Facial Segmental Infantile Haemangioma: A Cutaneous Mosaicism Quandary.

Mosaicism has long been considered the underlying mechanism of segmental infantile hemangiomas (SIH). This was a prospective pilot case-control study conducted with the objective to quantify the percentage overlap of silhouettes of facial SIH with those of Blaschko lines (the most well studied archetypical pattern of mosaicism on face) as compared to other mosaic disorders on face. Lesional silhouettes of 8 patients with SIH (Group A) and 6 patients with other facial dermatosis known to have blaschkoidal distribution (Group B), were overlapped on a standardized template with Blaschkoidal lines on the frontal view of face. The alignment was done via the auto align tool of Photoshop and the percentage of overlap was calculated with an online image comparison software (IMGonline.com.ua). There was a significant difference in mean overlap in Group A (72.92 ± 15.6 %) as compared to Group B (90.1 ± 4.3%; P=0.018). Hence, we concluded that facial SIH do not follow lines of Blaschko.

Gharat: an environment friendly livelihood source for the natives of western Himalaya, India.

The native people of the Himalayan region use an indigenously designed watermill locally called as 'Gharat'. The 'Gharat' is made from the natural resources that are easily available in the area and operates through the water force coming from glaciers, perennial streams and rivulets. Thus, it is an environment friendly source of livelihood for the 'Gharatis' (the owner of Gharat). In the present communication, an effort was made to document the status of Gharats in the high-altitude Himalayan regions of Pangi and Tissa of district Chamba. Field surveys were conducted in a total of twenty remote villages of Pangi and Tissa regions (ten in each site). Information was collected on pre-designed semi-structured questionnaires from local people that primarily consisted of Gharatis, elderly informants and the inhabitants living in the vicinity of Gharats. All possible reasons responsible for the increase and reduction in the number of Gharats in the study areas were also documented by interviewing the informants. The study revealed that in the Pangi region number of Gharats have increased and are in good condition whereas there is a drastic decline in the number of Gharats in Tissa. The informants in Pangi gave maximum responses favouring motivational factors while in Tissa maximum responses were obtained for demotivational factors. Based on these responses, we prioritized the corrective measures for improving the efficiency of Gharats and restoration of this age-old indigenous and environment friendly technology. This will surely enhance livelihood security of the inhabitants besides optimum utilization of the natural resources in the Himalayan region. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10668-021-01455-4.

Ethnobotanical knowledge among the semi-pastoral Gujjar tribe in the high altitude (Adhwari's) of Churah subdivision, district Chamba, Western Himalaya.

