Journal of Chromatography & Separation Techniques
Open Access

Abstract

Several years of experience performing columns chromatography have been condensed into simple guidelines useful for translating thin layer chromatography (TLC) results into either isocratic- or gradient-flash chromatography. The present study describes the use of such protocol with a silica Biotage SNAP KP-Sil 10 g cartridge (21 mm × 55 mm, 50 μm particle size), to purify, in a single step, four (4) peptides from crude proteins extract of germinated Amaranthus hybridus seeds. The best solvent system used was a combination of n-hexane (non-polar solvent B from 5% to 0% final concentration) and a mixture of polar solvents (solvent A) composed of acetonitrile, n-butanol, glacial acetic acid and water in 0.9:2:1:1 ratio, respectively. The elution was done in normal-phase with a linear gradient. All the flash chromatography procedure took at most twenty-five minutes (25 min), and less than a liter (750 mL) of total solvents was used. The purified peptides named AhPA, AhPB, AhPC and AhPE showed apparent homogeneity on TLC plates, and purification yields of about 11.15%, 10.38%, 14.50% and 17.25%, respectively. This innovative technique provides an efficient alternative to researchers’ in peptides purification and participates in reducing the waste of solvents, gels, time and thus, money.

Author Info

Citation: Bedikou EM, Niamke SL, Logesh R, Dhanabal P (2020) A rapid technique for plant peptides isolation based on their polarity by using BIOTAGE Isolera One flash chromatography. J Chromatogr Sep Tech. 11:427. DOI: 10.35248/2157-7064.20.11.427

Received: 07-Apr-2020 Accepted: 21-Apr-2020 Published: 28-Apr-2020

Copyright: © 2020 Bedikou EM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.