86290870, E-mail: ewuwei@sicau.edu.com
Supported by:
The Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant No. 20115103110009) and “211” Project Double-Support Plan of Sichuan Agricultural University (Grant No. 03570313) and Modernization of Chinese Traditional Medicines in Hainan Province (Grant No. ZY201410).

摘要/Abstract

摘要：

核苷和碱基具有多种生理生化活性, 定量测定瓦布贝母的含量对其质量控制也很重要。本文建立了高效液相色谱-二极管阵列检测器联用技术(HPLC-DAD)同时测定瓦布贝母鳞茎中6种核苷(鸟苷、胞嘧啶核苷、胸苷、腺苷、尿苷、肌苷)和4种碱基(胞嘧啶、尿嘧啶、胸腺嘧啶、腺嘌呤)成分含量的测定方法。采用Zorbax 300 Å 300 Extend C₁₈色谱柱(150 mm×4.6 mm, 5 µm), 检测波长为260 nm, 以甲醇和水为流动相梯度洗脱, 以1.0 mL/min的流速在30 min以内能够实现所选用的核苷和碱基实现完全分离。采用优化色谱条件后, 所有10种待测成分具有良好的线性关系(R²≥0.9993), 精密度(RSD≤3.715%), 重复性(RSD≤3.748%),稳定性(RSD≤5.545%) 和加样回收率(97.688%-102.923%)。利用所建立的色谱方法成功地应用于不同生长年限瓦布贝母核苷和碱基的测定, 并且首次分析了不同生长年限(1–7年)瓦布贝母的10种核苷和碱基的含量变化。本研究将有助于对瓦布贝母的栽培时间选择、质量控制、药理研究和临床应用。

关键词: 定量检测, 核苷和碱基, 瓦布贝母, HPLC-DAD

Abstract:

Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis (FUW) due to their physiological and pharmacologicalactions. In the present study, we developed a sensitive and reliable HPLC-diode-array detection method to simultaneously determine ten nucleosides and nucleobases, including cytosine, uracil, cytidine, uridine, thymine, adenine, inosine, guanosine, thymidine and adenosine. Complete separation of all the analytes was achieved on a Zorbax 300 Å 300 Extend C₁₈ column with a gradient of methanol-ultrapure water at a ﬂow rate of 1 mL/min in less than 30 min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. The optimized method provided good linearity (R²≥0.9993 for all the analytes), satisfactory precision (RSD≤3.715%), good repeatability (RSD≤3.748%) and good recovery (RSD from 97.688% to 102.923%). In addition, the developed method was successfully applied to simultaneous determination of ten nucleosides and nucleobases from FUW, and their content changes of various cultivation time (1-7 years) were further analyzed for the ﬁrst time. Our findings were useful for ensuring the cultivation time choice of artificial cultivation, quality control, pharmaceutical studies and clinical efﬁcacy of FUW.

Key words: Quantitative determination, Nucleosides and nucleobases, Fritillaria unibracteata var. wabuensis, HPLC-DAD

中图分类号:

R917

Supporting:

潘峰, 吴卫, 董品利, 胡博, 官玲亮, 舒玲. HPLC-DAD法对不同生长年限瓦布贝母10种核苷和碱基含量的测定[J]. 中国药学(英文版), 2017, 26(5): 346-354.

Feng Pan, Wei Wu, Pinli Dong, Bo Hu, Lingliang Guan, Ling Shu. Simultaneous determination of 10 nucleosides and nucleobases from different cultivation years of Fritillaria unibracteata var. wabuensis by HPLC-DAD[J]. Journal of Chinese Pharmaceutical Sciences, 2017, 26(5): 346-354.

参考文献

[1] Wang, D.; Wang, S.; Chen, X.; Xu, X.; Zhu, J.; Nie, L.; Long, X. J. Ethnopharmacol. 2012, 139, 189-193.

[2] Ozler, H.; Pehlivan, S. Bangladesh J. Botan. 2007, 36, 111-120.

[3] Gill, B.D., Ph. D. thesis, University of Waikato. 2013.

[4] Brohi, R.O.Z.Z.; Khuhawar, M.Y.; Khuhawar, T.M.J. J. Anal. Sci. Technol. 2016, 7, 1.

[5] Qiu, X.L.; Xu, X.H.; Qing, F.L. Tetrahedron. 2010, 66, 789-843.

