高效毛细管电泳法同时测定桦褐孔菌中7种核苷类成分的含量

Simultaneous Determination of Seven Nucleosides in Inonotus obliquus by HPCE

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 simultaneous determination of seven nucleosides and nucleobases, including adenine, uracil, adenosine, hypoxanthine, uridine, guanosine, and inosine in Inonotus obliquus samples of different batches. METHODS Based on the mode of HPCE, uncoated fused silica capillary （75 μm×64.5 cm , 56 cm of effective length ） was used with separation voltage of 20 kV. The 55 mmol·L-1 sodium borate solution was selected as the running buffer solution （pH 9.5）. The detection wavelength was set at 260 nm. The sample was injected at 5 kPa×6 s and column temperature was maintained at 30 ℃. RESULTS Good linear relationship was obtained between the response peak areas and the concentrations of all the seven nucleosides （r〉0.998 8）. The average recoveries of the method were between 96.46%-102.12%. The contents of the seven nucleosides in the Inonotus obliquus samples of different origins were different. CONCLUSION The established method is reliable, accurate, and can be used for the quality control of Inonotus obliquus.