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KMID : 0903620040450060312
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Journal of the Korean Society for Horticultural Science
2004 Volume.45 No. 6 p.312 ~ p.317
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Sample Purification Using Polyvinylpyrrolidone for the HPLC Analysis of Salicylic Acid from Cucumber Leaf Extract
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Choi Seong-Jin
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Abstract
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Salicylic acid is known as an important signal substance that induces systemic acquired resistance against pathogens in plants. In this study, a method was developed, by which the sample of plant extract can be purified for the purpose of salicylic acid analysis by using a HPLC. Insoluble polyvinylpyrrolidone (PVP) is known to bind specifically with phenolic compounds. The salicylic acid, as a phenolic compound, can be absorbed on the PVP under acidic condition and desorbed from the PVP under alkaline condition. It is based on the association and dissociation of carboxyl group in its molecular structure according to the change of pH. The sample of the cucumber leaf extract was acidified and loaded on a PVP cartridge. After washing successively with acid solution and small amount of methanol, the bound salicylic acid was eluted from the PVP by the eluant of alkaline methanol. The performance of this PVP method was compared with that of the liquid-liquid extraction method. The recovery of the PVP method for salicylic acid was higher, ranging from 92.0 to 95.7%, than that of the liquid-liquid extraction method. Impurities that interfere with the reversed phase HPLC analysis of salicylic acid using a C₁? column and a UV detector were also effectively removed by this method. The salicylic acid content in cucumber leaves was determined as 52 or 53 ngㆍg?¹ fresh weight by using this PVP method.
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KEYWORD
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liquid-liquid extraction, solid-phase extraction
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