Abstract
A reverse phase high performance liquid chromatographic (RP-HPLC) method was developed for identification and estimation of 18-β-glycyrrhetinic acid (GA) in HepG2 cell line. The analysis was carried out using a JASCO HPLC system with a C-18 (3 μm) Supelco reversed phase column (150 x 4.7 mm) using a mobile phase of 80% CH3OH and 20% of CH3CN: tetrahydrofuran: water (10:80:10, v/v/v). The method was linear in the concentration range of 1.5–120 μg /mL (n = 5). The LOD and LOQ were determined based on standard deviation of the y-intercept and the slope of the calibration curve. The LOD and LOQ values were found to be 11.46 μg/mL and 34.72 μg/mL, respectively. The mean percentage recovery by standard addition experiments of GA is 92.4 % ± 5.2%. The intracellular GA concentration value, obtained as mean of five different determinations, was 45.8 ± 7.45 μg/mL. We have developed a HPLC-UV method for quantitative determination of GA inside cells, with advantages in the cost reduction and economy of the analytical process.
Highlights
The use of natural substances as adjuvants in many drug therapies is a new important trend in modern medicine, due to a satisfactory clinical efficacy and a low degree of toxicity [1,2,3]
We have developed a High-Performance Liquid Chromatography (HPLC)-UV method for quantitative determination of glycyrrhetinic acid (GA) inside cells, with advantages in the cost reduction and economy of the analytical process
Several studies [10, 11] have reported that inappropriate use of licorice can produce pseudoaldosteronism, by inactivating 11betahydroxysteroid-dehydrogenase [11-βHSD] and by binding to mineralocorticoid receptors. 11-βHSD catalyzes the oxidation of the active mineralocorticoid, cortisol, to the inactive cortisone and 11alpha-hydroxysteroiod-dehydrogenase is International Journal of Analytical Chemistry responsible for the reduction reaction
Summary
The use of natural substances as adjuvants in many drug therapies is a new important trend in modern medicine, due to a satisfactory clinical efficacy and a low degree of toxicity [1,2,3]. Liquorice is a perennial plant with well-known pharmacological properties that is largely employed in the cosmetic and pharmacological fields, due to its several biological effects: antimicrobial, antiulcer, immunomodulatory, anti-inflammatory, etc. GA exhibits corticosteroid and mineral-corticoid activity due to the presence of the α,β-unsaturated ketone group: GA is able to interact with mineral-corticoid and glucocorticosteroid receptors and exhibits anti-inflammatory properties [7]. Several studies [10, 11] have reported that inappropriate use of licorice can produce pseudoaldosteronism, by inactivating 11betahydroxysteroid-dehydrogenase [11-βHSD] and by binding to mineralocorticoid receptors. 11-βHSD catalyzes the oxidation of the active mineralocorticoid, cortisol, to the inactive cortisone and 11alpha-hydroxysteroiod-dehydrogenase is International Journal of Analytical Chemistry responsible for the reduction reaction. GA potentiates the anti-inflammatory activity of cortisol by inhibiting its intracellular inactivation. GA is involved in strengthen red blood cell membrane integrity against both oxidative and proteolytic damage [12]
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