Hellenia speciosa (J. Koenig) S. R. Dutta (Syn: Costus speciosus (J. Koenig) Sm.) is well known for its anti-diabetic properties and is in high demand for its steroidal sapogenin, diosgenin, which is used in the pharmaceutical industry for manufacturing steroidal drugs. The increasing demand for this plant has led to the depletion of its wild population. This study was conducted to reveal the genetic diversity of H. speciosa collected from various agro-ecological regions of India for use in devising conservational strategies, and cultivation and breeding programs. Twenty inter-simple sequence repeat markers yielded 304 products with 100

M. dioica Roxb. ex Willd. (Family: Cucurbitaceae) widely referred to as wild bitter gourd (Janglee karela), is well known plant used in Indian system of traditional medicine for the treatment of fever, diabetes, leprosy and snake bites. In this study, pharmacognostical standardization, proximate and pharmacological analysis was performed to validate its traditional claims as a food supplement. A validated method developed for HPTLC to determine the amount of caffeic acid in fruits. The chromatogram of the marker compound was developed using the tertiary mobile phase (Toluene: Ethyl acetate: Formic acid) ratio (6:4:0.5 v/v/v). Caffeic acid was detected at an Rf value 0.44 ± 0.04 and densiometric scanning was performed at 270 nm. The Caffeic acid content within the plant was found to be 0.0034% on dry weight basis. This method is highly accurate, linear and precise with respect to guidelines provided by ICH. The pharmacognostical parameters viz. moisture content, ash and extractive values of samples was found to be within the limits of standard. The phytochemical evaluation of metabolites through spectroscopy reveals the presence of flavonoid, phenolics, starch, sugar and tannin. Among them the highest value in starch estimation. The Pharmacological activities such as (Anti-diabetic, Anti-inflammatory & Anti-oxidant activity) were also analyzed. The findings showed that M.dioica have most potential and is useful for future standardization, quality control and validation of plant material for final product in herbal drug industry. Furthermore, the results show that the fruits of M.dioica are suitable for rich source of nutrients also as proteins, carbohydrates and lipids.

The natural population(s) of Gloriosa superba L. was mapped from 19 different locations of Western Himalayas, up to altitude of 1800 m and no significant morphotypic variation was observed. The major metabolite colchicine was quantified through HPTLC (validated) method and significant (p < 0.05) intraspecific variation was recorded. The colchicine content ranges from 0.023 to 0.256% and maximum was in NBG-03 from Jyolikot, Nainital (Uttarakhand, India). Total polyphenolics content was also significant among the collected samples. Cluster analysis based on colchicine and total polyphenolic content of the population’s results in identification of five elite chemotypes viz. NBG-03, NBG-54, NBG-57, NBG-65 and NBG-80. The in vitro antigout activity of elite germplasms reveals that NBG-57 was the most promising, although its colchicine content was in moderation. Therefore, it is suggested that the whole extract may exert more promising antigout potential due to the synergistic action of various phytomolecules. Hence, the study will promote the conservation and cultivation of targeted species in Western Himalayas for the availability of good quality raw material for industry. It will also aid in commercial cultivation of elite germplasm(s) which can be used in herbal product development as well as in the economic upliftment of local inhabitants.

Coleus forskohlii Briq. is an important medicinal herb, endowed with a wide range of medicinal properties against the variety of ailments. Seven germplasm of C. forskohlii collected from different phyto-geographical locations and identification of elite chemotype was performed with the help of high performance thin layer chromatography. Data of soil analysis correlated with the bioactive compounds and inhibitory potential of the species. Quantification of forskolin and its isomer (iso-forskolin) content were done in all the collected samples of C. forskohlii, which revealed a wide range of variations, varying from 1.15-0.004% and 0.0091 to 0.1077% per dry weights basic, respectively. Variation in the bioactive content may be due to the soil nature and environmental factors. Soil analysis of collected samples demonstrated that there is significant variation in available NPK and micronutrient content and may be reasoned for existing chemotypic variability. In vitro biological activity (antioxidant and antidiabetic) analyses were performed, which reveals that germplasms have a high amount of forskolin and iso-forskolin, both show more activity. The aim of this study was to elucidate the effect of elicitors and precursors on the production of bioactive compounds and identification of best elite germplasm among the populations, to provide basic lead to the industry for commercial exploitability including its location-specific commercial cultivation.

