New Analytical Methods for the Determination of New Anti-Viral Drug Favipiravir: A Potential Therapeutic Drug Against Covid-19 Virus, in Bulk and Dosage Forms.

Simple, accurate and robust analytical methods have been developed and validated for the determination of favipiravir (FVPR) by RP-HPLC and UV spectroscopy techniques as per the ICH guidelines. In the RP-HPLC method for FVPR determination, the mobile phase was ammonium acetate buffer pH 6.5 in pump Aand methanol in pump B. The C18 (Sunfire) 5 µm, 4.6 × 250 mm column was used as a stationary phase, and the detection wavelength was at 323 nm. Under these conditions, FVPR was eluted as a sharp peak at 2.65 min and the overall time taken for each injection was 10 min. In case of the UV spectroscopy method, standard FVPR solutions were prepared with pure ethanol and scanned from 250 to 400 nm and a flourishing spectrum was obtained at 323 nm. Hence, the wavelength of 323 nm was fixed for the whole process of validation in both techniques. The limit of detection (LOD) and limit of quantification (LOQ) in the RP-HPLC method were 1.0 and 3.5 µg/mL, respectively, and the linearity was established in the 10 to 50 µg/mL range. In the UV spectroscopy method, the LOD and LOQ values were found to be 3.5 and 12 µg/mL, respectively, and the linearity was established within 20 to 60 µg/mL range. The regression coefficient was found to exceed 0.999 in both methods. The proposed RP-HPLC and UV spectroscopy techniques are simple, accurate, rugged and robust.

Simple and Sensitive RP-HPLC and UV Spectroscopic Methods for the Determination of Remogliflozin Etabonate in Pure and Pharmaceutical Formulations

Simple, novel and selective reverse phase-high performance liquid chromatography (RP-HPLC) and ultraviolet (UV) spectroscopic methods have been developed and optimized for the determination of remogliflozin etabonate (RMZ) in bulk and dosage forms. In the HPLC method, the principal peak and internal standard peak were eluted separately at different retention times (RT) with the chromatographic conditions such as, mobile phase consisting of 0.02 M ammonium acetate buffer (pH was adjusted to 4.0 by 1.0 M ortho phosphoric acid), acetonitrile and tetrahydrofuran in the ratio 50:45:05, respectively (v/v) and the stationary phase used was C18, 5 µm, 4.6 mm x 250 mm kromasil column. The flow rate was 2.0 mL min-1, sample injection volume was 10 µL, and the wavelength of detection was fixed at 228 nm. In case UV spectroscopic method, the RMZ was diluted with pure ethanol. The RMZ showed a maximum absorbance at 228 nm. Hence throughout analysis 228 nm was used for the determination of RMZ. The RT of RMZ and internal standard, atorvastatin (ATST) were 6.2 min and 7.0 min, respectively. The resolution between the peaks was found to be more than 2.0. The total run time was fixed at 10 min. The linearity range for RP-HPLC method was found to be 10 µg mL-1 to 50 µg mL-1, at a fixed concentration of ATST. The linearity range for the UV spectroscopic method was found to be in the range of 100 to 250 µg mL-1. Regression coefficients (R2) were found above 0.999 for both of the techniques. The limit of detection and quantification for RMZ were found to be 1.0 µg mL-1 and 3.5 µg mL-1 respectively, in RP-HPLC method and 10.0 µg mL-1 and 40 µg mL-1, respectively, in UV spectroscopic method. The developed methods were found to be simple, accurate, reproducible, and precise. The RMZ can be analyzed in dual techniques, i.e., chromatographic and UV spectroscopic methods for its routine analysis.

Nickel and Oxidative Stress: Cell Signaling Mechanisms and Protective Role of Vitamin C.

BACKGROUND: Nickel activates the signaling pathways through the oxygen sensing mechanism and the signaling cascades that control hypoxia-inducible transcriptional gene expressions through oxidative stress. This review emphasizes on the recent updates of nickel toxicities on oxidant and antioxidant balance, molecular interaction of nickel and its signal transduction through low oxygen microenvironment in the in-vivo physiological system. DISCUSSION: Nickel alters intracellular chemical microenvironment by increasing ionized calcium concentration, lipid peroxidation, cyclooxygenase, constitutive nitric oxide synthase, leukotriene B4, prostaglandin E2, interleukins, tumor necrosis factor-α, caspases, complement activation, heat shock protein 70 kDa and hypoxia-inducible factor-1α. The oxidative stress induced by nickel is responsible for the progression of metastasis. It has been observed that nickel exposure induces the generation of reactive oxygen species which leads to the increased expression of p53, NF-kß, AP-1, and MAPK. Ascorbic acid (vitamin C) prevents lipid peroxidation, oxidation of low-density lipoproteins and advanced oxidation protein products. The mechanism involves that vitamin C is capable of reducing ferric iron to ferrous iron in the duodenum, thus the availability of divalent ferrous ion increases which competes with nickel (a divalent cation itself) and reduces its intestinal absorption and nickel toxicities. CONCLUSION: Reports suggested the capability of ascorbic acid as a regulatory factor to influence gene expression, apoptosis and other cellular functions of the living system exposed to heavy metals, including nickel.

