Simultaneous determination of lactic acid and pyruvic acid in tissue and cell culture media by gas chromatography after in situ derivatization-ultrasound-assisted emulsification microextraction.
Anal Bioanal Chem
; 411(3): 787-795, 2019 Jan.
Article
en En
| MEDLINE
| ID: mdl-30467769
ABSTRACT
Lactic acid and pyruvic acid are important metabolites in the tricarboxylic acid cycle that can reflect the cytoplasmic redox state and mitochondrial respiratory chain function. The combination of these acids is considered as a screening index for mitochondrial disorders. Due to their biological effects, a derivatization method was developed to simultaneously detect pyruvic acid and lactic acid in tissue and cell culture media using gas chromatography. In this work, the combined derivatization method with methoxyamine hydrochloride and isobutyl chloroformate was first proposed. To improve the efficiency of derivatization, in situ derivatization-ultrasound-assisted emulsification microextraction (USAEME) was used in this study. After optimizing the volume of reagents and reaction times, good linearity values were obtained from 50 to 1000 µmol/L and 1 to 100 µmol/L for lactic acid and pyruvic acid, respectively. The limits of detection (LODs) were 0.12 µmol/L for lactic acid and 0.29 µmol/L for pyruvic acid. The recoveries of the two analytes were between 93.60 and 102.80%, and the precisions were less than 6.20%. This method was successfully applied to quantify pyruvic acid and lactic acid in the animal and cellular hypoxia models which provided an auxiliary means for the diagnosis of mitochondrial diseases. Graphical abstract á
.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Sonicación
/
Cromatografía de Gases
/
Ácido Láctico
/
Ácido Pirúvico
/
Medios de Cultivo
/
Enfermedades Mitocondriales
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Anal Bioanal Chem
Año:
2019
Tipo del documento:
Article
País de afiliación:
China