Brij Lal

Simultaneous densitometric determination of shikonin, acetylshikonin, and β-acetoxyisovaleryl-shikonin in ultrasonic-assisted extracts of four Arnebia species using reversed-phase thin layer chromatography.

A simple, precise, and rapid high-performance thin-layer chromatographic (HPTLC) method for the simultaneous quantification of pharmacologically important naphthoquinone shikonin (1) together with its derivatives acetylshikonin (2), and beta-acetoxyisovalerylshikonin (3) in four species of genus Arnebia (A. euchroma, A. guttata, A. benthamii, and A. hispidissima) from the Indian subcontinent has been developed. In addition, the effect of solvents with varying polarity (hexane, chloroform, ethyl acetate, and methanol) for the extraction of these compounds was studied. HPTLC was performed on precoated RP-18 F(254S )TLC plates. For achieving good separation, mobile phase consisting of ACN/methanol/5% formic acid in water (40:02:08 v/v/v) was used. The densitometric determination of shikonin derivatives was carried out at 520 nm in reflection/absorption mode. The method was validated in terms of linearity, accuracy, precision, robustness, and specificity. The calibration curves were linear in the range of 100-600 ng for shikonin and acetylshikonin, and 100-1800 ng for beta-acetoxyisovalerylshikonin. Lower LOD obtained for compounds 1-3 were 18, 15, and 12 ng, respectively, while the LOQ obtained were 60, 45, and 40 ng, respectively.

Stratification of forest density and its validation by NDVI analysis in a part of western Himalaya, India using Remote sensing and GIS techniques

This study deals with the assessment of the status of forest density in the Kangra district of Himachal Pradesh, western Himalaya. A classified forest map of the area with an accuracy of 88.17% was produced using the hybrid classification method in Erdas Imagine. An IRS 1D LISS III satellite image was used for mapping and classification. Forest density was calculated in the ArcGIS environment by overlaying a mesh of uniform resolution cells (500 m×500 m) on a classified forest map. The forest density value of each cell was later used for the preparation of forest density contours. The forest density output was verified by Normalized Difference Vegetation Index (NDVI) analyses. The forest cover of the study area was found to be 34.3%. Baroh area had the highest (45.87%) forest density and Baijnath (18.65%) the lowest. Central and western regions of the district showed high‐value forest density contours (>50%). The derived NDVI values were compared against the forest density classes for assessing the accuracy of the results obtained. A positive correlation (r = 0.99) between NDVI values and forest density confirms the accuracy of the results.

Vascular plants, Kinnaur, Himachal Pradesh, India

In the present study, we provide a checklist of the vascular plants of Kinnaur district situated in the Himachal Pradesh state of India in the western Himalaya. This checklist includes 893 taxa ( viz ., species, subspecies and varieties) belonging to 881 species of angiosperms and gymnosperms distributed among 102 families and 433 genera, and 30 species of pteridophytes. Information about the growth habit, threat and endemicity status is also provided. Our results show that family Compositae is by far the most species rich family with 122 species, followed by Poaceae (69), Rosaceae (58), Leguminosae (49) and Lamiaceae (38). Among the genera, Artemisia is the most diverse genus with 19 species, followed by Potentilla (14), Saussurea (13), Polygonum (11), Astragalus (10), Lonicera (10) and Nepeta (10). Similar to other regions in the western Himalayan range, family-to-genera ratio was 1:4.25 and the genera-to-species ratio was 1:2.04. Out of 893 taxa, our checklist includes 606 herb species, 63 trees, 108 shrubs, 28 climbers, 67 graminoids and 21 sedges and rushes. Of all the species recorded, 108 (12.2%) are endemic to western Himalaya and 27 (3%) are placed under IUCN threatened categories. The present checklist on the flora of Kinnaur provides an important baseline data for further quantitative studies on the characteristics of plant communities in this region and will help in the identification of priority conservation areas.

Assessment of Bioaccumulation of Heavy Metal by Pteris Vittata L. Growing in the Vicinity of Fly Ash

Pteris vittata L. subsp. vittata, a potential arsenic hyperaccumulator fern, growing naturally in the vicinity of fly ash was analyzed for the concentration of nine heavy metals (Fe, Cu, Zn Ni, Al, Cr, Pb, Si, and As) from five different sites around of Kanti Thermal Power Station at Muzaffarpur in Bihar State, India. Metal accumulation in P. vittata was correlated with the level of pollution at five selected sampling sites. The results revealed significantly more accumulation of these metals in the above ground parts of the plant than the parts below ground. Statistical parameters such as the coefficient of variation (CV%) showed a higher for As, Cu, Cr, and a lower one for Fe, Ni, Al. There was high spatial variability in the total metal concentration at different sites. The present study confirmed that P. vittata is a heavy metals accumulator and that it is a highly suitable candidate for phytoremediation of metal contaminated wastelands.

