An optimal saponification and extraction method to determine carotenoids in avocado

Quantification of carotenoids in avocado fruit is a great challenge due to their co-extraction with high-oil concentration and the inherent nature of carotenoids to degrade and undergo cis/trans photoisomerization with prolonged extraction times and high temperatures. The study provides an optimised and validated methodology for quantification of carotenoids in the high-oil avocado matrix, with > 93% recovery of all carotenoids tested being significantly greater than previously published. Saponification with 15% KOH for 60 min was optimal for the avocado matrix. For the first time, this study identified that soap, produced during the saponification reaction, resulted in a significant reduction of carotenoid content from the avocado matrix, due to the production of micelles. A significantly higher carotenoid content (3.58 versus 2.0 mg/100 g DW) was able to be extracted after saponification with acidified phosphate buffer, instead of water as reported previously. Carotenoid profiles of five avocado cultivars were identified and quantified.