Costus speciosus is a rich source of commercially important compound Diosgenin, distributed in different regions of India. The present investigation was aimed to quantify diosgenin through High Performance Thin Layer Chromatography in 34 germplasms of Costus speciosus and also to identify the superior sources and to correlate the macronutrients of rhizospheric soil. The starch content varied in microscopic examination and correlated inversely (r=-0.266) with diosgenin content. Findings revealed that the extraction process with acid hydrolysis yielded higher diosgenin content (0.15-1.88 %) as compared to non-hydrolysis (0.009-0.368 %) procedure. Germplasms from Uttar Pradesh (NBCS-4), Jharkhand (NBCS-39) and Bihar (NBCS-2) were identified as elite chemotypes based on hierarchical clustering analysis. The phosphorous content of respective rhizospheric soil correlated positively (r=0.742) with diosgenin content. Findings of present study are useful to identify the new agrotechniques. The elite germplasms can also be used as quality planting material for large scale cultivation in order to assure a sustained supply to the herbal drug industry.

Cover Picture. Diosgenin is a commercially viable aglycone molecule and is therapeutically valuable as it is key precursor in corticosteroid synthesis at industrial scale. The preferred natural source of Diosgenin, worldwide is Dioscorea spp., and the species is continuously facing threat of biodiversity loss due to over exploitation. Alternatively, the yield of diosgenin from natural resources varies significantly from one population to other depending upon the phytogeographical determinants. In the present context, an alternative species C. speciosus as a source of diosgenin was explored and a comprehensive analysis of variation in its diosgenin content (rhizomes) was carried out in the germplasm(s) collected from different locations of the Indo-Gangetic Plain (India). In due course of study, a systematic collection strategy was adopted to cover the maximum possible variation existing in the natural population of species, its growth stage, season of collection etc. Further the extraction process also plays a very crucial role to set the diosgenin in free form (therapeutically active form) for quantification. The effect of soil micro/macronutrients on the diosgenin content has been established. The chemotaxonomic study on C. speciosus will result in identification of elite (diosgenin rich) germplasm(s) of species and its commercial cultivation at site specific locations will lead to the identification of good quality planting materials. This will ultimately ensure the sustainable availability of good quality raw material to the herbal drug industries and also decrease the burden on natural resources. Moreover, the development of suitable agro techniques for elite germplasm(s) of C. speciosus will uplift the availability of raw material and socio-economic upliftment of local inhabitants in area's having similar phytogeographical location, as reported by Sharad Srivastava et al. in their full paper at 10.1002/cbdv.202000977.

Gloriosa superba L. has economic significance due to colchicine, a bioactive compound used for gout. In present study metabolic and molecular variability in natural population of species was analyzed and correlated with edaphic and climatic factors. Thirty populations (wild) of G. superba were mapped from 10 different eco-regions of India at an elevation range of 10-1526 m, having no morphotypic variations. The two known biologically active alkaloids colchicine (ranged from 0.015-0.516%) and gloriosine (0.19-0.44%) were significantly varied (p < 0.05) among populations, leading to the identification of four elite chemotypes. Molecular variability from ISSR data divides the population in different sub clusters at intra-specific level, presenting the high similarity percentage with bootstrap value of 66-100%. Principal component analysis (PCA) revealed that elite chemotypes are related to temperature, precipitation and aridity gradient. The rhizospheric soil selenium was significantly correlated with colchicine content in G. superba.

Costus speciosus (Koen.) Smith (family Costaceae), is a perennial rhizomatous herb commonly known as “Crepeginger”.1 It is widely distributed in central parts of India, Sub-Himalayan tract, Karnataka, Western Ghats of Maharashtra and Kerala.2 The rhizome is useful to treat fever, cough, indigestion, asthma, helminthiasis, bronchitis and skin diseases. Diosgenin is the principle constituent of the rhizome used in the commercial production of steroidal hormones3 along with, tigogenin and saponin as other key ingredients.4