BACKGROUND: The wild plants not only form an integral part of the culture and traditions of the Himalayan tribal communities but also contribute largely to the sustenance of these communities. The tribal people use large varieties of wild fruits, vegetables, fodder, medicinal plants, etc. for meeting their day-to-day requirements. The present study was conducted in Churah subdivision of district Chamba where large populations of Muslim Gujjars inhabit various remote villages. These tribal people are semi-pastoralists, and they seasonally (early summers) migrate to the upper altitudes (Adhwari's) along with their cattle and return to permanent settlements before the onset of winters. A major source of subsistence of these tribal people is on natural resources to a wide extent, and thus, they have wide ethnobotanical knowledge. Therefore, the current study was aimed to report the ethnobotanical knowledge of plants among the Gujjar tribe in Churah subdivision of district Chamba, Himachal Pradesh. METHODS: Extensive field surveys were conducted in 15 remote villages dominant in Gujjar population from June 2016 to September 2017. The Gujjars of the area having ethnobotanical knowledge of the plants were interrogated especially during their stay at the higher altitudes (Adhwari's) through well-structured questionnaires, interviews, and group meetings. The data generated was examined using quantitative tools such as use value, fidelity, and informant consensus factor (Fic). RESULTS: This study reveals 83 plants belonging to 75 genera and 49 families that were observed to have ethnobotanical uses. Plants were listed in five categories as per their use by the Gujjars, i.e. food plants, fruit plants, fodder plants, household, and ethnomedicinal plants. The leaves, fruits, and roots were the most commonly used plant parts in the various preparations. The highest number of plants was recorded from the family Rosaceae followed by Polygonaceae and Betulaceae. On the basis of use value (UV), the most important plants in the study area were Pteridium aquilinum, Juglans regia, Corylus jacquemontii, Urtica dioica, Diplazium maximum, and Angelica glauca. Maximum plant species (32) were reported for ethnomedicinal uses followed by food plants (22 species), household purposes (16 species), edible fruits (15 species), and as fodder plants (14 species). The agreement of the informants conceded the most from the use of various plants used as food plants and fruit plants (Fic = 0.99), followed by fodder plants and household uses (Fic = 0.98) while it was least for the use of plants in ethnomedicine (Fic = 0.97). The fidelity value varied from 8 to 100% in all the use categories. Phytolacca acinosa (100%), Stellaria media (100%), and Urtica dioica (100%) were among the species with high fidelity level used as food plants, while the important species used as fruit plants in the study area were Berberis lycium (100%), Prunus armeniaca (100%), and Rubus ellipticus (100%). Some important fodder plants with high fidelity values (100%) were Acer caesium, Aesculus indica, Ailanthus altissima, and Quercus semecarpifolia. The comparison of age interval with the number of plant use revealed the obvious transfer of traditional knowledge among the younger generation, but it was mostly concentrated in the informants within the age group of 60-79 years. CONCLUSIONS: Value addition and product development of wild fruit plants can provide an alternate source of livelihood for the rural people. The identification of the active components of the plants used by the people may provide some useful leads for the development of new drugs which can help in the well-being of mankind. Thus, bioprospection, phytochemical profiling, and evaluation of economically viable products can lead to the optimum harnessing of Himalayan bioresources in this region.

Antifilarial activity of diterpenoids from Taxodium distichum.

BACKGROUND: Lymphatic filariasis caused by Wuchereria bancrofti, Brugia malayi and B. timori, is a debilitating disease with an adverse social and economic impact. The infection remains unabated in spite of treatment with existing antifilarial drugs diethylcarbamazine (DEC) and ivermectin which are chiefly microfilaricides. There is therefore, need for macrofilaricides, embryostatic agents and better microfilaricides. In the present study we explored the antifilarial potential of crude extract and its molecular fractions of the plant Taxodium distichum using in vitro assay systems and rodent models of B. malayi infection. METHODS: Ethanolic extract (A001) of aerial parts of T. distichum was solvent fractionated and sub-fractionated. Four molecules, 3-Acetoxylabda-8(20), 13-diene-15-oic acid (K001), Beta-sitosterol (K002), labda-8(20),13-diene-15-oic acid (K003) and Metasequoic acid A (K004) were isolated from the fractions and their structure determined by spectroscopic analysis. The extract, subfractions and molecules were evaluated for antifilarial activity against B. malayi by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction and motility assays in vitro and in two animal models, Meriones unguiculatus and Mastomys coucha, harbouring B. malayi infection. RESULTS: A001 was effective in killing microfilariae (mf) and adult worms in vitro. The diterpenoid K003 produced 100 % reduction in motility of both mf and adult worms and > 80 % inhibition in MTT reduction potential of adult female worms. In B. malayi-M. unguiculatus model, A001 killed all the adult worms in > 80 % of infected animals. K003 was embryostatic (> 95 %) in this model. In the B. malayi-M. coucha model, K003 killed ~54 % of adult worms (macrofilaricidal activity) and rendered > 36 % female worms sterile; it also stopped any further rise in microfilaraemia after day 42 post-initiation of treatment. CONCLUSION: Ethanolic extract of aerial parts of the plant T. distichum possesses potent antifilarial activity and the active principle was localised to K003 which showed significant macrofilaricidal activity and late suppression of peripheral microfilaraemia and some embryostatic activity. These findings indicate that labdane diterpenoid molecule(s) may provide valuable leads for design and development of new macrofilaricidal agent(s). To the best of our knowledge, this is the first report on antifilarial efficacy of products from the plant T. distichum.

