Thin Layer and High Performance Column Liquid Chromatographic Analysis of Selected Carboxylic Acids in Standards and from Helisoma trivolvis (Colorado Strain) Snails

Abstract

Separations and detections of acetic, fumaric, lactic, malic, pyruvic, and succinic acid standards were compared using previously reported thin layer chromatography (TLC) and high performance TLC (HPTLC) systems, which consisted mainly of silica gel or cellulose layers and acid‐base indicator or aniline‐aldose detection reagents. The best results were obtained with an HPTLC cellulose layer, triple development with n‐propanol‐2 M ammonium hydroxide mobile phase, and aniline‐xylose reagent, but resolution and sensitivity were not adequate for determination of these acids in Helisoma trivolvis (Colorado strain) snails.

Analysis of snail digestive gland‐gonad complex (DGG) samples was carried out using extraction with 50% Locke's solution, cleanup of the extract by anion exchange solid phase extraction (SPE), and ion exclusion high performance liquid chromatography with ultraviolet detection. Hemolymph was applied directly to the SPE column. Acetic, fumaric, malic, and pyruvic acids were detected at concentrations ranging from 24 to 200 ppm in the DGG and less than 154 to 16,000 ppm in hemolymph.

Keywords:

• Thin layer chromatography
• TLC
• Ion exclusion chromatography
• Carboxylic acids
• Digestive gland‐gonad complex
• Hemolymph
• Helisoma trivolvis snails
• Solid phase extraction

Acknowledgment

D. Massa was supported by a Camille and Henry Dreyfus Foundation Senior Scientist Mentor Initiative Program award to J. Sherma.