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Journal of Chinese Pharmaceutical Sciences ›› 2017, Vol. 26 ›› Issue (5): 346-354.DOI: 10.5246/jcps.2017.05.036

• Original articles • Previous Articles     Next Articles

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

Feng Pan1, Wei Wu1*, Pinli Dong2, Bo Hu1, Lingliang Guan3, Ling Shu1   

  1. 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).

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

CLC Number: 

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