Phyto-cosmeceutical gel containing curcumin and quercetin loaded mixed micelles for improved anti-oxidant and photoprotective activity.

Ultra Violet radiations induced skin damage and associated skin disorders are a widespread concern. The consequences of sun exposure include a plethora of dermal conditions like aging, solar urticaria, albinism and cancer. Sunscreens provide effective protection to skin from these damages. Besides FDA approved physical and chemical UV filters, phytoconstituents with their multi functionalities are emerging as frontrunners in Therapy of skin disorders. Objective of this study was to develop novel phyto-dermal gel (PDG) with dual action of sun protection and antioxidant potential using polymeric mixed micelles (PMMs) are nanocarriers. PMMs of Pluronic F127 and Pluronic F68 loaded with curcumin and quercetin were optimized by 32 factorial designs. Responses studied were vesicle size, SPF, entrapment efficiency of curcumin and quercetin and antioxidant activity. Droplet size ranged from 300 to 500 nm with PDI in between 0.248 and 0.584. Combination of curcumin and quercetin showed enhanced sun protection and antioxidant activity. Pluronics played a significant positive role in various parameters. In present studies vesicle size of factorial batches was found to be between 387 and 527 nm, and SPF was found to be between 18.86 and 28.32. Transmission electron microscopy revealed spherical morphology of micelles. Optimized micelles were incorporated into Carbopol 940. Optimized PDG was evaluated for pH, drug content, spreadability, rheology, syneresis, ex vivo permeation, and skin retention. Hysteresis loop in the rheogram suggested thixotropy of PDG. Syneresis for gels from day 0-30 days was found to be between 0% and 12.46% w/w. SPF of optimized PDG was 27±0.5. Optimized PDG showed no signs of erythema and edema on Wistar rats. PMMs thus effectively enhanced antioxidant and skin protective effect of curcumin and quercetin.

Quantification of ternary mixture of paracetamol, chlorzoxazone and ibuprofen present in tablet dosage form using ratio subtraction spectrophotometric approaches.

A sensitive and selective spectrophotometric approach comprising of successive ratio subtraction was developed for quantification and resolution of spectrum of mixture containing three components without prior separation. Three components, namely paracetamol, chlorzoxazone and ibuprofen were present in tablet dosage form. The linearity studies were carried out by recording zero order spectra and measuring absorbances at 285.0, 282.0 and 220.0 nm for paracetamol, chlorzoxazone and ibuprofen respectively. The drugs exhibited linear response in the concentration range of 6.0-18.0, 3.0-15.0 and 4.0-20.0 µg / mL for paracetamol, chlorzoxazone and ibuprofen respectively. The spectrum of paracetamol was most extended which was subsequently followed by moderately extended spectrum of chlorzoxazone and unextended spectrum of ibuprofen. The ratio spectra were manipulated successfully for quantification of paracetamol, chlorzoxazone, and ibuprofen. The developed method was validated as per ICH guidelines for the parameters of specificity, linearity, precision and accuracy. The percent recoveries for all the three drugs were in the range of 98.0-102.0 % with mean recovery of paracetamol, chlorzoxazone and ibuprofen were 99.72, 99.02 and 100.34 % respectively. Additionally the validity of the method is assured by analyzing marketed formulation.

Impact of protein deficient diet on the pharmacokinetics of glibenclamide in a model of malnutrition in rats.

