HPLC-MS/MS Analysis Of Carnitine And Acylcarnitines In Clinical Samples | 6030
Journal of Analytical & Bioanalytical Techniques
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Detailed analysis of carnitine and acylcarnitines is used for: 1) Follow-up to
positive tandem MS / newborn screening (TMS/NBS) results to eliminate false positives
and resolve isomeric acylcarnitines, 2) Monitoring with accurate values (as opposed to
a semi/pseudo quantitative screen), and 3) In support of clinical research, with accurate
and precise values, isotopomer analysis, and isomer separation.
Develop HPLC-MS/MS methods for the quantifi
cation of acylcarnitines
regardless of sample type, acyl chain length, or isomeric confi
1) Isolation of acylcarnitines, 2) Sample simplifi
cation, 3) Derivatization
without inadvertent hydrolysis of acylcarnitines, 4) Chromatography that resolves
isomeric acylcarnitines, 5) MRM detection, 6) Quantifi
cation using standard compounds
and class appropriate internal standards with multiple-point calibration curves, and 7)
Validation, demonstrating accuracy, precision, and robust performance.
Accuracy - acetylcarnitine add-back experiments to plasma, urine, and skeletal
muscle with values � 5% of expected values. Precision -acetylcarnitine batch-to-batch
percent relative standard deviations less than 5% (N=18). Robustness - quality control
values over a 5 month period had a percent relative standard deviation of � 6%, with
no individual value in excess of �15% of the expected value. Reference intervals were
determined from patient samples - 4000 plasma, 900 urine, and 500 skeletal muscle
A rigorously quantitative, accurate, and precise method for acylcarnitine
analysis in plasma, urine, and tissue samples is presented. Chromatographic separation
of isomeric acylcarnitines allows for detailed analysis of patient samples not possible by
Charles Hoppel completed his M.D. at St. Louis University School of Medicine and did his residency in Internal Medicine at the University of Kansas in Kansas City, Kansas. He then did postdoctoral studies at Western Reserve University School of Medicine. He is a Professor of Pharmacology and Medicine, and director of the Center for Mitochondrial Diseases at Case Western Reserve University School of Medicine He has published more than 250 papers in reputed journals.
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