Determination of betulinic acid, oleanolic acid and ursolic acid from Achyranthes aspera L. using RP-UFLC-DAD analysis and evaluation of various parameters for their optimum yield.

Achyranthes aspera L. is a well known herb commonly used in traditional system of Indian medicine to treat various disorders, such as cough, dysentery, gonorrhea, piles, kidney stone, pneumonia, renal dropsy, skin eruptions, snake bite, etc. Here, we used RP-UFLC-DAD method for determining triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from A. aspera. Optimum yield of these compounds were studied and evaluated using parameters viz., method of extraction, time of extraction, age of plant and plant parts (leaves, stem and roots). Linear relationships in RP-UFLC-DAD analysis were obtained in the range 0.05-100 µg/mL with 0.035, 0.042 and 0.033 µg/mL LOD for BA, OA and UA, respectively. Of the variables tested, extraction method and parts used significantly affected content yield. Continuous shaking extraction (CSE) at ambient temperature gave better extraction efficiency than exposure to ultra sonic extraction (USE) or microwave assisted extraction (MAE) methods. The highest content of BA, OA and UA were determined individually in leaf, stem and root extracts with CSE. Collective yield of these triterpenoids were higher in leaf part exposed to 15 min USE method. To best of our knowledge, the study newly reports UA from A. aspera and the same was confirmed using ATR-FT-IR studies. This study explains the distribution pattern of these major triterpenoids and optimum extraction parameters in detail.

Identification of potentially bioactive compounds from Blumea lacera essential oil by gas chromatography-mass spectroscopy and molecular docking studies for targeting inflammatory bowel disease.

Blumea lacera (Burm.f.) DC. (Asteraceae) is used in the traditional system of medicine for the treatment of inflammation or irritable bowel disease (IBD). In this study, B. lacera was collected from different geographical regions and oil was extracted by hydro-distillation and further chemo-profiled using GC-FID-MS. The major compounds identified were 2,5-dimethoxy-p-cymene (28.7-0.4%), ß-caryophyllene (25.5-0.5%), carvotanacetone (24.5-0.4%), chrysanthenone (21.9-9.8%) and 2,6-dimethyl phenol (11.4-1.8%). The constituents of B. lacera also showed marked qualitative and quantitative variations. The percent chemical similarity was observed to be in the range of 51.7% to 59.2% between the localities. Moreover, molecular modelling, membrane molecular dynamics simulations, target prediction were implemented to decipher the potential targets relevant to IBD. This inferred that all these major compounds could be potential drug moieties for treating IBD in terms of targeting h5HTR3A, thereby substantiating the traditional use of B. lacera for the treatment of IBD ailments.

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.

Elicitor-mediated enhancement of biomass, polyphenols, mangiferin production and antioxidant activities in callus cultures of Salacia chinensis L.

The present investigation aimed to improve callus biomass, polyphenolic content, biosynthesis of mangiferin and biological potential following application of different elicitor treatments for medicinally important Salacia chinensis L. The leaf-derived callus cultures were established on Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D: 2.0 mg/l) and 6-benzylaminopurine (BAP: 1.5 mg/l). These cultures were treated with different elicitors viz. jasmonic acid (JA), methyl jasmonate (MeJA) and yeast extracts (YE). The highest calli biomass (five-fold increase within 4 weeks) was achieved when callus was treated with JA (75 µM). The callus obtained on MS medium supplemented with 2,4-D (2.0 mg/l), BAP (1.5 mg/l) and treated with JA (75 µM) displayed augmented values for total phenolics, flavonoids and mangiferin contents. Besides, same treatment elicits the calli for antioxidant properties as evaluated by 2,2-diphenyl-2-picrylhydrazyl (DPPH), ferric-reducing antioxidant power (FRAP) and metal chelating assays. This is the first report on the elicitation study in genus Salacia and, therefore, the discoveries suggested that, S. chinensis calli might be a perfect source for large-scale production of industrially important secondary metabolites. Concurrently data provide accumulated information demonstrating its prominent antioxidant effect revealing its potential without disturbing natural resources.

