THE EFFECT OF METFORMIN ON NON-ENZYMATIC GLYCOSYLATION OF RECOMBINANT HUMAN SERUM ALBUMIN

Objective: The present research work was aimed to observe the effect of Metformin hydrochloride (MET) on non-enzymatic glucosylation of recombinant human serum albumin (rHSA).Methods: Albumin was incubated at 37 ° in 10 mM phosphate buffer (pH 7.4) using 0.2% sodium azide for 7 d with different concentrations of D-(+)-glucose and MET. Incorporation of D-(+)-glucose (5-12 mM) into rHSA was calculated in the presence of metformin hydrochloride (1-5 µg/ml) following extensive dialysis of the incubation mixture. The binding experiments were exercised at respective plasma concentration of rHSA, D-(+)-glucose and MET in corresponding to that of diabetic and non-diabetic population. Glycosylation of rHSA was quantified using 2-thiobarbituric acid (2-TBA). Whereas, metformin hydrochloride was quantified using a validated LC-MS/MS method with negative ion electrospray ionization using multiple reactions monitoring (MRM) mode. MET was eluted isocratically on X-Bridge BEH Phenyl column with a mobile phase consisting of mixture of water (containing 50 mM ammonium acetate) and acetonitrile in a ratio of 70:30.Results: The weighted (1/X2) calibration curve from 20 to 10, 000 ng/ml was employed for the calculation ofMET in sample with line equation of Y= 0.0020584x+0.0037475. The binding pattern ofMET to rHSA, was specific and pH dependent, possibly due to a change in ionization state of MET and change in conformation of rHSA during the binding process.Conclusion: All the described parameters were in accordance to the FDA Guideline (inside 85-115% for the accuracy and less than 15% for the precision), thus it can be concluded that the bioanalytical method is were fully validated as per USFDA guideline. These experiments showed the affinity ofMETtoward the rHSA proved to be higher than its affinity toward the glucose

Potential anti-diabetic activity of Bombax ceiba

Bombax ceiba bark extract was evaluated for its hypoglycemic and hypolipide-mic potential through normal and streptozotocin-induced diabetic rats administered with graded oral doses (200, 400, 600 mg/kg/day) for 21 days. The results showed that a dose of 600 mg/kg of B. ceiba extract is the most effective to cause significant (p<0.001) hypoglycemic and/or hypolipidemic effects on  streptozotocin-induced diabetic rats. This dose also significantly (p<0.001) lowered the total cholesterol and triglyceride level in severely diabetic rats. Phytochemical and GC-MS studies confirmed the presence of the triterpenoid compounds in the extract, which may account for its significant hypoglycemic activity. The present study thus provides a scientific rationale for the traditional use of this plant in the management diabetes

Development of validated HPLC-UV method for simultaneous determination of Metformin, Amlodipine, Glibenclamide and Atorvastatin in human plasma and application to protein binding studies

A simple, sensitive, fast, and economical HPLC method was developed and validated for simultaneous estimation of two fixed dose combinations frequently prescribed in diabetes (Metformin plus Glibenclamide) and hypertension with dyslipidemia (Amlodipine plus Atorvastatin) in Human plasma for the first time. The validated HPLC method was used to quantify the concentration of selected actives in ultrafiltrate. Optimum separation conditions were obtained with Water’s Novapack Phenyl (150 mm × 4.6 mm, i.d., 5.0 μm) column with mobile phase consisting of 0.1% Phosphoric acid (pH 3.0) and acetonitrile (ACN) in gradient mode with column oven temperature maintained at 30 °C and elution monitored by a UV detector at 227 nm. Protein precipitation was employed to extract the selected analyte form human plasma. The recoveries were more than 90% for all analytes in cold aqueous 10% trichloroacetic acid (TCA) and acetonitrile. The optimized HPLC-UV was validated in the calibration range of 10–10,000 ng mL−1 for Metformin, 25–5000 ng mL−1 for amlodipine, 50–10,000 ng mL−1 for glibenclamide and 10–5000 ng mL−1 for atorvastatin. The mean relative error was least when weighing of 1/×2 was applied for calibration curve. The accuracy of samples for six replicate measurements at LLOQ level was within limit. The precision and accuracy of samples for six replicate measurements at LLOQ level was within limit. The validated method was applied for quantitation of selected analytes in ultrafiltrate from protein binding experiments. A four to five fold increase in unbound fraction was observed when spiked to human serum albumin. Further the unbound fraction of highly albumin bound drugs was increased nearly to double when incubated with Gly-HSA as compare to HSA

DEVELOPMENT OF GASTRORETENTIVE OPTIMIZED ONCE A DAY FLOATING AND/OR BIOADHESIVE TABLET OF ALFUZOSIN

ABSTRACT: A novel extended release Alfuzosin hydrochloride (HCL) tablet formulation which possesses a unique combination of floatation and bioadhesion for prolonged residence in the stomach has been developed. Alfuzosin HCl is a selective α-adrenergic antagonist used against benign prostatic hyperplasia (BPH). The usual dose of immediate release Alfuzosin HCl tablet is 2.5mg thrice a day. Therefore, there is need to formulate once daily sustained release tablet of Alfuzosin HCl which is convenient for older patients. In this work floating and/or bioadhesive tablets were prepared by direct compression method which is economical and easy to scale up. Polymers like Hydroxy propyl methyl cellulose (HPMC K100M), Carbopol 934P, Sodium carboxyl methyl cellulose (Na CMC) &amp; Sodium bicarbonate in different ratios. Formulations were evaluated for in vitro drug release profile, swelling characteristics and in vitro bioadhesion property.  Sustained drug release with floating duration up to 24hrs and high bioadhesive strength was observed in case of Optimized formulation. Key-words: Alfuzosin, Gastro retentive, Optimized, Controlled release