scholarly journals Characterization of Green Pigment from Sauropus androgynus Shoot Cultures

2021 ◽  
Vol 3 (3) ◽  
pp. 128-137
Author(s):  
Oeke Yunita ◽  
Devi Veronica Wijayanto ◽  
Aulia Dwi Hapsari ◽  
Muji Lestari Wahyu Wilang Sari ◽  
Alfian Hendra Krisnawan
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Spectrophotometric Analysis ◽  
Human Consumption ◽  
Shoot Cultures ◽  
Green Pigment ◽  
Mother Plants ◽  
Dark Green ◽  
Tlc Analysis ◽  
Two Peaks ◽  
Layer Chromatography

The dark green leaves of Sauropus androgynus (Euphorbiaceae) have various nutritive values and are commonly used for human consumption as food, medicine, and natural dye substance in South-east Asia. Shoot cultures of this plant were established by adding various concentrations of kinetin (Kn) and benzyl adenine (BA) using nodal explants. The best results were recorded when Kn 0.1 mg/L was used with BA 1 mg/L (BA1Kn0.1). Spectrophotometric analysis showed two peaks of green pigment in shoot cultures, A pigment (λmax = 663.6 - 663.8 nm, absorbance 0.1111) and B pigment (λmax= 611.3 - 613.9 nm, absorbance 0.0390). Thin Layer Chromatography (TLC) analysis showed two green spots (Rf Y = 0.31 and Rf Z = 0.25) of shoot cultures on medium supplemented with BA1Kn0.1 for 10 days. Pigment profiles of shoot culltures were similar to their corresponding mother plants. Random Amplified Polymorphic DNA (RAPD) was used as a preliminary technique to evaluate the genetic similarity of the shoot cultures and their corresponding mother plants. It showed four similar DNA banding patterns to their leaves, ranging from 271-765 bp.

scholarly journals TLC analysis of some phenolic compounds in kombucha beverage

2004 ◽  
pp. 199-205 ◽  
Author(s):  
Radomir Malbasa ◽  
Eva Loncar ◽  
Ljiljana Kolarov
Keyword(s):  
Phenolic Compounds ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Green Tea ◽  
Black Tea ◽  
Wide Range ◽  
Tea Fungus ◽  
Natural Phenolic Compounds ◽  
Tlc Analysis ◽  
Layer Chromatography

Black and green tea contains a wide range of natural phenolic compounds Flavanoids and their glycosides, catechins and the products of their condensation, and phenolic acids are the most important. Kombucha beverage is obtained by fermentation of tea fungus on black or green tea sweetened with sucrose. The aim of this paper was to investigate the composition of some phenolic compounds, catechin, epicatechin, quercetin, myricetin, gallic and tanic acid, and monitoring of their status during tea fungus fermentation. The method used for this study was thin layer chromatography with two different systems. The main phenolic compounds in the samples with green tea were catechin and epicatechin, and in the samples with black tea it was quercetin.

scholarly journals Ekstraksi, Karakterisasi dan Uji Aktivitas Antioksidan Astaxanthin dari Produk Fermentasi Udang (Cincalok)

2021 ◽  
Vol 24 (3) ◽  
pp. 311-322
Author(s):  
Mauludia Mauludia ◽  
Thamrin Usman ◽  
Winda Rahmalia ◽  
Dwi Imam Prayitno ◽  
Siti Nani Nurbaeti
Keyword(s):  
Antioxidant Activity ◽  
High Pressure ◽  
Liquid Chromatography ◽  
High Pressure Liquid Chromatography ◽  
Aquatic Organisms ◽  
Dpph Method ◽  
Wet Weight ◽  
Tlc Analysis ◽  
Layer Chromatography

