Plant Biotechnology: Promises and Challenges

Development of procedures in cell biology to regenerate plants from single cells in any desired quantity provides the prerequisite for the practical use of plant tissue culture and genetic engineering in crop improvement. Such regenerating cell cultures are used for selection of mutants and for DNA transformation experiments. DNA transfer by means of engineered Ti and Ri plasmids has become an established technique for the rapidly growing list of dicotyledonous plants. Considerable success has also been achieved in making gene transfer techniques independent of cell culture methods. These techniques have given the opportunity to create, characterise and select plant cultivars which cannot be obtained by traditional breeding methods. The exploitation of plant cell cultures for production of pharmaceuticals, natural products of commercial importance and mass propagation of high-value crops by automation, have developed into an important industry with considerable potential for future. This paper discusses the recent advances and applications of plant biotechnology in agriculture and industry and the challenges the still exist

CACO-2 CELLS: AN OVERVIEW

The Caco-2 cell line is an immortalized line of heterogeneous human epithelialcolorectal adenocarcinoma cells. Human colonic adenocarcinoma cells that are able toexpress differentiation features characteristic of mature intestinal cells, such asenterocytes or mucus cells. These cells are valuable in vitro tools for studies related tointestinal cell function and differentiation. The Caco-2 cell line is widely used with invitro assays to predict the absorption rate of candidate drug compounds across theintestinal epithelial cell barrier. Caco-2 may also refer to a cell monolayer absorptionmodel. Cell-based functional assays, such as the Caco-2 drug transport model forassessing intestinal transport, are extremely valuable for screening lead compounds indrug discover

DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR THE ANALYSIS OF TOLPERISONE HYDROCHLORIDE IN PHARMACEUTICAL DOSAGE FORM

A simple, sensitive, rapid and precise high performance thin layer chromatographic method has been developed and validated for the estimation of Tolperisone hydrochloride in bulk and pharmaceutical dosage form. The stationary phase used was silica gel precoated aluminum plate 60F254 plates. The mobile phase used was a mixture of chloroform:acetone:toluene (6:2:2, v/v/v). The detection of spots was carried out at 265 nm. The method was validated in terms of specificity, linearity, precision and accuracy. The calibration curve was found to be linear between 50-600 ng/band. The developed method was subjected for forced degradation studies like acid, alkali, peroxide and thermal stress conditions were performed as per ICH guidelines. The proposed method was suitable for routine quality control analysis of Tolperisone hydrochloride in bulk and pharmaceutical formulation

Development of an In house Computer Code for the Simulation of Detonation Shock Dynamics in Underwater Explosion Scenario

Detonation Shock Dynamics (DSD), involved in an underwater explosion scenario is numerically simulated by an in-house computer code, ‘DSSDYN’. The simulation is based on Chapman-Jouguet (CJ) theory, formulated in Arbitrary Lagrangian Eulerian frame work. Specifically, the propagation of detonation front is simulated with‘Burn Fraction Model’. The classical burn fraction model is improved for achieving better computational efficiency. The simulation capability of DSS-DYN is demonstrated through a case study on explosion of PETN charge under the deep-water medium.Through this study, the salient features of DSD with better insight have been brought out. Besides, the physical parameters, such as work potential of PETN, are predicted efficiently.The apportionment of energy distributions indicates that about 70 % of chemical energy of explosive is transmitted to the surrounding water that is the major contribution of damage potential of the explosive. The predictions of peak velocity and peak pressure values by DSS-DYN and LS-DYNA show satisfactory comparison.DSS-DYN consumes lesser computational time (~1h), compared to LS-DYNA (~3h)

Numerical Simulation of Ballistic Impact on Armour Plate with a Simple Plasticity Model

Ballistic impact of a steel projectile on armour steel plate is examined by numerical simulations using 3-D nonlinear dynamic explicit finite element code ANSYS LS-DYNA. Simulations are attempted using a simple strain rate dependent plasticity model that can capture large strain, strain rate hardening and fracture encountered at high velocity ballistic impacts. Initial simulations are carried out for a cylindrical bullet with a semi-spherical nose shape impacting a military vehicle door at two different velocities as a test problem. This is then extended to simulate a real problem of armour piercing shot impact on a thick armour steel plate at ordnance velocity regime. The former is compared with results reported in published literature while the latter is assessed with the experimental findings. The deformation pattern generated in the deformed armour plate, residual projectile velocity and displacement of the projectile are taken as the necessary parameters for evaluating the results of simulation. The study presented in this paper demonstrates the effectiveness of the adopted simple plasticity model to simulate a highly nonlinear phenomenon to reasonably predict the physically measurable impact parameters.Defence Science Journal, Vol. 64, No. 1, January 2014, DOI:10.14429/dsj.64.452

DEVELOPMENT AND VALIDATION OF NEW STABILITY INDICATING REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR SIMULTANEOUS DETERMINATION OF METFORMIN HYDROCHLORIDE AND ERTUGLIFLOZIN IN BULK AND PHARMACEUTICAL DOSAGE FORM

