摘要
目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)法定量检测小鼠血清中吲哚乙酸(IAA),考察非酒精性脂肪肝病(NAFLD)小鼠血清中IAA水平。方法:采集对照组与NAFLD组小鼠的血清,UPLC-MS/MS法定量检测IAA含量。色谱条件:色谱柱为BEH C_(18)(2.1 mm×50.0 mm, 1.7μm),流动相为0.01%甲酸乙腈-0.01%甲酸水,梯度洗脱,流速为0.4 mL·min^(-1),电喷雾正离子模式,多反应监测(MRM)。结果:IAA质量浓度在5~1 000 ng·mL^(-1)(r^(2)=0.996 4)范围内线性关系良好,日间和日内精密度RSD均<15%,提取回收率为76.83%~98.8%,基质效应为90.2%~103.8%,稳定性良好。对照组小鼠血清中IAA浓度为(130.65±31.23) ng·mL^(-1),NAFLD组小鼠血清中IAA浓度为(28.72±4.83) ng·mL^(-1)。结论:本研究建立的方法简单,专属性高,稳定性好,可准确检测小鼠血清中IAA的含量。NAFLD组小鼠血清IAA表达显著低于对照组,IAA可能是NAFLD诊断的潜在标记物。
OBJECTIVE To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) for quantitatively detection of indole acetic acid(IAA) in murine sera and determine the serum level of IAA in sera of mice with non-alcoholic fatty liver disease(NAFLD).METHODS The serum samples were collected from control and NAFLD groups. IAA was quantitatively detected by UPLC-MS/MS. The chromatographic column was BEH C_(18)(2.1 mm×50.0 mm, 1.7 μm), mobile phase 0.01% formic acid acetonitrile-0.01% formic acid water, gradient elution, flow rate 0.4 mL·min^(-1), multiple reaction monitoring(MRM) and electrospray ionization positive ion mode were employed for monitoring.RESULTS IAA concentration had an excellent linear relationship in the range of 5^(-1) 000 ng·mL^(-1)(r^(2)=0.996 4), the inter-day and intra-day precision RSD both <15%, extraction recovery 76.83%-98.8%, matrix effect 90.2%^(-1)03.8% and stability decent. The serum concentration of IAA was(130.65±31.23) ng·mL^(-1) in control group and(28.72±4.83) ng·mL^(-1) in NAFLD group.CONCLUSION Simple, highly specific and stable, the above method may be employed for detecting IAA in murine sera. The serum expression of IAA is significantly lower in NAFLD group than that in control group. IAA may be a diagnostic marker of NAFLD.
作者
张靖悦
许秀秀
王园芝
李晓纯
闫晨
赵经文
袁恒杰
ZHANG Jing-yue;XU Xiu-xiu;WANG Yuan-zhi;LI Xiao-chun;YAN Chen;ZHAO Jing-wen;YUAN Heng-jie(Department of Pharmacy,General Hospital,Tianjin Medical University,Tianjin 300052,China;Department of Gastroenterology,General Hospital,Tianjin Medical University,Tianjin 300052,China)
出处
《中国医院药学杂志》
CAS
北大核心
2021年第19期1940-1944,1954,共6页
Chinese Journal of Hospital Pharmacy
基金
国家自然科学基金青年项目(编号:81102447)。
关键词
超高效液相色谱-串联质谱
吲哚乙酸
非酒精性脂肪肝病
高脂饮食
ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)
indole acetic acid
nonalcoholic fatty liver disease
high fat diet