Shrimp is one of the aquatic organisms that contain several active compounds, including astaxanthin. Cincalok is one of the fermented shrimp products containing astaxanthin. This study aims to determine the characteristics of astaxanthin extract from cincalok and its antioxidant activity. Extraction of astaxanthin from cincalok was carried out using the reflux method with acetone : cyclohexane (20:80 v/v) as a solvent. The identification and characterization of astaxanthin was carried out using thin-layer chromatography (TLC), UV-Vis spectrophotometry, and High-Pressure Liquid Chromatography (HPLC). Meanwhile, the antioxidant activity test was carried out using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method in one serial concentration (5; 15; 25 ppm). The results of TLC analysis showed that astaxanthin in cincalok extract has Rf value (0.32). The analysis using a UV-Vis spectrophotometer produced a spectrum with a maximum wavelength of 477 nm, which corresponds to the maximum wavelength of standard astaxanthin. The yield of astaxanthin extract from cincalok in this study was 1.47 mg/100 g wet weight. The chromatogram from the results of UHPLC analysis showed that the retention time of cincalok astaxanthin extract was 6.27 minutes with a purity of 18.03%. The antioxidant activity of cincalok astaxanthin extract was 568.32 ppm. Udang merupakan salah satu organisme air yang mengandung banyak senyawa aktif, termasuk astaxanthin. Cincalok merupakan salah satu produk hasil fermentasi udang yang mengandung astaxanthin. Penelitian ini bertujuan untuk mengetahui karakteristik ekstrak astaxanthin dari cincalok dan aktivitas antioksidannya. Ekstraksi astaxanthin dari cincalok menggunakan metode refluks dengan pelarut aseton:sikloheksan (20:80 v/v). Identifikasi dan karakterisasi astaxanthin dilakukan dengan menggunakan kromatografi lapis tipis (KLT), spektrofotometri UV-Vis, dan High Pressure Liquid Chromatography (HPLC). Sedangkan uji aktivitas antioksidan dilakukan menggunakan metode 1,1-difenil-2-pikrilhidrazil (DPPH) dengan memvariasikan konsentrasi larutan uji, yaitu 5; 15; 25 ppm. Hasil dari penelitian ini melaporkan astaxanthin pada ekstrak cincalok menunjukkan nilai Rf 0,32 pada kromatografi lapis tipis (KLT). Hasil analisis menggunakan spektrofotometer UV-Vis menghasilkan spektra dengan panjang gelombang maksimum 477 nm, yang sesuai dengan panjang gelombang maksimum astaxanthin standar. Randemen ekstrak astaxanthin dari cincalok pada penelitian ini adalah 1,47 mg/100 g berat basah. Kromatogram dari hasil analisis UHPLC menunjukkan waktu retensi ekstrak astaxanthin cincalok yaitu selama 6,27 menit dengan kemurnian sebesar 18,03%. Aktivitas antioksidan dari ekstrak astaxanthin cincalok diperoleh sebesar 568,32 ppm.  

Analysis of Dairy Products for Chlorinated Insecticide Residues by Thin Layer Chromatography

1966 ◽  
Vol 49 (4) ◽  
pp. 795-800
Author(s):  
William A Moats
Keyword(s):  
Aluminum Oxide ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Silica Gel ◽  
Color Development ◽  
Tlc Plates ◽  
One Step ◽  
Rf Values ◽  
Tlc Analysis ◽  
Layer Chromatography

Abstract Butterfat and milk samples were analyzed for chlorinated insecticides by thin layer chromatography (TLC) on aluminum oxide or silica gel plates containing a small amount of silver nitrate. The adsorbent was washed with distilled water before preparing the plates. A one-step cleanup on a partially inactivated Florisil column was performed prior to TLC analysis. For color development, the TLC plates were sprayed lightly with hydrogen peroxide to suppress possible interference from fat and then steamed before exposure to ultraviolet light to accelerate and intensify the color reaction. Rf values for a number of solvent systems on aluminum oxide and silica gel plates are given. With this procedure, 0.05 μg or less of insecticide can be detected in a 0.4 g butterfat sample or the extract from 10 ml milk.

scholarly journals Characterization and localization of a flagellar-specific membrane glycoprotein in Euglena.

1980 ◽  
Vol 86 (2) ◽  
pp. 424-435 ◽  
Author(s):  
A A Rogalski ◽  
G B Bouck
Keyword(s):  
Cell Surface ◽  
Minor Component ◽  
Membrane Vesicles ◽  
Surface Region ◽  
Membrane Glycoprotein ◽  
Low Concentrations ◽  
Tlc Analysis ◽  
A Minor ◽  
Layer Chromatography ◽  
Specific Membrane