[6] Schmidt, A.P.; Lara, D.R.; de Faria Maraschin, J.; da Silveira Perla, A.; Onofre Souza, D. Brain Res. 2000, 864, 40-43.

[7] Virág, L.; Szabo, C. FASEB. J. 2001, 15, 99-107.

[8] Ichikawa, S.; Matsuda, A. Expert Opin. Ther. Pat. 2007, 17, 487-498

[9] Isono, K. Pharmacol. Ther. 1991, 52, 269-286.

[10] Gudbjörnsson, B.; Zak, A.; Niklasson, F.; Hällgren, R. Ann. Rheum. Dis. 1991, 50, 669-672.

[11] Itoh, K.; Konno, T.; Sasaki, T.; Ishiwata, S.; Ishida, N.; Misugaki, M. Clin. Chim. Acta. 1992, 206, 181-189.

[12] Zheng, Y.F.; Yang, J.; Zhao, X.J.; Feng, B.; Kong, H.W.; Chen, Y.J.; Lv, S.; Zheng, M.H.; Xu, G.W. World J. Gastroenterol. 2005, 11, 3871-3876.

[13] Zheng, Y.F.; Kong, H.W.; Xiong, J.H.; Lv, S.; Xu, G.W. Clin. Biochem. 2005, 38, 24-30.

[14] Gutiérrez-Castrellón, P.; Mora-Magana, I.; Díaz-García, L.; Jiménez-Gutiérrez, C.; Ramirez-Mayans, J.; Solomon-Santibánez, G.A. Br. J. Nutr. 2007, 98, S64-S67.

[15] Kang, D.G.; Oh, H.; Cho, D.K.; Kwon, E.K.; Han, J.H.; Lee, H.S. J. Ethnopharmacol. 2002, 81, 49-55.

[16] Kang, D.G.; Sohn, E.J.; Lee, Y.M.; Lee, A.S.; Han, J.H.; Kim, T.Y.; Lee, H.S. J. Ethnopharmacol. 2004, 91, 51-56.

[17] Cao, X.W., Ph. D. thesis, Peking Union Medical College, 2008.

[18] Zhou, J.L.; Jiang, Y.; Bi, Z.M.; Li, P. Chin. Pharm. J. 2008, 43, 894-896.

[19] Chang, S.C.; Hsu, L.W. US 09/853,524, 2002.

[20] Hao, D.C.; Gu, X.J.; Xiao, P.G.; Peng, Y. Chin. J. Nat. Med. 2013, 11, 330-344.

[21] Huang, L.; Duan, B.; Wang, L.; Wei, D.; Lu, Q.; Chen, S. China J. Chin. Mater. Med. 2011, 36, 585-588.

[22] Wu, L.; Chen, L.; Selvaraj, J.N.; Wei, Y.; Wang, Y.; Li, Y.; Zhao, J.; Xue, X. Food Chem. 2015, 173, 1111-1118.

[23] Peng, R.; Ma, P.; Mo, R.; Sun, N. Acta Pharm. Sin. B. 2013, 3, 167-173.

[24] Zong, Y.; Wang, Y.; Li, H.; Li N.; Zhang, H.; Sun, J.; Niu, X.; Gao, X. Pharmacogn. Mag. 2014, 10, 391.

[25] Wang, X.Z.; Wu, H.; Li, N.; Cheng, Y.; Wen, H.M.; Liu, R.; Chai, C. Food Anal. Methods. 2015, 1-12.

[26] Yun, H.X.; Chen, Z. Lishizhen Med. Mater. Med. Res. 2010, 21, 1057-1058.

[27] Liu, R.M.; Zhang, X.J.; Li, X.F.; Xiao, J.H.; Zhong, J.J. Lat. Am. J. Pharm. 2015, 34, 283-290.

[28] Wen, Q.F.; Zhang, Y.F.; Zhang, J.Q.; Zhao, H. J. Chin. Pharm. Sci. 2016, 25, 906-913.

[29] Cao, X.W.; Li, J.; Chen, S.B.; Li, X.B.; Xiao, P.G.; Chen, S.L.; Yang, D.J. J. Sep. Sci. 2010, 33, 1587-1594.

[30] Zhang, D.Y.; Wang, S.; Li, Q.; Liu, Z.D.; Wang, C. W. China J. Pharm. Sci. 2010, 25, 725-726.

