Thin Layer Chromatography Test Research Articles

Qualitative study in vitro fruit and epicarpes Citrus Limetta Risso, Citrus Limon Burm and Citrus aurantiifolia ( Christm. ) Swingle Gharb of Morocco

Several epidemiological studies have provided good evidence of the inverse relationship associated with the consumption of citrus fruits and chronic diseases. After many years of research in the role of phytomicronutrients such as phenolic compounds, carotenoids and alkaloids, they are now widely studied and appreciated to use in the control of these degenerative diseases. These positive influences on human health have significantly increased production and consumption of citrus in recent decades. The lemon is a citrus fruit of the Rutaceae family which is rich in bioactive compounds. These could play a much more important protective role in the perspective of preventive nutrition. Indeed, these molecules have not revealed all their secrets until today. This is qualitative study based on a phytochemical screening and analytical technique for separating the thin layer chromatography (TLC ) extracts of fruits, epicarpes limetta Citrus , Citrus limon and Citrus aurantiifolia to highlight the existence of secondary metabolites that characterize them. The results of this study allowed us to reveal the existence of flavonoids, tannins, coumarins, alkaloids, carotenoids, quinones, sterols, terpenes, reducing compounds and iridoides. TLC tests confirm the presence of these secondary metabolites. Several epidemiological studies have provided good evidence of the inverse relationship associated with the consumption of citrus fruits and chronic diseases. After many years of research in the role of phytomicronutrients such as phenolic compounds, carotenoids and alkaloids, they are now widely studied and appreciated to use in the control of these degenerative diseases. These positive influences on human health have significantly increased production and consumption of citrus in recent decades. The lemon is a citrus fruit of the Rutaceae family which is rich in bioactive compounds. These could play a much more important protective role in the perspective of preventive nutrition. Indeed, these molecules have not revealed all their secrets until today. This is qualitative study based on a phytochemical screening and analytical technique for separating the thin layer chromatography (TLC ) extracts of fruits, epicarpes limetta Citrus , Citrus limon and Citrus aurantiifolia to highlight the existence of secondary metabolites that characterize them. The results of this study allowed us to reveal the existence of flavonoids, tannins, coumarins, alkaloids, carotenoids, quinones, sterols, terpenes, reducing compounds and iridoides. TLC tests confirm the presence of these secondary metabolites.

English

The species Staphylococcus aureus and Escherichia coli have shown increased antimicrobial resistance, which can be overcome with plant extracts, in view of their molecular complexity, for example extracts of Anacardium occidentale Linn (cashew). The objective was to evaluate the in vitro antimicrobial activity of cashew stem bark extract. Samples were collected in Santa Cruz / RN / Brazil, and after dried and crushed, the extract was prepared by maceration with ethanol: water solution (70:30 v / v) at a ratio (1:10 w / v ) for 7 days. The filtrate was reduced by rotary evaporator and the resulting frozen. Extracts were characterized by Thin Layer Chromatography (TLC) and tested for antimicrobial activity in various concentrations (200 to 6.25 mg / ml) by diffusion disk. TLC testing indicated that the plant extract has phenolics, tannins mainly responsible for the pharmacological properties. The minimum inhibitory concentration was determined at 12.5 mg / ml only for S. aureus. Key words: Cashew, antimicrobial activity, S. aureus.

TLC Screening and Evaluation of Antioxidant, Antibacterial Activity of Onopordon Macrocephalum by Bioautography Method

TLC screening and Evaluation of antioxidant, antibacterial activity of Onopordon macrocephalum by bioautography methodOnopordon known as scotch thistle is a medicinal plant of the Asteraceae family that is widely distributed in Europe and Asia, also it has many valuable medicinal properties as hypotensive and antitumor. Thin layer chromatography (TLC) is wildly used in natural product extract analysis as a finger print which can be used for identification and quality control of medicinal preparations. Performing Thin-layer chromatography TLC tests on the plant show the presence of flavonoids, coumarins and bitter principles. Planar chromatographic analysis hyphenated with the biological detection method is known as bioautography which is an effective and inexpensive technique for the phytochemical analysis of plant extracts. Thin-layer chromatography-direct bioautography links separation on the adsorbent layer with biological tests performed directly on it in order to identify antimicrobial and antioxidant activities. Bioautography revealed that Staphylococcus aureus was inhibited by most of the flavonoids separated on the TLC plates. Similarly, growth of the Bacillus cereus was also inhibited by one flavonoid band on TLC. Antioxidant bioautography shows the strong antioxidant activity of one flavonoid band, whereas no activity detected of bitter principles. Bioautography showed that the antimicrobial and antioxidant activity was probably due to flavonoids.

