Abstract
Background: Short-chain fatty acids (SCFAs), mostly found in colon feces, is an important group of gut microbial metabolites from anaerobic fermentation of indigestible carbohydrates. Objectives: To develope an HPLC method to determined SCFAs in rat feces treated with resistant starch. Materials and methods: Sample is the rat feces fed with acetate wheat starch and normal starch; fatty acid hydrazides are derived from SCFAs in feces and measured by HPLC. After validated as guidance of US-FDA, method is applied to identify SCFAs in rat treated two types of starch. Results: the HPLC condition was optimized as follow: Eclipse XDB–C8 (4.6mmx150mm, 5µm) column; mobile phase: methanol, acetonitrile and 0,057 mM acid triflouroacetic (pH 4.5) (0:13:87 – 10:20:70 – 0:13:87, v/v/v) in 40 mins;, examinized wavelength: 396 nm. Method was validated with parameters: system suitability, specificity, linearity, precision, accuracy and stability. The result showed amount ratio of SCFAs in feces of mice group treated with acetate wheat starch containing resistant starch higher than from the diet containing normal starch significantly. Conclusion: This method can be used to investigate SCFAs in the gastrointestinal segments of the living organism. Key words: SCFAs, HPLC, resistant starch, feces
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