Paper Chromatograms Research Articles

Quatitative and Qualitative Estimation of Free Amino Acids in the Haemolymph of parasitized bugs (Leptocoris augur).

Leptocoris augur is a pest of Kusum plant, (Schleichera oleosa), which in turn is a host of Lac insect. This bug is a gregarious feeder and by its desapping habit loss the viability of seeds. In L. augur a total of 16 amino acids were identified in the control bug’s haemolymph (both sexes) by paper chromatogram. Thus, free amino acids level of haemolymph is greatly affected by the parasitization. In this way the Quantitative level of amino acid in parasitized bug haemolymph significantly altered by mermithid parasitism. Keywords: Leptocoris augur, Haemolymph, Free amino acids, Parasitized bug, Quantitative level.

Flavonoid as chemotaxonomic markers in endemic/endangered species of Rauvolfia from Southern Western Ghats of India: A preliminary study

Preliminary analysis of flavonoid chromatographic migration profiles of endemic/endangered species of Rauvolfia L from Southern Western Ghats of India were carried out. Paper chromatogram showed maximum separation in the solvent system of forestral. In the paper chromatogram, number of flavonoid spots varied from 9 to 12 in the five taxa studied. The main aglycones detected in high performance liquid chromatography (HPLC) analysis were flavones apigenin and luteolin, flavonol kaempferol, myricetin, quercetin and anthocyanidins such as delphinidin and cyanidin. Flavonol Quercetin was detected in all the five species of Rauvolfia giving a chemotaxonomic significance to its presence at the generic level. The two species Rauvolfia serpentina and Rauvolfia tetraphylla could be regarded as the most primitive in the evolutionary line with respect to the flavonoid pattern. Rauvolfia densiflora has the most advanced pattern of flavonoids. The dendrogram generated by unweighted pair group method with arithmetic average (UPGMA) cluster analysis of chemo metric data showed a clear grouping of five species in three clusters. Flavonoid profiles were efficiently used for the identification of Rauvolfia beddomei, which due to morphological similarity, was erroneously suspected to be the medicinally significant species Rauvolfia micrantha. Flavonoid profiling using paper chromatography, in the solvent system of forestral could suggest an easy and quick procedure for identifying adulteration by substitution in Rauvolfia species.

Textile dyes Disperse Orange 1 and Yellow 3 contain more than one allergen as shown by patch testing with thin-layer chromatograms.

It is known that some patch-test preparations containing disperse dyes contain impurities with unknown relevance for the development or elicitation of contact allergy. To evaluate the significance of the impurities found in the commercial dyes Disperse Orange 1 (DO1) and Disperse Yellow 3 (DY3) regarding contact allergy in patients with known sensitivity to them. Ten patients allergic to DY3 and/or DO1 were tested with a dilution series of commercial and purified DY3 and DO1 (with water-soluble parts prepared from the commercial dyes) and with naphthalene sulfonate. Nine patients were additionally tested with thin-layer chromatograms (TLCs) made from the commercial DO1 and DY3 and with paper chromatograms made from the water-soluble part of these dyes. Eight of nine and three of six patients tested positively to the TLCs of DO1 and DY3, respectively. Among them, 4 of 8 and 2 of 3 patients, respectively, were positive also to another spot on the TLCs. One patient was positive to the paper chromatogram from the water-soluble part of DO1. None of the tested patients reacted to naphthalene sulfonate. The results of our study suggest that there are more relevant allergens in the fat-soluble and water-soluble fractions of the commercial disperse dyes.

The chemistry of anti-pernicious anaemia factors; the ninhydrin-reacting hydrolytic fragment of vitamin B12.