Streptomyces: isolation, optimization of culture conditions and extraction of secondary metabolites

The objectives of this study were to isolate and identify Streptomyces from soil sediments as well as to optimize cultural growth conditions for maximum antibacterial productivity. A total of fifty soil sediments were collected from Red Sea, Sudan. The soil sediments were pretreated and cultivated on agar medium. Promising Streptomyces spp. were isolated by agar overlay method using indicator organisms. Optimization of chemical and physical culture conditions was carried out. The later was judged by assessment of antibacterial activity. Ethyl acetate was used to extract the secondary metabolite compounds. The separation of the active ingredients was performed using both thin layer chromatography (TLC) and gas chromatography-mass spectrometer (GC-MS). The results revealed nine strains of Streptomyces. Of them two (PS1 and PS28) isolates exhibited high activity against pathogenic bacteria. The optimum growth conditions were pH 7.5, temperature at 30°C, soyabean concentration 2.5 g/l, incubation period in 7 days, MgSO4.7H2O conc. 1g/l and K2HPO4 conc. 2.5g/l. TLC test showed three and two fragments from metabolites of PS1 and PS28 respectively, while the GC-MS analysis revealed eight and eleven compounds with antibacterial activity of PS1 and PS28 respectively. It is concluded that marine is promising source of secondary metabolites.Khattab et al., International Current Pharmaceutical Journal, February 2016, 5(3): 27-32

Synthesis of Polymerizable Ionic Liquid Monomer and Its Characterization

The polymerizable monomer of 1-(6-hydroxyhexyl)-3-methylimiadazolium bromide ionic liquid (IL) was synthesized. The thin layer chromatography tests were conducted at each step to verify the reactions. It was observed that the ionic liquid compound formed after 48 hours though confirmed the presence of starting materials. A high viscous transparent liquid was extracted and believed to be formed of 99% yielding of ionic liquid. Apart from that, the synthesis of ionic liquid was also performed using microwave treatment and heating to stimulate the reaction. Surprisingly, it was observed that the reaction was taking place within a fraction of second when the synthesis process was conducted with microwave treatment. In the second step, esterification IL with methacryloyl chloride was done to obtain monomer of polymerizable ionic liquid (m-PIL). The final product was a high viscous yellowish liquid.

Cytotoxic Effect and Constituent Profile of Alkaloid Fractions from Ethanolic Extract of Ficus septica Burm. f. Leaves on T47D Breast Cancer Cells.

The study aimed to investigate the profile of alkaloids in two ethyl acetate soluble fractions, namely fractions A and B from an ethanolic extract of Ficus septica leaves and cytotoxic effect on T47D breast cancer cells. Preparation of both fractions involved maceration of leaves with 70% (v/v) ethanol, filtration with Al2O3, precipitation with 0.1 N HCl, Mayer reagent, and 0.1 N NaOH, and also partition with ethyl acetate. Qualitative thin layer chromatography (TLC) was conducted to determine the profile of alkaloids in the two fractions, using alkaloid specific reagents such as Dragendorff, sodium nitrite, and Van Urk-Salkowski. Cytotoxic effects of both fractions on T47D cells were evaluated using MTT assay with a concentration series of 1.56; 3.12; 6.25; 12.5; 25 and 50 μg/mL. The TLC test showed that fractions A and B contained alkaloids with Rx values of 0.74 and 0.80 for fraction A and 0.74, 0.84, 0.92 for fraction B with regard to yohimbine using the mobile phase of n-buthanol:glacial acetic acid:distilled water (3:1:1 v/v/v). Moreover, an indole alkaloid was detected with Rx values of 0.80 and 0.84, respectively. Fractions A and B exhibited high cytotoxic effects on T47D cells with IC50 values of 2.57 and 2.73 μg/mL, respectively. In conclusion, overall the results of this study showed that fractions of Ficus septica contain alkaloids including indole alkaloid or its derivatives and possess a cytotoxic effect on T47D cells. This research supports the idea that alkaloids in F. septica have anticancer activity.

