Comparative analysis of vitamin E extraction methods in <i>Cleome viscosa</i> L. and the role of key biosynthesis genes

Vitamin E is a lipid-soluble, potent radical-scavenging antioxidant that inhibits lipid peroxidation, which damages cell membranes, proteins, and DNA in the human body. Currently, it is widely utilized in dietary supplements and cosmetic products and is primarily sourced from plant-derived oils. However, Cleome viscosa L. (Asian spider flower) an ethnobotanical species that is not as widely recognized, could provide a different natural source of tocopherols, especially in the early stages of seed development. The present study aimed to evaluate which extraction methods produced the highest vitamin E contents by comparing different extraction techniques. A method was developed in which different solvents and extraction durations were tested to analyze the vitamin E and antioxidant contents in Cleome viscosa L. seed from 2-5 weeks after fruit set (WAF). Two methods were used, the Soxhlet method and maceration. The highest amount of gamma-tocopherol was present at 5 WAF and a concentration of 1946.78 µg/100 g fresh weight with an IC50 value of 32.02 ± 2.02 µg/ml using the Soxhlet method with ethanol for 6 hours. To gain information on the relationship between vitamin E content and the expression of four genes (VTE1-4) involved in vitamin E biosynthesis was examined. The higher expression of the VTE1 and VTE4 genes from the early seed stage to the mature seed stage. Furthermore, the maximum amount of alpha-tocopherol, 898.44 µg/100 g fresh weight, with an IC50 value of 7.46 ± 2.20 µg/ml, was produced by the maceration method using ethanol solution at 3 WAF, and the VTE4 gene was more highly expressed compared to other tocopherol biosynthesis genes.