Journal of Chinese Pharmaceutical Sciences ›› 2018, Vol. 27 ›› Issue (3): 201-208.DOI: 10.5246/jcps.2018.03.021

• Original articles • Previous Articles     Next Articles

Development and validation of an HPLC-UV method for the separation of entecavir optical isomers

Dan Huang1, Daolin Zhang2, Nana Wang3, Xinhui Jiang1*   

  1. 1. School of pharmacy, Chongqing Medical University, Yuzhong District, Chongqing 400016, China
    2. Chongqing Pharmaceutical Research Institute Co., Chongqing Engineering Research Center for Chemical Pharmaceutical, Nan'an District, Chongqing 400016, China
    3. Luohe Third People's Hospital of Henan Province, Luohe, Henan 462000, China
  • Received:2017-12-13 Revised:2018-01-10 Online:2018-03-31 Published:2018-02-26
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A simple and effective high-performance liquid chromatography with ultraviolet detection (HPLC-UV) method was established to determine entecavir optical isomers. With 3 chiral carbon atoms, entecavir had 7 optical isomers, including 6 diastereoisomers and 1 enantiomer. The separations were performed on a Symmetrix ODS-AQ C18 column (4.6 mm×250 mm, 5 µm) and a Daicel CHIRALPAK AD column (4.6 mm×250 mm,10 µm), respectively. The detection wavelength was set at 254 nm.The limit of detection (LOD) and the limit of quantification (LOQ) of diastereoisomers (diastereoisomer-1, diastereoisomer-2, diastereoisomer-3) were 0.0371, 0.0376, 0.0377, and 0.124, 0.125, 0.126 µg/mL, respectively. The LOD and LOQ of enantiomer were 0.14 and 0.46 µg/mL, respectively. The precision was within 0.36%, 0.44%, 1.04%, and 0.67% for diastereoisomer-1, diastereoisomer-2, diastereoisomer-3, and enantiomer, respectively. The recoveries of enantiomers ranged from 98.4% to 100.5%. The method could be applied to control the quality of entecavir.

Key words: Entecavir, Chiral separation, Diastereoisomers, Enantiomer, HPLC-UV

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