Assessing the quality of anti-malarial drugs from Gabonese pharmacies using the MiniLab®: a field study.

BackgroundRecent studies alluded to the alarming scale of poor anti-malarial drug quality in malaria-endemic countries, but also illustrated the major geographical gaps in data on anti-malarial drug quality from endemic countries. Data are particularly scarce from Central Africa, although it carries the highest burden of malaria. The aim of this medicine quality field survey was to determine the prevalence of poor-quality anti-malarial drugs in Gabon.MethodsA field survey of the quality of anti-malarial drugs in Gabonese pharmacies was conducted using the Global Pharma Health Fund Minilab® tests, following the Medicine Quality Assessment Reporting Guidelines. Anti-malarial drugs were purchased randomly from selected pharmacies in Gabon. Semi-quantitative thin-layer chromatography (TLC) and disintegration testing were carried out to measure the concentration of active pharmaceutical ingredients (APIs). The samples failing the TLC test were analysed by high-performance liquid chromatography. Following the collection of anti-malarial drugs, a street survey was conducted to understand where people purchase their anti-malarial drugs.ResultsA total of 432 samples were purchased from 41 pharmacies in 11 cities/towns in Gabon. The prevalence of poor-quality anti-malarial drugs was 0.5% (95% CI 0.08–1.84%). Two out of 432 samples failed the MiniLab® semi-quantitative TLC test, of which a suspected artemether-lumefantrine (AL) sample was classified as falsified and one sulfadoxine-pyrimethamine (SP) sample as substandard. High performance liquid chromatography with ultraviolet photo diode array detection analysis confirmed the absence of APIs in the AL sample, and showed that the SP sample did contain the stated APIs but the amount was half the stated dose. Of the people interviewed, 92% (187/203) purchased their anti-malarial drugs at a pharmacy.ConclusionUsing the GPHF Minilab®, the prevalence of poor-quality anti-malarial drugs is far lower than anticipated. The findings emphasize the need for randomized and robust sampling methods in order to collect representative data on anti-malarial drug quality.Trial registration: NTR4341 (Dutch Trial Registry)Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0795-z) contains supplementary material, which is available to authorized users.

Quality of the antibiotics--amoxicillin and co-trimoxazole from Ghana, Nigeria, and the United Kingdom.

Little is known about the quality of antibiotics despite being in high demand globally. Thirty five samples (27 brands) of the antibiotics amoxicillin (N = 20; 16 brands) and co-trimoxazole (N = 15; 11 brands), manufactured in six countries (China, Ghana, India, Ireland, Nigeria, and United Kingdom), were purchased in Ghana, Nigeria, and the United Kingdom. Their quality was assessed using German Pharma Health Fund (GPHF) MiniLab® as the screening tool—two capsules of amoxicillin (10%) and two tablets of co-trimoxazole (20%) failed the thin-layer chromatography (TLC) test. Definitive drug quality was measured using high-performance liquid chromatography–photodiode array detection (HPLC-PDA) for content of the stated active pharmaceutical ingredients (APIs) and bioavailability was determined with in vitro dissolution testing. All the samples of amoxicillin complied with U.S. Pharmacopeia (USP) tolerance limits, but 60% tablets of co-trimoxazole (purchased in Ghana and Nigeria) did not. There was disparity in the results obtained for co-trimoxazole and amoxicillin samples using the MiniLab® TLC tests. This highlights the need to invest in techniques such as HPLC-PDA and dissolution testing alongside the screening tests for assessing drug quality.

Processing of bitumens modified by a bio-oil-derived polyurethane

Castor oil (CO) functionalized by isocyanate groups (–NCO) is proposed as a novel bio-based reactive polyurethane (PU) for bitumen modification. This work presents a comparative analysis conducted on blends of bitumen and 2wt.% of a PU prepolymer prepared by NCO-functionalization of castor oil. Four preparation procedures were evaluated, which resulted from the combination of two processing times (1h or 24h, at 90°C) followed by two different post-treatments (water addition or ambient curing for up to 6months). It was found that the degree of modification attained after post-treatment depends on the previous processing conditions. Thus, short processing times are required if the binder is further subjected to ambient curing. Instead, the success of the water-addition modification falls on a previous long processing step. As revealed by rheological tests, ambient curing was seen to be by far a more efficient way of modification if compared to direct addition of water, and makes clear that the resulting binder evolves towards a better performance when in-service. In that sense, Thin Layer Chromatography tests, Modulated DSC and AFM images demonstrated a more complex microstructure characterized by the presence of a larger content of molecules with higher polarity, size and molecular weight.