Observations on traditional usage of ethnomedicinal plants in humans and animals of Kangra and Chamba districts of Himachal Pradesh in North-Western Himalaya, India.

ETHNOBOTANICAL RELEVANCE: Medicinal plants are frequently used by Gaddi and Gujjar tribes of Kangra and Chamba districts of Himachal Pradesh, India to cure various ailments in humans and livestock. Therefore, extensive field work was conducted to document the traditional use of ethnomedicinal plants by these tribes. MATERIALS AND METHODS: Direct interviews of 208 informants were conducted. The data generated through interviews was analysed using quantitative tools such as use-value (UV), factor informant consensus (Fic) and fidelity level (Fl). RESULTS: A total of 73 plant species in 67 genera and 40 families were observed to be medicinal and used to cure 22 ailment categories. The highest number of ethnomedicinal plants was recorded from the family Asteraceae followed by Lamiaceae, Apiaceae, Acanthaceae, Caesalpiniaceae, Polygonaceae, Ranunculaceae, Rosaceae and Rutaceae. Leaves were the most frequently used plant part used to treat various ailments followed by whole plant and roots or rhizomes. Ajuga parviflora, Berberis lycium, Viola canescens, Vitex negundo and Zanthoxylum armatum were the most important medicinal plants used for treating human diseases, whereas Achyranthes bidentata, Aloe sp., Cassia fistula, Podophyllum hexandrum and Pogostemon benghalensis were the most important medicinal plants used for treating animal diseases as per use value. The important ailment categories classified on the basis of factor informant consensus were gastrointestinal and respiratory disorders. CONCLUSION: The present study revealed that people of the study area are extensively using the ethnomedicinal plants to cure various ailments. Plants with high use value and fidelity level should be subjected to pharmacological investigation for scientific validation.

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

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.

Chemical Composition and Biological Activity of Essential Oils from Wild Growing Aromatic Plant Species of Skimmia laureola and Juniperus macropoda from Western Himalaya.

The Himalayan region is very rich in a great variety of medicinal plants. In this investigation the essential oils of two selected species are described for their antimicrobial and larvicidal as well as biting deterrent activities. Additionally, the odors are characterized. Analyzed by simultaneous GC-MS and GC-FID, the essential oils' chemical compositions are given. The main components of Skimmia laureola oil were linalool and linalyl acetate whereas sabinene was found as the main compound for Juniperus macropoda essential oil. Antibacterial testing by agar dilution assay revealed highest activity of S. laureola oil against all tested bacteria, followed by J. macropoda oil. Antifungal activity was evaluated against the strawberry anthracnose causing plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Juniperus macropoda essential oil indicated higher antifungal activity against all three pathogens than S. laureola oil. Both essential oils showed biting deterrent activity above solvent control but low larvicidal activity.

Chemical prospection of important ayurvedic plant Tinospora cordifolia by UPLC-DAD-ESI-QTOF-MS/MS and NMR.

A rapid, sensitive, and accurate ultra-performance liquid chromatography coupled with mass spectrometric method (UPLC-MS) was developed and validated for simultaneous determination of four bioactive compounds, syringin (3), cordifolioside A (4), magnoflorine (6) and tinocordiside (10) in the stem of Tinospora cordifolia. The analysis was performed using an Acquity C18 column and gradient elution of 0.05% formic acid in water and acetonitrile at a detection wavelength of 267 nm in 5 min. A high correlation coefficient (r2 > 0.998) indicated good correlation between investigated compounds concentration and their peak area within the test ranges. The LODs for compounds 3, 4, 6 and 10 were 1.95, 0.97, 3.90 and 0.97 ng/mL, respectively, and LOQs were 6.64, 3.20, 12.87 and 3.20 ng/mL, respectively. The overall intra- and inter-day variations of the four compounds were less than 1%. The variation of these four bioactive compounds in T. cordifolia hosted on fifteen different trees was also determined. The compounds (3, 4, 6 and 10) were found in high amount in the T. cordifolia hosted on Azadirachta indica and Mangifera indica as compared with other plants. Twelve compounds were identified on the basis of their mass and UV-vis spectra. The NMR fingerprinting of the extract revealed the presence of alkaloids, fatty acid methyl esters, polysaccharides and marker components of T. cordifolia.