Purpose: The present investigation deals with the impact of protein energy malnourished condition on the pharmacokinetic profile of glibenclamide. Protein energy malnourished condition leads to malnutrition related diabetes mellitus (MRDM), Fibrocalculus pancreatic diabetes mellitus (FCPD) or Lean body mass diabetes mellitus (LBMDM). Method: In the present study, malnutrition was developed in female wistar rats using a modified protein deficient diet (0.5%). The experiment was performed on 12 animals, each group containing 6 female wistar rats. The control group animals were fed with standard pellet diet (AIN 93 G diet) while group 2 received the low protein diet (0.5%) for 75 days. Glibenclamide (Gli) suspension (30 mg/kg) was administered orally to these rats on 75 days and kinetic parameters were evaluated by HPLC analysis.The pharmacokinetic interpretation done by pksolver software version 2.0, statistical comparison done by applying student T test. Results: The results of body weight and hematological parameters indicated a significant decreased in the body weight in protein deficit rats to 124.1 ± 6.2 g compared to 235.5 ± 8.4 g (p < 0.01) control rats; whereas a decrease in the hemoglobin to 5.8 ± 0.6 g/dL, total blood protein level to 6.9 ± 0.6 g/dL and blood albumin levels to 2.7 ± 0.4 g/dL in protein deficit rats compared to 15 ± 0.7 g/dL(p < 0.05), 8.1 ± 0.4 g/dL(p < 0.05), and 4.5 ± 0.2 g/dL(p < 0.05), respectively in control rats. All these findings reflect the malnourished condition and weight loss due to a protein deficit diet in experimental animals. There was an increase in the fasting blood glucose levels up to 150 ± 17.4 mg/dL in the protein deficit diet group as compared to 98.7 ± 14.1 mg/dL(p < 0.05) in control rats reflect the prediabetes state in malnourished animals. The results of the pharmacokinetic study reflect a significant lowering of half-life (T½) of glibenclamide to 96.8 ± 0.8 min. in malnourished rats compared to 166.7 ± 0.74 min. (p < 0.001) in control rats. The maximum concentration (Cmax) of glibenclamide in the malnourished rats was significantly higher 20.74 ± 0.65 µg/mL and also took double time i.e. about 180 min. to reach maximum concentration (Tmax) compared to the control rats values 7.9 ± 0.84 µg/mL (p < 0.001) and 90.0 ± 0.24 min. (p < 0.001) respectively. The area under the plasma concentration-time curve [AUC(0-∞)] in malnourished rats increased 4439.1 ± 40.6 µg/ml*min as compared to 1235.9 ± 55.8 µg/ml*min (p < 0.001) in control rats. There was a lowering in the total body clearance (CL) to 0.4 ± 0.02 L/hr and volume of distribution (Vd) to 1.75 ± 0.07 L of glibenclamide in the protein deficit group compared to 1.4 ± 0.3 L/hr (p < 0.001) and 3.14 ± 0.8 L (p < 0.01), respectively in the control rats. Conclusion: From this study it concludes that there is an increase in the T½, Cmax, Tmax and AUC(0-∞) of glibenclamide in malnourished rats while the total body clearance and volume of distribution is lowered. Therefore this study proposes to conduct an adequate pharmacokinetic study in malnourished patients to decide whether the standard glibenclamide dose should be adapted according to the nutritional status of the individual.

A methodology employing retention modeling for achieving control space in liquid chromatography method development using quality by design approach.

This study reports the application of retention modeling and quality by design practices for reverse-phase liquid chromatographic method development of a new chemical entity. Prior to the retention modeling, preliminary screening experiments were performed for the selection of stationary phase, organic modifiers, and method parameters. Based on the results of preliminary method conditions, tG-T (gradient time - temperature) 2-D modeling with 4 input runs, and tG-T-tc (gradient time-temperature-ternary composition) 3-D modeling with 12 input runs were designed to build a model for achieving the optimized separation. Modeling of reverse phase separations was based on the measurement of both retention times and peak areas. A design space with appropriate input variables and control strategy was established prior to optimization and robustness evaluation following the quality by design framework. DryLabⓇ was used to predict the optimized gradient profile and separation temperature. The robustness evaluation was carried out using the multiple factors at a time approach and the control space was established. The interdependence of control space and the control strategy was demonstrated by evaluating method robustness using two levels of system suitability criteria. The predictive accuracy of the retention modeling was established through experimental verification of the in-silico predictions. The quality by design based method development approach demonstrated the in-silico optimization as an integral component of reverse-phase chromatographic method development to evaluate the interplay of factors such as organic modifiers, separation temperature and gradient time, which greatly integrated and enhanced method robustness during method development.

Rapid and efficient chiral method development for lamivudine and tenofovir disoproxil fumarate fixed dose combination using ultra-high performance supercritical fluid chromatography: A design of experiment approach.