Traditional Horticulture Practices Increase the Production of Selected Withanolides in Withania Somnifera (L.) Dunal-A RP-UFLC Analysis.

The study evaluates the effect of two traditional horticulture treatments mentioned in Vrikshayurveda, a text from ancient India on the science of plant life, namely Kunapa jala (KJ) and Pancha gavya (PG) on the production of Withaferin A (WFA), withanolide A (WIA) and Withanolide B (WIB) in Withania somnifera (L) Dunal. Leaves and roots of W. somnifera were collected from different treated groups viz. control, KJ, PG, farmyard manure (FYM) and inorganic fertilizer (NPK). Reverse phase ultra-flow liquid chromatography (RP-UFLC) method was developed, validated for simultaneous detection of WFA, WIA and WIB. Statistical analysis of data was performed by ANOVA and tested for significance by the Dunnett multiple comparison test and data were expressed as mean ± standard deviation (SD). Results revealed, leaves possessed highest WFA content and roots possessed highest content of WIA and WIB. PG treated leaves were observed highest WFA (18.29 mg/g) and roots were observed highest WIA (19.63 mg/g) and WIB (1.36 mg/g). Conclusively, RP-UFLC method for simultaneous detection of withanolides has been developed and validated to evaluate the effect of traditional horticulture treatments. It is concluded that the enhanced production of withanolides can be achieved by the application of PG when compared to NPK application.

Nutritional Quality and Antioxidant Activity of Wheatgrass (Triticum aestivum) Unwrap by Proteome Profiling and DPPH and FRAP assays.

Precious contribution of plants in the field of medicine is very well known. Wheat (Triticum aestivum) seeds and seedlings are an important source of food and feed due to the presence of various health-promoting compounds. Proteomic analysis of three seed developmental stages (0, 8, and 16 days after germination [DAG]) of wheat was carried out using liquid chromatography-mass spectrometry. A total of 297 proteins were identified and their functional annotation revealed that a majority of them were involved in preventing many diseases, oxidative stress, primary metabolism, storage, and energy related mechanisms. Particularly to mention, peroxidases, superoxide dismutases, and cytochromes are abundantly present in wheatgrass. In the ferric-reducing antioxidant power assay, antioxidant activity was increased by 1.55 times after 16 DAG as compared to 0 DAG, however it was decreased after 8 DAG. The antioxidant activity of the plant extracts by DPPH had an increasing trend after all the three time points. The percent radical scavenging activity of extract by DPPH was 15, 22, and 30 after 0, 8, and 16 DAG, respectively. Observations obtained revealed that antioxidant power of the plants is directly proportional to the age of seedlings. Data attained on wheatgrass showing that it can be a strong antioxidant agent due to its free radical scavenging activity and could be used in stress and nourishing human health. PRACTICAL APPLICATION: Wheatgrass contains minerals, phytochemicals, active enzymes, and vitamins that can be easily absorbed. The consumption of wheatgrass juice can give better health benefits. Information about beneficial properties of wheat grass juice is clearly mentioned in this work. Proteins found in wheatgrass are known to be involved in preventing many diseases, oxidative stress, primary metabolism, storage, and energy-related mechanisms. Results of this work revealed that Triticum aestivum seedlings can act as an antioxidant agent due to their free radical scavenging activity and can be constructive to control or treat many health complications. From all these results we believed that wheatgrass can be used for the nourishment of humans.

Extraction efficiency, phytochemical profiles and antioxidative properties of different parts of Saptarangi (Salacia chinensis L.) - An important underutilized plant.

