Determination of Perillic Acid in Bioconversion Supernatants by Gas Chromatography
| Autor: | Marcelo Raul Romero Tappin, Elba P. S. Bon, Maria Antonieta Ferrara, Igor Cunha cardoso, Virginia G. Correia, Eliane G. Carvalho, Antonio Carlos Siani |
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| Rok vydání: | 2017 |
| Předmět: | |
| Zdroj: | American Journal of Analytical Chemistry. :636-644 |
| ISSN: | 2156-8278 2156-8251 |
| Popis: | Perillic acid can be obtained from microbial oxidation of the exocyclic methyl group of limonene. Due to the pharmacological potential of such a metabolite, the biotransformation processes leading to its synthesis have been approached in recent studies. A robust analytical method is needed to assess the performance of such studies. An analytical method was developed and validated to determine perillic acid in the supernatants of a yeast-induced bioconversion of limonene, involving gas chromatography (GC) and an acid-induced precipitation during the sample preparation. GC analysis was performed using a column with polyethylene glycol as stationary phase (HP-Innowax) which resulted in higher loads and better peak shape. The sample preparation involved the supernatant initial filtration and precipitation with 0.6 M HCl followed by centrifugation and dissolution in ethyl acetate. GC analysis conditions were oven from 50°C to 250°C at 20°C·min-1, and then held 5 min (total runtime 15 min). Injector was set at 280°C, and detector at 300°C. Helium was the carrier gas at 1 ml·min-1. Injections of 1.0 μl were at the split ratio 25:1. The method was validated: Linearity with R2 of 0.9992, Accuracy of 98.3% in the range 190 - 950 μg·ml-1; Limit of detection of 10.4 μg·ml-1; Repeatability of 2.1% RSD. Thus, a complete methodology to determine perillic acid in a bioconversion supernatant was developed and validated. This overall approach may be useful for bioconversions of monoterpenes by other microorganisms that metabolize limonene. |
| Databáze: | OpenAIRE |
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