Summary and Conclusions Our previous conclusion that acid hydrolysis of vitamin B12 leads to the formation of a “ninhydrin-reacting” substance is confirmed. The behaviour of this “ninhydrin-reacting” substance and of 2-aminopropanol on paper chromatograms has been examined. The results appear to show that these two substances are identical. Evidence has been obtained for the presence in acid hydrolysates of vitamin B12 of material showing selective absorption with bands and inflections at 2850, 2768, 2690, 2585 and 2500 å (referred to as “the 285 component“). This “285 component” is resolved into at least two structurally related substances by chromatography employing n-butyl alcohol-acetic acid as the irrigation solvent. Formation of ammonia occurs during acid or alkaline hydrolysis of vitamin B12. The authors thank Mr. R. E. Rodway for technical assistance in the isolation of crystalline vitamin B12 from a concentrate kindly supplied to the Research Department by Mr. A. W. Davidson. The ultra-violet absorption data were kindly determined by Dr. E. R. Holiday (M.R.C. Spectrographic Unit The London Hospital, E.1***), who also carried out the examinations with the Corning filter. The authors thank the Directors of The British Drug Houses, Ltd., for permission to publish these results.

A furfural reagent of high specificity for the detection of carbamates on paper chromatograms.

Abstract A spot reaction with high specificity for the detection of carbamates, and its application to paper chromatograms, is described. Chromatographic separation methods are given, together with results obtained on a number of currently used carbamate drugs. The incorporation of the reagent in a normal screening procedure for acidic and neutral drugs is suggested

A note on the factors affecting R-F values on citrate buffered paper chromatograms.

Abstract Experiments have been made to determine the effect of changes in experimental conditions on the speed of running of citrate/butanol systems on paper chromatograms.

SOME FACTORS AFFECTING THE RF VALUES OF SYMPATHOMIMETIC CATECHOLAMINES.

Abstract The behaviour of three catecholamines on paper chromatograms is reported as an investigation of the effects of variables on RF values. The use of these values as a means of locating and identifying catecholamines on paper is misleading as they are dependent on too many different factors.

THE INFLUENCE OF HYDROCHLORIC ACID ON THE CHROMATOGRAPHIC BEHAVIOUR OF SYMPATHOMIMETIC CATECHOLAMINES.

Abstract The behaviour of three catecholamines on paper chromatograms is shown to be markedly influenced by hydrochloric acid. Double spot formation exhibited by noradrenaline acid tartrate, for example, results from the use of hydrochloric acid in the developing solvent, but the formation of the second spot is prevented when the amine is applied to the paper from solution in dilute hydrochloric acid. Retention of amine at the application area and additional multiple spot phenomena are exhibited by all of the three amines investigated when chromatographed from concentrated hydrochloric acid. The significance of these and related observations is discussed in relation to the use of hydrochloric acid during the separation of catecholamines from biological tissues and fluids.

SOME POSSIBLE CAUSES OF PHARMACOLOGICAL ACTIVITY IN BLANK ELUATES FOLLOWING THE SEPARATION OF SYMPATHOMIMETIC CATECHOLAMINES BY PAPER CHROMATOGRAPHY.

Abstract Eluates prepared from paper chromatograms at Rf values and under experimental conditions that were expected to eliminate the possibility of contamination with catecholamines were shown to inhibit the rat uterus and to cause a fall in the mean arterial blood pressures of anaesthetised rats. These isoprenaline-like actions have been shown to be due to inadequate removal of the phenolic developing solvent and to the use of sodium dihydrogen phosphate solutions as eluants. Attempts to establish qualitative and quantitative relationships between the chromatography blank and isoprenaline have failed, and it is concluded that these factors could not alone be responsible for the isoprenaline-like substance reported to occur naturally in various species of mammal.

50 years in insect toxicology: 1956–2006

50 years in insect toxicology: 1956–2006

Antibacterial Activity and Property of the Fermentation Product of Marine Streptomyces sp. GB-2

Marine Streptomyces GB-2, isolated from marine samples collected from the intel tidal zone of Lianyungang, is found to produce antibacterial substance which has exhibited significant inhibitory effects on 11 Gram-positive bacteria and 4 Gram-negative bacteria. The antibacterial substance was proved to be neutral and water-soluble according to paper chromatogram analysis, whose production was significantly associated with artificial seawater. The results of stability studies of the fermentation broth of Streptomyces GB-2 showed that it was very stable at pH1 and pH12 under 121°C and had only little change after ultraviolet treatment. The potential use of substance produced by strain GB-2 in bio-control, food, and medical application was predicted.