Fixed dose combination (FDC) of tenofovir disoproxil fumarate (TDF) and lamivudine (3TC) is one of the most preferred FDC for the treatment of acquired immunodeficiency syndrome (AIDS)/human immunodeficiency virus (HIV) infection. To the best of authors' knowledge there are no reported methods for chiral purity estimation of both drugs simultaneously from a FDC. The current study was focused on the development of a single chiral method uisng supercritical fluid chromatography (SFC) for separation of stereoisomers of TDF and 3TC combination employing design of experiment (DoE) approach. Method development was planned in three steps by using different experimental designs for each step. I-optimal, Taguchi orthogonal array and face-centred central composite designs (CCD) were employed for primary parameter selection, secondary parameter screening and final method optimization, respectively. All six stereoisomers were separated in a 10 minute run on Chiralpak IA column with carbon di-oxide /methanol (containing 0.5 % v/v n-butylamine) as mobile phase at 1.5 mL/min in gradient mode. The optimized method was verified for performance through establishing specificity, precision, linearity, accuracy, limit of quantification, and solution stability. Resolution between each isomeric pair was more than 1.5. The method was found to be linear from 1.5 µg/mL to 7.5 µg/mL for 3TC and 7.5 µg/mL to 37.5 µg/mL for TDF stereoisomers. The R2 values for all the linearity curves for undesired isomers were greater than 0.995. The method proved to be rapid, reproducible and efficient to quantify stereoisomers of both drugs in a single run.

High-performance thin-layer chromatographic determination of etoricoxib and thiocolchicoside in combined tablet dosage form.

A new, simple HPTLC method for determination of etoricoxib (ETO) and thiocolchicoside (THIO) in combined tablet dosage form has been developed and validated. The pharmaceutical dosage form used in this study was Nucoxia-MR tablets. Sample solutions were prepared at concentrations of 25 and 20 microg/mL for ETO and THIO, respectively. The separation was carried out on 20 x 10 cm Merck aluminum sheets precoated with a 250 microm layer of silica gel 60F254 using ethyl acetate-methanol (8 + 2, v/v) as the mobile phase. The calibration curve was linear over a range of 50-250 and 100-500 ng/band for ETO and THIO, respectively. Quantitative determination was done by densitometric scanning of bands at 290 nm. LOD and LOQ values were 10.993 and 33.314 ng/band, respectively, for ETO and 25.133 and 76.161 ng/band, respectively, for THIO. The method was validated with respect to linearity, accuracy, precision, and robustness in accordance with the International Conference on Harmonization guidelines. The method has been successfully applied to the analysis of drugs in the pharmaceutical formulation.

Antioxidant, antinociceptive and anti-inflammatory activities of atorvastatin and rosuvastatin in various experimental models.

The present study was planned to investigate the antioxidant, antinociceptive, and anti-inflammatory activities of atorvastatin and rosuvastatin (1, 3 and 10 mg/kg, p.o.) in various animal models. The antinociceptive effect was assessed by chemically- (formalin, acetic acid) and thermally- (hot plate) induced nociception, while anti-inflammatory effect was evaluated using carrageenan-, formaldehyde-induced paw oedema and cotton pellet-induced granuloma. The effect of atorvastatin and rosuvastatin on liver antioxidant enzymes like superoxide dismutase, glutathione, LPO, CAT along with the effect on lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) was evaluated in the cotton pellet-induced granuloma model. Atorvastatin and rosuvastatin showed significant decrease (p < 0.05) in carrageenan- and formaldehyde-induced rat paw oedema and reduced granuloma formation in the cotton pellet-induced granuloma method (p < 0.01) while the levels of LDH and ALP were also significantly decreased (p < 0.05). The liver antioxidant enzyme levels were found to be restored (p < 0.05). Atorvastatin and rosuvastatin also showed antinociceptive activities (p < 0.05 and p < 0.01) in the acetic acid- and formalin-induced nociception in mice, while there was no significant activity in the hot plate method. The present findings suggest that atorvastatin and rosuvastatin possess dose-dependent antioxidant, analgesic, and anti-inflammatory activities.

HPTLC determination of cefuroxime axetil and ornidazole in combined tablet dosage form.

