Extraction optimization and UHPLC method development for determination of the 20-hydroxyecdysone in Sida tuberculata leaves
| Autor: | Elisa Helena Farias Jandrey, Andreas Sebastian Loureiro Mendez, Marí Castro Santos, Vanderlei Folmer, Amélia T. Henriques, Mariana Koetz, Hemerson Silva da Rosa |
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| Rok vydání: | 2017 |
| Předmět: |
Clinical Biochemistry
01 natural sciences Biochemistry Endocrinology Limit of Detection Maceration (wine) Molecular Biology Malvaceae Chromatography High Pressure Liquid Mathematics Pharmacology Detection limit Peak area Chromatography biology 010405 organic chemistry 010401 analytical chemistry Organic Chemistry Extraction (chemistry) Reproducibility of Results biology.organism_classification Method development 0104 chemical sciences Plant Leaves Ecdysterone Linear Models Surface response methodology |
| Zdroj: | Steroids. 132 |
| ISSN: | 1878-5867 |
| Popis: | Sida tuberculata (ST) is a Malvaceae species widely distributed in Southern Brazil. In traditional medicine, ST has been employed as hypoglycemic, hypocholesterolemic, anti-inflammatory and antimicrobial. Additionally, this species is chemically characterized by flavonoids, alkaloids and phytoecdysteroids mainly. The present work aimed to optimize the extractive technique and to validate an UHPLC method for the determination of 20-hydroxyecdsone (20HE) in the ST leaves. Box-Behnken Design (BBD) was used in method optimization. The extractive methods tested were: static and dynamic maceration, ultrasound, ultra-turrax and reflux. In the Box-Behnken three parameters were evaluated in three levels (−1, 0, +1), particle size, time and plant:solvent ratio. In validation method, the parameters of selectivity, specificity, linearity, limits of detection and quantification (LOD, LOQ), precision, accuracy and robustness were evaluated. The results indicate static maceration as better technique to obtain 20HE peak area in ST extract. The optimal extraction from surface response methodology was achieved with the parameters granulometry of 710 nm, 9 days of maceration and plant:solvent ratio 1:54 (w/v). The UHPLC-PDA analytical developed method showed full viability of performance, proving to be selective, linear, precise, accurate and robust for 20HE detection in ST leaves. The average content of 20HE was 0.56% per dry extract. Thus, the optimization of extractive method in ST leaves increased the concentration of 20HE in crude extract, and a reliable method was successfully developed according to validation requirements and in agreement with current legislation. |
| Databáze: | OpenAIRE |
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