The study aimed to evaluate extraction efficiency, detection and quantification of phytochemicals, minerals and antioxidative capacity of different parts of Salacia chinensis L. Continuous shaking extraction, steam bath assisted extraction, ultrasonic extraction and microwave assisted extraction with varied time intervals were employed for extraction of phenolics, flavonoids, and antioxidants. Preliminary screening revealed the presence of wide array of metabolites along with carbohydrates and starch. Steam bath assisted extraction for 10 min exposure was found most suitable for extraction phenolics (46.02 ± 2.30 mg of gallic acid equivalent per gram of dry weight and 48.57 ± 2.42 mg of tannic acid equivalent per gram of dry weight) and flavonoids (35.26 ± 1.61 mg of quercetin equivalent per gram of dry weight and 51.60 ± 2.58 mg of ellagic acid equivalent per gram of dry weight). In support, reverse phase-high performance liquid chromatography- diode array detector confirmed the presence of seven pharmaceutically important phenolic acids. Antioxidant capacity was measured by 1, 1- diphenyl-1-picryl hydrazyl (DPPH), ferric reducing antioxidant power (FRAP), 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) scavenging (ABTS) and N, N-dimethyl-p-phenylenediamine (DMPD) assays and represented as trolox equivalent antioxidant capacity (TEAC) and ascorbic acid equivalent antioxidant capacity (AEAC). Antioxidant capacity ranged from 121.02 ± 6.05 to 1567.28 ± 78.36 µM trolox equivalent antioxidant capacity and 56.62 ± 2.83 to 972.48 ± 48.62 µM ascorbic acid equivalent antioxidant capacity. Roots showed higher yields of illustrated biochemical parameters, however fresh fruit pulp was found a chief source of minerals. Gas chromatography-mass spectroscopic analysis revealed the presence of a vast array of phytoconstituents associated with different plant parts. The present study revealed the amounts of minerals and diverse phytoconstituents in various parts of S. chinensis and confirmed its medicinal and nutritional implications.

Reinvestigation of carvotanacetone after 100 years along with minor terpenoid constituents of Blumea malcolmii Hook. F. essential oil.

Reinvestigation of essential oil constituents of Blumea malcolmii Hook. F. (Asteraceae) was carried out after 100 years using gas chromatography equipped with flame ionisation detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). Eighteen constituents, representing 99.2% of the total oil, were identified. The major compound was identified by GC/MS and NMR ((1)H and (13)C) as carvotanacetone (92.1%). The minor compounds of essential oil have also been identified for the first time in B. malcolmii. The oil was found to be rich in oxygenated monoterpene-type constituents (95.0%).

Storing of Extracts in Polypropylene Microcentrifuge Tubes Yields Contaminant Peak During Ultra-flow Liquid Chromatographic Analysis.

BACKGROUND AND AIM: This study was designed to understand the effect of storage in polypropylene microcentrifuge tubes and glass vials during ultra-flow liquid chromatographic (UFLC) analysis. MATERIALS AND METHODS: One ml of methanol was placed in polypropylene microcentrifuge tubes (PP material, Autoclavable) and glass vials (Borosilicate) separately for 1, 2, 4, 8, 10, 20, 40, and 80 days intervals stored at -4°C. RESULTS: Contaminant peak was detected in methanol stored in polypropylene microcentrifuge tubes using UFLC analysis. The contaminant peak detected was prominent, sharp detectable at 9.176 ± 0.138 min on a Waters 250-4.6 mm, 4 µ, Nova-Pak C18 column with mobile phase consisting of methanol:water (70:30). CONCLUSION: It was evident from the study that long-term storage of biological samples prepared using methanol in polypropylene microcentrifuge tubes produce contaminant peak. Further, this may mislead in future reporting an unnatural compound by researchers. SUMMARY: Long-term storage of biological samples prepared using methanol in polypropylene microcentrifuge tubes produce contaminant peakContamination peak with higher area under the curve (609993) was obtained in ultra-flow liquid chromatographic run for methanol stored in PP microcentrifuge tubesContamination peak was detected at retention time 9.113 min with a lambda max of 220.38 nm and 300 mAU intensity on the given chromatographic conditionsGlass vials serve better option over PP microcentrifuge tubes for storing biological samples. Abbreviations used: UFLC: Ultra Flow Liquid Chromatography; LC: Liquid Chromatography; MS: Mass spectrometry; AUC: Area Under Curve.

Pharmacognostic Screening of Piper trichostachyon Fruits and its Comparative Analysis with Piper nigrum Using Chromatographic Techniques.

