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目的建立高效毛细管电泳法(HPCE)同时测定不同产地桦褐孔菌中腺嘌呤、尿嘧啶、腺苷、次黄嘌呤、尿苷、鸟苷和肌苷等7种核苷类成分含量的方法。方法采用未涂渍标准熔融石英毛细管(75μm×64.5 cm,有效长度56 cm)为分离通道,55 mmol·L~(-1)硼砂(p H 9.5)为运行缓冲液,分离电压为20 k V,检测波长为260 nm,毛细管温度为30℃,压力进样为5 k Pa×6s。结果 7种核苷类成分的响应峰面积与其相应浓度的线性关系良好(r>0.998 8);加样回收率为96.46%~102.12%。实验表明,不同产地桦褐孔菌中7种核苷类成分的含量有所差异。结论该方法准确、可靠,重复性较好,可用于桦褐孔菌内在质量的评价和控制。
OBJECTIVE To establish a high performance capillary electrophoresis (HPCE) method for the simultaneous determination of seven nucleosides, including adenine, uracil, adenosine, hypoxanthine, uridine, guanosine and inosine in different habitats. Methods Uncoated standard fused silica capillary (75 μm × 64.5 cm, effective length 56 cm) was used as separation channel, and 55 mmol·L -1 borax (p H 9.5) was used as running buffer. The separation voltage was 20 kV , The detection wavelength is 260 nm, the capillary temperature is 30 ℃ and the pressure is 5 kPa × 6s. Results The linear relationship between the peak areas of seven nucleoside components and their corresponding concentrations was good (r> 0.998 8). The recoveries of samples ranged from 96.46% to 102.12%. Experiments show that, the content of seven nucleosides in different habitats of Inonotus obliquus varies. Conclusion The method is accurate, reliable and reproducible. It can be used to evaluate and control the intrinsic quality of Inonotus obliquus.