ANTIOXIDANT AND CYTOTOXIC ACTIVITIES OF ARECA CATECHU SEED EXTRACT IN SWISS ALBINO MICE USING EAC CELL LINE IN DIFFERENT CULTURE MEDIUM

Objective: Areca nut is the dried ripe seeds of Areca catechu, belonging to family Palmae. Areca nut contains a number of alkaloids, belonging to pyridine and piperidine groups, derived from amino acid lysine. Arecoline a nicotinic acid-based alkaloid present which exerts sialagogue property. But the habit of chewing marketed gutka may cause oral leukoplakia, sometimes lead to squamous cell carcinoma. However, Based on the phytochemical compounds present, it is predicted that it must show antioxidant and may show cytotoxic activities. Methods: The antioxidant activity (nitrite scavenging and hydrogen peroxide scavenging) and cytotoxic activity in two different medium were checked in EAC cell line using the swiss albino mice model. Results: The study was also postulated an idea about the qualitative and quantitive analysis of Areca catechu. The plant extract showed good Nitrite and Hydrogen peroxide scavenging activity. The cytotoxicity study conducted in swiss albino mice, cell viability and IC50 value was checked. The cytotoxic activity of different concentrations of different fractions of plant extract was checked in two different medium, i.e., PBS and RPMI 1640. IC50 values for following fractions which were studied in PBS; as, 91.73±73 (µg/ml), 183±36.24 (µg/ml), 53.74±1.562 (µg/ml) for crude ethanolic extract, alkaloid fraction and flavonoid fraction respectively. IC50 value in RPMI 1640 medium obtained as; 44.18±1.09µg/ml, 54.27±0.2279µg/ml and 51.24±2.461µg/ml for crude ethanolic extract, Alkaloid and Flavonoid fraction, respectively. Conclusion: Areca nut extract showed good scavenging activity depending on concentration. Relatively RPMI 1640 medium showed better cytotoxic activity than other mediums

BIOEQUIVALENCE STUDY OF AZELNIDIPINE 16 MG TABLET TO EVALUATE PHARMACOKINETIC PROFILE OF SINGLE DOSE IN HEALTHY, ADULT, HUMAN VOLUNTEERS UNDER FASTING CONDITION

Objective: The present study's objective is to conduct a comparative bioavailability study with a special emphasis on the test product's bioequivalence using a standard reference product as a comparator. Methods: Before initiating the bioequivalence study, the plasma sample analysis method was developed and validated by using LC-MS/MS method. The entire study was conducted as a single-dose crossover randomized bioequivalence study with open-label, two treatment, two-period, and two sequences on 24 healthy volunteers under fasting condition. With proper informed consent process the oral dose of the Reference product (R) or Test product (T) was administered on healthy volunteers at 0 h during each period of the study. After the drug's oral administration, a certain quantity of blood sample was collected, and the plasma sample was separated using a cold centrifuge. The plasma samples were analysed by using the validated LC-MS/MS method. The pharmacokinetic parameters, statistical data and ANOVA of the test and reference product were evaluated. Results: The Cmax, Auc0-t, AUC0-∞ and tmax of the test product were found to be 6.29 ng/ml, 117.0 ng. h/ml, 161.67 ng. h/ml and 3.33 h. respectively. And the Cmax, Auc0-t, AUC0-∞ and tmax of reference product were found 6.59 ng/ml, 123.21 ng. h./ml, 172.20 ng. h/ml and 3.38 h respectively. Relative bioavailability was found 94.96%. The overall results show that the 90% confidence intervals (Log-Transformed and Untransformed) for Cmax, AUC0-t and AUC0-∞ for Azelnidipine were within the acceptable limit of 80%-125%. Conclusion: The entire study's conclusion can be drawn as the test product was bioequivalence with the reference product's comparator

AN LC-MS/MS BASED BIOANALYTICAL APPROACH TO RESOLVE PHARMACOKINETIC INVESTIGATION OF ACOTIAMIDE HYDROCHLORIDE AND ITS APPLICATION TO BIOEQUIVALENCE STUDY

