ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF ERLOTINIB HYDROCHLORIDE BULK AND IN PHARMACEUTICAL DOSAGE FORM

A simple, precise, and accurate RP – HPLC method was developed and validated for the  determination of Erlotinib Hydrochloride in bulk and tablet dosage form. Gradient elution at a flow rate of 1.0 mL/min was employed on zorbax XDB C18 (150 X 4.6 I.D., 5µm particle size) at ambient temperature. The mobile phase consisted of Acetonitrile and 0.02M ammonium acetate, adjusted to pH 3.3 with acetic acid. Acetonitrile and water(50:50) was used as a diluents. The UV detection wavelength was 247nm and 20µL of sample was injected. The retention times of Erlotinib Hydrochloride was found to be 4.576 min. The linearity was obtained in the range of 50 – 150 µg/mL. The % RSD for precision and accuracy of the method was found to be less than 1%.  The method was validated as per the ICH guideline. The proposed method was suitable for the analysis of Erlotinib Hydrochloride in tablet formulation for quality control purpose. Key Words: Erlotinib Hydrochloride, RP – HPLC, UV, %RSD, Method validatio

Interspecific hybridization between Cajanus cajan (L.) Millsp. and C. lanceolatus (WV Fitgz) van der Maesen

Cultivated pigeonpea has a narrow genetic base. Wild relatives play an important role in the efforts to broaden its genetic base. In this report, we present a successful wide-cross between the cultivated pigeonpea and Cajanus lanceolatus, a wild relative from the secondary gene pool, native to Australia, with desirable traits such as frost and drought resistance. A range of F1 progeny were obtained and the resultant F1 hybrid plants set mature pods and seeds. The hybrids had intermediate morphology, sharing the traits of both the parents. All the F1 hybrids flowered profusely. Some of the hybrids were completely male sterile and some were partially fertile with pollen fertility ranging from 35 to 50 %. Meiotic analysis of the fertile F1 hybrids revealed a high degree of meiotic chromosome pairing between the two parental genomes. Meiotic analysis of the sterile F1 hybrids revealed that the breakdown of microsporogenesis occurred at the post-meiotic stage after the formation of tetrads. Fertile plants formed regular bivalents with normal disjunction, except for occasional asynchrony at meiotic II division

Health and Wellness Product from Mangosteen (Garcinia mangostana L.) Rind: Bioactive Potentials

Mangosteen rind (MSR) (Garcinia mangostana L.) is a predominant component of the fruit contributing to 62% of the whole fruit. However, utilization of the same for the preparation of health products was not explored due to its sensorially less acceptable parameters. Differential extraction in different polarity solvents of MSR was done and evaluated their acceptability for product preparation.Current study thus is a detailed investigation on bioactivity profiling of MSR fraction and utilization of the same for health product preparation. Among various extracts, 70% ethanol (70%AE) yielded the maximum (15g/100g). Xanthone:Phenolic ratio was 1: 2.8, in 70%AE as opposed to hot water extract – HWE and 50% AE, which contained Xanthone:Phenolic ratio of 1:1.4/5. Higher the phenolic content obviously reduces the bitterness of Xanthones. 70% AE contained phenolics 60.08± 0.213 mg/g and xanthones 22.56± 0.317 mg/g. HPLC analysis revealed a spectrum of phenolic acids such as gallic, chlorogenic, caffeic, epicatechin, catechin and ferulic acids at various levels. Potent Free Radical Scavenging (FRS) activity, cytoprotectivity, DNA protectivity, H+K+ATPase inhibitory (PPAI) activities were observed in 70% AE. Gallic/tannic acid appear to contribute to antioxidant activity; while ferulic acid was responsible for PPAI activity in 70%AE. Among xanthones, although α- mangostin was the dominating component, gartanin, 8 deoxygartanin and 3-isomangostin contributed to FRS activity. The products were prepared from 70%AE which are sensorially acceptable. Data thus for the first time delineate the specific health beneficial role of both phenolic and xanthone constituents in MSR particularly with higher abundance of phenolics than xanthones

Research for practice in small human service organisations: doing and disseminating smallscale research

A series of novel alkynyl substituted 3,4-dihydropyrimidin-2(1H)-one (DHPM) derivatives were designed, synthesized and evaluated in vitro as potential inhibitors of chorismate mutase (CM). All these compounds were prepared via a multi-component reaction (MCR) involving sequential I2-mediated Biginelli reaction followed by Cu-free Sonogashira coupling. Some of them showed promising inhibitory activities when tested at 30 μM. One compound showed dose dependent inhibition of CM with IC50 value of 14.76 ± 0.54 μM indicating o-alkynylphenyl substituted DHPM as a new scaffold for the discovery of promising inhibitors of CM