Functional Characterization of PmHS1, a Pasteurella multocida Heparosan Synthase

Heparosan synthase 1 (PmHS1) from Pasteurella multocida Type D is a dual action glycosyltransferase enzyme that transfers monosaccharide units from uridine diphospho (UDP) sugar precursors to form the polysaccharide heparosan (N-acetylheparosan), which is composed of alternating (-alpha4-GlcNAc-beta1,4-GlcUA-1-) repeats. We have used molecular genetic means to remove regions nonessential for catalytic activity from the amino- and the carboxyl-terminal regions as well as characterized the functional regions involved in GlcUA-transferase activity and in GlcNAc-transferase activity. Mutation of either one of the two regions containing aspartate-X-aspartate (DXD) residue-containing motifs resulted in complete or substantial loss of heparosan polymerizing activity. However, certain mutant proteins retained only GlcUA-transferase activity while some constructs possessed only GlcNAc-transferase activity. Therefore, it appears that the PmHS1 polypeptide is composed of two types of glycosyltransferases in a single polypeptide as was found for the Pasteurella multocida Type A PmHAS, the hyaluronan synthase that makes the alternating (-beta3-GlcNAc-beta1,4-GlcUA-1-) polymer. However, there is low amino acid similarity between the PmHAS and PmHS1 enzymes, and the relative placement of the GlcUA-transferase and GlcNAc-transferase domains within the two polypeptides is reversed. Even though the monosaccharide compositions of hyaluronan and heparosan are identical, such differences in the sequences of the catalysts are expected because the PmHAS employs only inverting sugar transfer mechanisms whereas PmHS1 requires both retaining and inverting mechanisms.

CYTOTOXICITY ASSAY OF ACTIVE COMPOUNDS OF Gynura procumbens (Lour) Merr TO SEVERAL CELL LINE

The leaves of Dewa (Gynura procumbens (Lour) Merr), is utilized in traditional medicine as anticancer, had been reported to contain flavonoid derivatives and supposed to have cytotoxic properties. Hence, these particular substances might be useful to be developed as anticancer in the future. Research on ethanol extract of Gynura procumbens (Lour) Merr leaves as anticarcinogen has been carried out, using toxicity assay model to vero, myeloma and lymphocyte cells. This research applied varied concentrations and adriamycin as a positive control. Observation to the cell growth inhibition was carried out for 4 days. The result showed that the dominant substance (the third spot of paper chromatogram) inhibit the growth of vero and myelome cells but lymphocyte cell. The LC50 of this substance to myelome, vero and lymphocyte cells were 2650 g/ml, respectively, whereas the LC50 of adriamycin to the cell lines were < 0.015 g /ml. Therefore, substances obtained from Gynura procumbens (Lour) Merr leaves might be developed against cancer. Key word : Gynura procumbens (Lour) Merr, mieloma