Effects of Some Indigenous Plants of North Karnataka (India) on Cardiovascular and Glucose Regulatory Systems in Alloxan-Induced Diabetic Rats.

BACKGROUND: Kenaf (Hibiscus cannabinus Linn, Pundi), Chick pea (Cicer arietinum Linn, Chana) and Prickly lettuce (Lactuca scariola Linn, Hattaraki) leaves are a few of indigenous plants which are routinely consumed by the people of north Karnataka in the diet. Studies on these plants showed some potential anti-diabetic efficacies. OBJECTIVES: To examine the effect of leaves extracts of Hibiscus cannabinus Linn, Cicer arietinum Linn and Lactuca scariola Linn on cardiovascular integrity, glucose homeostasis and oxygen sensing cell signaling mechanisms in alloxan induced diabetic rats. METHOD: In vitro and in vivo tests on glucose regulatory systems and molecular markers such as - NOS3, HIF- 1α and VEGF were conducted in alloxan induced diabetic rats supplemented with all the three plant extracts. Electrophysiological analysis (HRV, LF: HF ratio, baroreflex sensitivity, BRS) and histopathogy of myocardial tissues and elastic artery were evaluated in diabetic rats treated with L. scariola linn. RESULTS: Out of these three plant extracts, Lactuca scariola Linn supplementation showed significant beneficial effects on glucose homeostasis and oxygen sensing cell signaling pathways in alloxaninduced diabetic rats. Furthermore, effects of sub chronic supplementation of Lactuca scariola Linn aqueous extracts showed significant improvement in sympatho-vagal balance in diabetic rats by increase of Heart Rate Variability (HRV) and regaining of Baroreflex Sensitivity (BRS). These results were also corroborated with myocardial and elastic artery histopathology of Lactuca scariola Linn supplemented diabetic rats. CONCLUSION: These findings indicate an adaptive pathway for glucose homeostasis, oxygen sensing cell signaling mechanisms and cardio protective actions in alloxan - induced diabetic rats supplemented with Lactuca scariola Linn extracts.

Groundwater fluoride contamination and its possible health implications in Indi taluk of Vijayapura District (Karnataka State), India.

Groundwater fluoride concentration and fluoride-related health problems were studied in twenty-two villages of Indi taluk of Vijayapura district, Karnataka, India. Present study (2015) was also used to compare groundwater fluoride concentration in same 22 villages with previous government report (2000). Groundwater fluoride concentrations of 62 bore wells of 22 villages were analyzed by using an ion-sensitive electrode. A total of 660 adults and 600 children were screened for fluorosis symptoms and signs. Sixty clinically suspected fluorosis patients' urine samples were further analyzed for fluoride. The mean value (1.22 ± 0.75 mg/L) of fluoride concentration of 62 bore wells and 54.83 % bore wells with ≥1.0 mg/L of fluoride concentrations in Indi taluk indicates higher than the permissible limit of drinking water fluoride concentration recommended for India. Clinical symptoms like arthritis, joint pains, gastrointestinal discomfort and lower limb deformities with high urinary fluoride concentrations in some subjects suggest fluorosis. Results also showed an increase in groundwater fluoride concentration of the same 22 villages between previous and present study. Preliminary arthritis symptom of the villagers could be due to drinking fluoride-contaminated water. Increase in fluoride concentration with time to the bore wells definitely indicates future danger.

Anti-diabetic effects of aqueous prickly lettuce (Lactuca scariola Linn.) leaves extract in alloxan-induced male diabetic rats treated with nickel (II).

BACKGROUND: Hattaraki pallye or prickly lettuce (Lactuca scariola Linn.) is one among several green leafy plants that grow in north Karnataka; it is usually consumed by the people of this region and is found to be antidiabetic in nature. The objective of this study is to evaluate hypoglycemic activities of supplementation with aqueous extract of prickly lettuce (L. scariola) leaves in vivo in acute and subchronic exposure with or without nickel (II) along with its glucose reduction capabilities with or without nickel (II) at pH 7.0 and 9.0 in vitro. METHODS: Percentage glucose reduction (in vitro) was determined by glucose oxidase-peroxidase enzymatic method at pH 7.0 and pH 9.0 using UV-Vis spectrophotometer. Hypoglycemic activities of L. scariola were carried out in alloxan-induced male diabetic rats at both acute and subchronic exposure. RESULTS: The results showed a significant alteration in the λmax value of Ni (II) in combination with L. scariola leaves extracts at both pH 7.0 and 9.0. The aqueous extract also produced a significant reduction in the glucose concentration at pH 7.0 and pH 9.0 even in presence of Ni (II) in vitro. Lactuca scariola leaves in either acute or subchronic supplementation showed a greater glucose tolerance and hypoglycemic regulation of blood sugar in diabetic rats with or without nickel (II) treatments. CONCLUSIONS: Lactuca scariola leaves can be a substitute for synthetic drugs to treat diabetic patients.