Analyze the Energy Charge of the Rats' Liver Using a Modified HPLC Method

改良HPLC方法分析大白鼠肝臟的能量積

常見有關adenosines的hplc分析法，均僅附標準品於緩衝液或體外分析的圖譜，這些方法似乎不適用於分析肝細胞萃取液。我們萃取大白鼠肝細胞內的adenosines，並在較少分解率的pH 6.0環境下以chloroacetaldehyde反應生成1,N6-ethenopurines螢光衍生物，再用配備胺基分離管的hplc系統，定性定量分析肝萃取液內腺核苷三磷酸或其代謝物。本法所用胺基分離管的固定相，似乎具有逆相和離子交換的雙重特質。使用本hplc分析方法，大白鼠肝臟萃取液內的腺核苷三磷酸或其代謝物、或類同品可以被成功的單離和定性、定量分析，依此結果以Atkinson的方法計算得大白鼠肝臟能量績是8.3 ± 0.4。此分析方法將有助於解析肝細胞凋亡和能量代謝的關係。

Most reported methods for HPLC have shown chromatography of biological adenosines in clean buffers, indicating a defect in the method used. In the present research an amine column equipped-HPLC system together with a fluorescent derivatization of 1,N6-ethenopurines using chloroacetaldehyde, and conditioned at pH 6.0 to decrease adenosines degradation, it were tested for the purpose of biological energy charge analysis in the liver. The fluorescent derivatives of ATP, adenosine metabolites, or its analogues taken from a rat’s liver were prepared, and specifically resolved using HPLC. The amine column seems to have both the stationary merits of the reverse phase and ion exchange characteristics. This HPLC system enables us to analyze the energy charge in the liver that represents mitochondria functions. The achievement of this study may help us to interpret the relationships of energy charge on hepatocytes and its apoptosis.