Chemical composition and biological activity of essential oils of Dracocephalum heterophyllum and Hyssopus officinalis from Western Himalaya.

The essential oils of two representatives of the Lamiaceae, Dracocephalum heterophyllum Benth. and Hyssopus officinalis L., are described for their antifungal, antibacterial, larvicidal and inect biting deterrent activities. Additionally, the chemical compositions of the essential oils, analyzed by simultaneous GC-MS and GC-FID, and odor descriptions are given. The main components of H. officinalis oil were pinocarvone, cis-pinocamphone, and ß-pinene. Citronellol was found as the main compound of D. heterophyllum essential oil. Antibacterial testing by agar dilution assay revealed greater activity of D. heterophyllum against Staphylococcus aureus compared with H. officinalis. D. heterophyllum essential oil also showed promising antifungal activity against Colletotrichum species and was more toxic to Aedes aegypti larvae in a larvicial bioassay. Both essential oils showed high activity in the biting deterrent bioassay.

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.

Chemical composition and biological effects of Artemisia maritima and Artemisia nilagirica essential oils from wild plants of western Himalaya.

Artemisia species possess pharmacological properties that are used for medical purposes worldwide. In this paper, the essential oils from the aerial parts of Artemisia nilagirica and Artemisia maritima from the western Indian Himalaya region are described. The main compounds analyzed by simultaneous GC/MS and GC/FID were camphor and 1,8-cineole from A. maritima, and camphor and artemisia ketone from A. nilagirica. Additionally, the oils were evaluated for their antibacterial, antifungal, mosquito biting deterrent, and larvicidal activities. A. nilagirica essential oil demonstrated nonselective antifungal activity against plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides, whereas A. maritima did not show antifungal activity. Both Artemisia spp. exhibited considerable mosquito biting deterrence, whereas only A. nilagirica showed larvicidal activity against Aedes aegypti. Antibacterial effects assessed by an agar dilution assay demonstrated greater activity of A. maritima essential oil against Staphylococcus aureus and Pseudomonas aeruginosa compared to A. nilagirica.

Zephgrabetaine: a new betaine-type amaryllidaceae alkaloid from Zephyranthes grandiflora.

Zephgrabetaine (1), a new betaine type Amaryllidaceae alkaloid, along with seven known alkaloids, lycorine, galanthine, lycoramine, hamayne, haemanthamine, tortuosine, and ungeremine were isolated from the bulbs of Zephyranthes grandiflora and their structures elucidated by spectroscopic data analysis. The isolated alkaloids were tested for in vitro cytotoxic activities against two cell lines, C-6 (rat glioma cells) and CHO-K1 (Chinese hamster ovary cells). A dose dependent cytotoxic effect was exhibited by all the alkaloids on these two cancer cell lines with prominent activity of lycorine and haemanthamine.

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 offly 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.

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-CO(2)) 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-CO(2) extracted oils. Nineteen constituents, which accounted for 89.32% of total constituents in HD oil, represented by limonene (4.31%), (Z)-ß-ocimene (3.69%), terpinolene (22.24%), neryl acetate (36.19%), nerol (9.51%) and p-cymene-8-ol (2.61%) were identified. In SC-CO(2) 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)).

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-CO(2) 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-CO(2) extraction, 31 in HD and 17 in HS analysis were identified. Constituents in SC-CO(2) and HD oils were identified by gas chromatography mass spectrometry analysis. SC-CO(2) extraction was carried out at 40°C and 140 bar pressure and the oil represented by major constituents as ß-caryophyllene (5.96%), α-humulene (4.06%) and p-menthadiene-2,9-diol (7.28%); in HD, oil limonene (11.26%), ß-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%), ß-phellandrene (3.44%) and ß-ocimene (7.15%) were present as major constituents.