Purified flagella from Euglena yield a unique high molecular weight glycoprotein when treated with low concentrations of nonionic detergents. This glycoprotein termed "xyloglycorien" cannot be extracted from other regions of the cell, although a minor component that coextracts with xyloglycorien does have a counterpart in deflagellated cell bodies. Xyloglycorien is tentatively identified with a flagellar surface fuzzy layer that appears in negatively stained membrane vesicles of untreated flagella but not in similar vesicles after Nonidet P-40 extraction. The localization of xyloglycorien is further confirmed to be membrane associated by reciprocal extraction experiments using 12.5 mM lithium diiodosalicylate (LIS), which does not appreciably extract xyloglycorien, visibly solubilize membranes, or remove the fuzzy layer. Rabbit antibodies directed against the two major flagellar glycoproteins (xyloglycorien and mastigonemes) to some extent cross react, which may in part be caused by the large percentage of xylose found by thin-layer chromatography (TLC) analysis to be characteristic of both antigens. However, adsorption of anti-xyloglycorien sera with intact mastigonemes produced antibodies responding only to xyloglycorien, and vice versa, indicating the nonidentity of the two antigens. Antibodies or fragments of these antibodies used in immunofluorescence assays demonstrated that xyloglycorien is confined to the flagellum and possibly the adjacent reservoir and gullet. Binding could not be detected on the cell surface. The sum of these experiments suggests that, in addition to mastigonemes, at least one major membrane glycoprotein in Euglena is restricted to the flagellar domain and is not inserted into the contiguous cell surface region.

scholarly journals PRODUCTION AND PARTIAL CHARACTERIZATION OF PIGMENTS PRODUCED BY KOCURIA SP BRI 36: INFLUENCE OF HEAVY METALS

2017 ◽  
Vol 9 (10) ◽  
pp. 137 ◽  
Author(s):  
Anuradha Mulik ◽  
Priyanka Kumbhar ◽  
Rama Bhadekar
Keyword(s):  
Heavy Metals ◽  
Antimicrobial Activity ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Pigment Production ◽  
Orange Pigment ◽  
Maximum Stability ◽  
Tlc Analysis ◽  
Physical And Chemical ◽  
Layer Chromatography

Objective: To study the production of pigments by Kocuria sp. BRI 36, their characteristics and influence of heavy metals on pigments.Methods: The effects of various physical and chemical parameters on pigments production by Kocuria sp. BRI 36 were examined. Pigments were extracted and partially characterised by Thin Layer Chromatography (TLC) and Fourier Transform Infrared Spectroscopy (FTIR). The effects of heavy metals such as Pb2+, Cd 2+, Ni2+ and Cr3+ were studied on pigment production. Antimicrobial activity and stability studies of crude pigment were also conducted.Results: Kocuria sp. BRI 36 isolated from cold oceanic region maximally produced red-orange pigment in presence of glucose (5% w/v) and protease peptone (0.2% w/v) at pH 7.5, 10±1 °C. Thin layer chromatography (TLC) analysis revealed the occurrence of three different compounds in the crude pigment belonging to carotenoid and xanthophyll group. Metals like Ni2+ and Cr3+ adversely affected pigment production while Pb2+and Cd2+enhanced the yield. The significant features of Kocuria sp. BRI 36 pigment are i) antimicrobial activity against Gram-positive and Gram-negative bacteria, ii) maximum stability at pH 7.5 and 10±1 °C and iii) ~38% color loss at 50±1 °C in 5 h.Conclusion: Our results suggest application potential of Kocuria sp. BRI 36 pigments in various biotechnological fields.

scholarly journals Characterization of an Alkaline GH49 Dextranase from Marine Bacterium Arthrobacter oxydans KQ11 and Its Application in the Preparation of Isomalto-Oligosaccharide

Marine Drugs ◽  
2019 ◽  
Vol 17 (8) ◽  
pp. 479 ◽  
Author(s):  
Hongfei Liu ◽  
Wei Ren ◽  
Mingsheng Ly ◽  
Haifeng Li ◽  
Shujun Wang
Keyword(s):  
Escherichia Coli ◽  
Marine Bacteria ◽  
Recombinant Expression ◽  
Bifidobacterium Longum ◽  
Lactobacillus Rhamnosus ◽  
Km Value ◽  
Tlc Analysis ◽  
Layer Chromatography ◽  
Arthrobacter Oxydans