Assessment of native plant species for phytoremediation of heavy metals growing in the vicinity of NTPC sites, Kahalgaon, India

ABSTRACT The present investigation was carried out to screen native plants growing in fly ash (FA) contaminated areas near National Thermal Power Corporation (NTPC), Kahalgaon, Bihar, India with a view to using them for the eco-restoration of the area. A total number of 30 plant species (5 aquatic and 25 terrestrial including 6 ferns) were collected and their diversity status and dominance were also studied. After screening of dominant species at highly polluted site, 8 terrestrial and 5 aquatic plants were analyzed for heavy metals (Fe, Zn, Cu, Ni, Si, Al, Pb, Cr, and Cd). Differential accumulations of various heavy metals by different species of plants were observed. Typha latifolia was found to be most efficient metal accumulator of Fe (927), Cu (58), Zn (87), Ni (57), Al (67), Cd (95), and Pb (69), and Azolla pinnata as Cr (93) hyper-accumulator among aquatic species in µg g−1. In terrestrial species the maximum levels of Fe (998), Zn (81), Ni (93), Al (121), and Si (156) were found in Croton bonplandium. However, there was high spatial variability in total metal accumulation in different species indicated by coefficient of variation (CV%). These results suggest that various aquatic, some dominant terrestrial plants including fern species may be used in a synergistic way to remediate and restore the FA contaminated wastelands.

Screening of bioconstituents and in vitro cytotoxicity of Clematis gouriana leaves

Clematis gouriana (Ranunculaceae), a perennial herb, is used by the local inhabitants of the western Himalayan region for its medicinal properties. Major bioconstituents of C. gouriana leaves using different solvent extracts were obtained and analysed. The results revealed promising contents of phenolics (from 18.19 ± 0.10 to 22.17 ± 0.10 mg g− 1) as gallic acid and flavonoids (from 2.83 ± 0.01 to 6.52 ± 0.08 mg g− 1) as quercetin equivalent in different extracts. Aqueous acetone extract showed higher antioxidant activity with IC50 value of 129.11 and 25.35 μg mL− 1 against DPPH and ABTS free radicals, respectively. Antioxidant yield ranged from 16.87 ± 0.27 to 24.48 ± 0.13 mg g− 1 of Trolox equivalent in different extracts as measured by the FRAP assay. Furthermore, ethylacetate extract exhibited strong in vitro cytotoxicity against Chinese hamster ovary and glioma cell lines. Proximate composition (proteins, fats, ash and minerals) of C. gouriana leaves was also assessed. Results demonstrated the potential of C. gouriana bioconstituents as nutraceuticals.

Differences in tree species diversity and soil nutrient status in a tropical sacred forest ecosystem on Niyamgiri hill range, Eastern Ghats, India

We have quantitatively analyzed the tree species diversity with respect to soil nutrient status in three sites of a sacred forest ecosystem of Niyamgiri hill range, Eastern Ghats, India. Extensive field surveys and sampling were conducted in 3 sites of the hill range: Site 1 Pterocarpus dominated forest (PTF) (19°40′02.2″ N and 83°21′23.1″ E), Site 2 Mangifera dominated forest (MAF) (19°40′02.8″ N and 83°21′40.8″ E) and Site 3 Mixed forest (MIF) (19°36′47.1″ N and 83°21′02.7″ E). A total of 28 families, 42 genera, 46 tree species, and 286 individual trees were recorded on an area of 0.6 ha. Tree density varied between 470 and 490 individuals ha−1 and average basal area between 3.16 and 10.04 m2 ha−1. Shannon Index (H′) ranged from 2.34 to 4.53, Simpson’s Index ranged from 0.07 to 0.09, and equitability Index ranged from 0.7 to 1.34. The number of individuals was highest in the girth at breast height (GBH) class of 50–70 cm. The soil nutrient status of the three forest types was related to tree species diversity. The soil pH value of the three sites reflected the slightly acidic nature of the area. Species diversity was positively correlated with organic carbon and phosphorus and negatively with nitrogen, EC and pH. The results of the current study may be helpful to further develop a conservation plan for tree species in tropical sacred forest ecosystems.

Comparative studies of volatile oil composition of Rhododendron anthopogon by hydrodistillation, supercritical carbon dioxide extraction and head space analysis

Volatile oil composition of the leaves of Rhododendron anthopogon (Ericaceae) growing wild in alpine Western Himalaya was studied using different extraction techniques including SC-CO2 extraction and hydrodistillation (HD). Results from different extraction methodologies were compared with headspace analysis (HS) and evaluated for the effectiveness of techniques in characterisation of various terpene categories and to assess their influence on the yield and composition of volatiles. Variability in constituents and in quantitative yields was observed. The results varied with different extraction methods. A total of 27 constituents in SC-CO2 extraction, 31 in HD and 17 in HS analysis were identified. Constituents in SC-CO2 and HD oils were identified by gas chromatography mass spectrometry analysis. SC-CO2 extraction was carried out at 40°C and 140 bar pressure and the oil represented by major constituents as β-caryophyllene (5.96%), α-humulene (4.06%) and p-menthadiene-2,9-diol (7.28%); in HD, oil limonene (11.26%), β-caryophyllene (11.62%), α-humulene (7.22%), and E-nerolidol (5.83%) dominated the oil and in HS analysis, limonene (24.14%), γ-terpinene (40.73%), α-terpinene (4.92%), β-phellandrene (3.44%) and β-ocimene (7.15%) were present as major constituents.

