In vitro and ex vivo approach for anti-urolithiatic potential of bioactive fractions of gokhru with simultaneous HPLC analysis of six major metabolites and their exploration in rat plasma. (1st January 2017)

• Record Type: Journal Article
• Authors: Sharma, Ikshit; Khan, Washim; Ahmad, Sayeed
• Is Part Of: Pharmaceutical biology. Volume 55:Number 1(2017)
• Page Start: 701
• Page End: 711
• Publication Date: 2017-01-01
• Subjects: Tribulus terrestris -- fractionations -- aggregation -- turbidity -- nucleation -- HPLC quantification; Pharmacognosy -- Periodicals; Materia medica, Vegetable -- Periodicals; 615.321
• DOI: 10.1080/13880209.2016.1266671
• Languages: English
• ISSNs: 1388-0209

Abstract

Abstract: Context: Tribulus terrestris L. (Zygophyllaceae) fruits have long been used in traditional systems of medicine for the treatment of various urinary diseases including urolithiasis. Objective : To explore the anti-urolithiatic potential of gokhru and to develop an analytical method for quantitative estimation of metabolites for its quality control. Materials and methods: Aqueous extract of gokhru fruit was prepared through maceration followed by decoction to produce a mother extract, which was further used for polarity-based fractionations. In vitro and ex vivo anti-urolithiatic activity of mother extract and fractions at different concentration (100–1000 μg/mL) were carried out using aggregation assay in synthetic urine and in rat plasma, however, nucleation assay for 30 min was done using confocal microscopy. A simultaneous HPLC method has been developed for quantification of diosgenin, catechin, rutin, gallic acid, tannic acid and quercetin in mother extract and in fractions. Results: The extraction resulted in 14.5% of w/w mother extract, however, polarity-based fractionation yielded 2.1, 2.6, 1.5, 1.3 and 6.1% w/w of hexane, toluene, dichloromethane (DCM), n -butanol and water fractions, respectively. In vitro and ex vivo studies showed a significant anti-urolithiatic potential of n -butanol fraction. Further, HPLC analysis revealed significantly ( p < 0.01) higher content of quercetin (1.95 ± 0.41% w/w), diosgenin (12.75 ± 0.18% w/w) and tannic acid (9.81 ± 0.47% w/w) in n -butanol fraction as compared to others fractions. Discussion and conclusion: In vitro and ex vivo studies demonstrated potent anti-urolithiatic activity of n -butanol fraction which can be developed as new phytopharmaceuticals for urolithiasis. HPLC method can be used for quality control and pharmacokinetic studies of gokhru.