Background: Costus speciosus known as "insulin plant" for its anti-diabetic potential. It has commercial significance due to the presence of industrially viable metabolite diosgenin. Objective: The identification of elite chemotypes of C. speciosus (Rhizome) through high-performance thin-layer chromatography (HPTLC) from the Eastern Ghats (India). Materials and Methods: A validated HPTLC method for the quantification of diosgenin was developed in accordance with the International Conference on Harmonization Guidelines. Results: In total, 11 populations of species were collected from their natural habitat with all the Global Positioning System (GPS) coordinates. The method was developed on HPTLC pre-coated silica gel 60 F-254 plates under a binary solvent system of n-hexane and ethyl acetate (7:2 v/v). The linearity was established at concentration range of 0.1-0.9 mu g/spot having regression equation, 0.010x + 0.002. The limit of detection and limit of quantification were 0.907 and 2.751 with a regression coefficient of 0.999. The diosgenin content varies significantly (P < 0.05) from 0.002% to 0.076% and NBCS-06 from Patiya, Bhubaneswar, was identified as elite chemotype. Conclusion: The validation data confirm that developed HPTLC method was precise, accurate, robust, reproducible, and reliable in nature. The study resulted in the identification of elite chemotype of C. speciosus through validated HPTLC method from the Eastern Ghats of India. It will promote site-specific commercial cultivation of high metabolite yielding germplasm for good quality raw material to meet the industrial demand and in turn income generation of local inhabitants. The developed method will also aid in the regulation of quality standard and batch consistency of diosgenin-containing formulations in industry.

Sphaeranthus indicus is an important medicinal plant in Ayurveda which grows as weed in rice fields throughout India, Sri Lanka, Australia and Africa. The present study aimed for the pharmacognostical evaluation and pharmacological validation of this high-value medicinal species. All the pharmacognostical parameters were done as per the API guideline. A simple, rapid, sensitive and reproducible method was also developed for simultaneous HPTLC quantification of two bioactive compounds eugenol and β-sitosterol in S. indicus. The HPTLC was performed on silica gel 60 F254 by using toluene/ethyl acetate/glacial acetic acid (8:2:0.2) as a mobile phase for eugenol and β-sitosterol at Rf value 0.64 and 0.48, respectively. The content of eugenol (0.03%) and β-sitosterol (0.013%) was detected in dry weight per gram. In vitro activities viz. antioxidant, antidiabetic and anti-inflammatory were done to evaluate the pharmacological potential of S. indicus. Four antioxidant models viz. DPPH, ferric reducing power, 2-deoxyribose assay and antioxidant capacity were used to determine the free radical scanning. Antidiabetic and anti-inflammatory potential were determined by using starch–iodine colour assay and inhibition of protein denaturation model. IC50 value of S. indicus extract in DPPH and 2-deoxyribose assay was found (328.96 ± 0.003 µg/ml) and (30.36 ± 0.004 µg/ml), respectively. The present study will be helpful for quality check of the raw material and monitoring batch-to-batch consistency of herbal drugs, wherein S. indicus is used as an ingredient.

The variation in alkaloid metabolites (colchicine and gloriosine) was found significant in the nine germplasms of G. superba (L.), collected from Central India. The maximum content of colchicine and gloriosine was in NBG-15 (Chitrakoot, M.P) and NBG-13 (Bheraghat, M.P). The phenolic acids viz. quercetin and kaempferol was first ever quantified in G. superba tuber. Cluster analysis on chemical variability (colchicine and gloriosine content) results in the identification of three elite germplasm(s). The radical scavenging potential was also found promising in the selected elite germplasm viz. NBG-13, NBG-14 and NBG-15. Further, the protein denaturation potential of elite chemotypes was found at par with standard colchicine. The study will aid in site specific exploration of high metabolite yielding chemotype(s) with validated pharmacological action to meet out the industrial demands. This will also promotes the commercial cultivation of species for socio economical upliftment in the area having similar phyto geographical conditions.