BACKGROUND: Piper trichostachyon is a wild, endemic Piper species from Western Ghats of India. The folklore healers of Belagavi region use this plant, similar to Piper nigrum. AIMS: The present study investigates the comparison between P. nigrum and P. trichostachyon using pharmacognostic parameters. MATERIALS AND METHODS: Pharmacognostic evaluation was carried out in terms of morphological, microscopic characters, and phytochemical analysis using standard methods. Comparative physicochemical analysis between P. trichostachyon and P. nigrum was also carried out through estimation of micro-macro nutrients, high-performance thin layer chromatography (HPTLC) investigation and using piperine as a marker compound for reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique. RESULTS: P. trichostachyon grows in the forests, and the fruits are morphologically similar to P. nigrum fruits, so the name in Kannada "Kaadu Kalu menasu" (wild/forest black pepper). The microscopy revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells, and yellowish brown pigment layer, parenchymatous cells. The presence of alkaloids, oil, and tannins were observed in P. trichostachyon fruits. The HPTLC studies visibly indicated differences among two species with 12 peaks and varied banding pattern. RP-UFLC results showed less amount of piperine in P. trichostachyon (0.05 ± 0.002 mg/g) than in P. nigrum (16.14 ± 0.807 mg/g). CONCLUSION: The study reports on pharmacognostic parameters of P. trichostachyon for the 1(st) time and will be useful for the identification and authentication. The comparative HPTLC and RP-UFLC studies resolve the differentiation impasse among two species. However, further biological efficacy studies are required to establish its use in traditional medicine. SUMMARY: Piper trichostachyon grows in the forests, and the fruits are morphologically similar to Piper nigrum fruitsThe microscopy of P. trichostachyon revealed the presence of stone cells, starch grains, oil cells and globules, beaker cells and yellowish brown pigment layer, parenchymatous cellsThe high-performance thin layer chromatography studies visibly indicated differences among two species with varied banding patternReversed phase-ultra flow liquid chromatographic results showed less amount of piperine in P. trichostachyon than in P. nigrum. Abbreviation used: HPTLC: High Performance Thin Layer Chromatography, RP-UFLC: Reversed phase-ultra flow liquid chromatographic analysis, DST: Length of line, Maj: Length of large half axis for ellipse RDS - radius for circle, Rf: Retention Factor, TS: Transverse Section, TLC: Thin Layer Chromatography.

RP-HPLC analysis of seco-iridoid glycoside swertiamarin from different Swertia species.

Genus Swertia is valued for its great medicinal potential; mainly Swertia chirayita (Roxb. ex Fleming) H. Karst. is used in traditional medicine for a wide range of diseases. Seco-iridoid glycosides like swertiamarin is referred with enormous pharmacological potentials. The aim of the study was to identify a suitable substitute to S. chirayita by quantifying seco-iridoid swertiamarin from five different Swertia species endemic to the Western Ghats. The reverse-phase high-performance liquid chromatography diode array detector analyses were performed and chromatographic separation was achieved on a Lichrospher 100, C18e (5 µm) column (250-4.6 mm). A mobile phase consisting of acetonitrile and water (25:75) was used for separation. Results indicated that the concentration of the marker compound has been found to vary largely between and within the species from different localities. The content of swertiamarin was the highest in S. chirayita compared to the other species studied herein, advocating the use of Swertia minor as an alternate source to S. chirayita.

Reverse Phase-ultra Flow Liquid Chromatography-diode Array Detector Quantification of Anticancerous and Antidiabetic Drug Mangiferin from 11 Species of Swertia from India.

