Extraction and rapid quantification of pure bixin from Bixa orellena L.

Pattanaik, Lopa ; Pradhan, Subhalaxmi ; Shaoo, Narendra Kumar; Naik, Satya Narayan


Bixin (9Z-6, 6’-diapocarotene-6, 6’-dioate), one of the carotenoid-based colour extracted from the seeds of the tropical tree Bixa orellana L., used primarily as a natural colouring agent in the food industry. In the present study, different solvents were used in both hot and cold conditions to extract bixin from B. orellana seeds, collected from two different geographical locations (Chhattisgarh and Uttaranchal) of India. The crude annatto extract was characterized by Thin Layer Chromatography (TLC). The major component, bixin was identified by TLC and further isolated from the crude extract by Preparative TLC (PTLC) method with a purity of 80%. Analysis of purified bixin contents isolated from samples was quantified by both High-Performance Liquid Chromatography (HPLC) and spectrophotometer. The results obtained from both the analytical techniques were comparable and lead to the conclusion that the highest amount of bixin was obtained from Chhattisgarh variety of annatto seed (21.13%), extracted by hot extraction method using acetone as a solvent. As compared to HPLC, the spectrophotometric analysis is an easy, rapid and cost-effective alternate route for the quantitative determination of isolated and purified bixin.


Bixin; HPLC; Preparative TLC; Solvent extraction; Spectrophotometer.

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