A new simple high-performance thin layer chromatographic method for determination of cefuroxime axetil and ornidazole in combined tablet dosage form is developed and validated. Cefuroxime axetil is second-generation cephalosporin used to treat or prevent infections that are proven or strongly suspected to be caused by bacteria. Ornidazole is used to cure protozoan infections. The separation is carried out on Merck precoated silica gel aluminium plate 60 F(254) using toluene-n-butanol-triethylamine (8.5:2:0.5, v/v/v) as mobile phase. Quantitative determination of drugs is carried out by densitometric scanning of plates at 285 nm. The retention factor for ornidazole and cefuroxime axetil is found to be 0.51 +/- 0.007 and 0.67 +/- 0.009, respectively. The method is validated with respect to linearity, accuracy, precision, and robustness. Response found to be linear in the concentration range of 100-500 ng/band for both cefuroxime axetil and ornidazole. The method has been successfully applied for the analysis of drugs in pharmaceutical formulation. The % assay is found to be 102.36 +/- 0.775 and 101.00 +/- 1.192 for cefuroxime axetil and ornidazole, respectively.

High-performance thin-layer chromatographic determination of rabeprazole sodium and domperidone in combined dosage form.

A new, simple, high-performance thin-layer chromatographic method for determination of rabeprazole sodium (RAB) and domperidone (DOM) in combined tablet dosage form has been developed and validated. The mobile phase was toluene-acetone-methanol (4.5 + 4.5 + 0.5, v/v/v) with UV detection at 285 nm. The retention factors for RAB and DOM were found to be 0.53 +/- 0.12 and 0.32 +/- 0.20. The method was validated with respect to linearity, accuracy, precision, and robustness. Beer's law was obeyed in the concentration range of 50-800 ng/band for both RAB and DOM. The method has been successfully applied for the analysis of drugs in a pharmaceutical formulation.

Simultaneous spectrophotometric determination of Cefpodoxime proxetil and Potassium clavulanate.

Two accurate, precise, sensitive and economical procedures for simultaneous estimation of Cefpodoxime proxetil and Potassium clavulanate in tablet dosage form have been developed. The methods employed were absorbance correction method (I) and first order derivative spectroscopic method (II). The first method employs wavelength 288 nm for direct estimation of Cefpodoxime proxetil where Potassium clavulanate shows nil absorbance. Estimation of Potassium clavulanate is carried out after correction for absorbance of Cefpodoxime proxetil at 218 nm. The second method is based on first order derivative spectroscopy. Wavelengths 235.6 nm and 308.2 nm were selected for the estimation of the Potassium clavulanate and Cefpodoxime proxetil, respectively. Both the drugs obey Beer's law in the concentration range 5-50 microg/ml. The results of analysis have been validated statistically and by recovery studies. The percentage assay was found to be 99.54 +/- 0.285 for Cefpodoxime proxetil and 98.53 +/- 0.760 for Potassium clavulanate (Mean +/- SD) by method I and 99.93 +/- 0.270 for Cefpodoxime proxetil and 99.40 +/- 0.723 for Potassium clavulanate (Mean +/- S.D) by method II respectively.

Simultaneous estimation of Cefixime and Erdosteine in capsule dosage form by spectrophotometric method.

Two accurate, precise, rapid and economical methods viz. Absorption correction method and Dual wavelength method were developed for the estimation of Cefixime (CEF) and Erdosteine (ERDO) in capsule dosage form. In both the methods linearity was observed in the concentration range of 2-25 microg/ml for Cefixime and 3-37.5 microg/ml for Erdosteine. The results of the analysis have been validated statistically and by recovery studies. The percentage assay was found to be 100.03 +/- 0.68 for Cefixime and 99.5 +/- 1.0 for Erdosteine (Mean +/- S.D) by method A and 99.54 +/- 0.84 for Cefixime and 100.54 +/- 1.3 for Erdosteine (Mean +/- S.D) by method B respectively.

Column reversed-phase high-performance liquid chromatographic method for simultaneous determination of rabeprazole sodium and domperidone in combined tablet dosage form.

