Adsorption Column Chromatography Research Articles

Chemical constituents from leaves of Jatropha curcas

ObjectiveTo investigate the chemical constituents from the leaves of Jatropha curcas and evaluate their inhibition on lipopolysaccharide (LPS)-activated BV-2 microglia cells. MethodsThe n-BuOH extract of the leaves of J. curcas was isolated by macroporous adsorption resin, silica gel, ODS, column chromatography and semi-preparative HPLC. The structures of the compounds were identified by MS, NMR, ECD, and other spectroscopic methods. In addition, anti-neuroinflammatory effects of isolated compounds were evaluated by measuring the production of nitric oxide (NO) in over-activated BV-2 cells. ResultsSeventeen compounds, including (7R,8S)-crataegifin A-4-O-β-D-glucopyranoside (1), (8R,8′R)-arctigenin (2), arctigenin-4′-O-β-D-glucopyranoside (3), (-)-syringaresinol (4), syringaresinol-4′-O-β-D-glucopyranoside (5), (-)-pinoresinol (6), pinoresinol-4′-O-β-D-glucopyranoside (7), buddlenol D (8), (2R,3R)-dihydroquercetin (9), (2S,3S)-epicatechin (10), (2R,3S)-catechin (11), isovitexin (12), naringenin-7-O-β-D-glucopyranoside (13), chamaejasmin (14), neochamaejasmin B (15), isoneochamaejasmin A (16), and tomentin-5-O-β-D-glucopyranoside (17) were isolated and identified. Compounds 2, 4 and 8 significantly inhibited the release of NO in BV-2 microglia activated by LPS, with IC50 values of 18.34, 29.33 and 26.30 μmol/L, respectively. ConclusionCompound 1 is a novel compound, and compounds 2, 3, 8, 14–17 are isolated from Jatropha genus for the first time. In addition, the lignans significantly inhibited NO release and the inhibitory activity was decreased after glycosylation.

INVESTIGATION OF THE COMPOSITION AND PROPERTIES OFAKSHABULAK OIL

Annotation. With the help of modern methods of physico-chemical analysis, the physicochemical properties, origin and fractional compositionof the Akshabulak oil field were studied. Using adsorption column chromatography, the oil was purified from asphaltenes and resins contained in it. The full composition of oil was determined by chromatography-mass spectrometry and the content of compounds in it was calculated by internal normalization. The NIST-08 database was used as a library of Mass spectra. The origin of the oil was discovered, studied by the ratio of 2,6,10,14-tetramethylpentadecane to 2,6,10,14-tetramethylhexadecane on the obtained chromatogram. The proven oil was formed at the seabed off the coast by origin. The fractional composition of oil and the products obtained from it was studied by the method of simulated distillation. The heat of combustion of the products obtained and the octane number of gasoline obtained from oil were determined. The results obtained as a result of the study can be used in oil refining.

The composition of flavonoids in the inflorescences of spreading marigold (<i>Tagetes Patula</i> L.)

Aim to isolate and identify individual compounds of flavonoid nature from the inflorescences of spreading marigold (T. patula L.).
 Material and methods. The research material was the inflorescences of T. patula L. of the variety "Mandarin", collected in August 2018 in the Botanical Garden of the Samara University during the mass flowering and fruiting period. We isolated individual substances by adsorption column chromatography on silica gel L 40/100 (Czech Republic) with subsequent recrystallization. For the identification of the isolated compounds, we used the UV, 1H-NMR, 13C-NMR spectroscopy and mass spectrometry data.
 Results. The dominant and diagnostically significant flavonoids of the inflorescences of the spreading marigold, patulitrin and patuletin, were isolated and identified. For the first time, the 6-methoxykaempferol and 7-O--D-glucopyranoside of 6-methoxykaempferol were isolated from this type of plant raw material. Quercetin was obtained in minor quantities. The results of the study can be used for developing the methods of identification and standardization of the inflorescences of Tagetes patula L.