Ethnopharmacological relevance: Gloriosa superba L. (Colchicaceae) is used in the treatment of gout and rheumatism as a traditional medicine dates back to 1810. It has also been used as ethnobotanical and folklore medicine to induce abortion/vaginal poison. Aim of Study: The present study was carried out to identify the chemical variation existing in the major alkaloid metabolite (colchicine) in a threatened species, Gloriosa superba L. and is correlated with invitro antigout activity. Material and method: The samples (tuber) were collected from their natural locations in Gangetic plain of India. HPLC-PDA quantification of colchicine was done on C-18 column at 245 nm and invitro antigout activity was analyzed by inhibition of protein denaturation, DPPH and Hydroxyl radical scavenging assay. Results: The colchicine content within the 29 samples ranges from 0.021 to 0.665% and the maximum contents was in NBG-10 from Kanth (U.P). Such high colchicine (0.665%) containing natural population of G. superba is reported for the first time in Indian population. Four chemotypes viz. NBG-10, NBG-120, NBG-126 and NBG-88 were selected on the basis of colchicine content for invitro antigout activity. NBG-10 was separated from rest of the population exhibiting the most promising activity with high colchicine content. Conclusion: The outcomes will be helpful in the identification of elite chemotype for herbal product development and quality check of metabolites in raw material. The study will also support the site-specific commercial cultivation to meet out the industrial demand as well as income generation to farmers.

Objective: Pharmacognostical study along with the development of a quantitative HPTLC method for Crinum latifolium and evaluation of its traditional claims.Methods: Quantification of three marker compounds oleanolic acid, linoleic acid, and lupeol was done through HPTLC. In vitro antioxidant activity was determined by six different models, namely total phenolic and total flavonoid content, DPPH radical scavenging assay, ferric reducing power, antioxidant capacity and hydroxyl radical scavenging assay. In vitro antidiabetic activity was evaluated by α-amylase inhibition assay based on starch iodine and DNS method.Results: The content of oleanolic acid, linoleic acid, and lupeol were found to be higher in aerial parts like 0.015%, 0.048%, and 0.028% respectively, while in root extract 0.006%, 0.027% and 0.025% respectively on a dry weight basis. Free radical scavenging activity was done by DPPH assay, showing the IC50 value of 410±1.105 µg/ml in roots and 441.95±1.788 in aerial parts. In vitro antidiabetic potential of both the parts were assessed by starch iodine color assay and DNS method of alpha-amylase inhibition model. In 3,5 DNS assay, IC50 of extract from aerial parts was 282.21±2.151µg/ml whereas in root extract it was 193.33±2.45µg/ml. Iodine-starch assay of C. latifolium (aerial part) shown the IC50 value of 340.81±0.49 µg/ml and C. latifolium (root) of 74.64±1.28 µg/ml.Conclusion: The results indicate that the aerial parts of the plant possess more antidiabetic potential in comparison to the root. Thus, the aerial part can be used to get better results as a drug and roots can be used as an alternative.

Background: Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. Objective: A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Materials and Methods: Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Results: Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at Rfof 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300–1200 ng/spot with the regression coefficient (R2) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. Conclusion: The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop.

Two elite germplasm(s) ofC. forskohlii viz.NBC-24 (0.728%) and NBC-16 (0.641%) were obtained as the highest accumulator of forskolin with high genetic variability (92%).

Plant metabolite varies with season and geographic conditions. The present study is aimed at the identification of the potential chemotypes of Coleus forskohlii, available in the natural habitat of Nilgiri hills and adjoining area, in order to provide a basic lead for the industry concerning commercial exploitability, including the location-Specific commercial cultivation of the plant. The effect of intra-Specific variability in the forskolin content among the populations was estimated using high-performance thin-layer chromatography (HPTLC)—densitometric method. The roots of fourteen naturally occurring populations from the entire hill range were collected, covering the wide topography from foot hills up to the highest peak. The method developed for the quantification of forskolin was validated and found to be linear, Specific, and accurate with precision and accuracy. The limit of detection (LOD) and limit of quantification (LOQ) were 1.04 and 3.16 ng spot−1. Precision studies (both inter-day and intra-day) were within the standard limit of relative standard deviation (RSD) (%) less than 3%. The quantification of forskolin within the population revealed that it varied from 0.0046 ± 0.0005 (NBC-36) to 1.156 ± 0.003% (NBC-46). The analysis of variance (ANOVA) suggested that there are significant differences in forskolin content among the populations. A positive correlation (Karl Pearson) was found between the altitude and the forskolin content. The cluster analysis of the population on forskolin content suspected the presence of two chemotypes. The study suggests the presence of chemotaxonomic variation among the populations which can be due to the change in phyto-geographical factors.