Objective: Acotiamide, a prokinetic drug used to treat Functional Dyspepsia, which acts by modulating gastric motility. However, in this present study, a simple and accurate bioanalytical method was developed for the estimation of Acotiamide in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and validated according to US-FDA guideline. Methods: The method was developed in blank human blood plasma; propranolol was used as internal standard (IS). Protein Precipitation technique was followed for the extraction of the drug from the plasma sample. In liquid chromatography, the C18 analytical column (50 x 3 mm, particle size-5 μm) was used; as a mobile phase, 0.1% formic acid in Mili Q water, and ACN with methanol (1:1) used, at 0.50 ml/min flow rate. Detection was done by positive electrospray ionization (ESI) with a run time of 7 min in multiple reaction monitoring (MRM) mode. Eight calibration concentrations were taken, ranging from 1.5625-200 ng/ml for Acotiamide. Different stability studies were performed and obtained results found within the acceptable range. Moreover, a comparative pharmacokinetic analysis was done in 24 healthy human volunteers in a single dose, randomized, crossover study. Results: The precursor to production reaction was; m/z 451.200 → 271.200 for Acotiamide and m/z 260.300 → 116.100 m/z for IS. The obtained calibration curve was linear, with a mean r2value 0.9953. Among the pharmacokinetic parameters, Cmax and Tmax were 25.71±2.31,23.61±2.32 ng/ml; 2.54±0.12, 2.43±0.21 h for reference and test samples, respectively. Conclusion: No major adverse events were noted in the clinical phase, the developed method was accurate and linear; obtained pharmacokinetic parameters hence represented

DETERMINATION OF METFORMIN AND SITAGLIPTIN IN HEALTHY HUMAN VOLUNTEERS' BLOOD PLASMA AND ITS BIOEQUIVALENCE STUDY UNDER FASTING CONDITION

Objective: Metformin hydrochloride and sitagliptin are the oral anti-hyperglycemic medications used to treat type 2 diabetes and are used in combination to treat patients. In this work, we have developed a bioanalytical method for simultaneous estimation of both the drugs form some formulation and subsequently the validation of the developed method metformin and sitagliptin in human plasma. Methods: The stability studies were done as per USFDA and EMA guidelines. The sample extraction approach presented here was a straightforward liquid extraction. The linearity range of metformin was 11.72 ng/ml to 3000 ng/ml, and sitagliptin was 4.68 ng/ml. to 1200 ng/ml. For metformin, the LOD was 1.0 ng/ml, and LLOQ was 11.72 ng/ml. and for sitagliptin, the LOD was 0.75 ng/ml, and LLOQ was 4.68 ng/ml. LC-ESI-MS/MS was used to develop and validate this method using the Phenomenex Kinetex C18 column. Milli-Q water containing 10 mmol Ammonium Acetate (pH =3.6) and Acetonitrile containing 0.1% Formic Acid (pH =2.4) as solvent systems for the estimation of Sitagliptin in a single dose. Metoprolol is used as an Internal Standard. Results: The total chromatographic run time was only 7.0 min, and the elute time of metformin and sitagliptin was 3.94 min and 3.97 min, respectively. Relative Bioavailability was found at 101.14% for Metformin and 96.96% for Sitagliptin. The overall results show that the Cmax, AUC0-t, and AUC0-∞ for metformin and sitagliptin were within the acceptable limit of 80%-125%. Conclusion: This bioanalytical method was successfully applied in the bioequivalence study. The study design was a randomized, open-label, two treatment, two-period, two sequences, single-dose, crossover bioequivalence study under fasting conditions

Formulation development and optimization of sustained release matrix tablet of Itopride HCl by response surface methodology and its evaluation of release kinetics

AbstractThe objective of this present investigation was to develop and formulate sustained release (SR) matrix tablets of Itopride HCl, by using different polymer combinations and fillers, to optimize by Central Composite Design response surface methodology for different drug release variables and to evaluate drug release pattern of the optimized product. Sustained release matrix tablets of various combinations were prepared with cellulose-based polymers: hydroxy propyl methyl cellulose (HPMC) and polyvinyl pyrolidine (pvp) and lactose as fillers. Study of pre-compression and post-compression parameters facilitated the screening of a formulation with best characteristics that underwent here optimization study by response surface methodology (Central Composite Design). The optimized tablet was further subjected to scanning electron microscopy to reveal its release pattern. The in vitro study revealed that combining of HPMC K100M (24.65 MG) with pvp(20mg)and use of LACTOSE as filler sustained the action more than 12h. The developed sustained release matrix tablet of improved efficacy can perform therapeutically better than a conventional tablet