Nutritional content and the activities of l-Dopa (L-3,4-dihydoxyphenyalanine) from Mucuna pruriens L. DC seeds of Central Java accession

Mucuna pruriens (L.) DC (Velvet beans) is a tropical bean containing large amounts of l-Dopa (L-3,4-dihydroxyphenylalanine) and is the most important natural remedy for Parkinson’s disease. This plant has been used in Ayurveda medicine because it has a pharmacological activity, one of which is an antibacterial activity obtained from its seed extract. M. pruriens seeds fulfill the basic nutritional requirements, so they can be consumed. However, their utilization as food is limited due to the presence of some anti-nutritional factors. The objective of the research was to determine the nutritional content of seeds of M. pruriens Central Java accession by proximate analysis, isolation, and characterization of l-Dopa, determine the toxicity value ( LC50), against Artemia salina L. larvae by BSLT (brine shrimp lethality test), and determine its antibacterial activity against Streptococcus mutans ATCC 35668, and Escherichia coli ATCC 25,922 by agar disk diffusion method. The isolate of l-Dopa was prepared by maceration using methanol 80%, followed by extraction using water, then, the isolated product was analyzed by analytical TLC to determine the suitable mobile phase. The sample was then run on preparative TLC. The spot that corresponds to the l-Dopa standard was collected and further purified using adsorption column chromatography. Then, the isolate was characterized by HPLC and FTIR. The antioxidant activity was determined by the DPPH radical method. Finally, anti-Parkinson's activity was determined to the rotenone-induced behavior of white rats (Rattus norvegicus) by the classic labyrinth test method. The result showed that the strongest antibacterial activity against S. mutans ATCC 35,668 was shown by crude extract. Based on the HPLC analysis, the isolated l-Dopa showed a retention time of 5.345 min that is relatively similar to the l-Dopa standard and had a high purity. l-Dopa content in M. pruriens seed of Central Java accession was 0.49 mg/g dry weight. The l-Dopa isolate from M. pruriens Central Java accession has antioxidant activity with IC50 of 8.92 ± 0.03 ppm (highly active) and could significantly reduce the non-motoric Parkinsonism symptoms (p < 0.001) compared to the negative control.

Isolation and Characterization of Gibberellin (GA-3) from Banana Blossom of Musa balbisiana Colla and Effect of The Isolate Addition on The Growth of Soybean (Glycine max) Plant

There is a type of banana that is unique because the flesh is filled with black seeds locally known as “pisang batu” (Musa balbisiana Colla). The objective of research was directed to determine the content of GA3 in banana blossom of M. balbisiana Colla and its effect on the growth of soybean (Glycine max) plant in vegetative and generative phase. GA3 was prepared by maceration followed by extraction using ethyl acetate and further purified using adsorption column chromatography and preparative TLC. The isolated product was then characterized using FTIR and HPLC. Sprouts of soybeans with uniform height were planted on polybags containing a mixture of planting media (soil: husk: manure = 3:2:1). The sprout was divided into five groups. The first group is the control only spraying with water, while others groups were treated by spraying GA3 isolate, crude extract, commercial Biostimulant and 1% NPK fertilizer. The soybean was determined for chlorophyll content of the leaf, proximate content and α-amylase activity. The results of HPLC and FTIR on gibberelline isolates showed a retention time and wave number which is relatively similar to the GA3 standard. GA3 levels in banana blossom of M. balbisiana Colla were 2.55 mg/g dry weight. The results of research showed that the GA3 isolate, crude extract and commercial biostimulant have significant effects on the growth of soybean plant both in the vegetative and generative phases for each parameter determined, proximate content, chlorophyll content and α-amylase activity compared to the control and NPK fertilizer.