AKTIVITAS ANTIFUNGI EKSTRAK ETANOL BATANG BINAHONG (Anredera cordifolia(Tenore) Steen.) TERHADAPCandida albicansSERTA SKRINING FITOKIMIA

Anredera cordifolia is a plantused as antifungal empirically. This study aims to find out the antifungal activity of ethanol extract of A. cordifolia stem against C. albicans and to identify the chemical subtance groups of the ethanol extract of A. cordifolia. The A. cordifolia stem was extracted by maceration method using ethanol 70 % as solvent. The antifungal assay was done using liquid dilution method with various concentrations of (85; 86; 87; 88; 89; 90% w /v). The mixtures of extract and suspension of C. albicans in CYG (Casein Yeast Glucose) medium and the media were incubated at 37oC for 18-24 hours to determine Minimum Inhibitor Conceration (MIC). Theculturewas streaked on SDA (Sabouraud Dextrose Agar) medium to determine Minimum Fungicidal Conceration (MFC). Chromatographic test and tube test were done to identify the chemical substances of A. cordifolia extract. The result showed that the MIC could not be determined because the extract was brownish and turbid while the MFC of A. cordifolia stem ethanolic extract was 86% w /v. The result of the phytochemical screening with tube test and the thin layer chromatography test showed that the extract of A. cordifolia stem contained flavonoids, polyphenols, and saponin.

English

Urtica angustifolia is a traditional medicinal material which has hypoglycemic function. The objectives of this study were to isolate and identify hypoglycemic active ingredients from it. Three groups of mice were gavaged with the decoction ofUrtica root, stem and leaf for 14 days at a dose of 20 mg/(kg·d). Area under the curve (AUC) values were 24.62±5.24, 24.33±4.66 and 25.51±4.25, respectively. The AUC in leaf decoction group was significantly lower than the other two groups (P < 0.05), which indicated the leaf extracts did have significant hypoglycemic action. Four kinds of extractions were obtained from leaves and fed to mice. AUC values were 33.44±3.58, 34.24±3.64, 30.44±2.57 and 9.93±1.98, respectively. AUC in group U3 (P < 0.05) and U4 (P < 0.01) were significantly lower than in the group of U1 and U2, which indicated that extraction U3 and U4 obviously decreased the blood sugar. Also, U3 was flavonoid and U4 was daucosterol, which was identified by thin-layer chromatography test, IR and ESI-MS spectral analysis.   Key words: Urtica angustifolia, active component, glucose tolerance test, hypoglycemic.

Factitious Diarrhea Induced by Stimulant Laxatives: Accuracy of Diagnosis by a Clinical Reference Laboratory Using Thin Layer Chromatography

Surreptitious ingestion of laxatives can lead to serious factitious diseases that are difficult to diagnose. Most cases involve ingestion of bisacodyl or senna. Thin layer chromatography (TLC) of urine or stool is the only commercially available test for these laxatives. Such testing is considered highly reliable, but its accuracy in clinical practice is unknown. Our aim was to evaluate the reliability of TLC laxative testing by a clinical reference laboratory in the United States. Diarrhea was induced in healthy volunteers by ingestion of bisacodyl, senna, or a control laxative (n = 11 for each laxative group). Samples of urine and diarrheal stool were sent in blinded fashion to the clinical reference laboratory for bisacodyl and senna analysis. TLC testing for bisacodyl-induced diarrhea revealed a sensitivity of 73% and specificity of 91% when urine was tested and sensitivity and specificity of 91% and 96%, respectively, when stool was analyzed. When diarrhea was induced by senna, the TLC assay for senna failed to identify even a single urine or stool specimen as positive (zero% sensitivity). Considering the expected prevalence of surreptitious laxative abuse in patients with chronic idiopathic diarrhea (2.4%-25%, depending on the clinical setting), TLC of urine or stool for bisacodyl by this reference laboratory would often produce misleading results, and testing for senna would have no clinical value. The major problems are false-positive tests for bisacodyl and false-negative tests for senna.

