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Open Access
Structure-based virtual high-throughput screening for ligands of ecdysone receptor
(CCSHAU, Hisar, 2020) Gagan Rani; Sudhir Kumar
Insects occupy more than 70% of entire kingdom Animalia and are the most successful group o among the organisms dwelling on earth, a set of injurious insects referred as pests. Pests/insects can harm agricultural plants, devour and/or harm harvested crops or transmit diseases to people and animals. These insect pests have several enzymes. hormones receptors, these hormone receptor binds with specific hormone and thus regulates development and also plays a key role in their life processes. If the normal functioning of these enzymes and the receptors are disturbed, their normal life cycle may also be affected thereby causing death of the insect. In the present study we focused only on ecdysone receptor, which is a type of nuclear receptor steroids hormone. The ecdysone receptor (EcR) is a nuclear transcription factor depends upon ligands found in arthropods. The receptor binds to ecdysteroids hormones, which play a major role in reproduction and regulate vital processes such as growth, molting and metamorphism. For activation of gene transcription, a non-covalent heterodimerization of EcR and USP (ultra-spiracle protein) is necessary, that occurs after binding of an agonist to ecdysone receptor. Because of the importance of this hormone receptor in growth, reproduction and metamorphosis of insects, it was considered to be excellent targets for pest control agents and chemical insecticides. In the presents study virtual high-throughput of two libraries of 705,632 (on the basis of substructure) and 4591,276 (clean leads) were carried out for ecdysone receptor. At the same time vHTS of 21 known inhibitors for ecdysone receptor also performed. vHTS results showed that 1,85,785 and 5,15,112 leads had binding energy within the range or lesser than the binding energy of known inhibitors (from both libraries respectively). These selected leads were then subjected to ADME-Tox study, from this 264 and 509 leads were predicted to be non-toxic.
Open Access
Effect of extraction conditions on phytochemicals and antioxidant potential of Giloy (Tinospora cordifolia) stem and Kair (Capparis decidua) fruit
(CCSHAU, Hisar, 2020-06-26) Promila; Sushila Singh
Tinospora cordifolia (Giloy) and Capparis decidua (Kair) are important medicinal plants which are known to possess antioxidant, immunomodulatory, antibacterial and hepatoprotective properties. This study was aimed to determine the proximate and chemical composition as well as the effect of different pH levels of water and herbal extraction conditions (viz. infusion 5 & 10 min, decoction and strong decoction) on phenolics, flavonoids and antioxidant potential of stem & leaves of Giloy and aerial parts, fruits & flowers of Kair. Total phenolics was calculated by Folin-Ciocalteu reagent, flavonoids content by Aluminium chloride colorimetric assay. Antioxidant activities were tasted using DPPH and phosphomolybdenum assay. Amount of bitter principles in Giloy stem was 2.74% (w/w). Alkaloids and saponin content in Kair fruits were 2.43 and 8.46% (w/w), respectively. The highest amount of total phenolics (mg GAE/g) in Giloy stem (7.43) & Kair aerial parts (11.51) was at pH 2; in Giloy leaves (25.63), Kair fruits (17.49) & flowers (22.09) was at pH 5. The highest concentration of total flavonoids (mg CE/g) in stem & leaves of Giloy and aerial parts & flowers of Kair were- 0.68, 2.39, 1.07 and 2.18 at pH 2, respectively. In Kair fruits, flavonoids content was highest (1.09) at pH level 11. IC50 values (ppm) as inversely related to DPPH free radical scavenging activity followed the order: pH 5 (865.91) < pH 7 (915.13) < pH 11 (919.50) < pH 2 (1451.22) in Giloy stem; pH 11 (304.48) < pH 2 (343.82) < pH 7 (380.04) < pH 5 (409.51) in Giloy leaves; pH 7 (332.74) < pH 2 (553.47) < pH 5 (622.42) < pH 11 (788.35) in Kair aerial parts; pH 5 (389.96) < pH 7 (584.86) < pH 11 (690.35) < pH 2 (973.99) in Kair fruits; pH 7 (454.03) < pH 2 (579.10) < pH 5 (635.72) < pH 11 (710.72) in Kair flowers. Total antioxidant capacity (mg AAE/g) of stem of Giloy (34.38) and aerial parts (51.26), fruits (75.00) & flowers (90.45) of Kair was maximum at pH 5 and of Giloy leaves (67.45) was maximum at pH 2. Total phenolic content (mg GAE/g) of stem (4.84) & leaves (19.38) of Giloy and aerial parts (9.54), fruits (14.84) & flowers (17.48) of Kair was highest in strong decoction (90 min). Total flavonoids content (mg CE/g) of Giloy stem (0.75) & leaves (2.71) was