BACKGROUND: Genus Swertia is valued for its great medicinal potential, mainly Swertia chirayita (Roxb. ex Fleming) H. Karst. is used in traditional medicine for a wide range of diseases. Mangiferin one of xanthoids is referred with enormous pharmacological potentials. OBJECTIVE: The aim of the study was to quantify and compare the anticancerous and antidiabetic drug mangiferin from 11 Swertia species from India. The study also evaluates hierarchical relationships between the species based on mangiferin content using multivariate analysis. MATERIALS AND METHODS: The reverse phase-ultra flow liquid chromatography-diode array detector analyses was performed and chromatographic separation was achieved on a Lichrospher 100, C18e (5 µm) column (250-4.6 mm). Mobile phase consisting of 0.2% triethylamine (pH-4 with O-phosphoric acid) and acetonitrile (85:15) was used for separation with injection volume 20 µL and detection wave length at 257 nm. RESULTS: Results indicated that concentration of mangiferin has been found to vary largely between Swertia species collected from different regions. Content of mangiferin was found to be highest in Swertia minor compared to other Swertia species studied herein from the Western Ghats and Himalayan region also. The same was also evident in the multivariate analysis, wherein S. chirayita, S. minor and Swertia paniculata made a separate clade. CONCLUSION: Conclusively, the work herein provides insights of mangiferin content from 11 Swertia species of India and also presents their hierarchical relationships. To best of the knowledge this is the first report of higher content of mangiferin from any Swertia species. SUMMARY: The present study quantifies and compares mangiferin in 11 species of Swertia from India. The study also evaluates hierarchical relationships between the species based on mangiferin content using multivariate analysis. The mangiferin content was highest in S. minor compared to the studied Swertia species. To the best of our knowledge this is the first report of higher content of mangiferin from Swertia species. Abbreviations used: LOD: Limit of detection, LOQ: Limit of quantification, RP-UFLC-DAD: Reverse phase-ultra flow liquid chromatography-diode array detector, RSD: Relative standard deviation, SAN: Swertia angustifolia, SAP: Swertia angustifolia var. pulchella, SBI: S. bimaculata, SCH: S. chirayita, SCO: S. corymbosa, SDE: S. densifolia, SDI: S. dialatata, SLA: S. lawii, SMI: S. minor; SNE: S. nervosa, and SPA: S. paniculata.

Simultaneous determination of vasicine and vasicinone by High-performance liquid chromatography in roots of eight Sida species.

INTRODUCTION: Sida L. is a medicinally important genus widely used in conventional systems of medicine in India. AIM: The present study aims toward simultaneous determination of two bioactive compounds vasicine and vasicinone in root extracts of eight Sida spp. from Western Ghats, India. MATERIALS AND METHODS: Determination of vasicine and vasicinone was undertaken in methanolic root extracts (10% w/v) of Sida acuta, Sida cordata, Sida cordifolia, Sida rhombifolia, Sida spinosa, Sida indica, Sida retusa and Sida mysorensis by high performance liquid chromatography (HPLC) method. The standards were prepared with the concentration of mg/mL. Data were expressed as mean values of three reading and relative standard deviations. The separation was achieved on a Waters, Nova-Pack, C18 (250 mm × 4.6 mm, 5 µ) column, with acetonitrile - 0.1 M phosphate buffer-glacial acetic acid (15: 85: 1, v/v/v) as solvent system at a flow-rate of 1.0 mL/min. The effluent was monitored using ultraviolet detection at a wavelength of 300 nm. RESULTS: Both calibration curves of standard showed good linear regression (R2 > 0.994). The limit of detection and the limit of quantification for vasicine was 0.110 and 0.333 µg/mL and for vasicinone was 0.059 and 0.179 µg/mL respectively. The vasicine content was highest in S. cordifolia (9.891 ± 0.495 µg/100 mg) and vasicinone content was rich in S. cordata (33.013 ± 1.651 µg/100 mg.) The content of vasicinone was higher than vasicine. CONCLUSION: HPLC method provides simple, accurate, and reproducible quantitative analysis for simultaneous determination of vasicine and vasicinone. Among the selected Sida species, S. cordifolia and S. cordata were found to be rich in the vasicine and vasicinone contents, respectively.

Monitoring seasonal variation of epicatechin and gallic acid in the bark of Saraca asoca using reverse phase high performance liquid chromatography (RP-HPLC) method.

