RP-HPLC法测定急性溴苯腈中毒小鼠肝组织腺苷酸含量

摘要

Objective To develop a new RP-HPLC method for determination of adenosine content and to evaluate the effect of bromoxynil on liver energy metabolism in mice.Methods A C18ODS column was used in RP-HPLC with acetonitrile phosphate buffer as mobile phase,the detection wavelength was 254 nm,the column temperature was 16℃ and the flow rate was 1.1 ml/min.Adenosine (ATP,ADP and AMP) contents in the liver tissue of mice were measured.Results The average recovery rates of ATP,ADP and AMP were (93.63 ± 8.02)％,(87.02 ± 8.28)％ and (94.99 ± 6.97)％,respectively.The relative standard deviations (RSDs) of within-day were 0.79％,1.02％ and 1.05％;and the RSDs of between-days were 2.09％,1.75％ and 2.17％,respectively.Conclusion The RP-HPLC method is simple,rapid and sensitive in analysis of the adenosine contents in mouse liver.Bromoxynil can cause hepatic injury and disturb liver energy metabolism in mice.%目的 建立测定小鼠肝组织腺苷酸含量的反相高效液相色谱(RP-HPLC)法,并观察溴苯腈中毒对小鼠肝组织能量代谢的影响.方法 将24只SD大鼠随机分为急性中毒组予溴苯腈(97％溴苯腈原药以10％二甲亚砜为溶媒配成10mg/ml溶液)200mg/kg灌胃染毒和正常对照组(予10％二甲亚砜溶液20ml/kg灌胃),每组12只.采用RP-HPLC法(C18ODS色谱柱,流动相为乙腈磷酸缓冲液,检测波长254nm,柱温16℃,流速1.1 ml/min)测定小鼠肝组织三磷酸腺苷(ATP)、二磷酸腺苷(ADP)、磷酸腺苷(AMP)含量.结果 APT、ADP和AMP浓度在1～ 200mg/L范围内线性关系良好;3者平均回收率分别为(93.63±8.02)％、(87.02±8.28)％、(94.99±6.97)％;日内峰面积变异系数分别为0.79％、1.02％和1.05％,日间峰面积变异系数分别为2.09％、1.75％和2.17％;溴苯腈中毒组ATP明显低于正常对照组,AMP明显高于正常对照组.结论 该方法可简便、快速、准确测定小鼠肝组织ATP、ADP和AMP含量;溴苯腈对小鼠肝组织能量代谢有明显抑制作用.