Purification of Artepillin C (ARC) with Anticancer Activities from Supercritical CO2-Ethanol Extracts of Brazilian Green Propolis by Using Adsorption Column Chromatography

Phenolic compounds with varying phenolic structures are present in both terrestrial and marine organisms. Artepillin C (ARC) is a prenylated phenolic compound produced specifically in Brazilian green propolis. Earlier studies have demonstrated the potential of ARC as a dietary supplement for cancer prevention due to its ability to suppress tumor growth and metastasis. Here we report the development of an effective adsorption chromatography combined with supercritical CO2 extraction for the massive purification of ARC. A mixture of supercritical CO2 and ethanol extract of Brazilian green propolis was passed through a stainless chromatographic column packed with protruding metals at 4,000 psi and 50 °C to remove wax from the column’s base. ARC mixed with flavonoids collected from the top of the column was applied to another stainless column filled with silica gel under similar working conditions. After flavonoids were removed, strongly adsorbed ARC at the top portion of the column was collected by ethanol elution. The purity of ARC isolated using this method was between 90% and 95%, as indicated by HPLC chromatograms. Purified ARC strongly inhibited cancer cell division based on its cytotoxic effects on HCT116 human colon cancer cells, resulting in a significantly increased distribution of cells in the S phase. ARC also blocks the motility of HCT116 cells, indicating its ability to suppress tumor metastasis. The developed chromatographic purification protocol can be used to isolate prenylated phenolic compounds from marine organisms.

Characterization of Molecular Species and Anti-Inflammatory Activity of Purified Phospholipids from Antarctic Krill Oil.

The phospholipids (PLs) from Antarctic krill oil were purified (>97.2%) using adsorption column chromatography. Forty-nine PL molecular species were characterized by ultrahigh-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Most of molecular species contained eicosapentaenoic acid (EPA, 20:5), docosahexaenoic acid (DHA, 22:6), docosapentaenoic acid (DPA, 22:5), and arachidonic acid (AA, 20:4). Notably, a special species PC (20:5/22:6) (1298.17 nmol/g) and many ether PLs were detected. The Antarctic krill PL liposome (IC50 = 0.108 mg/mL) showed better anti-inflammatory activity than crude Antarctic krill oil (IC50 = 0.446 mg/mL). It could block NF-κB signaling pathway via suppression of IκB-α degradation and p65 activation and dose-dependently reduce the cellular content of inflammatory mediators including nitric oxide (NO), reactive oxygen species (ROS), and inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In addition, it can suppress carrageenan-induced mouse paw swelling. Results from the present study could provide a reference for better evaluation of nutritional and medicinal values of Antarctic krill oil.

Chromatographic recovery and purification of natural phytochemicals from underappreciated willow bark water extracts

Willow (Salicaceae) bark contains a variety of antioxidant phytochemicals depending mainly on the hybrid. In this study, hot water extraction of willow bark was coupled with a sequential preparative scale chromatography to purify the crude extracts to a well-separated and highly concentrated phytochemical fraction. The pre-equilibrated hydrophilic weak cation exchange resins (with acrylic matrix in Na+ ions form) achieved a good separation of phytochemicals from monosaccharide fractions. Secondly, the individual phytochemical can be purified based on their size and hydrophobicity differences, using gel permeation chromatography and adsorption column chromatography. Surprisingly, the G-types of sephadex demonstrated an efficient separation of not only monosaccharides but a complete separation between (+)-catechin and triandrin. Picein and salicin can be further separated according to their hydrophobicity differences. The purified compounds can be quickly quantified using high performance liquid chromatography, gas chromatography and high-performance anion-exchange chromatography. The purified picein and triandrin were characterized by one- and two-dimensional nuclear magnetic resonance spectroscopy techniques. Additionally, the authentic compounds were applied to confirm the identification, quantification and purity. The technique described here might be a fast, economical, and highly efficient way in the large-scale isolation and purification of the phytochemical compounds from willow bark.

Investigation of photochemical conversion processes in aromatic hydrocarbons of Balakhani oil

In this paper, Balakhani oil, as the case study, was separated into 4 groups of aromatics by adsorption column chromatography. With the aid of GC-MS and UV/vis spectrometers, it was proved that the composition of the separated groups is mainly composed of mono-, di-, tri- and a trace amount of tetracyclic aromatics. The conversion process of separated aromatics under photo-irradiation was investigated by IR, UV, 1H NMR and GC-MS spectroscopies. It was found that the components of aromatic groups undergo photo-conversion processes at different rates depending on their composition. In the 3rd and 4th groups, the mentioned process occurs slowly, so their use as petroleum luminophores (PLs) may be recommended. It was found that the PLs based on these groups have variable luminescence properties, and the variation range of their spectral-luminescence properties was also determined. Finally, the role of singlet (1O2) and triplet (3O2) oxygens in reversible and irreversible photooxidation reactions with aromatics was investigated as well as, the significance of molecular and chain-radical reactions in this process was also determined.

