Makhdoom Humera Shafi, Khan Mohammad Dilawar, Chughtai Omar Rasheed, Abbas Zeerak, Azad Areeba, Batool Hijab
Departments of Chemical Pathology and Toxicology, Chughtai Healthcare, Lahore, Pakistan.
J Coll Physicians Surg Pak. 2023 Mar;33(3):254-260. doi: 10.29271/jcpsp.2023.03.254.
To optimize and validate a specific, sensitive and fast liquid Chromatography coupled to triple quadrupole Mass Spectrometric (LC-MS / MS) technique for accurate detection of serum α-tocopherol (Vitamin E) levels.
An experimental based study.
The Clinical and Forensic Toxicology section of Chughtai Lab, Jail Road Lahore, from April to September 2022.
Methanol was used to deproteinize serum samples. The chromatographic separation was achieved using an Agilent Infinity-Lab Poroshell 120EC-C18 column, Agilent 6470 LC-MS/MS (equipped with an Electron Spray Ionization source) in gradient elution mode using 0.1% LCMS grade formic acid in water and LCMS-grade methanol as mobile phases. Hexa-deuterated α-tocopherol was employed as internal standard to minimise matrix interferences.
The retention time of α-tocopherol was 3.0 ± 0.1 minutes. The linear concentrations obtained were ranged from 0.05-2 mg/dL with ≥0.985% coefficient of linearity. Detection and lower quantification limits determined were 0.025mg/dL and 0.05mg/dL, respectively. Recovery ranged from 96.5 to 99.8% and ionization suppression was -15.2% and -15.9% at high and low concentrations of α-tocopherol in serum. Intra-day and inter-day coefficient variation values were 4.2-4.9% and 5.0-5.9%, respectively.
An efficient and reliable tandem mass spectrometric technique for vitamin E analysis in serum was optimized, validated, and applied to 80 patient samples. This method has usefulness in clinical application for the accurate determination of vitamin E without potential matrix interferences.
Vitamin E, LC-MS/MS, Tocopherol, Internal standard, Validation.
优化并验证一种特定、灵敏且快速的液相色谱-三重四极杆质谱联用（LC-MS / MS）技术，用于准确检测血清α-生育酚（维生素E）水平。
基于实验的研究。
2022年4月至9月，拉合尔监狱路Chughtai实验室临床与法医毒理学部门。
使用甲醇对血清样本进行去蛋白处理。采用安捷伦Infinity-Lab Poroshell 120EC-C18色谱柱进行色谱分离，安捷伦6470 LC-MS/MS（配备电喷雾电离源），以0.1% LCMS级甲酸水溶液和LCMS级甲醇为流动相，采用梯度洗脱模式。使用六氘代α-生育酚作为内标，以最小化基质干扰。
α-生育酚的保留时间为3.0±0.1分钟。获得的线性浓度范围为0.05 - 2mg/dL，线性系数≥0.985%。测定的检测限和定量下限分别为0.025mg/dL和0.05mg/dL。回收率在96.5%至99.8%之间，在血清中高、低浓度的α-生育酚下，电离抑制分别为-15.2%和-15.9%。日内和日间变异系数值分别为4.2 - 4.9%和5.0 - 5.9%。
优化、验证了一种用于血清中维生素E分析的高效可靠的串联质谱技术，并应用于80份患者样本。该方法在临床应用中对于准确测定维生素E且无潜在基质干扰具有实用性。
维生素E；LC-MS/MS；生育酚；内标；验证