Identification of the antibacterial component of an ethanolic extract of the Australian medicinal plant, Eremophila duttonii.

Activity-guided fractionation was used to determine the antibacterial component of an ethanolic extract of the leaves of an Australian native medicinal plant, Eremophila duttonii F. Muell. (Myoporaceae). The extract, previously shown to have activity against Gram positive bacteria, was shown to have activity against additional Gram positive bacteria, including Clostridium perfringens, C. sporogenes and Listeria monocytogenes. Thin layer chromatography (TLC) was used to separate the extract into seven coloured fractions in visible light, one of which was shown by bioautography to contain antibacterial activity. Recovery of the component from the TLC plate and testing for antibacterial activity using a plate-hole diffusion assay supported this result. The purity of the component was verified by high-performance liquid chromatography and a time-kill experiment indicated that the purified component showed identical bactericidal activity to the whole extract. TLC spray reagents indicated that the component was a sterol, terpene or sugar but not a flavonoid, while the pigmented nature suggested a carotenoid.

Mixed iron-nickel complexes in Versatic 10 solutions

Iron and nickel Versatic solutions were prepared by solvent extraction with 33% Versatic 10 in Escaid 110. Mixed iron-nickel Versatic solutions with different iron:nickel ratios were prepared by mixing single metal solutions. The infrared spectra of the single metal and mixed metal Versatic solutions were recorded at 25 and 160°C. Iron Versatic showed absorption bands at 1685, 1575 and 1418 cm −1, probably due to solvating acid, bidentate chelation and bidentate bridging, respectively. Nickel Versatic absorbed at 1665 and 1590 cm −1; the 1590 cm −1 band has been assigned to monodentate coordination. All mixed iron-nickel solutions showed absorption bands at 1610 and 1585 cm −1. The relative intensities of the bands changed with solution composition, but not the frequencies. The mixtures showed none of the absorption bands of iron-only or nickel-only Versatic solutions; thus all the metal ions in solution are assumed to be present as mixed complexes. The average Versatic to total metal charge ratio in the complexes at 160°C was 1.1–1.3 for relatively high metal concentrations. Thin layer chromatography tests were carried out at 25°C; the retention factors of all mixed iron-nickel solutions were the same, and different from those of iron-only or nickel-only Versatic. This gives further evidence for the existence of mixed-metal complexes.

Thin Layer Chromatography for Rapid Detection of Carbohydrate Utilization by Bacteroides Strains

19 weakly saccharolytic Bacteroides strains of different species were tested by thin-layer chromatography (TLC) for their fermentative abilities for fructose, sucrose, lactose, galactose, and raffinose. Conventional fermentation tests were run parallel. In general, a good agreement between both methods was recorded. Two strains, however, showed a degradation in the TLC test without an acidification. With some strains, sucrose as a substrate yielded a fructose spot, lactose a galactose spot, and raffinose a melibiose spot, indicating an incomplete degradation of these carbohydates.

Thin Layer Chromatographic Method for Analysis and Chemical Confirmation of Sterigmatocystin in Cheese

A semi-quantitative method is described for the analysis of sterigmatocystin in cheese. The method is based on extraction of cheese with MeOH--4% KCl (9 + 1), followed by Florisil and polyamide column cleanup and 2-dimensional thin layer chromatography (TLC). Visualization of sterigmatocystin on the TLC plates is enhanced by an AlCl3 spray reagent. The identity of sterigmatocystin is confirmed by a 2-dimensional TLC test based on reaction with trifluoroacetic acid (TFA) on the plate after first development. The reaction product formed is visualized by spraying with AlCl3. The method allows detection and confirmation of sterigmatocystin in cheese at concentrations as low as 5 micrograms/kg. The method has been applied to cheese samples ripening in warehouses and naturally molded with Aspergillus versicolor.

Some observations on the behavior of rancid groundnut oil with regard to the thin layer chromatography test for the detection of castor oil

AbstractRefined groundnut oils were autoxidized to varying degrees as indicated by the peroxide values. Some autoxidized oils gave spots around the same Rf as that attributed to castor oil in the thin layer chromatography test prescribed by ISI. The molybdic acid test was negative.