Screening CZE–UV Green Method for Acetate, Propionate, Butyrate, and Valerate Analysis in Human Stool for Complementary Gut Dysbiosis Diagnosis

ABSTRACT Short‐chain fatty acids present relevant function in the human body as fermentation metabolites pathways in the natural intestine microbiota activity. Their evaluations can be useful for indication as biomarkers for gut dysbiosis, provided an important health assessment to physicians by specialized laboratories. In the present work, a capillary zone electrophoresis with indirect ultraviolet (CZE–UV) detection method for simultaneous separation of acetate, propionate, butyrate, and valerate in human stool samples was performed. The background electrolyte consisted of 15 mmol L −1 of tris(hydroxymethyl)aminomethane (TRIS), 12 mmol L −1 of 3,5‐dinitrobenzoic acid (3,5‐DNB) and 0.5 mmol L −1 of cetyltrimethylammonium bromide (CTAB) as buffer at pH 7.5, within run time of 7.5 min. The proposed method resulted in an accuracy of 105.5%, 90.12%, 95.9%, and 86.6% and precision of 12.2%, 8.96%, 8.84%, and 18.3% relative standard deviation (RSD) for acetate, propionate, butyrate, and valerate, respectively. The optimized CZE–UV method was successfully applied as potential complementary analytical tool for gut dysbiosis diagnostic assay at 98 stool samples from patients under investigation of intestine‐related anomalies, combining eco‐friendly, quickness, automated, cost‐effective, and high‐throughput aspects. Furthermore, the method offers educational value by exemplifying how sustainable analytical practices can be integrated into modern laboratory training and the teaching of Green Chemistry principles.