• Extractives Analysis
    At Celignis Analytical

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Publications on Extractives By The Celignis Team

Shweta Shinde, Archana Zambare, Vibha Parashere; Vasudeo Zambare (2018) In Vitro studies on antimicrobial activity and phytochemical analysis of Tinospora cordifolia and use as chocolate supplement, International Journal of Advanced Biotechnology and Research 9(4): 632-639

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Background and Objectives: Day by day the antibiotic resistance is increasing and become a serious problem. Hence its a todays need to find some naturally occurring bioactive compounds such as plant bioactive compounds. The objective of the study was to evaluate the antimicrobial activity of Tinospora cordifolia root extracts against human pathogens and T. cordifolia extract were tested for phytochemical examination. A chocolate product development with T. cordifolia extract was also developed. Materials and Methods: Antimicrobial efficiency of Tinospora cordifolia, a medicinal plants were examined using isopropyl alcohol, water, methanol, and acetone, as solvents and tested against four human pathogens like Escherichia coli, Salmonella sp., Staphylococcus aureus, Pseudomonas auruginosa using agar well diffusion method and minimum inhibitory concentration. The phytochemical analysis carried out for qualitative testing methods and T. cordifolia water extract was used as a supplement in chocolate preparation. Results: Water extract showed more extraction yield of 2.68%, followed by methanol (1.6%), isopropyl alcohol (1.13%) and acetone (0.58%). All the extracts showed significant activity against all pathogens, but the isopropyl alcoholic extract of T. cordifolia showed maximum zone of inhibition against all the microorganisms. Also, S. aureaus was inhibited by all extracts. Acetone extract was less potent to inhibit pathogens except S. aureus. The minimum inhibitory concentration of isopropyl alcohol, water, methanol and acetone extracts showed 3.48 mg, 8.04 mg, 3.9 mg and 1.74 mg, respectively. The phytochemical analysis carried out revealed the presence of alkaloids, carbohydrates, terpenoids, proteins, flavonoids, steroids, glycosides in most of the extracts of T. cordifolia. A chocolate preparation containing water extract of T. cordifolia was developed and can be available as source of medication for kids. Conclusion: The Spectrum of activity observed in the present study may be indicative of the present study extracts of T. cordifolia plants could be a possible source to obtain new and effective herbal medicines to treat infections, hence justified the ethnic uses of T. cordifolia against various infectious diseases.

Ahire V, Zambare A, Zambare V. (2017) Extraction, purification and characterization of protease from latex of Plumeria Sp, International Journal of Advanced Biotechnology and Research 8(2): 1349-1353

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Protease was isolated from the latex of Plumeria sp. using ammonium sulphate precipitation (60% saturation) method and purified by a dialysis followed by DEAE cellulose column chromatography. DEAE Cellulose chromatographic method showed 9.86-fold purification which is about 57.36% yield as compared to crude latex protease. Purified plumerian protease was showing optimum activity at pH 7 and temperature 500C. It was activated by 10mM Calcium chloride and betamercaptoethanol, however inhibited by 10mM iodoacetamide, indicated the presence of sulfhydryl as an essential group for its activity. The enzyme kinetic with casein substrate showed km and Vmax of 1.66 mg/ml and 333U/mg, respectively. Plumeria sp. showed a single protein band on SDS-PAGE and molecular weight was of 80 kDa. Thus, protease from the latex of Plumeria sp. was purified and characterized and it may be explored further to study its impact in medical science as an effective anti-inflammatory agent.

Padul MV, Patil MT, Chouguale AD, Zambare VP, Patil RM, Ghule RB, Naikwade SV, Garad AS, Shaikh FK, Gadge PP, Shinde KD, Dama LB and Salve AN. (2013) In vitro screening of proteinase inhibitors (trypsin, chymotrypsin and helicoverpa gut proteinase inhibitors) in different plant tissue extracts, Trends in Biotechnology Research 1(1): 7-14