EFFECTS OF ANTI ANXIETY DRUG MIDAZOLAM AS PRETREATMENT THERAPY FOR ANXIETY OF ZEBRA FISHES INDUCED BY THE EXPOSURE OF UNFAMILIAR ENVIRONMENT OF AQUATIC WHITE/BLACK MAZE

Objectves: The objective of our present research investigation was to evaluate the effectiveness of the aquatic white/black plus maze model toevaluate the efficacy of different treatment drugs on zebra fish behavior studies.Methods: Animals were divided into three groups each containing six animals. The first two groups were pretreated with lower and higher doses ofmidazolam and then they were exposed to unfamiliar environments of aquatic white/black plus maze. The last group of animals was only immersedinto the unknown familiar in the form of aquatic white/black plus maze without any pretreatment of the drugs.Results: The results illustrated that the pretreatment drugs certainly improved the behavior status of the animal in the form of releasing anxiety andstress, but the doses of different pretreatment have no significant effects on improvement of behavioral status.Conclusion: In conclusion, it can be described that this model is highly effective to find the proper drug for the treatment of anxiety but it is not soeffective to find the proper dose of treatment.Keywords: Midazolam, Zebra fish, White/black maze, Anti anxiety, Pretreatment therapy

Design of multi-particulate "Dome matrix" with sustained-release melatonin and delayed-release caffeine for jet lag treatment

10.1016/j.ijpharm.2020.119618INTERNATIONAL JOURNAL OF PHARMACEUTICS58

LC-MS/MS simulatneous determination of itopride hydrochloride and domperidone in human plasma

A rapid, simple, sensitive and specific liquid chromatography–tandem mass spectrometry method was developed and validated for simultaneous quantification of itopride hydrochloride and domperidone in human plasma. Both drugs were extracted by liquid–liquid extraction with ethyl acetate and saturated borax solution. The chromatographic separation was performed on a reversed-phase C18 column with a mobile phase of water–methanol (2:98, v/v) containing 0.5% formic acid. The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The assay exhibited linearity over the concentration range of 3.33–500 ng mL−1 for itopride hydrochloride and 3.33–100 ng mL−1 for domperidone in human plasma. The precursor to product ion transitions of m/z 359.1–72.3 and 426.0–147.2 were used to measure itopride hydrochloride and domperidone respectively. The method was found suitable for the analysis of plasma samples collected during phase 1 pharmacokinetics study of itopride HCl 50 mg and domperidone 20 mg in 12 healthy volunteers after single oral doses of the combination drug

LC-MS-MS development and validation for simultaneous quantitation of metformin, glimepiride and pioglitazone in human plasma and its application to a bioequivalence study

A simple, precise and reproducible liquid chromatography–tandem mass spectrometry method has been developed and validated according to the Food and Drug Administration guidelines for the simultaneous quantitation of antidiabetic drugs metformin, glimiperide and pioglitazone in human plasma using glipizide as an internal standard. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in multiple reaction monitoring and positive ion mode. Inter-batch and intra-batch coefficient of variation across four validation runs for the quality control samples was less than 7%. The accuracy determined at quality control levels was within 92.81–105.13%. The method was applied to a bioequivalence study

Advanced Dome cellulose/alginate/chitosan composite matrix design with gastric and intestinal co-targeting capacities

Dome matrix was designed with gastric and intestinal targeting capacities using melatonin and caffeine as model drugs, and alginate, chitosan and cellulose as composite materials. The melatonin, caffeine and intermediate hydroxypropylmethylcelluose-based dispersible modules were prepared through compaction. Caffeine piled module was capped at both ends with melatonin void modules via intermediate dispersible modules into Dome matrix. Dispersion of intermediate module detached melatonin module from Dome matrix and had it floated in stomach providing a more complete melatonin release due to favorable pH-pKa relationship of dissolution medium and drug. With reference to the caffeine module, the detachment of melatonin module facilitated its gastrointestinal transit as a reduced size matrix, with majority of caffeine delivered in colon. The dual site -targeted and-release Dome matrix is applicable as reference oral carrier for pharmaceutical, nutraceutical, functional food and veterinary medicine where a complex formulation and performance in vivo are required