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KMID : 0985420210430030162
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Laboratory Medicine and Quality Assurance
2021 Volume.43 No. 3 p.162 ~ p.165
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Implementation and Validation of a Gas Chromatography-Mass Spectrometry Method for Pristanic Acid and Phytanic Acid Quantification in Plasma Specimens
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Yi Ah-Ram
Lee Jun-Hyung
Yoo Ga-Hyun
Lim Hye-Jin
Park Eun-A
Han Jung-Sun
Kim Geun-Young
Cho Sung-Eun
Lee Sang-Gon
Lee Eun-Hee
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Abstract
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We validated a gas chromatography-mass spectrometry (GC-MS) method for quantifying pristanic acid (PrA) and phytanic acid (PhA) in plasma specimens. We developed a GC-MS method based on the analysis of 20 μM 2H3-pristanic acid (PrA-IS) and 80 μM 2H3-phytanic acid (PhA-IS) as internal standards. The GC-MS was fitted with a 30 m×0.25 mm×0.25 μm HP-5MS (Agilent, USA) column. The mass spectrometer was operated through the transitions from the precursor to the product ions (m/z [mass-to-charge ratio] 355, 369, 358, and 372 for PrA, PhA, PrA-IS, and PhA-IS, respectively). The retention times of PrA, PhA, PrA-IS, and PhA-IS were 19.33, 20.39, 19.31, and 20.38 minutes in a 26.83-minute analysis, respectively. Linearity, recovery, precision, and carry-over were evaluated to validate the method. The GC-MS method yielded a linear response from 0.032 to 9.951 μmol/L for PrA (R2=0.9999) and from 0.127 to 39.432 μmol/L for PhA (R2=0.9998). The limits of quantification by the methods were 0.032 μmol/L for PrA and 0.127 μmol/L for PhA. The recovery of PrA and PhA GC-MS method measurements were within ±10% when evaluated with external quality assessment materials. The within-batch and total coefficients of variation were all below 6% for both PrA and PhA test results. Twenty-interday imprecision (%) were all below 5% for both PrA and PhA test results. Carry-over was found to be 0.001% for PrA and ?0.008% for PhA. The GC-MS PrA and PhA assay showed adequate recovery, precision, sensitivity, and linearity. Hence, it is suitable for routine clinical work.
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KEYWORD
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Pristanic acid, Phytanic acid, Gas chromatography-mass spectrometry, Method validation
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