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Abstract
ABSTRACT This work aimed to optimise and validate the vortex-assisted extraction with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) method to quantify phorbol esters (PEs) in Jatropha leaves. Additionally, to evaluate the correlation between PEs content in leaves and seeds. The results of PEs content were expressed as equivalent of the major Jatropha phorbol ester, 12-deoxy-16-hydroxyphorbol-4'-[12',14'-butadienyl]-6'-[16',18',20'-nonatrienyl]-bicyclo[3.1.0]-hexane-(13-O)-2'-[carboxylate]-(16-O)-3'-]8'-butenoic-10']ate (DHPB) . One-variable-at-time strategy and 25-1V factorial fractional design were employed to determine the effects of solvent type, volume, stirring frequency, extraction time and cycles on yield. The quantification was performed using HPLC-UV. The optimum conditions were three extractions with 2% tetrahydrofuran:dichloromethane (1:1) in methanol (THF:DCM/MeOH). The sample:solvent ratio was 1 mg:25 μL, and the vortex stirring was 2200 rpm for 5 minutes. The method showed good linearity (R2 0.999), recoveries (97 to 105%), selectivity and repeatability (RSD 1.93-7.95%). In addition, good limits of detection and quantification of 2.19 and 6.65 ng μL"1, respectively, were noted. DHPB content in leaves and seeds ranged from 0.01 to 0.38 mg g"1 and 0.41 to 1.82 mg g"1, respectively. There is a statistically significant and positive linear relationship between leaves and seeds for DHPB content. These results may have practical application in analysing and predicting the amount of PEs in Jatropha leaves for environmental safety studies and the selection of better varieties for genetic improvement. Keywords: correlation, Jatropha leaves; Jatropha seeds; phorbol esters; vortex-assisted.
Highlights
Jatropha is a non-edible, drought-resistant, multipurpose oil crop grown in many tropical and subtropical countries.[1]
Solvent selection for extraction of phorbol esters (PEs) from Jatropha leaves The crucial step for efficient extraction and analysis of PEs from leaves is the selection of a suitable solvent
The best DHPB recovery was obtained using MeOH and tetrahydrofuran:dichloromethane (1:1) in methanol (THF):DCM/MeOH; these solvents were selected for the experimental design
Summary
Jatropha is a non-edible, drought-resistant, multipurpose oil crop grown in many tropical and subtropical countries.[1]. Several Jatropha germplasms have been established worldwide, through which breeding programs have been performed to turn Jatropha into a profitable industrial crop.[4,5] Intensification of cultivation campaigns of Jatropha has led to a significant increase of contact between humans and the Jatropha plant. This contact imminently leads to occupational health risks due to toxic tigliane diterpenes (referred to as phorbol esters (PEs)) in the whole plant.[6] These compounds are analogues of diacylglycerol and act by activating the protein kinase C, which is involved in various effects, including apoptosis, platelet aggregation, cell differentiation, and tumours.[7] Studies of Devappa et al.[8] revealed that PEs exhibited severe alterations towards reconstructed human epithelium and human corneal epithelium, are highly toxic in contact with eyes and skin
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