A method for lactate and pyruvate determination in filter-paper dried blood spots.
Lactic acidemia is commonly associated with severe diseases in pediatric patients. Quantitation of blood lactate and pyruvate is important for the diagnosis and clinical management. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using dried blood spots (DBS) was developed and could be used for simultaneous quantification of blood lactate and pyruvate. The applicability of the developed method was tested and confirmed by the regression analysis between LC-MS/MS method and enzymatic assay. Lactate and pyruvate were extracted from DBS obtained from 580 full-term, 120 pre-term infants (gestations ranging from 24 to 36 weeks), and 65 patients with suspected lactic acidemia, with methanolic internal standard (IS) solutions of sodium L-lactate-(13)C(3) and pyruvate-(13)C(3). An API-2000 LC-MS/MS system with multiple reaction monitoring (MRM) mode was applied. The within-run and between-run precisions (CV%) were determined and the results were 1.9% and 3.9% for lactate (n=20) and 5.7% and 7.3% for pyruvate (n=20). The linearity of lactate (r=0.9986) and pyruvate (r=0.9973) based on the IS was excellent. The parameter r squared (r(2)) of linear regression between LC-MS/MS method and enzymatic assay was 0.9405 for lactate and 0.9447 for pyruvate, respectively, and the agreement between these methods was consistent and acceptable. The stability of lactate and pyruvate on DBS was also confirmed. The LC-MS/MS method we developed is a specific, sensitive, and reproducible method for measuring blood lactate and pyruvate concentrations. The use of DBS in this method makes it particularly attractive for pediatric patients.
Chuang
Chih-Kuang
CK
Division of Genetics and Metabolism, Department of Medical Research, Mackay Memorial Hospital, No. 92, Sec. 2, Chung-Shan N. Rd., Taipei 10449, Taiwan.
Wang
Tuen-Jen
TJ
Yeung
Chun-Yan
CY
Lin
Dar-Shong
DS
Lin
Hsiang-Yu
HY
Liu
Hsuan-Liang
HL
Ho
Hsin-Tsung
HT
Hsieh
Wen-Shyang
WS
Lin
Shuan-Pei
SP
D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D057075:Enzyme Assays; D005374:Filtration; D006801:Humans; D007231:Infant, Newborn; D019344:Lactic Acid / Q000097:blood*; D010209:Paper; D019289:Pyruvic Acid / Q000097:blood*; D012015:Reference Standards; D012016:Reference Values; D012044:Regression Analysis; D053719:Tandem Mass Spectrometry / Q000379:methods*
D019344:Lactic Acid; D019289:Pyruvic Acid