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Most of the plant protection strategies are focused on selection and application of the natural proteinase inhibitors (PIs) against insect pests. In addition, PIs also play a vital role in medicine for treatment of immunity related diseases. PI activity exists mainly in seeds, leaves and flowers of plants. In search of novel PIs, 135 different plant tissue extracts (leaf, flower and seed) were screened for their PI (trypsin, chymotrypsin and Helicoverpa gut proteinase inhibitors) activities by using dot-blot assays. Most of the plant tissues screened revealed moderate PI activity, few showed low PI activity and very few of them showed strong PI activity against trypsin, chymotrypsin and Helicoverpa gut proteinases. The inhibitory potency of positive samples was further determined by solution assays. Five plants namely Arachis hypogaea, Vigna sinensis, Dolichos lablab, Phaseolus aureus and Cassia siamea showed higher activity which ranged from 22.91 to 58.33 %. Higher activities recorded in the seed as compare to leaf and flower tissues. Dolichos lablab showed highest PI activity (58.33 %) followed by Cassia siamea (52.08 %). PI activity was found to be distributed unequally in ammonium sulfate (NH 2 SO 4) fractions. INTRODUCTION Proteolytic enzymes catalyzing the hydrolytic cleavage of specific peptide bonds in target proteins are called as proteinases. These proteolytic enzymes are widely distributed in nearly all plants, animals and microorganisms (Christeller, 2005; Joanitti et al., 2006). In higher organisms proteinases play key roles in many biological processes. The proteolytic events catalyzed by these enzymes serve as mediators of signal initiation, transmission and termination in many of the cellular events such as inflammation, apoptosis, blood clotting and hormone processing pathways (Ivanov et al., 2006). But they may be potentially damaging when present in higher concentrations. For this reason their activities need to be strictly regulated and controlled.

Gadge PP, Madhikar SD, Yewle JN, Jadhav UU, Chougale AD, Zambare VP, Padul MV (2011) Biochemical Studies of Lipase from Germinating Oil Seeds (Glycine max), American Journal of Biochemistry and Biotechnology 7(3): 141-145

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Problem statement: Lipase is one of the important enzymes in food, pharmaceutical, detergent and biofuels industries. Search for the lipase with distinct features, possibly from germinating seeds, is of interest for industrial applications. Approach: The lipase produced by soybean oil seeds was partially purified and characterized in terms of the optimal pH and temperature for activity as well as substrate specificity. Results: The lipase was extracted and partially purified from germinating soybean seeds using chilled acetone and ammonium sulfate precipitation. Partially purified and dialyzed enzyme profile was observed on native-Polyacrylamide Gel Electrophoresis (PAGE). The lipase was optimally active at pH 8 and temperature of 24C. In the presence of Ca2+ and Mg2+ enhance the activity at low concentration, while the Hg2+ and Ethylene Diaminotetracetic Acid (EDTA) showed inhibitory effect. The enzyme was found to be metalloenzyme. Enzyme kinetics with olive oil emulsion substrate showed km and vmax of 7.67 mg and 0.0125 m mL min-1, respectively. Conclusion: The mettaloenzyme enzyme was able to attack specifically on oil in seeds to generate free fatty acids as the major end product. This understanding may help in devising efficient methods to overcome the problem of soybean seed oil in stability.

Vasudeo Zambare, Archana Zambare, Lew Christopher (2010) Antioxidant and antibacterial activity of extracts from lichen Xanthoparmelia somloensis, native to the Black Hills, South Dakota, USA, International Journal Medical Science and Technology 3(7): 46-51

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The present study was carried out to evaluate the antioxidant and antibacterial activity of lichen Xanthoparmelia somloensis, native to the Black Hills in South Dakota, USA. The antioxidant activity of lichen extracts was assessed using the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging assay. The lipid peroxidation reaction of acetone and methanol extracts was inhibited 85% and 81%, respectively A free radical scavenging activity of 77% (acetone extract) and 65% (methanol extract) was determined. The antibacterial activity was assayed against four clinical strains using the agar well diffusion method. Except for Escherichia coli, both extracts were found inhibitory to Streptomyces aureus, Streptococcus pyogenes,and Steptococcus agalactiae with minimum inhibitory concentration values of 0.7-0.9 mg/ml. It was demonstrated that both the antioxidant and antibacterial activities correlated well with the protein to polysaccharide ratio rather than the polyphenol content of the lichen extracts. To the best of our knowledge, this is the first literature report on antibacterial activity from the lichen X.somloensis. The results reported here warrant further investigations to establish the usefulness of X.somloensis in biomedical applications such as treatment of respiratory and urinary tract infections.

