高效液相色谱法测定大鼠脑组织中腺苷与肌苷的含量

Determination of Adenosine and Inosine in Rat Brain Tissues by High Performance Liquid Chromatography

目的建立大鼠脑组织中腺苷与肌苷的高效液相色谱检测方法。方法脑组织用提取液(含0.1mol·L-1高氯酸溶液和0.2mmol·L-1乙二胺四醋酸二钠溶液)匀浆提取后进样。色谱柱用Agilent HC-C18(4.6mm×250mm);流动相为甲醇-水(含磷酸二氢钠50mmol·L-1、磷酸氢二钠4mmol·L-1,pH值5.8)(12:88),流速:1.0m L·min-1;检测波长:254nm,柱温40℃。结果腺苷与肌苷的标准曲线在一定浓度内线性关系良好,方法加样回收完全,日内及日间测定RSD<3.87%。结论该方法简便、快速、准确,可用于脑组织中腺苷与肌苷测定。

Objective To develop a method for determining adenosine and inosine in rat brain tissues by HPLC. Methods The brain tissues were homogenized in 0.1 mmol·L^-1 HClO4. The Agilent TC-C18 column was used in HPLC assay and the mobile phrase was consisted of methanol and water （ containing NaH2 PO4 50 mmol·L^-1, Na2 HPO4 4 mmol·L^-1, pH 5.8 ） （12 ： 88）. The wavelength of UV detector was 256 nm and the column temperature was 40 ℃. Results The calibration curves of adenosine and inosine were linear within certain concentration ranges. The added samples were fully recovered, inerday and intra-day RSD lowser than 3.87%. Conclusion The method is simple, fast and accurate, which can be used to determine adenosine and inosine in rat brain tissues.