HPLC-DAD method for the quantitative determination of short-chain fatty acids in meconium samples

Abstract

The purpose of this study was to develop and validate a high-performance liquid chromatography with a diode-array detector (HPLC-DAD) method for the determination of short-chain fatty acids (SCFAs) such as: acetic, propionic, butyric, valeric and isovaleric acids in meconium. Additionally, the proposed analytical procedure was used to determine SCFAs in meconium obtained from 17 neonates. All determined organic acids in meconium samples were separated with good resolution. The HPLC system was characterized by high precision (the coefficient of variance ≤ 2.5%), high determination coefficients of calibration curves (R2>0.997) and low limits of detection (LOD) and quantification (LOQ). For all investigated organic acids, LOD ranged from 0.01 to 0.80 mmol/ kg and LOQ values ranged between 0.04 and 2.64 mmol/ kg. Recovery of the HPLC assay for individual analytes comprised the range of 90±2 - 106±2%. Concentrations of acids in meconium collected from healthy children varied in wide ranges (CV within the range 0.79–2.47%) and no regularity was found in the molar ratios of the individual acids. The validation parameters obtained in this study, made the HPLC-DAD method reliable and useful tool for the determination of short-chain fatty acids in meconium samples.