Composition and antimicrobial activity of the essential oil of Heracleum thomsonii (Clarke) from the cold desert of the western Himalayas.

Volatile oil composition of hydro-distilled (HD) and supercritical carbon dioxide (SC-CO2) essential oil of freshly collected aerial parts of Heracleum thomsonii (Umbeliferae) from the western Himalayas was studied by GC-FID and GC-MS. Results revealed qualitative and quantitative dissimilarity in the composition of hydro-distilled and SC-CO2 extracted oils. Nineteen constituents, which accounted for 89.32% of total constituents in HD oil, represented by limonene (4.31%), (Z)-β-ocimene (3.69%), terpinolene (22.24%), neryl acetate (36.19%), nerol (9.51%) and p-cymene-8-ol (2.61%) were identified. In SC-CO2 extracted oil, 24 constituents representing 89.95% of total constituents were identified. Terpinolene (5.08%), germacrene D (2.17%), neryl acetate (51.62%), nerol (9.78%), geranyl acetate (2.06%), α-bisabolol (2.48%) and 1-nonadecanol (4.96%) were the dominating constituents. In vitro antimicrobial activity of hydro-distilled oil was conducted against microrobial strains including two Gram-positive (Staphylococcus aureus and Bacillus subtilis) and five Gram-negative (Burkholderia cepacia, Escherichia coli, Enterobacter cloacae, Klebseilla pneumoniae and Pseudomonas aeruginosa) bacteria as well as seven fungi (Candida albicans, Issatchenkia orientalis, Aspergillus flavus, Aspergillus niger, Aspergillus parasiticus, Aspergillus sydowii and Trichophyton rubrum) using broth microdilution method. The results of bioassay showed that the oil exhibited moderate to high antimicrobial activity against fungi C. albicans (MIC 625 µg ml−1), A. parasiticus (MIC 312.5 µg ml−1), A. sydowii (MIC 312.5 µg ml−1), T. rubrum (MIC 625 µg ml−1), Gram-positive bacteria B. subtilis (MIC 625 µg ml−1) and Gram-negative bacteria P. aeruginosa (MIC 312.5 µg ml−1).

Comparison of headspace analysis of volatile constituents with GCMS analysis of hydrodistilled and supercritical fluid extracted oil of Capillipedium parviflorum

Volatile constituents of the aerial parts of Capillipedium parviflorum were studied by two different extraction procedures, hydrodistillation (HD), supercritical CO2 extraction (Sc-CO2) and compared with headspace analysis (HS). Sc-CO2 extraction was carried out using nine differential optimization extractions (DOX) at different temperatures (°C) and pressures (MPa), i.e. 35/9, 60/9, 85/9, 35/17, 60/17, 85/17, 35/25, 60/25 and 85/25, to assess the influence on the yield and composition of volatile constituents. In DOX experiments, oil yields varied between 0.01–0.18% and maximum yield (0.18%) was obtained using temperatures and pressures 35/17 and 85/25. The oil is characterized by distinctive presence of very high content of non-terpenes (90.7%) identified in HD oil, 84.0% in Sc-CO2 oil and 92.5% in HS analysis. Monoterpenes were represented in low concentrations in HD oil and were totally absent in Sc-CO2 as well as in HS analysis. Very little variability was observed in the oxygenated monoterpenes in both Sc-CO2 and HD oil representing a low percentage of constituents. Sesquiterpene hydrocarbons were represented between 1.5% and 3.1% in Sc-CO2 extraction, which was higher in comparison with HD and HS techniques. Oxygenated sesquiterpenes were represented in higher percentage in Sc-CO2 than in HD oil and HS analysis. Variability among the major constituents of 4-nonanone, 4-nonanol, 4-undecanone and 4-undecanol was observed in Sc-CO2 extraction carried out under varied temperatures and pressures. Their percentage also varied in HD and HS analysis. 4-Nonanone (2.5%, 60/9 in Sc-CO2; 6.4% in HD and 4.3% in HS), 4-nonanol (21.5% in Sc-CO2, 31.7% in HD and 29.5% in HS), 4-undecanone (33.3%, 85/9 in Sc-CO2; 35.3% in HD and 44.5% in HS) and 4-undecanol (16.5%, 84/17, 60/25 in Sc-CO2; 14.6% in HD and 10.3% in HS) dominated the volatile constituents.