ISOLATION OF MIRICETINE-CONTAINING FRACTIONS FROM LINOSYRIS VILLOSA PLANT AND THEIR APPLICATION AS ANTIANEMIC AGENT

With the development of chemistry, natural substances were actively supplanted from human life by chemical products. However, despite the enormous progress in this area, in recent decades, the problems of serious side effects of synthetic chemicals on the environment have become more and more pronounced. Methods of two-dimensional and one-dimensional paper chromatography, column adsorption chromatography, gas chromatography, and IR spectroscopy were used. The purpose of the article is to study the chemical composition of the Linosyris villosa plant by physicochemical methods of analysis. Creation of dosage forms based on it. The problem of the article is the search and creation of drugs based on the flora of Northern Kazakhstan. The scientific novelty lies in the fact that the chemical composition and biological activity of the Linosyris villosa plant, growing in the territory of Northern Kazakhstan, is being studied for the first time. According to the results of the analysis, in 13% of patients the treatment effect is insignificant, and in 87% of patients there was an improvement in blood values (increase in hemoglobin, an increase in the number of red blood cells, almost all patients returned color index of blood), the number of white blood cells increased to normal, and the WBC differential improved. Therefore, the drug “Vitin”, an aqueous-alcoholic extract of the Linosyris villosa, is an effective drug for the treatment of patients with anemia of varying severity. The practical significance of the work is defined as follows: biologically active compounds from the Linosyris villosa plant were obtained for the purpose of studying and applying them in medicine and agriculture.

The study of the composition of phenolic compounds of the populus nigra leaves

This paper presents the findings of the study of the composition of phenolic compounds in black poplar leaves (Populus nigra L.). For the first time flavonoid compounds such as сalendoflavobioside (quercetin-3-O-neohesperidoside), rutin (quercetin-3-O-rutinoside), and ferulic acid (phenylpropanoid) have been extracted from the leaves of the black poplar by the methods of extraction and the following adsorption column chromatography on silica gel and on polyamide sorbent. The obtained substances were characterized with the use of UV-, 1H-NMR-, 13C-NMR-spectroscopy and mass spectrometry and also by means of the results of acid hydrolysis. The flavonoids are the dominant phenolic components of the raw material of this plant.

In vitro and in silico antioxidant and antiproliferative activity of rhizospheric fungus Talaromyces purpureogenus isolate-ABRF2

The present study evaluated the potential biological activities of rhizospheric fungi isolated from the Achanakmar Biosphere Reserve, India. Fungus, Talaromyces purpureogenus isolate-ABRF2 from the soil of the Achanakmar biosphere was characterized by using morphological, biochemical and molecular techniques. Fungus was screened for the production of secondary metabolites using a specific medium. The metabolites were extracted using a suitable solvent and each fraction was subsequently evaluated for their antioxidant, antimicrobial, antiproliferative and anti-aging properties. The ethanolic extract depicted the highest antioxidant activity with 83%, 79%, 80% and 74% as assessed by ferric reducing power, 2,2-diphenyl 1-picrylhydrazyl, 2,2′-azino-bis3-ethylbenzthiazoline-6-sulfonic and phosphomolybdenum assays, respectively. Similarly, ethanolic extracts depicted marked antimicrobial activity as compared with standard antibiotics and antifungal agents as well as demonstrated significant antiproliferative property against a panel of mammalian cancer cell lines. Furthermore, different fractions of the purified ethanolic extract obtained using adsorption column chromatography were evaluated for antiproliferative property and identification of an active metabolite in the purified fraction using gas chromatography–mass spectroscopy and nuclear magnetic resonance techniques yielded 3-methyl-4-oxo-pentanoic acid. Thus, the present study suggests that the active metabolite 3-methyl-4-oxo-pentanoic acid extracted from Talaromyces purpureogenus isolate-ABRF2 has a potential antiproliferative, anti-aging, and antimicrobial therapeutic properties that will be further evaluated using in vivo studies in future.