A new, simple column reversed-phase high-performance liquid chromatographic (HPLC) method for simultaneous determination of rabeprazole sodium (RAB) and domperidone (DOM) in a combined tablet dosage form has been developed and validated. Determination was performed using a Jasco HPLC system with a HiQ SiL octadecylsilane (C18) column (250 x 4.6 mm id), acetonitrile-0.1 M ammonium acetate (50 + 50, v/v) mobile phase, and paracetamol as an internal standard. The detection was performed using a UV detector set at 280 nm. The method was validated with respect to linearity, accuracy, precision, and robustness. Beer's law was obeyed in the concentration range of 1.0-10.0 and 0.5-5.0 microg/mL for RAB and DOM, respectively. The method has been successfully applied for the analysis of drugs in a pharmaceutical formulation.

Spectrophotometric and reversed-phase high-performance liquid chromatographic methods for simultaneous determination of escitalopram oxalate and clonazepam in combined tablet dosage form.

Simple, accurate, precise, and sensitive ultraviolet spectrophotometric and reversed-phase high-performance liquid chromatographic (RP-HPLC) methods for simultaneous estimation of escitalopram oxalate (ESC) and clonazepam (CLO) in combined tablet dosage form have been developed and validated. The spectroscopic method employs an absorbance correction method using 238.6 and 308 nm as 2 wavelengths for estimation with methanol and water as solvents. Beer's law is obeyed in the concentration range of 10.0-50.0 and 0.5-3.0 micro/mL for ESC and CLO, respectively. The RP-HPLC method uses a Jasco HPLC system with HiQ SiL C18 column (250 x 4.6 mm id) acetonitrile-0.005 M tetrabutylammonium hydrogen sulfate (55 + 45, v/v) as the mobile phase, and satranidazole as an internal standard. The detection was carried out using an ultraviolet detector set at 287 nm. For the HPLC method, Beer's law is obeyed in the concentration range of 10.0-60.0 and 0.5-3.0 microg/mL for ESC and CLO, respectively. Both methods have been successfully applied for the analysis of the drugs in a pharmaceutical formulation. Results of analysis were validated statistically and by recovery studies.

Simultaneous determination of cefuroxime axetil and ornidazole in tablet dosage form using reversed-phase high performance liquid chromatography.

A simple, accurate and sensitive reversed-phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous determination of cefuroxime axetil and ornidazole in combined tablet dosage form has been developed. The method was performed with a HiQ-SiL C18 column (250 mm x 4.6 mm) and photodiode array (PDA) detector, using 0.01 mol/L potassium dihydrogen orthophosphate-methanol (56 : 44, v/v) as the mobile phase and tinidazole as the internal standard. Beer's law obeys in the concentration ranges of 5 - 25 microg/mL and 10 - 50 microg/mL for cefuroxime axetil and ornidazole, respectively. The method has been successfully validated statistically and applied for the analysis of the drugs in pharmaceutical formulation.

Spectrophotometric simultaneous determination of rabeprazole sodium and itopride hydrochloride in capsule dosage form.

A new simple, economical, rapid, precise and accurate method for simultaneous determination of rabeprazole sodium and itopride hydrochloride in capsule dosage form has been developed. The method is based on ratio spectra derivative spectrophotometry. The amplitudes in the first derivative of the corresponding ratio spectra at 231nm (minima) and 260nm were selected to determine rabeprazole sodium and itopride hydrochloride, respectively. The method was validated with respect to linearity, precision and accuracy.

Clubbed thiazoles by MAOS: a novel approach to cyclin-dependent kinase 5/p25 inhibitors as a potential treatment for Alzheimer's disease.

A novel clubbed triazolyl thiazole series of cdk5/p25 inhibitors, potentially useful for the treatment of Alzheimer's disease, is disclosed. Evaluation of the SAR of substitution within these series has allowed the identification of a range of compounds which significantly reduce brain cdk5/p25 and thus have potential as possible treatments for Alzheimer's disease.

Simultaneous determination of rabeprazole sodium and itopride hydrochloride in capsule dosage form by spectrophotometry.

Three methods viz. Absorbance Ratio Method (I), Dual Wavelength Method (II) and First Order Derivative Spectroscopic Method (III) for simultaneous estimation of Rabeprazole sodium and Itopride hydrochloride have been developed. The drugs obey Beer's law in the concentration range 2-20 microg/ml for RAB and 5-75 microg/ml for ITO. The results of analysis of drugs have been validated statistically and by recovery studies.