液相色谱-串联质谱法检测维生素A、25羟维生素D_(3)和维生素E方法的建立

Establishment of LC-MS/MS method in simultaneous detection of serum vitamin A,25(OH)D3 and vitamin E

目的:基于临床实验室建立一种高特异性液相色谱-串联质谱法(LC-MS/MS)技术方法,以快速准确检测不同年龄人群的血清维生素A、25羟维生素D_(3)[25(OH)D_(3)]和维生素E水平。方法:采用液-液萃取前处理,建立LC-MS/MS法检测血清中维生素A、25(OH)D_(3)和维生素E的方法,并通过基质效应、线性、精密度测定、加样回收率等对此方法进行评价。收集健康体检人群标本542例,分析不同年龄组即青少年(1~18岁组)、中年(19~59岁组)和(老年≥60岁组)血清维生素A、25(OH)D_(3)和维生素E水平,并依据卫生行业标准(WS/T553-2017和WS/T677-2020)判断不同年龄段维生素A、25(OH)D_(3)和维生素E营养状况。结果:采用LC-MS/MS技术同时检测血清维生素A、25(OH)D_(3)和维生素E的总检测时间为6.5 min,未见明显干扰因素。标准曲线相关系数r^(2)>0.99。维生素A、25(OH)D_(3)和维生素E批内不精密度均<6.0%,批间不精密度<7.5%。加标回收率均在100.5%~112.9%。不同年龄组中1~18岁组未见到维生素A缺乏;7~18岁组维生素A不足占8.96%;25(OH)D_(3)缺乏和不足分别占10.12%和38.10%;维生素E不足占5.06%;19~59岁组维生素A和E较少见到缺乏,25(OH)D_(3)缺乏和不足分别占20.90%和46.33%;≥60岁组维生素A、25(OH)D_(3)和维生素E缺乏和不足总和分别占16.36%、87.3%和12.72%。结论:建立的LC-MS/MS技术方法能够快速准确地检测血清维生素A、25(OH)D_(3)和维生素E水平。在1~18岁和≥60岁人群中存在一定程度的维生素A、25(OH)D_(3)和维生素E的缺乏,各年龄段均显示25(OH)D_(3)缺乏和不足。

Objective:To establish a highly specific liquid chromatography-tandem mass spectrometry(LC-MS/MS)method based on clinical laboratory,so as to quickly and accurately detect serum vitamin A,25 hydroxyvitamin D_(3)[25(OH)D_(3)]and E of population with different ages.Methods:Liquid-liquid extraction pre-process was adopted to establish LC-MS/MS method to detect serum vitamin A,25(OH)D_(3) and vitamin E.The method was evaluated by matrix effect,linear evaluation,precision measurement and sample recovery rate.A total of 542 specimens were collected from population who received physical examination,and the serum vitamin A,25(OH)D_(3) and vitamin E levels of different age groups,which included adolescents(1-18 years old group),middle-aged(19-59 years old group)and elderly(≥60 years old group),were analyzed.The nutritional statuses of vitamin A,25(OH)D_(3) and vitamin E of different populations were judged according to health industry standards(WS/T 553-2017 and WS/T 677-2020).Results:The total detection time was 6.5min when LC-MS/MS method was adopted to simultaneously detect the serum vitamin A,25(OH)D_(3) and vitamin E,and the obvious interfering factors did not were found.The correlation coefficient(r^(2))of standard curve was larger than 0.99.The intra batch imprecisions of this method in detecting vitamin A,25(OH)D_(3) and vitamin E were less than 6.0%,and the inter batch imprecisions were less than 7.5%.The recovery rate was between 100.5%-112.9%.There was no Vitamin A deficiency in 1-18 years old group.In 7-18 years old group,the rate of Vitamin A insufficiency was 8.96%,and the rates of 25(OH)D_(3) deficiency and insufficiency were respectively 10.12%and 38.10%,and the rate of Vitamin E sufficiency was 5.06%.In 19-59 years old group,the deficiencies of Vitamin A and E were less,and the rates of 25(OH)D_(3) deficiency and insufficiency were respectively 20.90%and 46.33%.In the“≥60 years old”group,the sums of deficiencies and insufficiencies of vitamin A,25(OH)D_(3) and vitamin E were respectively 16.36%,87.3%and 12.72%.Conclusion:The established LC-MS/MS technique method can quickly and accurately detect serum vitamin A,25(OH)D_(3) and vitamin E.There are a certain vitamin A,25(OH)D_(3) and vitamin E deficiencies in 1-18 years old population and≥60 years old population,and all age groups appear 25(OH)D_(3) deficiency and insufficiency.