Simultaneous determination of nucleosides and nucleobases in Cervi Cornu Pantotrichum of sika deer (Cervus nippon) with different processing methods by UPLC / 中草药

ABSTRACT

Objective To establish the method of simultaneous determination of the content of 13 nucleosides and nucleobases, including cytosine, uracil, adenine, guanine, 6-hydroxypurine, 2,6-dihydroxypurine, uridine, thymine, inosine, guanosine, adenosine, 2′-deoxyguanosine (2′-dG), beta-thymidine, in Cervi Cornu Pantotrichum (CCP) of sika deer (Cervus nippon) by UPLC, and compare the distribution differences of nucleosides and nucleobases in different zones of the CCP with different processing methods. Methods The nucleosides and nucleobases in CCP were extracted by water with assistance of ultrasound. Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) was used as chromatographic column to separate the nucleosides and nucleobases. Thirteen target compounds were eluted with acetonitrile 100% (eluent A) and water plus 0.006% formic acid (eluent B) at a flow rate of 0.3 mL/min. The column temperature was 30 ℃, and the injection volume was 3 μL and the detection wavelength was 260 nm. Results A total of 13 nucleosides and nucleobases basically reached the baseline separation with a good linearity within linear range (r > 0.999 6). The nucleosides and nucleobases content in wax slices, powder slices, gauze slices of CCP without and with blood were respectively 4.47, 3.95, 2.68 g/kg and 4.14, 3.44, 2.51 g/kg. And those three parts of CCP with boiling and freeze-drying processing were respectively 4.60, 2.95, 2.74 g/kg and 5.06, 4.24, 2.31 g/kg. Conclusion As far as the total content of nucleosides and nucleobases were concerned, the wax slices, powder slices and gauze slices of CCP without blood were all higher than those of CCP with blood. The wax slices, powder slices of CCP with freeze-drying processing were more than those of CCP with boiling processing, while the gauze slices of which were less than those of CCP with boiling processing.