maximum in decoction (30 min) while of Kair aerial parts (0.69), fruits (1.81) & flowers (1.69) was highest in strong decoction (90 min). IC50 values (ppm) as inversely related to DPPH free radical scavenging activity followed the order: infusion (5 min) (711.57) < decoction (30 min) (790.10) < infusion (10 min) < strong decoction (90 min) (1568.53) in Giloy stem; infusion (10 min) (61.21) < decoction (30 min) (183.69) < strong decoction (90 min) (272.24) < infusion (5 min) (311.51) in Giloy leaves; infusion (10 min) (169.72) < decoction (30 min) (515.93) < strong decoction (90 min) (658.60) < infusion (5 min) (754.67) in Kair aerial parts; infusion (10 min) (346.45) < decoction (30 min) (377.38) < infusion (5 min) (438.36) < strong decoction (90 min) (636.06) in Kair fruits; and decoction (30 min) (343.45) < infusion (10 min) (423.96) < strong decoction (90 min) (647.56) < infusion (5 min) (788.79) in Kair flowers. Total antioxidant capacity (mg AAE/g) of Giloy stem (82.55) was maximum in infusion (5 min) and of Giloy leaves (96.52), Kair aerial parts (79.04), Kair fruits (130.95) and Kair flowers (203.28) was maximum in decoction (30 min). Present research studies indicated that pH levels of water and herbal extraction conditions significantly affected the extraction of phytochemicals from stem & leaves of Giloy and aerial parts, fruits & flowers of Kair and their antioxidant potentials.
Open Access
Synthesis and characterization of metallic nanoparticles, antimicrobial and antioxidant potential of Triphala powder and its fruit constituents
(CCSHAU, Hisar, 2020) Tamanna; Madan, V. K.
The present study was undertaken to estimate the phytochemical and nutritional parameters; to determine the antioxidant potential of promising extracts of Triphala powder and its fruit constituents: Amla, Bahera and Harad using DPPH free radical scavenging assay and Phosphomolybdate assay; by making use of these bioactive components in plant extracts and their bio efficacies to synthesize the metallic nanoparticles; compare the antimicrobial activity of as synthesized NPs and their crude extracts. Moisture, fat, ash, protein, crude fibre and total carbohydrates contents were estimated in different parts viz. seeds, aerial parts & roots of Amla, Bahera and Harad. All parts of these medicinal plants were found to be very rich sources of bioactive components/antioxidants. The fruit pulp of these plants also contained different minerals viz. Fe, Cu, Zn and Mn. Among these plant extracts: total phenolics, flavonoids, DPPH free radical scavenging activity and total antioxidant capacity were higher in herbal extracts of fruits in comparison to herbal extracts of aerial parts and roots. Hence, these medicinal plants are important source of good antioxidant potential and antimicrobial activity which may be due to the presence of various phytochemicals; therefore, play an important role in antioxidant defence system against free radicals and thus aid in human health. The total phenolics contents, total antioxidant capacity and thus antimicrobial activity followed the order as: Amla > Triphala > Bahera > Harad. The total flavonoids content varies as Bahera > Triphala > Amla > Harad. DPPH free radical scavenging assay follows the order as Bahera > Amla > Harad > Triphala. Aqueous extracts were better in all sorts of quantifications of antioxidants and their antioxidant potential. Amongst these natural therapeutic agents, Amla showed the highest antibacterial activity followed by Triphala powder, Bahera and Harad. In case of antifungal activity, it was Bahera which showed the highest activity against the test organisms taken; followed by Amla, Harad and Triphala powder. Green synthesis of metallic nanoparticles using plant derived bio-active components is an economical, eco-friendly and simple method in the synthesis route. Plant extracts contain diverse chemical compounds such as proteins, carbohydrates, alkaloids, tannins, phenolics, oils and saponins which have medicinal value and the same can act as reducing and capping agent for the nanoparticles synthesis. Bimetallic NPs possessed better antimicrobial activity as compared to monometallic NPs. Gram negative (E. coli and P. aeruginosa) bacteria were more susceptible to gram positive (S. aureus and B. subtilis) bacteria. The order for antibacterial efficacies was as follows: Ag@AuNPs > AgNPs > AuNPs. In case of antifungal activities, AuNPs were better than AgNPs i.e. the order was Ag@AuNPs > AuNPs > AgNPs. A. niger was most susceptible as compared to A. flavus and C. albicans to the action of these plant derived therapeutic agents.
