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本实验建立了同时测定天麻药材中天麻素及8种核苷和碱基类成分的高效液相色谱法:Agilent Zorbax BonusRP(4.6 mm×250 mm,5μm)色谱柱,甲醇-(0.04%冰乙酸)水溶液为流动相,梯度洗脱,柱温36℃,流速1.0 m L·min-1,检测波长254 nm,进样量20μL。该方法分离度良好,天麻素、胞嘧啶、胞苷、尿嘧啶、腺嘌呤、尿苷、胸腺嘧啶、鸟苷和腺苷等的质量浓度在2.04~262.00,0.20~24.67,0.18~23.75,0.20~25.83,0.20~26.67,0.16~20.00,0.22~27.71,0.20~24.29,0.24~30.58 mg·L-1内,相关系数r在0.998 9~0.999 9,具有良好的线性。天麻素和8种核苷及碱基平均加标回收率在96.4%~99.6%,RSD均小于2.7%(n=6)。测定了7个西藏栽培天麻药材中上述9种成分的含量。结果表明,7个天麻样品中天麻素的含量均高于2.0 mg·g-1,均含有较高的腺苷、鸟苷、尿苷和胞嘧啶,而胞苷、尿嘧啶、腺嘌呤和胸腺嘧啶含量较低。该方法简便、准确,重复性好,适用于天麻药材中天麻素和腺苷等8种核苷及碱基类成分的含量测定。
In this study, a gas chromatogram was established for the simultaneous determination of gastrodin and eight nucleosides and bases in Gastrodia elata Blume: Agilent Zorbax Bonus RP (4.6 mm × 250 mm, 5 μm) column with methanol- (0.04% glacial acetic acid ) As mobile phase and gradient elution.The column temperature was 36 ℃, the flow rate was 1.0 m L · min-1, the detection wavelength was 254 nm, and the injection volume was 20 μL. The method was well separated and the mass concentrations of gastrodin, cytosine, cytidine, uracil, adenine, uridine, thymine, guanosine and adenosine were in the range of 2.04-262.00,0.20-24.67,0.18-23.75,0.20 ~ 25.83,0.20 ~ 26.67,0.16 ~ 20.00,0.22 ~ 27.71,0.20 ~ 24.29,0.24 ~ 30.58 mg · L-1, the correlation coefficient r was between 0.998 9 and 0.999 9, with good linearity. The average recoveries of gastrodin and eight nucleosides and bases ranged from 96.4% to 99.6% with RSDs less than 2.7% (n = 6). The contents of the above nine components in seven Tibetan cultivated Gastrodia elata Blume were determined. The results showed that the content of gastrodin in 7 samples of Tianma was higher than 2.0 mg · g-1, both of them contained high adenosine, guanosine, uridine and cytosine, but cytidine, uracil, adenine and thymus Pyrimidine content is lower. The method is simple, accurate and reproducible, and is suitable for determining the content of eight nucleosides and base components of gastrodin and adenosine in Gastrodia elata Blume.