PJB-2019-130
COMPARATIVE ANALYSIS OF LYCOPENE CONTENT IN TOMATO USING A MICROPLATE READER AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY
Shiva Ram Bhandari
Abstract
Lycopene, a major carotenoid in tomato (Solanum lycopersicum L.), is a potent natural antioxidant and the main target of many tomato genetic and breeding programs. Crop improvement for increased fruit lycopene content requires a rapid and accurate method of lycopene content quantification. Here, we aimed to determine the lycopene content in tomato fruits at different maturity stages using a micro-plate reader that reduces the time and cost from that needed for high-performance liquid chromatography (HPLC). For this, lycopene from tomato was extracted using three different solvent systems: chloroform/methanol: 1/1 (v/v), n-hexane/ethanol: 1/1 (v/v), and n-hexane/acetone/ethanol: 2/1/1 (v/v/v). It was then centrifuged and syringe filtered, and the absorbance of aliquot was measured using HPLC. Lycopene content measured via HPLC and the micro-plate reader method at 470 and 503 nm was compared. The results showed a higher extraction efficiency for chloroform/methanol solvents (741-942 mg/kg) than for the different solvent systems (535-818 mg/kg). Both the 470 and 503 nm wavelengths showed similar lycopene content using HPLC. However, the 470 nm wavelength was more suitable than the 503 nm, as lycopene showed a higher response at 470 than at 503 nm. Similarly, lycopene measurement at 470 and 503 nm using 96-well microplate showed good linearity (y = 0.9959x) and a significant positive correlation (r = 0.9962), but the values at the early stages (green, breaker and pink) were overestimated at 470 nm. Comparative analysis of lycopene using HPLC at 470 nm and the 96-well micro-plate at 503 nm in tomatoes regardless of maturity stages showed good linearity, with a positive correlation coefficient (r = 0.9968). This simple, rapid, and inexpensive method for measuring lycopene content in tomato fruits shows promise for widespread screening of the tomato germplasm in breeding programs for the selection of functional genotypes.
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