Open Access
Virtual High-throughput Screening of Acetylcholinesterase and Chitin Synthase Inhibitors
(CCSHAU, 2019) Gupta, Neha; Sudhir Kumar
Insects are most diverse and abundant of all terrestrial animals. Insects dominated the earth by successfully adapting to wide range of ecosystem. As a part of the ecosystem, they compete for food with other biota and in turn they cause various infections to vegetation, livestock and humans. These insect pests have several enzymes and each of these enzymes plays a key role in their life processes. If the normal functioning of these enzymes is disturbed, their normal life cycle may also be affected thereby causing death of the insect. In the present study we focused only on two enzymes which are acetylcholinesterase (AChE) and Chitin synthase. Acetylcholinestearse is one of the key enzymes belonging to the family of serine hydrolase. Principle function of AChE is to terminate the impulse transmission by rapidly hydrolyzing the neurotransmitter ACh to choline and acetate. Due to its crucial role it acts as target for variety of chemical agents. Chitin synthase catalyzes the last step in the chitin biosynthetic pathway. It is a major component of exoskeletal structure of arthropods, including crustaceans and insects as well as mollusks, nematodes and worms. To allow growth and development, insects periodically replace their old cuticle with a new one during molting (ecdysis). Because of the importance of both the enzymes in growth of insects, these were assumed to be excellent targets for pest control agents and chemical insecticides. In the present study virtual high-throughput screening of 4,591,276 clean leads was carried out for both the enzymes. At the same time vHTS of known inhibitors of AChE and CHS was also performed. vHTS results showed that for AChE 227,036 leads and for CHS 977,130 leads had binding energy within the range or lesser than the binding energy of known inhibitors. These leads were then subjected to ADME-Tox study and 217 and 864 leads were predicted to be non-toxic, for AChE and CHS respectively.
Open Access
Synthesis of bioactive heterocyclic compounds using green synthetic methods and their bioevaluation
(CCSHAU, 2019) Suman; Rajvir Singh
Green chemistry is a sustainable approach to explain latest research topics and expresses an area of scientific discoveries about pollution awareness. Green chemistry is the future chemistry which is simple, efficient and benign in all steps of particular synthesis processes. Coumarins, chalcones, Pyrazoles and Imidazoles are versatile classes of heterocycles and intermediates for the preparation of other organic compounds due to their effective biological activities. For the preparation of these compounds and their derivatives, different organic solvents and reagents are being used which are quite hazardous to environment and human health due to their volatile and toxic nature. Since last few years attempts are being made for ecofriendly synthesis viz. biocatalysis and synthesis by using green catalyst. Biocatalyst is a magical intermediate which is helpful to overcome the negative environmental effects due to various hazardous and toxic chemicals used in synthetic pathways.Therefore an attempt has been made to synthesize above mentioned classes of compounds by using various green catalyst. Thesecompounds were characterized by spectroscopic technique viz.1HNMR, IR, Elemental analysis and other analytical data. The green catalysts were examined by SEM. Synthesized compounds were screened for their antifungal, herbicidal and antioxidant activities at four different concentrations. Compounds 12 and 10 were found more activeagainst the tested fungi Rhizoctonia solani and Aspergillus niger which have EC50values75.03 and 77.53μg/ml respectively. Compounds 11 and 60 were found more active against Raphanus sativus L. (root) and (shoot) exhibited growth inhibition 82.86, 91.66, 92.12, 97.05% and 43.81, 58.00, 67.01, 89.07% respectively at50, 100, 150, 200μg/ml all concentrations.Compound 84 was found most active with least IC50 36.30μg/ml and showed highest DPPH free radical scavenging capacity at 25, 50, 75, 100μg/ml.
Open Access
Design, synthesis and bioevaluation of some novel bioactive ingredients
(CCSHAU, 2019) Sheetal; Rajvir Singh
Coumarins, Chalcones, Propranolols, Proterenols, and Pyrazoles are versatile classes of heterocycles and intermedients for the preparation of other organic compounds due to their various biological activities. Many natural and synthetic drugs, dyes, pesticides are heterocyclic in nature. These compounds were synthesized by conventional method. These compounds were characterized by spectroscopic technique viz. 1NMR, IR, Elemental analysis and other analytical data. Synthesized compounds were screened for their Fungi toxicity by poison food technique at four different concentrations. Compounds 13 and 25 were found more active against the tested fungi Rhizoctoniasolani and Aspergillusniger which haveEC50values 52.91, 71.88 and 48.00, 78.60μg/ml respectively. Compounds 34 and 105were found more active against the tested fungi Rhizoctonia solani and Aspergillus niger which have EC50 values 40.51, 60.49 and 57.25, 56.52μg/ml respectively. Synthesized compounds were screened for their antibacterial activity by zone inhibition method at four different concentrations. Compounds 17 and 27 were found more active against Bacillus species at all the test concentrations. Compounds 79 and 132 were found more toxic against the Bacillus species at all the test concentrations. Compound 179 was found more toxic against the Bacillus species at all the test concentrations. Synthesized compounds were also screened for their herbicidal activity against growth of weed roots and shoots of Raphanussativus L. in vitro by percentage inhibition technique. Compounds 13 and 67 were found more toxic in % growth inhibition of both roots and shoots at all the tested concentrations. Compounds 92 and 118 were found more toxic in % growth inhibition of both roots and shoots at all the tested concentrations. Compounds 132 and 153 were found more toxic in % growth inhibition of both roots and shoots at all the tested concentrations.