Chromatographic study of the component composition and antioxidant activity of polyphenol complex in biological raw materials

Recently, the attention of many researchers has been attracted by raw materials that contain flavonoids. The purpose of the research is to summarize the results of studies of the composition of flavonoids of a non-fat sea buckthorn meal, which is a secondary raw material for the production of sea buckthorn oil. In this case, the sum of the five identified flavonoid compounds was isolated by extraction and subsequent adsorption column chromatography. At this stage, the actual problem of identifying and determining individual isolated flavonoids was solved using chromatographic (TLC, HPLC) and spectroscopic methods in comparison with reliably known samples. During the study, the flavonoids were identified: isorhamnetin, kaempferol, myritin, quercetin and its glycoside rutin, quercetin and isorhamnetin predominate. Ultimately, the effective parameters of the method for the improvement of the competitiveness of dietary supplements are established.

Progress in separation and purification of nattokinase and its enzyme activity determination

In this paper, the separation and purification of nattokinase have been reviewed by primarily focusing on solvent precipitation, adsorption column chromatography, magnetic microspheres, expanded bed, reverse micelle, and three-phase separation methods. The different methods for determination of the enzyme activity have been discussed and compared. Finally, the feasibility that the nucleic acid-based identification techniques can be used for nattokinase purification and enzyme activity assay has been proposed.

Occurrence of organochlorine and organophosphorus pesticide residues in poultry feeds, raw and cooked eggs from selected farms in Ilala and Kibaha Districts, Tanzania

This study assessed the levels of seventeen pesticides and metabolites residues in chicken feeds and raw eggs as well as the effects of processing methods on the levels in eggs in samples obtained from six poultry farms in Ilala and Kibaha districts, Tanzania. Extraction was performed by solid dispersion method and the extracts were cleaned-up by adsorption column chromatography. The analytes were determined by Gas Chromatography–Mass Spectrometry (GC–MS). The highest mean concentrations of the contaminants in feeds and eggs were as follows, respectively: aldrin 0.62 and 2 mg/kg, dieldrin 0.71 and 1.3 mg/kg, total DDT 6.68 and 8.14 mg/kg, total endosulfan 3.53 and 3.74 mg/kg, total HCHs 0.91 and 1.21 mg/kg, chlorpyrifos 12.2 and 0.59 mg/kg, fenitrothion 4.9 and 0.64 mg/kg and pirimiphos methyl 22.11 and 1.6 mg/kg. Chicken feeds were found to be the most contaminated followed by raw eggs and finally cooked eggs. Most of the concentrations were above the maximum residue limits (MRLs) indicating risks and concerns. Proper selection and preparation of poultry feeds could reduce the levels in the feeds and eggs. Keywords : Pesticides, Chicken, Feed, Eggs, Tanzania

Residues of pesticides and metabolites in chicken kidney, liver and muscle samples from poultry farms in Dar es Salaam and Pwani, Tanzania

The concentrations of organochlorine and organophosphorus pesticides and metabolites were investigated in chicken kidney, liver and muscle samples obtained from chickens collected from four poultry farms in Dar es Salaam and Pwani regions in Tanzania. The samples were extracted by solid dispersion using cyclohexane:ethyl acetate. The extracts were cleaned by adsorption column chromatography. The analytes were determined using gas chromatography-mass spectrometry (GC–MS). The concentrations of total DDT, total endosulfan and total HCHs in the samples ranged from 0.71 to 26, 0.3 to 7.9 and 0.02–10.4 mg/kg lipid weight (lw), respectively. The highest concentrations of aldrin, dieldrin, chlorpyrifos, fenitrothion and pirimiphos methyl were 5.5, 4.8, 9.7, 5.6 and 7.8 mg/kg lw, respectively. The highest concentrations of the contaminants were found in the kidney and liver samples. The sites in Dar es Salaam showed the highest concentrations of the compounds. Most of the concentrations were above the maximum residue limits (MRLs) indicating risks and concerns for livestock and public health.

