高效液相色谱-质谱法快速测定大鼠血清雌二醇的研究

目的建立测定大鼠血清中雌二醇含量的高效液相色谱-质谱（HPLC-MS）方法。方法确定实验条件并进行方法学研究,以对羟基联苯为内标,丹酰氯进行衍生化处理。色谱条件：色谱柱为Agilent Zorbax SB C18柱（4.6mm×250.0mm,5μm）,流动相为甲醇-10mmol/L醋酸铵溶液-醋酸（97∶3∶1）,流速为1ml/min;质谱条件：离子源为大气压化学电离离子源,监测模式为选择性阳离子监测,干燥气流速为7L/min,干燥气温度为350℃,雾化室压力为344.5kPa。结果 HPLC-MS法测定大鼠血清雌二醇的含量为14.44～1.68pg/ml,回收率为92.43%。结论经丹酰氯进行衍生化处理,采用HPLC-MS法可以快速测定大鼠血清中雌二醇含量,具有一定的应用价值,值得采用。

Rapid Monitoring of Estradiol in Rat Serum with HPLC-MS

Objective To establish a rapid monitoring method of HPLC-MS of estradiol in rat serum. Methods The para-hydroxybiphenyl was used as internal standard, and dansyl chloride was used as derivatization reagents. Chromatographic column was Agilent Zorbax SB C18 （4. 6 mm × 250.0 mm, 5 μm） , mobile phase was methanol - 10 mmol/Lammonium acetate solution - acetic acid （97： 3： 1 ） , flow rate was 1 ml/min; ionization ion source was atmospheric pressure chemical ionization ion source, monitoring mode was selective cation ion monitoring mode, drying gas flow was 7 L/min, drying gas temperature was 350℃ , neb pressure was 345.5 kPa, and the validation of the method was analyzed. Results The trace estradiol in rat serum was determined, the contents was 14.44 - 1.68 pg/ml, and the recovery rate was 92.43%. Conclusion With the derivatization of dansyl chloride, the estradiol in rat serum can be rapidly determined with HPLC-MS, which is of certain application value.