邻苯二甲醛柱前衍生化高效液相色谱法测定组织中肌肽含量

目的建立测定小鼠各组织肌肽含量的高效液相色谱法。方法采用C18色谱柱,以甲醇和0.05 mol/L磷酸盐缓冲液(pH 3.5)为流动相,用邻苯二甲醛为柱前衍生剂,在柱温38℃下采用二元梯度洗脱,对小鼠各组织中的肌肽含量进行了测定。结果肌肽在0.63~10.00μg/mL时峰面积与进样质量浓度有良好的线性关系,其相关系数为0.999 6。回收率在98.2%~99.7%之间。检测限为0.05μg/mL(S/N=10)。结论该法灵敏可靠、专属性强、重现性好。小鼠心脏、肝脏、肾脏、大脑皮层和海马中肌肽含量分别为123.67,22.86,158.86,76.57,81.86 mg/g。

Determination of carnosine by reversed phased high performance fiquid chromatography with precolumn O-phthalaidehyde derivation

Purpose To establish HPLC method for the determination of carnosine in tissue.Methods The sample was derived with O-phthalaldehyde(OPA),and then the derivative sample was separated on Kromail C18 column at 38 ℃.A linear gradient elution of A(methanol) and B(0.05 mol/L phosphate buffer,pH 3.5) was used.The flow rate was 1 mL/min,and detected by fluorescence detector with excitation wavelength(λex) at 338 nm and emission wavelength(λem) at 425 nm.Results The calibration curve was liner within the ranges of 0.63-10.00 μg/mL.The correlation coefficient was 0.999 6.The recoveries were between 98.2%-99.7%.Detection limit of carnosine was 0.05 μg/mL(S/N=10).Conclusion The result shows that this method is simple,rapid,specific,accurate and reproducible.The contents of Carnosine in rat′s heart,liver,kidney,cortex and hippocampus were 123.67,22.86,158.86,76.57 and 81.86 mg/g(wet weight).