BACKGROUND: Saraca asoca (Roxb.) Wilde (Fabaceae) is a high valued but vulnerable medicinal plant of Western Ghats region. This plant is mainly known for its use in various gynecological disorders. OBJECTIVE: The objective of the present study was to investigate seasonal variation of the polyphenolic compounds viz., epicatechin and gallic acid in the bark of S. asoca by using Reverse Phase High Performance Liquid Chromatography-Diode Array Detector (RP-HPLC-DAD) method. MATERIALS AND METHODS: The bark was collected in six different Ritu (season) viz. Varsha (monsoon), Sharad (autumn), hemant (early winter), Shishir (winter), Vasanta (spring), and Grishma (summer) mentioned in Ayurveda. RESULTS: The RP-HPLC-DAD analysis indicated that levels of epicatechin and gallic acid in the bark of S. asoca vary seasonally. The highest concentration of epicatechin was observed in Shishir Ritu (3315.19 ± 165.76 mg/100g) and gallic acid during Hemant Ritu (211.90 ± 10.60 mg/100 g). CONCLUSIONS: In present study, the ability to synthesize and accumulate both the compounds in bark of S. asoca varied greatly throughout the seasons. It was also observed that the compound epicatechin was present abundantly as compared to gallic acid throughout the seasons.

Preliminary pharmacognostic screening of Achyranthes coynei stem.

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes(®) CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick-walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High-performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material.

Quantitative determination of three pentacyclic triterpenes from five Swertia L. species endemic to Western Ghats, India, using RP-HPLC analysis.

Aim of this study was to identify pentacyclic triterpenoids betulinic acid (BA), oleanolic acid (OA) and ursolic acid (UA) from five Swertia species endemic to Western Ghats, which are used as substitutes as well as adulterants to Swertia chirayita. Our results indicate that the concentration of active compound vary largely among and within the species from different localities. OA was found in a higher amount in all species compared to BA and UA. From the results, it was clear that BA, OA and UA are present in the endemic species collected from Western Ghats, thus advocating the use of these species as alternate sources to S. chirayita. This in due course may release pressure of exploitation from natural resources of S. chirayita and help to bring it out from an endangered category from conservation point of view.

Total polyphenolic contents and in vitro antioxidant properties of eight Sida species from Western Ghats, India.

BACKGROUND: Sida L., is a medicinally important genus, the species of which are widely used in traditional systems of medicine in India. Pharmacologically, roots are known for anti-tumor, anti-HIV, hepatoprotective, and many other properties. Phenolic antioxidants help in reducing oxidative stress occurring during treatment of such diseases. OBJECTIVE: The study aimed to evaluate and compare polyphenol contents and antioxidant properties of eight selected species of Sida from Western Ghats, India. MATERIALS AND METHODS: Methanolic root extracts (10% w/v) of Sida species, viz., S. acuta, S. cordata, S. cordifolia, S. indica, S. mysorensis, S. retusa, S. rhombifolia, and S. spinosa were analyzed. RESULTS: Sida cordifolia possessed highest total phenolic content (TPC: 1.92 ± 0.10 mg Caffeic Acid Equivalent/g and 2.13 ± 0.11 mg Tannic Acid Equivalant/g), total flavonoid content (TF: 2.60 ± 0.13 mg Quercetin Equivalent/g) and also possessed highest antioxidant activities in 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging (51.31 ± 2.57% Radical Scavenging Activity, (RSA); Trolox Equivalent Antioxidant Capacity: 566.25 ± 28.31µM; Ascorbic acid Equivalent Antioxidant Capacity: 477.80 ± 23.89 µM) and Ferric Reducing Antioxidant Power assays (TEAC: 590.67 ± 29.53 µM; AEAC: 600.67 ± 30.03 µM). Unlike DPPH and Ferric Reducing Antioxidant Power (FRAP) activity, 2, 2'-Azinobis (3-ethyl Benzo Thiazoline-6-Sulfonic acid) ABTS(+) antioxidant activity was highest in S. indica (TEAC: 878.44 ± 43.92 µM; AEAC 968.44 ± 48.42 µM). It was significant to note that values of AEAC (µM) for all the antioxidant activities analyzed were higher than that of TEAC. CONCLUSION: The high contents of phenolic compounds in the root extracts of selected Sida species have direct correlation with their antioxidant properties. Conclusively, roots of S. cordifolia can be considered as the potential source of polyphenols and antioxidants.

Evaluating Nothapodytes nimmoniana population from three localities of Western Ghats using camptothecin as phytochemical marker and selection of elites using a new-content range chart method.