Molecular genetic relationships among Arachis diogoi and A. chiquitana accessions

Seventeen SSR markers were selected randomly for molecular characterization/fingerprinting of two accessions of Arachis diogoi and three accessions of A. chiquitana to ascertain whether the morphologically different A. chiquitana accession ICG 11560 may be an accession of A. diogoi (=A. chacoense). Two accessions of A. chiquitana, ICG 13181 and ICG 13241, formed a distinct group. A. chiquitana accession ICG 11560 did not group closely with the other A. chiquitana accessions, but showed closer relationship with them than with the A. diogoi accessions ICG 4983 and ICG 8962. The two accessions of A. diogoi, ICG 4983 and ICG 8962, grouped together and were separated from the group formed by section Procumbentes members. These results show that A. chiquitana accessions in general and ICG 11560 in particular are not related to accessions of A. diogoi. In another experiment, similar results were obtained when RAPDs (random amplified polymorphic DNAs) were used to distinguish wild Arachis species

Protease inhibitors of Cajanus conferring resistance to pod borer of pigeonpea (Cajanus cajan L. Millsp.).

Pigeonpea is susceptible to pod borer damage with resistance lacking in its primary gene pool. Many Cajanus species harbor high levels of resistance. Host plant resistance can play an important role in minimizing the extent of losses due to insects and pests as well as the use of insecticides/pesticides and thus protect the environment. A major initiative was undertaken to tap the defence genes from wild relatives of secondary and tertiary gene pool through wide hybridization and thereby introgress resistance to pod borer. A range of interspecific derivatives derived from C. lanceolatus, C. cajanifolius, C. volubilis and C. platycarpus along with their parents were screened for the pod borer resistance under unprotected field conditions at ICRISAT, Patancheru, India. Biochemical basis of resistance was also identified by studying the levels of defence proteins active against bovine pancreatic trypsin, chymotrypsin and trypsin-like enzymes of H. armigera mid-gut proteases. Protease inhibitor profiles of parents and interspecific derivatives differed in terms of activity units, number and intensities of activity bands visualized on gelatin-PAGE. As the protease inhibitors are anti-nutritional factors, parents and interspecific derivatives, which resulted in high levels of Helicoverpa gut protease inhibitor (HGPI) units were screened for Human pancreatic trypsin inhibitor (HPTI) activity levels. Samples with high ratio of HGPI/HPTI represent less or no effect on human pancreatic trypsin and high effect on insect gut proteases

Purification and characterization of Bowman-Birk and Kunitz isoinhibitors from the seeds of Rhynchosia sublobata (Schumach.) Meikle, a wild relative of pigeonpea

Rhynchosia sublobata, a wild relative of pigeonpea, possesses defensive proteinase/protease inhibitors (PIs). Characterization of trypsin specific PIs (RsPI) separated from seeds by column chromatography using 2-D gel electrophoresis and Edman degradation method identified R. sublobata possessed both Bowman-Birk isoinhibitors (RsBBI) and Kunitz isoinhibitors (RsKI). A quick method was developed to separate RsBBI and RsKI from RsPI based on their differential solubility in TCA and acetate buffer. N-terminus sequencing of RsBBI and RsKI by MALDI-ISD ascertained the presence of Bowman Birk and Kunitz type isoinhibitors in R. sublobata. RsBBI (9216 Da) and RsKI (19,412 Da) exhibited self-association pattern as revealed by western blotting with anti-BBI antibody and MALDI-TOF peptide mass fingerprint analysis, respectively. RsBBI and RsKI varied significantly in their biochemical, biophysical and insecticidal properties. RsBBI inhibited the activity of trypsin (Ki = 128.5 ± 4.5 nM) and chymotrypsin (Ki = 807.8 ± 23.7 nM) while RsKI (Ki = 172.0 ± 9.2 nM) inhibited the activity of trypsin alone, by non-competitive mode. The trypsin inhibitor (TI) and chymotrypsin inhibitor (CI) activities of RsBBI were stable up to 100 °C. But, RsBBI completely lost its TI and CI activities on reduction with 3 mM DTT. Conversely, RsKI lost its TI activity on heating at 100 °C and retained >60% of its TI activity in presence of 3 mM DTT. CD spectroscopic studies on RsBBI and RsKI showed their secondary structural elements in the following order: random coils > β-sheets/β-turns > α-helix. However, RsKI showed reversible denaturation midpoint (Tm) of 75 °C. Further, the significant inhibitory activity of RsBBI (IC50 = 24 ng) and RsKI (IC50 = 59 ng) against trypsin-like gut proteases of Achaea janata (AjGPs) and Helicoverpa armigera (HaGPs) suggest them as potential biomolecules in the management of A. janata and H. armigera, respectively