[31] Damaraju, D.; Damaraju, V.L.; Brun, M.; Mowles, D.; Kuzma, M.; Berendt, R.C.; Sawyer, M.B.; Cass, C.E. Biochem. Pharm. 2008, 75, 1901-1911.

[32] Kakuda, T.N. Clin. Ther. 2000, 22, 685-708.

[33] Wagner, H.; Ulrich-Merzenich, G. Evidence and Rational Based Research on Chinese Drugs. New York: Springer-Verlag Wien. 2013, 489-516.

HPLC-DAD法对不同生长年限瓦布贝母10种核苷和碱基含量的测定

Simultaneous determination of 10 nucleosides and nucleobases from different cultivation years of Fritillaria unibracteata var. wabuensis by HPLC-DAD

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	金祥飞, 程闽玥, 钟斯卉, 徐云根, 沈卫阳. 棕榈酸曲安奈德中未知杂质的鉴定[J]. 中国药学(英文版), 2022, 31(4): 270-278.
[2]	崔晓雨, 王攀, 莫伊点, 丁旭阳, 陈世忠, 郭洪祝, 胡琴, 李中军. HPLC-ELSD法测定菊粉中菊粉型果寡糖的含量[J]. 中国药学(英文版), 2022, 31(1): 23-30.
[3]	仝令坤, 王弘, 张笑天, 郭宁宁, 韩雅, 王岱东, 陈世忠. 中药罗布麻叶中化学成分的LC/MS分析及其最佳采收期的研究[J]. 中国药学(英文版), 2018, 27(4): 251-262.
[4]	程玲, 汪小涵, 申宝德, 徐平华, 沈成英, 郑娟, 廖魏波, 徐荣, 袁海龙. HPLC-DAD法同时测定六味五灵片中7种活性成分含量[J]. 中国药学(英文版), 2017, 26(4): 304-311.
[5]	曹规划, 付庆荣, 张藏曼, 王弘, 陈世忠. HPLC-DAD-ESI-MSn技术对橘叶(福橘叶)中96个化学成分的快速分离和鉴定[J]. 中国药学(英文版), 2016, 25(2): 91-110.
[6]	曹晨, 王静哲, 王璐, 尚明英, 刘广学, 徐风, 王璇, 蔡少青. 高效液相色谱法同时测定细辛中7种主要成分的含量[J]. 中国药学(英文版), 2015, 24(8): 530-537.
[7]	原红霞, 李慧峰, 裴妙荣. HPLC-DAD-ELSD法同时测定小柴胡汤正丁醇有效部位中7种成分的含量[J]. 中国药学(英文版), 2015, 24(4): 236-240.
[8]	范辉, 郭姣^*, 陈媛媛. HPLC-DAD-ELSD法同时测定复方贞术调脂胶囊15个活性成分[J]. , 2013, 22(1): 47-54.
[9]	赵明波, 魏荷琳, 李军, 屠鹏飞^*. 高效液相色谱法测定北刘寄奴药材中木犀草素和毛蕊花糖苷的含量[J]. , 2012, 21(4): 333-337.
[10]	李晓娜, 于宁, 张建美, 林文斯, 凌笑梅^*, 富戈, 李润涛, 崔景荣. 在线SPE HPLC-DAD方法测定大鼠血浆环磷酰胺浓度[J]. , 2012, 21(2): 156-161.
[11]	杨琳, 郭晓宇^, 王齐放, 陈颖, 车亦馨, 车庆明^. 大鼠血清中黄芩素代谢产物的鉴定[J]. , 2011, 20(3): 275-281.
[12]	王楠, 李春凤, 张斗胜, 胡昌勤^*. 比阿培南及杂质质控HPLC分析方法的建立[J]. , 2011, 20(2): 171-180.
[13]	范莉, 濮润, 赵海誉, 刘璇, 马超, 王宝荣, 果德安^*. HPLC-DAD-ESI-MSn法研究红花注射液中羟基红花黄色素A和脱水红花黄色素B的稳定性及降解过程[J]. , 2011, 20(1): 47-56.
[14]	宋丽军, 谭晓梅, 赵文昌, 罗佳波^*. HPLC-DAD法同时定量分析葛根芩连微丸中12种活性成分[J]. , 2010, 19(6): 464-470.
[15]	薛彬彬, 郭晓宇, 车庆明^* . 大鼠灌胃灵芝总三萜后血浆中灵芝酸类成分的检测[J]. , 2008, 17(3): 236-240.