A Novel Method for the Determination of Synthetic Colors in Ice Cream Samples

A simple method has been developed for the extraction, separation, and determination of synthetic colors in ice cream samples. The process involves the breakdown of emulsion by neutral detergents (Triton X-100 and Tween 20) followed by extraction with petroleum ether for removal of fat. The aqueous colored solution obtained is treated with 5% acetic acid, and the uptake of color is carried out by a wool-dyeing technique. The color is eluted from the wool with 5% ammonia solution, the solution is evaporated to dryness, and the residue is dissolved in 60% ethanol for paper chromatography using trisodium citrate-ammonia-water (2 + 5 + 95, w/v/v) as the mobile phase. The colored spots from the paper chromatogram are cut and eluted with 60% ethanol, and the absorbance is measured at the respective lambda maximum corresponding to the Rf value of the appropriate standard. The recoveries of 6 colors, including sunset yellow FCF (SSYFCF), tartrazine, carmoisine, ponceau 4R, brilliant blue FCF (BBFCF), and fast green FCF from spiked samples with either detergent were found to be >90%. However, recoveries of erythrosine were 21 and 65% with Triton X-100 and Tween 20, respectively. Indigo carmine could not be recovered at all because of its fugitive property in 5% ammonia solution, which is used to strip the color from the wool. The sensitivity of the method with the use of Tween 20 is 1 ppm (1 microg/g) for the colors in spiked ice cream samples. With this method, we analyzed samples of 20 branded colored ice cream. The results showed the presence of tartrazine (8.4-43.3 ppm), SSYFCF (23.5-117.6 ppm), carmoisine (traces-53.2 ppm), erythrosine (3.5 ppm), and BBFCF (4.1 ppm) in the ice cream samples. Apart from 2 samples of tuttifruity, all of the ice cream samples showed the presence of permitted synthetic colors below the permissible level of 100 ppm established by the Prevention of Food Adulteration Act of India.

Evaluation of stability of compost prepared with Korean food wastes

Food wastes were composted with saw dust, dried paper mill sludge, and decayed wood dust to reduce the volume of wastes and to produce a stable organic soil amendment. To determine the stability for agricultural utilization, the compost maturity was evaluated using physical, chemical, and biological parameters. Temperature of the inner compost heap remained high at over 50°C during the 80 d of composting, but the chemical and physical parameters apparently changed between 30 and 50 piling d: pH was a weakly acidic of the initial stage and neutral after 30 d of composting. The C / N ratio decreased to less than 12 after 65 d of composting, but the C / N ratio of the products was less than 0.5 time that of the initial value after 35 d of composting. Reducing sugar contents changed significantly between 40 and 45 d of composting, and the Y value that indicated color changes of the water extracts decreased and became stabilized at around 3 after 50 d of composting. Ring types and color on circular paper chromatograms showed an apparent difference before and after 40 d of composting. The content of sodium (Na), which might be one of the most harmful elements in Korean food wastes for agricultural utilization, increased from 12 to 17 g kg−1 with time and Na mostly occurred in a water-soluble form. Acetic acid was the main component of volatile organic acids, and total organic acids were produced at a very high rate (500–700 mg kg−1) during the first 5–30 d of composting and then the rate decreased rapidly to below 300 mg kg−1 after 35 d of compo sting. Germination index of Chinese cabbage in water extracts fluctuated with compost pile turning to a value below 50 by 40 d of composting, due to the effects of high contents of organic acids and Na. The value exceeded 50 after 45 d of compo sting but did not increase further with continuous composting, which might be due to the high content of Na. Lettuce, a sensitive species, was scarcely germinated in water extracts of the food compost. Consequently, the high content of Na in Korean food waste compost, in spite of maturity, could be a limiting factor for agricultural utilization. The effects of Korean food waste compost on plant growth and soil conditions should be evaluated further at the field level.

The pattern of ClO 2 stabilized by Na 2CO 3/H 2O 2

In this paper, the existing patterns of carbonate and ClO 2 in the so-called “stabilized chlorine dioxide” solution which is stabilized by stabilizers Na 2CO 3 and H 2O 2 are analyzed. Meanwhile, thermostability, UV absorption spectrum, specificity of the paper chromatogram, microstructure and ionic chromatogram of this solution were studied, and contrasted with pure NaClO 2 and ClO 2. The results show that ClO 2 in “stabilized chlorine dioxide” solution exists in the form of chlorite ClO 2 −, carbonate exists in the form of bicarbonate. Therefore, it is considered that “stabilized chlorine dioxide” solution is mixed solution of ClO 2 −and HCO 3 −, its pH being 8.5.

