Effects of starvation for 4-20 days on the amino acid content of Biomphalaria glabrata as determined by cellulose HPTLC-densitometry with confirmation of identity using silica gel and RP-18 W plates with two mobile phases causing inversion of the separation mechanism

Abstract

ABSTRACTThe effects of starvation at 4, 7, 10, 14, and 20 days on the amino acid content of Biomphalaria glabrata was determined by high performance thin layer chromatography (HPTLC)-densitometry. Cultures of control and starved snails were prepared for each duration of time. Samples were extracted using ethanol-water (70:30), and supernatant was collected after multiple centrifugations. Samples and standards were applied as bands using a CAMAG Linomat 4 onto cellulose HPTLC plates and developed in a 2-butanol-pyridine-acetic acid-water (39:34:10:26) mobile phase before spraying with ninhydrin detection reagent and scanning with a CAMAG 3 scanner to quantify ornithine (Orn), glycine (Gly), alanine (Ala), valine (Val), and leucine (Leu). Amino acid identification was confirmed by HPTLC on silica gel plates and RP-18 W plates using two different mobile phases causing inversion of the separation mechanism. Starvation led to an overall decrease in amino acid content in comparison to the control snails, however, the amount of amino acids in the starved snails had no correlation with the duration of starvation.