A GH49 dextranase gene DexKQ was cloned from marine bacteria Arthrobacter oxydans KQ11. It was recombinantly expressed using an Escherichia coli system. Recombinant DexKQ dextranase of 66 kDa exhibited the highest catalytic activity at pH 9.0 and 55 °C. kcat/Km of recombinant DexKQ at the optimum condition reached 3.03 s−1 μM−1, which was six times that of commercial dextranase (0.5 s−1 μM−1). DexKQ possessed a Km value of 67.99 µM against dextran T70 substrate with 70 kDa molecular weight. Thin-layer chromatography (TLC) analysis showed that main hydrolysis end products were isomalto-oligosaccharide (IMO) including isomaltotetraose, isomaltopantose, and isomaltohexaose. When compared with glucose, IMO could significantly improve growth of Bifidobacterium longum and Lactobacillus rhamnosus and inhibit growth of Escherichia coli and Staphylococcus aureus. This is the first report of dextranase from marine bacteria concerning recombinant expression and application in isomalto-oligosaccharide preparation.

scholarly journals Steroidal saponin concentrations in Brachiaria decumbens and B. brizantha at different developmental stages

2008 ◽  
Vol 39 (1) ◽  
pp. 279-281 ◽  
Author(s):  
Karine Bonucielli Brum ◽  
Mitsue Haraguchi ◽  
Mirella Biasoli Garutti ◽  
Fernanda Nogarol Nóbrega ◽  
Beneval Rosa ◽  
...  
Keyword(s):  
Thin Layer ◽  
Developmental Stages ◽  
Ethanolic Extract ◽  
Spectrophotometric Analysis ◽  
Steroidal Saponin ◽  
Steroidal Saponins ◽  
Brachiaria Decumbens ◽  
Aerial Parts ◽  
Seed Fall ◽  
Layer Chromatography

Brachiaria species contain steroidal saponins and are involved in outbreaks of hepatogenous photosensitization. This research presents the levels of a steroidal saponin, protodioscin, in the seeds and aerial parts of B. brizantha and B. decumbens during different developmental stages (growth, bloom, fructification and seed fall). The butanolic fraction of the ethanolic extract of each stage was submitted to thin layer chromatography (TLC) and spectrophotometric analysis through the Ehrlich reagent in 515nm. The chromatograms in TLC of the butanolic fraction of B. brizantha and B. decumbens showed similar spots as the protodioscin standard. The estimated level of protodioscin isomers in B. brizantha and B. decumbens ranged from 0.5% to 2.1%, having the highest level at the end of their developmental stages during seed falling comparison with the previous one. Protodioscin was not detected in the seeds. Outbreaks of Brachiaria spp. poisoning in central Western Brazil are frequently observed in pastures that had been more than 30 days without animals grazing, and also during the growing or blooming stage of the pastures. Other saponin determinations in toxic and non toxic pastures are necessary to determine the saponin concentrations that cause intoxication.

scholarly journals Sulfation of Nod Factors via nodHPQ Is nodD Independent in Rhizobium tropici CIAT899

1998 ◽  
Vol 11 (10) ◽  
pp. 979-987 ◽  
Author(s):  
Jorge Luis Folch-Mallol ◽  
Hamid Manyani ◽  
Silvia Marroquí ◽  
Carolina Sousa ◽  
Carmen Vargas ◽  
...  
Keyword(s):  
Phaseolus Vulgaris ◽  
Leucaena Leucocephala ◽  
Regulatory Region ◽  
Constitutive Expression ◽  
Nod Factors ◽  
Rhizobium Tropici ◽  
Independent Manner ◽  
Macroptilium Atropurpureum ◽  
Tlc Analysis ◽  
Layer Chromatography

A cosmid from the Rhizobium tropici CIAT899 symbiotic plasmid, containing most of the nodulation genes described in this strain, has been isolated. Although this cosmid does not carry a nodD gene, it confers ability to heterologous Rhizobium spp. to nodulate R. tropici hosts (Phaseolus vulgaris, Macroptilium atropurpureum, and Leucaena leucocephala). The observed phenotype is due to constitutive expression of the nodABCSUIJ operon, which has lost its regulatory region and is expressed from a promoter present in the cloning vector. Thin-layer chromatography (TLC) analysis of the Nod factors produced by this construction shows that it is still capable of synthesizing sulfated compounds, suggesting that the nodHPQ genes are organized as an operon that is transcribed in a nodD-independent manner and is not regulated by flavonoids. Se ha aislado un cósmido del plásmido simbiótico de Rhizobium tropici CIAT899 que contiene la mayoría de los genes de nodulación descrito para esta estirpe, menos el gen regulador nodD. La introducción de este cósmido en una estirpe curada del plásmido simbiótico de R. tropici CIAT899 permite la nodulación en las plantas ensayadas (Phaseolus vulgaris, Macroptilium atropurpureum, y Leucaena leucocephala). El fenotipo observado se debe a la expresión constitutiva del operón nodABCSUIJ bajo el promotor del gen de resistencia a la kanamicina, que lleva el vector donde se ha clonado el fragmento de ADN. Análisis por cromatografia de capa fina demuestran que esta construcción es capaz de sulfatar el extremo reductor del factor Nod. Estas evidencias sugieren que los genes nodHPQ constituyen un operón, y que su expresión es independiente del gen regulador nodD.

