A simple, sensitive, inexpensive, and rapid stability indicating high performance liquid chromatographic method has been developed for determination of gemcitabine in injectable dosage forms using theophylline as internal standard. Chromatographic separation was achieved on a Phenomenex Luna C-18 column (250 mm × 4.6 mm; 5μ) with a mobile phase consisting of 90% water and 10% acetonitrile (pH 7.00 ± 0.05). The signals of gemcitabine and theophylline were recorded at 275 nm. Calibration curves were linear in the concentration range of 0.5–50 μg/mL. The correlation coefficient was 0.999 or higher. The limit of detection and limit of quantitation were 0.1498 and 0.4541 μg/mL, respectively. The inter- and intraday precision were less than 2%. Accuracy of the method ranged from 100.2% to 100.4%. Stability studies indicate that the drug was stable to sunlight and UV light. The drug gives 6 different hydrolytic products under alkaline stress and 3 in acidic condition. Aqueous and oxidative stress conditions also degrade the drug. Degradation was higher in the alkaline condition compared to other stress conditions. The robustness of the methods was evaluated using design of experiments. Validation reveals that the proposed method is specific, accurate, precise, reliable, robust, reproducible, and suitable for the quantitative analysis.
A simple, economic, robust, reproducible, selective, and precise high-performance liquid chromatography (HPLC) method for estimation of cordifolioside A in both 60% methanolic extract of Tinospora cordifolia and marketed formulation is developed and validated in the present study. The mobile phase composed of acetonitrile-water (25:75%, v/v) give a sharp and well-defined peak of cordifolioside A at the retention time of 9.52 +/- 1.03 min. HPLC estimation of cordifolioside A is carried out at wavelength of 210 nm with flow rate of 1.0 mL/min. The linear regression analysis data for the calibration curve shows good linear relationship with correlation coefficient of 0.997 in the concentration range of 0.5-50 microg/mL. The linear regression equation is y = 29716x - 4417.4. The limit of detection and limit of quantification are 0.18 and 0.55 microg/mL, respectively. The developed method is validated for accuracy, precision, reproducibility, and robustness as per ICH guidelines. The proposed method with high degree of precision and accuracy is employed for the estimation of cordifolioside A in methanolic extract and in formulation. Statistical analysis proved that the method is precise, reproducible, selective, and accurate for the estimation of cordifolioside A.
Hyperglycemia‐associated oxidative stress leads to various pathophysiological complications in diabetes mellitus. Here, the effects of Glycyrrhiza glabra (G. glabra) root extract of streptozotocin (STZ)‐induced diabetic changes and the associated free iron‐mediated oxidative reactions were investigated. The animals were divided into five group, Group 1: Control (NC received buffer); Group 2: STZ‐induced (DC); Group 3: Control treated with G. glabra root extract (NT, 60 mg/Kg b.w daily for 1 month); Group 4: Diabetic treated with the extract (60 mg/Kg b.w daily for 1 month); Group 5: Diabetic treated with glibenclamide (DTG, 8.6 mg/Kg b.w for 1 month). STZ (i) induced hyperglycemia, abnormal intraperitoneal glucose tolerance test (IPGTT), increased HbA1c and decreased plasma insulin levels (ii) hyperlipidemia (iii) lowered antioxidant enzyme activities (iv) diminished RBC membrane fluidity (v) enhanced hemoglobin glycation‐induced iron release and associated free radical reactions. Treatment with the extract resulted in significant reversal of hyperglycemia (DC: 205.0 ± 7.0 mg/dl vs. DT: 87.5 ± 4.5 mg/dl, p < .05); HbA1c (DC: 11.5 ± 2.0 vs. DT: 7.5 ± 0.8 vs. DT: 7.5 ± 0.8, p < .05); insulin (DC: 0.3 ± 0.06 vs. DT: 1.25 ± 0.15 μgm/L, p < .05); free iron (DC: 150.4 ± 7.07 vs. DT: 98.8 ± 7.7 μgm/gm of Hb, p < .05); TBARS (DC + H2O2: 24.62 ± 11.30 vs. DC + H2O2: 9.82 ± 2.56 mmoles/h, p < .05); carbonyl (DC: 40.40 ± 1.57 vs. DT: 25.50 ± 1.12 mmoles/g of Hb, p < .05) levels and β‐cell count/pancreatic islet (DC: 85 ± 15 vs. DT: 125 ± 20, p < .05). Thus, G. glabra extract is quite effective against hyperglycemia and the associated free iron‐mediated oxidative stress.
Practical applications
Chronic use of oral hypoglycemic synthetic drugs may produce side effects and drug resistance. Recently, various plant extracts are being researched to explore their antihyperglycemic potential. Here, the effects of this alcoholic powdered root extract on STZ‐induced diabetic changes and associated oxidative stress, including hemoglobin‐induced free iron‐mediated oxidative reactions were examined. The STZ‐induced diabetic changes and hemoglobin‐glycation‐induced free iron‐mediated oxidative reactions were alleviated in the Wistar rats after 1‐month of treatment with the extract. We have also reported previously that glycyrrhizin, a bioactive constituent of Glycyrrhiza glabra root inhibits peroxidase, esterase activities of hemoglobin and hemoglobin‐mediated oxidative damage without affecting oxygen‐binding capacity of the protein. This preclinical work further substantiates the potential therapeutic use of the G. glabra whole root extract in the treatment of diabetes mellitus.
A new, simple, sensitive, selective, and precise high-performance liquid chromatography (HPLC) method for analysis of berberine in crude plant material, herbal extract, and ayurvedic dosage forms was developed and validated. The stationary phase was inert sil C18 column The mobile phase consisting of acetonitrile (HPLC Grade) and potassium dihydrogen phosphate buffer (pH 2.5) in a gradient flow was used.The column was equilibrated with the mobile phase (flow rate 1.0 ml/min); elution was monitored at 346 nm. The linear regression analysis data for the calibration plots showed good linear relationship, with r2= 0.9942 in the concentration range of 16380–30420 μg/ml with respect to the peak area. The method was validated for specificity, precision, recovery, and linearity according to the International Conference on Harmonization guidelines. Statistical analysis of the data showed that the method is reproducible and selective for the estimation of berberine.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.