BACKGROUND: Nothapodytes nimmoniana (Grah.) Mabb. is a high valued medicinal plant endemic to Western Ghats of India, distributed in fragmented populations. The plant is valued for potent anticancer drug camptothecin (CPT). OBJECTIVE: The study compares and expounds variation in CPT content from leaves and stems of N. nimmoniana obtained from three populations of Western Ghats, India. The study also describes a method for categorizing these populations using content range chart (CRC) method for percent yield of CPT. MATERIALS AND METHODS: A total of 60 samples were investigated including ten each of leaves and stems from three localities. Micro-extraction method was implemented to extract CPT. reversed phase ultra-performance liquid chromatography photo diode array technique was used to quantify CPT. RESULTS: Leaf samples of an individual collected from Joida, yielded lowest CPT content (0.002 ± 0.000 g/100 g), whereas a stem sample from Amgaon, yielded highest CPT content (0.123 ± 0.006 g/100 g). The findings suggest great variation in individuals producing and accumulating CPT. Using this data along with earlier published work, five categories of CPT yielding plants were made viz. I: Very low: <0.020, II: Low: 0.021-0.039, III: Moderate: 0.040-0.059, IV: High: 0.060-0.079 and V: Very high: >0.080. Based on CPT content in leaves, majority of individuals were under very low category (I(st)) and on the other hand stem samples were in 'II' category. Besides, very few individuals were observed in category 'V'. CONCLUSION: The study expounds use of CRC method for identifying elite population and suggests the need for its conservation.

Accumulation and trends in distribution of three triterpenoids in various parts of Achyranthes coynei determined using RP-UFLC analysis.

BACKGROUND: Achyranthes coynei Sant. (Family: Amaranthaceae) is a rare endemic medicinal plant used by local traditional practitioners to treat various diseases. The plant has been reported for promising antibacterial and antioxidant activities. However, the plant is not explored for its phytocompounds, especially triterpenoids. OBJECTIVE: TO STUDY THE ACCUMULATION AND TRENDS IN DISTRIBUTION OF TRITERPENOIDS: betulinic, oleanolic and ursolic acids (BA, OA and UA) in leaf, stem, root and inflorescence of A. coynei. MATERIALS AND METHODS: Extraction was achieved using refluxing and reversed phase-ultra flow liquid chromatographic (RP-UFLC) technique was employed for determination. Separation of triterpenoids was achieved on a Hibar 250-4.6 mm, 5 µ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase (pH adjusted to 5.0 using GAA) in an isocratic mode. RESULTS: Oleanolic acid was higher in leaf (0.172 ± 0.009%) followed by stem (0.035 ± 0.002%) and root (0.028 ± 0.001%). Ursolic acid was accounted to be highest in the inflorescence (0.099 ± 0.005%). The contents of BA and UA were lower than OA in leaf and stem while it was remained undetected in roots of A. coynei. CONCLUSION: THE TRITERPENOIDS: BA, OA and UA were detected, quantified and reported for the first time from A. coynei. In the present study leaves were found to be the major source of BA and OA, whereas inflorescence was for UA.

Compound Specific Extraction of Camptothecin from Nothapodytes nimmoniana and Piperine from Piper nigrum Using Accelerated Solvent Extractor.

Effects of varying temperatures with constant pressure of solvent on extraction efficiency of two chemically different alkaloids were studied. Camptothecin (CPT) from stem of Nothapodytes nimmoniana (Grah.) Mabb. and piperine from the fruits of Piper nigrum L. were extracted using Accelerated Solvent Extractor (ASE). Three cycles of extraction for a particular sample cell at a given temperature assured complete extraction. CPT and piperine were determined and quantified by using a simple and efficient UFLC-PDA (245 and 343 nm) method. Temperature increased efficiency of extraction to yield higher amount of CPT, whereas temperature had diminutive effect on yield of piperine. Maximum yield for CPT was achieved at 80°C and for piperine at 40°C. Thus, the study determines compound specific extraction of CPT from N. nimmoniana and piperine from P. nigrum using ASE method. The present study indicates the use of this method for simple, fast, and accurate extraction of the compound of interest.