Zheng Li-hong, Huang Li-ying, Chen Yu, Lin Shou-er, Huang Bing-lan
Zhong Yao Cai. 2015 Nov;38(11):2269-73.
To establish a method for simultaneous determination of nucleosides and nucleobases in natural, cultured and tissue culture Anoectochilus roxburghii by high performance liquid chromatography-electrospray ionization/ion trap mass spectrometry (HPLC-ESI/MS).
The separation was performed on a Welch Ultimate XB-C18 column (250 mm x 4.6 mm,5 μm). 20 mmol/L ammonium acetate solution and methanol were adopted as the mobile phase with gradient elution. The flow rate was 1.0 mL/min. The injection volume was 20 μL. The column temperature and UV wavelength were set at 30 degrees C and 260 nm, respectively.
Cytosine, uracil, cytidine, uridine, hypoxanthine, adenine, inosine, guanosine,fl-thymidine and adenosine were identified in natural, cultured and tissue culture Anoectochilus roxburghii. The total content of nucleosides and nucleotides in Anoectochilus roxburghii were 1.6639, 1.8568 and 2.2013 mg/g,respectively.
The contents of nucleosides and nucleobases in herb are affected by its growth pattern. The total content of nucleosides and nucleotides was tissue culture herb > cultured herb > natural herb. This investigation would provide the theoretic basis for quality standards and applications of Anoectochilus roxburghii in clinical research.
建立高效液相色谱 - 电喷雾电离/离子阱质谱法（HPLC - ESI/MS）同时测定天然、栽培及组培金线莲中核苷和碱基的方法。
采用Welch Ultimate XB - C18柱（250 mm×4.6 mm，5μm）进行分离。以20 mmol/L醋酸铵溶液和甲醇为流动相，梯度洗脱。流速为1.0 mL/min。进样量为20μL。柱温设定为30℃，紫外波长设定为260 nm。
在天然、栽培及组培金线莲中鉴定出胞嘧啶、尿嘧啶、胞苷、尿苷、次黄嘌呤、腺嘌呤、肌苷、鸟苷、β - 胸腺嘧啶核苷和腺苷。金线莲中核苷和核苷酸的总含量分别为1.6639、1.8568和2.2013 mg/g。
药材中核苷和碱基的含量受其生长方式的影响。核苷和核苷酸的总含量为组培药材＞栽培药材＞天然药材。本研究将为金线莲质量标准及临床研究应用提供理论依据。
