Nilesh V. Pawar

Free amino acid profiling in grain amaranth using LC-MS/MS.

Reliable and reproducible techniques for identification and quantification of amino acids usually require derivatization. However, techniques such as LC-MS/MS may perhaps sideline the derivatization with significant accuracy. Understanding the interaction between free amino acids and several physiological processes apparently requires that the pool be studied in the seed itself. Data on free amino acid content of grain amaranth is very inadequate. The present analysis revealed a higher concentration of essential amino acids especially Threonine and Tryptophan than non essential amino acids. The study constitutes first report of use of LC-MS/MS method for analysing the free amino acid composition of amaranth grain.

Chemical Characterization, Mineral Analysis, and Antioxidant Potential of Two Underutilized Berries (Carissa carandus and Eleagnus conferta) from the Western Ghats of India

Diversity needs to be “conserved through use.” Owing to the global need, the present study includes two underutilized berries, Carissa carandus and Eleagnus conferta, to explore their potential as supplementary food by evaluating their antioxidant activity and approximate chemical composition. Berries of E. conferta are a richer source of carotene, ascorbic acid, protein, and magnesium, whereas berries of C. carandus are rich in sugars, lipids, iron, and magnesium. The antioxidant properties of the berries and a comparative account on their performance using DPPH and FRAP assay was studied. Both assays revealed high activity in the peel extract of berries of C. carandus. The LC-MS/MS profile of C. carandus reveals the presence of eleven different polyphenolic compounds while E. conferta had only four of these compounds.

Seasonal variation in content of camptothecin from the bark of Nothapodytes nimmoniana (Grah.) Mabb., using HPLC analysis

Objective: To study and compare seasonal variation in camptothecin (CPT) content from bark samples of Nothapodytes nimmoniana obtained from geographically and climatologically isolated populations. Methods: A standard High Performance Liquid Chromatography methodology was used to analyze and quantify CPT from bark samples of N. nimmoniana. Results: Sample collected from Amboli yielded highest CPT content 1.337 g/100 g dry bark powder during the monsoon compared to other localities in study. Monsoon (August) showed to accumulate higher levels of CPT in barks of N. nimmoniana as compared to summer (May). Amboli averaged highest accumulation of CPT compared to other localities under study. Conclusion: These findings indicate season to have control over accumulation of CPT. Locality Amboli has highest CPT content in all seasons and were the elite population during the study. The study also suggests the need for further investigation in lights of biosynthesis in the plant.

First report of Aspergillus niger as a plant pathogen on Zingiber officinale from India.

Zingiber officinale Rosc., belonging to the Zingiberaceae family, is medicinally important. It is commonly known as ginger and has been extensively cultivated for many centuries for use as a spice and traditional medicine in India. During a survey in September of 2007, leaves of Z. officinale showed a necrotic leaf spot disease on a plant growing in Kolhapur, India. Symptoms of the disease appeared as small (10 to 15 mm), rectangular to irregular, yellow spots on the leaves that covered a major area of the leaf when severe. The infection caused defoliation. Symptomatic leaves were collected and isolations from infected leaves were made on Czapeks Dox agar supplemented with streptomycin sulfate (30 mg/l). Plates were incubated at 28 to 30°C for 6 to 7 days. The fungal colonies were colorless to pale on the reverse side and covered with a dense layer of dark brown-to-black conidial heads. Conidia were globose to subglobose (3.5 to 5.0 μm in diameter), dark brown to black, and rough walled. 16S rRNA of isolates was amplified and sequenced (EMBL Accession No. AM941157) and compared with sequences of known Aspergillus species obtained from GenBank. The closest matches (99% identity) were with A. niger. On the basis of morphological and molecular approaches, the pathogen was confirmed as A. niger. Of five isolates, AN-5 was used for pathogenicity study. Kochs postulates were satisfied after reisolating the fungus from leaves inoculated with a conidial suspension that showed symptoms (6 days after inoculation) similar to the lesions observed on the leaves collected from Z. officinale. The tests were repeated three times in the greenhouse. Healthy, potted Z. officinale plants were grown in isolation after artificial inoculation with a suspension of spores (10,000 conidia per ml), which was prepared in 100 ml of sterile distilled water and then sprayed on the abaxial and adaxial surface of leaves. Noninoculated plants served as controls. These results confirmed the pathogenicity of A. niger on Z. officinale. To our knowledge, this is the first report of A. niger causing a disease on ginger (1). Reference: (1) G. N. Dake. J. Spices Aromatic Crops. 4:40, 1995.

RP-HPLC Analysis of an Alkaloid–Methyllycaconitine from Mutagenic Delphinium malabaricum (Huth) Munz

A systematic search of some phytochemicals in Delphinium malabaricum will be undertaken to evaluate the therapeutic quality of D. malabaricum. The populations of D. malabaricum were subjected to analysis of alkaloids by Reversed phase HPLC technique. Among the various alkaloids present in Delphinium species methyllycaconitine is a principle alkaloid. Therefore, the populations of D. malabaricum were assessed for Methyllycaconitine (MLA) content in the mutants and their control. The results revealed that the MLA content was highest with 0.01% concentration of EMS (0.78 mg/g plant material) as compared to the control (0.76 mg/g plant material). The concentration of MLA decreased with an increase in dose/concentration of the mutagens. Less concentration of MLA in the roots of D. malabaricum indicates that it is less toxic and a further decrease in concentration after mutagenic treatment minimizes its toxic effects. As these populations are growing in the same environmental and edaphic conditions, observed variability can be related to genetic makeup altered due to mutation. The RP-HPLC analysis results could form the basis of a more detailed study of root alkaloids and any marked similarity/dissimilarity in their alkaloid pattern may draw reasonable conclusions of the difference between the mutants and the parent cultivar.