Zambare V. P, Bhoyte S. A. (2009) Antibacterial activity of tea, Biomedical and Pharmacology Journal 2(1): 173-175

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Antibacterial activities of tea extracts in various solvents were tested against six organisms, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Serracia sp., and Bacillus subtilis using agar-well method. Petroleum ether and chloroform extracts of tea showed strong antibacterial activities against P. aeruginosa and B. subtilis while other extracts were less active. The minimum inhibitory concentration (MIC) of chloroform extract of tea was found to be 25g/mL. This study may establish the need for daily use of this product for medicinal purposes.

M. V. Padul and V. P. Zambare (2008) In vitro antibacterial activity of Enicostema littorale plant extracts, Journal of Pure and Applied Microbiology 2(1): 245-247

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The antibacterial activity of aqueous, ethanolic, methanolic, ether, acetone, chloroform and hexane extracts from Enicostema littorale plant has been evaluated, in vitro, against Serrasia sp:, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. All extracts of Enicostema littorale exhibited highest antibacterial activity against Sarrasia sp and P. aeruginosa followed by very less activity against E. coli and S. aureus. The results indicate that Enicostema littorale plant may be a good candidate as antimicrobial agent.

V. P. Zambare and A. A. Bhole (2008) In vitro antioxidant activity of Plumbago zeylanica, Research Journal of BioTechnology 3(3): 36-37

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The total antioxidant activity, total phenolic content and reducing power of ethanol, methanol and water extracts of roots of plant Plumbago zeylanica were determined in vitro. Ethanol, methanol and water extracts of P. zeylanica roots showed the good antioxidant activity. However, there was no correlation between antioxidant activity and total phenolic content of the extracts. Although the methanol extract of P. zeylanica had the highest total phenolic contents, it exhibited low antioxidant activity. In contrast, there was a strong correlation between reducing power and total antioxidant activity of the extracts. The highest reducing power was determined for the methanol extract of Plumbago zeylanica.

Zambare V. P. , M. V. Padul, Yadav A. A, Shete T. B. (2008) Vermiwash: biochemical and microbiological approach as eco-friendly soil conditioner, ARPN Journal of Agricultural and Biological Sciences 3(4): 1-5

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Vermiwash was found to contain enzyme cocktail of proteases, amylases, urease and phosphatase. Microbiological study of vermiwash revealed that it contains nitrogen-fixing bacteria like Azotobactrer sp., Agrobacterium sp. and Rhizobium sp. and some phosphate solublizing bacteria. Laboratory scale trial showed effectiveness of vermiwash on Cowpea plant growth

Zambare V.P. and Bhoyte S. A (2008) Antibacterial activity of ginger (Zingiber officinalis), Journal of Pure and Applied Microbiology 2(2): 591-593

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V. P. Zambare, P. S. Kothari, M. V. Kulkarni (2007) Plasmid curing of plumbagin and its application in bacterial antibiotic resistance, Journal of Pure and Applied Microbiology 1(2): 285-288

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Plumbago zeylanica, a medicinal plant, contain plumbagin a naphthoquinonic compound. Partially purified plumbagin was tested for its plasmid curing activity. E. coli, isolated from pathological samples showed multiple antibiotic resistance. Plumbagin showed antibacterial activity at MIC value; 80 ?g ml -1. Antibiotics and plumbagin together decreased the antibiotic resistance pattern, suggesting plasmid curing by plumbagin. Thus, plumbagin can be used as an adjuvant to antibiotic therapy for treating infectious diseases caused by antibiotic resistant organisms.

S. S. Nilegaonkar, V. P. Zambare and P. P. Kanekar (2004) Antibacterial activity in vitro of plumbagin isolated from Plumbago zeylanica, Biotechnological Approaches for Sustainable Development, Allied Publishing Pvt. Ltd.

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