Open Access
Studies on phytochemicals, nutritional parameters and antioxidant potential of Chicory (Cichorium intybus L.) and Garden cress (Lepidium sativum L.)
(CCSHAU, 2018) Jangra, Satya Shree; Madan, V. K.
The present study was undertaken to estimate the phytochemical and nutritional parameters; comparison of microwave assisted and conventional extraction techniques and effect of herbal extracts (infusion, decoction and strong decoction) on total phenolics, flavonoids as well as on the antioxidant potential of the extracts of different plant parts of Chicory and Garden cress (Hisar and Solan regions). Moisture, fat, ash, protein, crude fibre and total carbohydrates were estimated in different parts viz. seeds, aerial parts & roots of Chicory and Garden cress (both regions). All parts of Chicory and Garden cress (both regions) were found to be very rich sources of energy on the basis of their calorific values. Seeds, aerial parts & roots of Chicory and Garden cress (both regions) also contained minerals viz. Fe, Cu, Zn and Mn. Different parts of Chicory and Garden cress (both regions) were found to contain ascorbic acid, starch, tannins, total sugars, reducing sugars and non-reducing sugars also in varying amounts. Oil content in seeds of Chicory, Garden cress (Hisar region) and Garden cress (Solan region) were 18.05, 22.42 and 25.52%, respectively. Major constituents in Chicory and Garden cress (both regions) seed oil were palmitic acid, oleic acid, linoleic acid and linolenic acid. Total phenolics (6.90 - 10.78 mg GAE/g), flavonoids (1.55 - 6.33 mg CE/g) and total antioxidant capacity (63.15 - 223.70 mg AAE/g) were highest in whole plant and seeds extracts of both crops prepared by soxhlet extraction technique followed by column chromatography, microwave assisted and homogenization techniques. DPPH free radical scavenging activity of whole plant and seeds (IC50: 410.6 - 647.8 μg/mL) was also highest of extracts of both crops prepared by soxhlet extraction technique followed by column chromatography (IC50: 411.9 - 666.7 μg/mL), microwave assisted (IC50: 419.1 - 668.0 μg/mL) and homogenization (IC50: 437.4 - 701.1 μg/mL) techniques. Among plant parts of Chicory, total phenolics, flavonoids, DPPH free radical scavenging activity and total antioxidant capacity were higher in whole plant extracts in comparison to seeds extracts; whereas in Garden cress of both regions, total phenolics, flavonoids, DPPH free radical scavenging activity and total antioxidant capacity of seeds extracts were higher in comparison to whole plant extracts. Strong decoctions of aerial parts and roots of both crops had highest total phenolics (1.83 - 19.62 mg GAE/g), flavonoids (0.53 - 8.53 mg CE/g) and total antioxidant capacity (11.93 - 80.31 mg AAE/g) in comparison to their corresponding decoctions and infusions. Strong decoctions of aerial parts and roots also exhibited highest DPPH free radical scavenging activity (IC50: 372.7 - 736.1 μg/mL) in comparison to corresponding decoctions (IC50: 385.8 - 739.2 μg/mL) and infusions (IC50: 396.1 - 744.0 μg/mL). Among different plant parts of Chicory and Garden cress (both regions); total phenolics, flavonoids, DPPH free radical scavenging activity and total antioxidant capacity were higher in herbal extracts of aerial parts in comparison to herbal extracts of roots. Hence, Chicory and Garden cress possess good antioxidant potential which may be due to the presence of various phytochemicals; therefore, play an important role in antioxidant defence system against free radicals and thus aid in human health.