Qualitative detection of selenium in fortified soil and water samples by a paper chromatographic–carboxyl esterase enzyme inhibition technique

Thevetia peruviana seed carboxyl esterase was employed as a biosensor for the detection of selenium compounds by an enzyme inhibition technique on paper chromatograms. The selenium compounds (sodium selenite and selenium dioxide) appeared as white spots on a magenta background due to the inhibition of Thevetia peruviana seed carboxyl esterase (substrate 1-naphthyl acetate, coupling reagent Fast blue B salt). The minimum detectable amounts were about 5 μg of sodium selenite and 5 μg of selenium dioxide. Many other animal and plant carboxyl esterases gave no inhibition spot under the same conditions. Soil and water samples were fortified with sodium selenite and selenium dioxide. A procedure for preparing test solutions and conditions for paper chromatography was established.

Fatal attraction evaded. How pathogenic bacteria resist cationic polypeptides.

Fatal attraction evaded. How pathogenic bacteria resist cationic polypeptides.

Paving the path

The Path of Carbon in Photosynthesis evolved in the minds of Sam Ruben and Martin Kamen as they followed incorporation of radioactive 11CO2 in their many experiments, 1938 to 1942. With the concepts and collaboration of C. B. van Niel it became clear that reduction of CO2 can occur in the dark and may require processes similar to bacterial systems. It could not involve the century-old A. von Baeyer theory's photochemical reduction of CO2 adsorbed on chlorophyll which had guided decades of painstaking efforts, of R. Willstatter and A. Stoll and many others, in vain searches for formaldehyde. In 1942 Sam Ruben gave me all of his home-made 14CO2 to look for the first steps of the path of carbon in photosynthesis. Measurements were tedious and activities low but progress was real. Liquid-liquid partition of the radioactivity indicated chemical properties of the products and their derivatives. Interrupted by the war and Sam Ruben?s tragic accidental death, I accepted Melvin Calvin's invitation to continue The Path and we established our lab in the Old Radiation Laboratory next to Ernest Lawrence's cyclotron. Pre-illuminated algae rapidly reduced 14CO2 in the dark! Simple 'partition'-based separations on paper chromatograms extended the earlier methodology. Sedoheptulose and ribulose made surprise appearances. Soon it became clear that Nature had selected ribulose bisphosphate as its universal CO2 acceptor molecule. Finally, our isolation of carboxydismutase in 1954 revealed its identity with Sam Wildman's Fraction 1 protein. The Path was paved with one of this planet's major proteins!

Enzyme inhibition XI: glutamate decarboxylase activity relationship with the reaction products as determined by the colorimetric and radioisotopic methods

The relationship of glutamate acid decarboxylase (GAD) activity with the reaction products was developed. It was based on incubating sodium glutamate substrate (S) with GAD enzyme (E) when the enzyme-substrate-complex (ES) product was obtained along with gamma aminobutyric acid (GABA) and unreacted sodium glutamate. The reaction products were separated by paper chromatography. The ES, GABA and S products were sprayed with ninhydrin reagent when ninhydrin-colored-complex (NCC) was formed on the paper chromatogram. The products were extracted with 75% ethanol containing 0.5% cupric sulfate. The NCC absorption readings of ES and S products were measured by a spectrophotometer. A standard curve was prepared by plotting absorption readings against different concentrations of sodium glutamate. This curve was the basis of determining GAD activity of E. coli and C. welchii. It was observed that NCC absorption of ES and S products was directly related with the enzyme activity. The qualities of ES and S products in the reaction mixture increased as the enzyme activity increased with the incubation time. On the other hand, some products in the reaction mixture decreased in the presence of an inhibitor of GAD activity. The relationship of reaction products with GAD activity was also established by the radioisotopic method. The results obtained by the chromatographic separation of products followed by the spectrophotometric method of determining GAD activity is a simple, safe and less expensive compared to the other methods.