Toxicity tests in eight species of Chrysochromulina (Haptophyta)

1997 ◽  
Vol 75 (1) ◽  
pp. 129-136 ◽  
Author(s):  
Susanne Simonsen ◽  
Øjvind Moestrup
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Artemia Salina ◽  
Toxicity Tests ◽  
Haemolytic Activity ◽  
Prymnesium Parvum ◽  
Unialgal Culture ◽  
Tlc Analysis ◽  
Layer Chromatography ◽  
Chloroform Methanol

Blooms of the marine flagellate Chrysochromulina have resulted in mortality of marine organisms in Scandinavian waters, including fish in aquaculture. Eight species of Chrysochromulina, namely C. apheles, C. brevifilum, C. ericina, C. hirta, C. leadbeateri, C. parva, C. polylepis, and C. simplex, isolated into unialgal culture, were examined for haemolytic activity and toxicity to the brine shrimp, Artemia salina. Haemolytic fractions were obtained from all species, but only C. polylepis cells were toxic to Artemia. Thin-layer chromatography (TLC) analysis in chloroform –methanol–water (75:25:4) and in chloroform–methanol (9:1) yielded up to six haemolytic spots. Except for one spot, these all occurred in extracts of the species examined, including Isochrysis sp., which was used as a control, C. polylepis, and the well-known fish killer Prymnesium parvum. The single unique haemolytic spot (Rf values 0.45 and 0.16 in solvents I and II, respectively) occurred in the extract from C. polylepis. When isolated by TLC, the contents of the single spot were toxic to Artemia. Key words: Chrysochromulina, toxicity, haemolytic, Artemia, thin-layer chromatography (TLC).

Pectic enzymes produced by Pseudomonas fluorescens, an organism associated with “pink eye” disease of potato tubers

1972 ◽  
Vol 50 (12) ◽  
pp. 2479-2488 ◽  
Author(s):  
S. S. Hagar ◽  
G. A. McIntyre
Keyword(s):  
Pseudomonas Fluorescens ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Pectin Methylesterase ◽  
Viscosity Reduction ◽  
Cellulose Acetate Electrophoresis ◽  
Pectic Enzymes ◽  
Ph Range ◽  
Two Peaks ◽  
Layer Chromatography

No pectin methylesterase (PME) activity was observed in crude or dialyzed extracts from macerated potato tuber tissue inoculated with Pseudomonas fluorescens; however, pectic lyase (syn. polygalacturonic acid transeliminase, PATE) activity was observed. Two PATE enzymes (peaks 1 and 2) were eluted from a pH 9.4 DEAE-cellulose column using a gradient of pH 7.6 Tris-HCl buffer (0.01–0.1 M). Enzyme in peak 1 was about 6 times more active than enzyme in peak 2 based on reducing group assays, and 10 times more active in viscosity reduction of 1% Na-polypectate (NaPP) at pH 8.5. No increase in absorbancy was observed at 515 nm of clarified reaction mixtures, indicating that saturated oligouronides did not accumulate. Other properties of the two peaks: optimum pH range was 8.5–9.5, substrate preference was NaPP vs. pectin, addition of Ca2+ (0.001 M) enhanced activity while EDTA (0.001 M) decreased activity to [Formula: see text], cellulose acetate electrophoresis revealed one band of protein per peak, and heat of inactivation was 51–60C. Thin-layer chromatography of hydrolytic products from NaPP revealed unsaturated uronides and pectic fragments after 2 h hydrolysis; after 96 h hydrolysis only unsaturated uronides were observed. Molecular weight estimations by Sephadex G-200 gel filtration were about 18 000 (peak 1) and 22 500 (peak 2). Enzyme in either peak macerated 400-μ sections of potato tissue.

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