Chemical fingerprint analysis of phenolics of Albizia chinensis based on ultra-performance LC-electrospray ionization-quadrupole time-of-flight mass spectrometry and antioxidant activity.

Albizia species have been shown to have anti-inflammatory and anti-allergic properties. However, efficient analytical methods for identification of their active constituents are still lacking. Ultra-performance liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) was used to study the phenolic composition of the ethanolic extracts of different parts (flowers, leaves, pods and bark) of A. chinensis. In addition, the antioxidant activity of the ethanolic extracts was evaluated by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) free-radical and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical cation scavenging methods. Four compounds were isolated from the ethanolic extract of the flowers and characterized by 1H and 13C NMR spectroscopy as quercetin-3-O-rhamnoside, quercetin, quercetin-3-O-arabinofuranoside, and myricetin-3-O-rhamnoside. Separation and quantification of the phenolics was accomplished using a reversed-phase BEH C18 column with the mobile phase of methanol-water (0.05% formic acid), and detection wavelengths of 360 and 254 nm.

Comparison of the volatile constituents of Elsholtzia fruiticosa extracted by hydrodistillation, supercritical fluid extraction and head space analysis.

Volatile constituents of Elsholtzia fruiticosa (D. Don) Rehder were studied by two different extraction techniques, supercritical fluid extraction (SFE) and hydrodistillation (HD), and the results were compared with head space analysis (HS). Thirty-five constituents were identified in both the SFE and HD oils and fourteen in the HS, accounting for 94.2%, 97.7% and 96.9% of the total identifications, respectively. A distinctive feature of the results was the very high content of non-terpenes (59.8%) in the HS, the high content of oxygenated monoterpenes (41.1%) in the HD oil and the high content of sesquiterpene hydrocarbons (21.8%) in the SFE oil. Monoterpene hydrocarbons were represented in HS (13.6%), HD (19.4%) and SFE (4.3%). In SFE sesquiterpene hydrocarbons formed 21.8% of the total, as compared to 6.6% in the HD and 1.1% in the HS Oxygenated sesquiterpenes represented 3.0% in SFE, 0.8% in HD and were absent in HS. Diterpenes were only present in the SFE oil (3.4%). Non-terpenes were represented by 24.5% in the SFE oil and 29.8% in the HD oil.

Simultaneous densitometric determination of shikonin, acetylshikonin, and beta-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.

Ethnomedicines used against four common ailments by the tribal communities of Lahaul-Spiti in western Himalaya.

The present study aimed to highlight the new or lesser known medicinal uses of plant bioresource along with validation of traditional knowledge that is widely used by the tribal communities to cure four common ailments in Lahaul-Spiti region of western Himalaya. The study area inhabited by Lahaulas and Bodhs (also called as Bhotias), is situated in the cold arid zone of the state of Himachal Pradesh (HP), India. During the ethnobotanical explorations (2002-2006), observations on the most common ailments like rheumatism, stomach problems, liver and sexual disorders among the natives of Lahaul-Spiti were recorded. Due to strong belief in traditional system of medicine, people still prefer to use herbal medicines prescribed by local healers. A total 58 plant species belonging to 45 genera and 24 families have been reported from the study area to cure these diseases. Maximum use of plants is reported to cure stomach disorders (29) followed by rheumatism (18), liver problems (15) and sexual ailments (9). Among plant parts used, leaves were found used in maximum herbal preparations (20) following flowers (12) and roots (11) respectively. Most of these formulations were prescribed in powder form, whereas juice and decoction forms were also used. Plants having more than one therapeutic use were represented with 24 species, however 34 species were reported to be used against single specific ailment. Validation of observations revealed 38 lesser known or new herbal preparations from 34 plant species, where 15 species were used to cure stomach disorders, 7 for rheumatism, 10 for liver disorders and 6 for sexual problems. Mode of preparation, administration and dosage are discussed along with the family and local names of plants and plant parts used.