高效液相色谱检测凡纳滨对虾血液中5-羟色胺的方法研究

Detection of 5-HT in Blood of Pacific White Leg Shrimp Litopenaeus vannamei by High Performance Liquid Chromatography

凡纳滨对虾样品经高氯酸去蛋白后,用Purospher STAR RP-18(150mm×4.6mm,5μm)色谱柱分离;流动相为0.1mol/L的磷酸二氢钾(pH 3.0)∶甲醇∶乙腈=70∶5∶25;在流速0.8mL/min,柱温25℃,进样量10μL的条件下,于荧光检测器激发波长280nm,发射波长340nm处进行检测。结果表明,该方法的线性范围为0.50～100.0μg/L,线性相关系数r2=0.9982,平均加标回收率为87.9%。

An experimental method was established to detect 5-hydroxy tryptamine （5-HT） in the blood of Pacific white leg shrimp Litopenaeus vannarnei using high performance liquid chromatography （HPLC）. After the protein in the samples was removed by high chlorine acid, the samples were detected by HPLC method using Purospher STAR RP-18 （150 min× 4.6 mm, 5 μm） column to separate, under the condition of mobile phases including potassium dehydrogenate phosphate of 0. 1 mol/L （pH 3.0）, methanol and acetonitrile（70 ： 5 ： 25）. The flow rate was maintained at 0.8 mL/min, and column temperature of 25 ℃, and the injection volume was 10 μL. The fluorescence detector under the condition of excitation wave- length 280 nm and emission wavelength 340 nm was utilized for testing. The results showed that the line- ar range of this method was ranged from 0.5 to 100.0μg/L and the linear correlation coefficient r2 was 0. 9982. ,with average recovery of 87.9%.