Journal of Bioanalysis & Biomedicine

Ultra-performance liquid chromatographic (UPLC) method, for ledipasvir and sofosbuvir determination in bulk mixtures, and in tablets was developed and validated. The determination of the drugs, ledipasvir and sofosbuvir was carried out applying BEH C18 column (50 mm × 2.1 mm i.d, 1.6 μm particle size), with UV detector at λ max of 220 nm. The mobile phase applied for the current study, composed of two solvents, A (0.01 N w/v potassium di-hydrogen orthophosphate buffer of pH 3.5 modified with dilute orthophosphoric acid) and B (acetonitrile) in the ratio of 50:50 v/v. The isocratic mobile phase was moved at a flow rate of 0.3 ml/min. The validation study with proportionate to selectivity, linearity, robustness, precision, accuracy, stability, (LOD) limit of detection and (LOQ) limit of quantification, was performed utilizing the ICH Guidelines. Ledipasvir and Sofosbuvir displaying a linear response between 22.5-135 μg/ml for Ledipasvir 100-600 μg/ml for Sofosbuvir, with correlation coefficient (R2) 0.9997 and 0.998 for Ledipasvir and Sofosbuvir respectively. The % recovery for Ledipasvir and Sofosbuvir was 99.72 ± 0.61% and 100.21 ± 0.68 respectively. The LOD and LOQ values for Ledipasvir and Sofosbuvir were obtained to be 0.31 and 0.95 μg/ml and 1.29 and 3.91 μg/ml, respectively. The method also exhibits good robustness for different chromatographic conditions like mobile phase, temperature and flow rate. So, the approach can be successfully implemented for the quantitative analysis of ledipasvir and sofosbuvir in the (Q.C) Quality Control of in-house developed tablets, and the same can be exercised for the industrial use.

Kidney diseases contributed to the worldwide burden of human diseases, with approximately more than ¾ million deaths annually. The exact reason (s) for the high prevalence of kidney and urinary tract diseases are unknown. The effects of 1 mL of methanolic fruit extract of L. breviflora administered orally, at 0.5 and 1.0 g/kg/body weight once daily for fourteen and twenty-eight days respectively on serum and tissue antioxidants status, electrolytes and the activities of some ‘marker’ enzymes of kidney toxicity in Wister rats was investigated. The control group received 0.9% physiology saline. Using established protocols, the results showed that the extract negatively altered (p<0.05) the electrolyte balance, alkaline phosphatase, gamma glutamyl transferase, Lactate dehydrogenase, antioxidant enzymes activities, glutathione and malondialdehyde status in dose and duration dependent manners when compared with the untreated group. Histological evidence revealed mild tubular and glomerular damage. Although, no significant change was observed in the kidney body weight ratio in all the extract treated groups, our results, suggest that extract of L. breviflora may be nephrotoxic at high dosage over a prolonged period.