Development of a new CMS system in pigeonpea utilizing crosses with Cajanus lanceolatus (WV Fitgz) van der Maesen

Cytoplasmic male sterility is an important biological tool which is now available to pigeonpea breeders to exploit heterosis/hybrid vigor. A variety of CMS systems have been developed when wild relatives of pigeonpea from different gene pools were crossed as the female parent with cultivated types as the male parent. This paper reports a second source of CMS developed by using the cultivated pigeonpea as the female parent and one of its wild relative Cajanus lanceolatus (WV Fitgz) van der Maesen as the pollen donor, as such the A5 CMS system derived from C. acutifolius. All the F1 hybrids were evaluated to confirm hybridity using 27 simple sequence repeat (SSR) markers. SSR marker analysis of parents provided 17 polymorphic markers from a total of 27 SSR markers used. Subsequently polymorphic SSRs were used to confirm the hybridity of the F1 plants. F1 hybrid plants were crossed with a range of pigeonpea cultivars to identify maintainers of male sterility. Morphology of the F1 and backcross generations, cytology of the sterile as well as fertile floral buds derived from the crosses between sterile F1 hybrids and unrelated pigeonpea cultivars were studied. An important observation made was that male sterility was a post meiotic process. Microsporogenesis was normal until the tetrad stage, but none of them formed pollen grains. Instead, they grouped together within the pollen mother cell wall and the tetrads did not separate into individual pollen grains

Effect of supplementing encapsulated microbial enzymes to corn-soybean meal based pelleted diets on performance and carcass variables in coloured broiler chicken

The experiment was conducted to determine the dietary supplementation of encapsulated (EC) or UEC enzymes on performance of colour broilers. The dietary supplementation of EC polysaccharide hydrolyzing enzymes and protease improved the performance compared to other groups. Feeding the birds with low nutrient-density diets did not affect the performance of the coloured broilers. The supplementation of EC enzymes resulted in a decrease in abdominal fat and increase in ready to cook yield compared to those diets with UEC or without enzymes

Expression of Pennisetum glaucum Eukaryotic Translational Initiation Factor 4A (PgeIF4A) Confers Improved Drought, Salinity, and Oxidative Stress Tolerance in Groundnut

Eukaryotic translational initiation factor 4A belong to family of helicases, involved in multifunctional activities during stress and non-stress conditions. The eIF4A gene was isolated and cloned from semi-arid cereal crop of Pennisetum glaucum. In present study, the PgeIF4A gene was expressed under the regulation of stress inducible Arabidopsis rd29A promoter in groundnut (cv JL-24) with bar as a selectable marker. The de-embryonated cotyledons were infected with Agrobacterium tumefaciens (LBA4404) carrying rd29A:PgeIF4A construct and generated high frequency of multiple shoots in phosphinothricin medium. Twenty- four T0 plants showed integration of both nos-bar and rd29A-PgeIF4A gene cassettes in genome with expected amplification products of 429 and 654 bps, respectively. Transgene copy number integration was observed in five T0 transgenic plants through Southern blot analysis. Predicted Mendelian ratio of segregation (3:1) was noted in transgenic plants at T1 generation. The T2 homozygous lines (L1-5, L8-2, and L16-2) expressing PgeIF4A gene were exhibited superior growth performance with respect to phenotypic parameters like shoot length, tap root length, and lateral root formation under simulated drought and salinity stresses compared to the wild type. In addition, the chlorophyll retention was found to be higher in these plants compared to the control plants. The quantitative real time—PCR results confirmed higher expression of PgeIF4A gene in L1-5, L8-3, and L16-2 plants imposed with drought/salt stress. Further, the salt stress tolerance was associated with increase in oxidative stress markers, such as superoxide dismutase accumulation, reactive oxygen species scavenging, and membrane stability in transgenic plants. Taken together our results confirmed that the PgeIF4A gene expressing transgenic groundnut plants exhibited better adaptation to stress conditions