Objective: The present study deals with the development, validation, and application of simple, precise, and accurate high-performance liquid chromatography (HPLC) method for the simultaneous estimation of metformin hydrochloride and ertugliflozin in pharmaceutical formulation and to validate.Methods: The analytical conditions were optimized on BDS C8 column (150 mm × 4.6 mm, 5 μm) at room temperature. The mobile phase consists of buffer: acetonitrile in 55:45 v/v ratio. Injection volume was 10 μl. The flow rate was maintained at 1.0 ml/min, and the analysis was carried out at 224 nm.Results: The method was found to be linear in the concentration range of 125–750 μg/ml and 1.875–11.25 μg/ml for metformin hydrochloride and ertugliflozin with regression coefficient r2 = 0.999. The method was found to be precise with percentage relative standard deviation below 2%. The limit of detection and limit of quantification were found to be within the limits. The percentage recovery of the developed method was 100.15%. All the validation parameters such as robustness, recovery, and precision were found to be within the limits. Degradation parameters such as acid, base, thermal and peroxide, light, temperature, and humidity were performed and found that the drugs are stable in all the extreme conditions.Conclusions: A simple, accurate, precise, and less time-consuming reversed-phase HPLC method for the simultaneous estimation of metformin hydrochloride and ertugliflozin has been developed and validated in accordance with the ICH guidelines

USE OF ION ASSOCIATION COMPLEX FORMATION FOR THE SPECTROPHOTOMETRIC DETERMINATION OF ITOPRIDE HCL IN BULK AND ITS PHARMACEUTICAL PREPARATIONS

Objective: The authors report two simple, accurate and economic spectrophotometric methods A and B for the determination of Itopride hydrochloride in bulk and dosage forms.Methods: The proposed methods are based on the formation of chloroform soluble ion-associates in the presence of acidic dyes namely BPB (Method A) and BCP (Method B) exhibiting lmax at 418 and 418 nm respectively.Results: Beer's law is found to be obeyed in the concentration range of 2.0-10.0 µg/ml and 2.0-10.0 µg/ml. The molar absorptivities are found to be 1.42x104 and 9.61x103L/mol. cm for methods A and B. These methods are successfully applied for the assay of Itopride hydrochloride in pharmaceutical formulations

A RAPID RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE QUANTITATIVE ESTIMATION RIBAVIRIN IN TABLETS

Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method and validate as per ICH guidelines for the quantitative estimation of Ribavirin (200mg) in tablets.Methods: The optimized method uses a reverse phase column, Enable Make KromasilC18 (250 X 4.6 mm; 5μ), a mobile phase of phosphate buffer (pH 4.2): acetonitrile in the proportion of 85:15 v/v, flow rate of 1.0 ml/min and a detection wavelength of 215 nm using a PDA detector.Results: The developed method resulted in Ribavirin eluting at 2.606 min. Ribavirin exhibited linear in the range 25-150μg/ml. The precision is exemplified by the relative standard deviation of 0.4%. Percentage Mean recovery was found to be in the range of 98â€102, during accuracy studies. The limit of detection (LOD) and limit of quantitiation (LOQ) was found to be 0.24ng/ml and 0.73ng/ml respectively.Conclusion: An accurate, precise and linear RP-HPLC method was developed and validated for the quantitative estimation of Ribavirin in VIRAZIDE (200mg) tablets as per ICH guidelines and hence it can be used for the routine analysis in various pharmaceutical industries.Â

STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF METFORMIN AND GLIPIZIDE

A novel, precise and accurate stability indicating RP-HPLC method was developed and validated for the simultaneous estimation of Metformin and Glipizide in combined pharmaceutical dosage form. Chromatographic separation was achieved on Microsorb-MV C18 column (250 × 4.6 mm, 5 μm) with UV detection at 257 nm. The mobile phase consists of acetate buffer (pH 4.0) and acetonitrile in the ratio of 60:40 v/v and at a flow rate of 1.0 mL/min. The method was linear over the concentration range of 60-140 μg/mL for Metformin and 10-50 μg/mL for Glipizide. The retention times for Metformin and Glipizide were found to be 2.434 min and 5.710 min respectively. The mean percentage recoveries of Metformin and Glipizide were found to be 100.42% and 100.39% respectively. The method was validated and was successfully employed for the routine quantitative analysis of pharmaceutical formulations containing Metformin and Glipizide in combined pharmaceutical formulation

Simultaneous determination of candesartan and hydrochlorothiazide in human plasma by LC-MS/MS

A simple, sensitive, rapid and highly efficient LC-MS/MS method was developed for the determination of Candesartan and Hydrochlorothiazide simultaneously in human plasma. The method employed Zorbax eclipse C18 (150 X 4.6 mm, 5µ) column using acetate buffer: acetonitrile (25:75%, v/v) as the mobile phase. The mobile phase flow rate is 1 mL/min which was delivered into the mass spectrometer electron spray ionization chamber. The Liquid/liquid extraction procedure was used in the method for the extraction of analytes. The chromatograph was attached to a negative ion mode tandem mass spectrometer and the method was validated for all the parameters as per the guidelines of US-FDA. The ions were detected in multiple reaction monitoring mode and the transitions are m/z 439.00®309.10 and 295.80®268.80 for candesartan and hydrochlorothiazide respectively. Isotopic standards were used as internal standards for effective recovery of the analytes. The drugs were analyzed over a calibration range of 1.027-302.047 ng/mL for candesartan and 1.044-306.945 ng/mL for hydrochlorothiazide respectively with regression coefficient greater than 0.99. The mean extraction recoveries are 96.95±5.61 and 100.55±4.82 for candesartan and hydrochlorothiazide respectively. The precision and accuracy values for all the studies were within the range of ≤15% and 85-115%. The performed stability studies indicate that the developed method is stable in plasma for 15 h at room temperature (bench top); 52 h (in injector); for 112 days at -70 ºC for long term stability; five successive freeze and thaw cycles. The developed method could be successfully employed for the determination of selected drugs in biological samples