Date (Phoenix dactylifera L.) palm stones as a potential new feedstock for liquid bio-fuels production

In the present investigation, Date (Phoenix dactylifera L.) stones were utilized in the production of different types of biodiesel in addition to bio-oil. The oil was extracted from the date stones with a yield of 10.50wt% and is used in the production of fatty acid methyl, ethyl and mixed methyl/ethyl esters via KOH- catalyzed transesterification process with methanol, ethanol and mixed methanol and ethanol, respectively. Properties of the obtained biodiesels, such as the density, kinematic viscosity, flash point, acid value, cloud and pour points, total and free glycerin contents and the refractive index were evaluated and found comparable to those of ASTM D 6751 biodiesel. The FTIR spectroscopy confirmed conversion of date stone oil to biodiesel. The date stones were also tested as new non-edible feedstock for bio-oil production by pyrolysis process in a semi batch reactor. The influence of the pyrolysis temperature, pyrolysis time and precursor particles size on the bio- oil yield was investigated. Maximum bio- oil yield (52.67%±1.50) was obtained at 500°C pyrolysis temperature with a pyrolysis time of 60min and feed particle size of 0.25mm. The ultimate analysis, FTIR spectroscopy and adsorption column chromatography were determined to characterize the chemical composition of the obtained bio- oil. Furthermore, several important fuel properties, like the density, kinematic viscosity, flash point, pour point and the acidity index of date stones bio- oil were also determined following ASTM standard test methods and found comparable to those documented for other bio- oils in literature. The chemical composition of the bio- oil produced through pyrolysis of date stones showed the potential of date stones as an important source of alternative fuel and chemicals as well.

Preliminary study of metalloporphyrins in some oil shales, red sea, Egypt

Occurence and distribution of metalloporphyrins were studied in asphaltene and maltene fractions of some Egyptian oil shales from the main producing mines (Abu-Shegeili, El-Beida, El-Nakheil and Abu-Tundub I, II) in red sea area (Fig. 1).Metalloporphyrins were extracted using adsorption column chromatography. The presence of Ni, Fe and Vo-porphyrins was monitored in each fraction by means of UV–Visible spectrometry, the result indicated that metalloporphyrins could be a mixture of Etio and DPEP types. Each fraction using UV–Visible spectrometry cannot differentiate between Ni and Fe porphyrins, so the extracted metalloporphyrins were subjected to purification and separation from each other by using thin layer chromatography (TLC).HPLC was used for fingerprinting Vo, Ni and Fe-porphyrins, it was found that the different porphyrins exhibited wide different chromatograms. HPLC technique could be used as a successful tool to characterize Fe and Ni-porphyrins.

Conversion of De-oiled Castor Seed Cake into Bio-oil and Carbon Adsorbents

Castor seed cake is the byproduct of castor seeds with cold pressing. This bio-waste was used as a raw material for the production of bio-oil and activated carbons through pyrolysis. De-oiled castor seed cake was pyrolyzed at various temperatures for various durations to find out the optimal temperature and duration for the pyrolysis. The highest bio-oil yield (63% w/w) was obtained at 400°C for 60 min. The ultimate analysis and the fuel properties for the produced bio-oil were determined. Fourier transform infra-red spectroscopy and fractionation using column adsorption chromatography for bio-oil were also conducted. The results showed that the properties of bio-oil were similar to those reported in the literature. The pyrolysis char was converted into activated carbons of micro-porous structure.

Sulfonated tea waste: A low-cost adsorbent for purification of biodiesel

ABSTRACTIn the present work, the crude biodiesel produced from spent fish frying oil through alkaline catalyzed transesterification was purified using a low-cost adsorbent viz. sulfonated tea waste. After separating the glycerol, the crude biodiesel was purified using the suggested adsorbent. Various methods of purification using the said adsorbent were applied such as purification using adsorption column chromatography and shaking methods. The results showed that purification using adsorption column chromatography exhibited the bst result. Properties of the purified fuels were determined and found conformed to those specified by the ASTM standards. For the sake of comparison, purification using zeolite and water washing method was also investigated. The result indicated that the suggested adsorbent was more successful on purification of the crude biodiesel compared to other methods.