Open Access
Studies on chemical constituents of Prosopis cineraria (L.) and interaction of phenolics with protein
(CCSHAU, 2018) Suman; Rajvir Singh
Prosopis cineraria (L.) Druce is native species of Indian desert, mainly known as “King of Desert”. Its all parts have various parmaceutical and medicinal importance. Plant materials i.e. bark, flowers, leaves and pods of Prosopis cineraria were collected and extracted with hot methanol by refluxing method. The respective extractives were divided into two parts. One major part was subjected to column chromatography which affored a total twenty-two compounds. Bark column contained four compounds namely, 24- methylenecycloartan-3-one, Lupeol, 5,7,4‟-trihydroxy-3‟-methoxy flavanone and β- Sitosterol while six compunds namely, Hexacosane, Tetracosyl tetracosanoate, 1- triacontanol, Patuletin, Bis (cis-13-docosenamido) methane and β- Sitosterol from flowers of P.cineraria. leaves afforded eight compounds namely, 1- Heptacosanol, 1- Nonadecanol, Stigmasterol, nonacosane-15-one, 1- tetracosanoic acid, β- Sitosterol, 3-O-methoxy-D-Glucose and 13- docosenamide. Four compounds namely, 1-heptatricontanol, 3,8,8-trimethoxy-3-piperidyl- 2,2‟binaphthalene-1,1‟,4,4‟-tetrone, β- Sitosterol and Catechin were isolated from pods of Prosopis cineraria. Various phytochemical activities like total phenolic content, total flavonoids, mineral contents and phenolics interaction with protein were evaluated from methanolic extract and their fractions. Total phenolic contents were found to be maximum in methanol fraction of pods of Prosopis cineraria while acetone fraction of pods contained maximum total flavonoid content. P. cineraria possessed mineral content in trend of N> P> K> Fe> Cu> Zn> Mn. Acetone fraction of pods part was found to be maximum protein binding capacity at highest concentraction (10μg/μl). It was significantly showed that P. cineraria has nutritionl and digestible value of various fractions by stepwise increased in the percentage activity at different concentrations.
Open Access
Efficacy of phenolics from agri-byproducts in vegetable oil
(CCSHAU, 2018) Nehra, Sukriti; Sushila Singh
The present study was undertaken to determine the efficacy of phenolics from pod coat of pulses in stabilising soybean oil. Pod coat of five pulse crops (cowpea, clusterbean, mungbean, Mothbean and pigeonpea) were extracted with three solvents (acetone, ethylacetate and chloroform). Phytochemical constituents (phenols, flavonoids, tocopherols and tannins) and DPPH free radical scavenging activity were estimated by using spectroscopic methods. Folin-Ciocalteu procedure and aluminium chloride colorimetric methods were used for evaluation of total phenolic content and flavonoid content respectively. Among the various extracts, highest yield was observed in acetone extract of clusterbean (5.44%) while lowest was observed in chloroform extract of cowpea (2.99%). Total phenolic content (mg GAE/g) varied from 7.24-24.18 in all of the extracts. Similarly, flavonoids (mg CAE/g) ranged from 3.30 to 10.77 in all of the extracts. Tocopherol content (mg/g) was highest in ethylacetate extract of pigeonpea (9.56) while lowest in chloroform extract of cowpea (3.41). Tannin content (mg TAE/g) was highest in acetone extract of clusterbean (2.48) while lowest in chloroform extract of mungbean (0.71). Results obtained by DPPH method showed that free radical scavenging capacities of various pod coat extracts from different species of legumes ranged from 54.95 to 78.21%. Their IC50 values (mg/mL) varied from 0.23- 0.83. Analogous to results obtained for total phenolics and flavonoids, free radical scavenging activities were highest in acetone extract of clusterbean. The high content of total phenolics and total flavonoids indicated that these bioactive metabolites contribute to synergistic effect for the antioxidant activity. Moreover, a highly significant correlation (p<0.05) between free radical scavenging efficiencies and phenolic contents as well as flavonoids was observed in most of the extracts under study. All extracts were added to crude soybean oil at two concentrations (1000 and 2000ppm). Soybean oil sample stabilized with various extracts were incubated at 50ºC for a period of 28 days. Effect of incorporation of these extracts on oxidative stability of crude soybean oil was measured in terms of various oxidative quality indices. Among the three extracts, acetone extract was found most effective in retarding oxidative degradation of soybean oil during incubation. Extracts were more effective at 2000ppm concentration in comparison to 1000ppm. Among the five crops, cluster bean extracts displayed highest inhibition towards oxidation of soybean oil. The results concluded that the pod coat extracts of these pulses may be valuable natural antioxidant sources and are potentially applicable in both medicine and food industry.

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