摘要
目的评估不同浓度的BHT对液相色谱串联质谱法检测脂溶性维生素样本稳定性的影响。方法收集北京航天总医院2024年1月至3月临床检测剩余血浆样本,制备成4个不同浓度的血浆池作为研究对象,对研究样本进行前处理,分别采用75%甲醇、含1mg/mL BHT的75%甲醇、含5mg/mL BHT的75%甲醇对提取后样本进行复溶,采用高效液相色谱串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)测定不同保存条件下样本中维生素A(vitamin A,VA)、25-羟基维生素D 3[25-hydroxyvitamin D 3、25(OH)D 3]、维生素E(α-vitamin E,α-VE)的含量,与即刻值比较,差异率超过总变化限(total change limit,TCL)被认为不可接受。结果采用75%甲醇、含1mg/mL BHT的75%甲醇、含5mg/mL BHT的75%甲醇复溶的样本在自动进样器4℃保存24h后VA的平均差异率分别为-4.97%、2.08%、2.99%,25(OH)D 3的平均差异率分别为2.61%、-0.47%、-3.33%,α-VE的平均差异率分别为-1.21%、3.49%、6.85%。自动进样器25℃保存24h后VA的平均差异率分别为-1.76%、1.60%、-3.87%,25(OH)D 3的平均差异率分别为3.95%、5.43%、5.37%,α-VE的平均差异率分别为-0.47%、-2.58%、-0.57%。冰箱2~8℃保存1周后VA的平均差异率分别为0.25%、-1.64%、-8.99%,25(OH)D 3的平均差异率分别为2.58%、0.76%、6.98%,α-VE的平均差异率分别为5.95%、-2.28%、-0.33%。冰箱-20℃保存1周后VA的平均差异率分别为5.74%、0.61%、-5.19%,25(OH)D 3的平均差异率分别为7.54%、-0.22%、6.28%,α-VE的平均差异率分别为4.20%、-1.87%、0.38%。冰箱-80℃保存1周后VA的平均差异率分别为-7.78%、-8.19%、-8.77%,25(OH)D 3的平均差异率分别为1.24%、1.95%、4.67%,α-VE的平均差异率分别为3.96%、4.97%、3.03%。结论提取后的VA﹑25(OH)D 3﹑α-VE样本在自动进样器中保存可稳定24h,冰箱保存可稳定1周,复溶液中是否添加BHT及BHT的添加浓度对处理后样本的稳定性没有影响。
Objective To evaluate the effect of different concentrations of BHT on the stability of fat-soluble vitamin samples by liquid chromatography-tandem mass spectrometry.Methods The remaining plasma samples from clinical tests in Beijing Aerospace General Hospital from January to March,2024 were collected and prepared as 4 plasma pools with different concentrations as the study objects.These samples were pre-treated and re-dissolved with 75% methanol,75% methanol containing 1mg/mL BHT and 75% methanol containing 5mg/mL BHT,respectively.High performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was applied to determine levels of vitamin A,vitamin D 3 and vitamin E.Compared to immediate values,a difference rate exceeding the total change limit(TCL)was considered as unacceptable.Results The average difference rates of VA of the re-dissolved samples with 75% methanol,75% methanol with 1mg/mL BHT and 75% methanol with 5mg/mL BHT were-4.97%,2.08% and 2.99%,respectively,after being stored at 4℃ in the automatic sampler for 24h.The average difference rates of 25(OH)D 3 were 2.61%,-0.47% and-3.33%and the average difference rates ofα-VE were-1.21%,3.49% and 6.85%,respectively.After the automatic samplers were stored at 25℃ for 24h,the average difference rates of VA were -1.76%,1.60%and-3.87%,the average difference rates of 25(OH)D 3 were 3.95%,5.43%and 5.37%and the average difference rates of α-VE were-0.47%,-2.58% and -0.57%,respectively.After storage at 2-8℃for one week,the average difference rates of VA were 0.25%,-1.64% and -8.99%,the average difference rates of 25(OH)D 3 were 2.58%,0.76% and 6.98% and the average difference rates ofα-VE were 5.95%,-2.28% and -0.33%,respectively.After one week of storage at -20℃,the average difference rates of VA were 5.74%,0.61%and-5.19%,the average difference rates of 25(OH)D3 were 7.54%,-0.22% and 6.28% and the average difference rates of α-VE were 4.20%,-1.87% and 0.38%,respectively.After one week of storage at-80℃,the average difference rates of VA were-7.78%,-8.19% and -8.77%,the average difference rates of 25(OH)D 3 were 1.24%,1.95% and 4.67% and the average difference rates ofα-VE were 3.96%,4.97% and 3.03%,respectively.Conclusion After extraction steps,VA,25(OH)D 3 andα-VE samples can remain as stable status for 24h in the automatic injector and one week in the refrigerator.Whether BHT is added to the resolution or not and the concentration of BHT have no effect on the stability of the samples after treatment.
作者
李敏
孟祥兆
郭盈盈
于洪远
李会强
LI Min;MENG Xiangzhao;GUO Yingying;YU Hongyuan;LI Huiqiang(Beijing Aerospace General Hospital,Beijing 100076,China;College of Laboratory Medicine,Tianjin Medical University,Tianjin 300203,China)
出处
《标记免疫分析与临床》
2025年第6期1237-1242,共6页
Labeled Immunoassays and Clinical Medicine
基金
国家自然科学基金面上项目(编号:81772259)
北京航天总医院创新基金项目(编号:2023-725)。
关键词
脂溶性维生素
液相色谱串联质谱
BHT
稳定性
Fat-soluble vitamin
Liquid chromatography-tandem mass spectrometry
BHT
Stability
