Paper Chromatographic Procedures Research Articles

Development and validation of a high-performance liquid chromatography-electrospray ionization-mass spectrometry assay for the determination of zaleplon in human plasma.

In this paper, a sensitive and rapid chromatographic procedure using a selective analytical detection method (electrospray ionization-mass spectrometry in selected-ion monitoring mode) in combination with a simple and efficient sample preparation step is first presented for the determination of zaleplon in human plasma. The separation of the analyte, internal standard, and possible endogenous compounds are accomplished on a phenomenex Luna 5-microm C8(2) column (250- x 4.6-mm i.d.) with methanol-water (75:25, v/v) as the mobile phase. In order to optimize the mass detection of zaleplon, several parameters such as ionization mode, fragmentor voltage, m/z ratios of ions monitored, type of organic modifier, and eluent additive in the mobile phase are discussed. An internal standard is selected to guarantee the quantitative accuracy. Each analysis takes less than 6 min. The calibration curve of zaleplon in the range of 0.1-60.0 ng/mL in plasma is linear with a correlation coefficient of > 0.9992, and the detection limit (s/n = 3) is 0.1 ng/mL. The within- and between-day variations (relative standard deviation) in the zaleplon plasma analysis are less than 2.4% (n = 15) and 4.7% (n = 15), respectively. The application of this method is demonstrated for the analysis of zeleplon plasma samples in a Phase-I human pharmacokinetic study.

A miniaturized rapid paper chromatographic procedure for quality control of technetium-99m sestamibi

A miniaturized rapid paper chromatographic technique (MRPC) for quality control of a technetium-99m sestamibi preparation was developed and compared with the manufacturer's and Hung et al. techniques. The MRPC system involves the use of 6.0x0.5cm Whatman 3MM paper strip developed in ethyl acetate. The procedure was completed within 3min while that of the manufacturer and Hung techniques took 30-35 and 4min respectively. The Rf range of 99mTc-sestamibi using MRPC was 0.55-0.75 while that of the other two techniques was 0.9-1.0. The results indicate that MRPC can be used to separate 99mTc-sestamibi from any 99mTc contaminant that migrates with the solvent front. The MRPC is a fast and effective chromatographic technique for routine quality control testing of 99mTc-sestamibi preparation.

A sensitive and simple paper chromatographic procedure for detecting neomycin phosphotransferase II (NPTII) gene expression

A sensitive and simple paper chromatographic procedure for detecting neomycin phosphotransferase II (NPTII) gene expression

Determination of radioactive proline and hydroxyproline by a simple paper-chromatographic procedure

A simple method to separate and determine radioactive proline and hydroxyproline by paper chromatography is described. The localization of the imino acids separation is achieved by direct nondestructing staining with 1-fluoro-2,4-dinitrobenzene dye. The imino acids are quantitated directly by liquid scintillation counting in the presence of paper strips. The method was applied to bone cultures with good reproducibility, sensitivity and linearity over a wide range of radioactivity. The procedure was also tested in fibroblast cultures. The results for hydroxyproline were in good correlation with the widely used method of Juva and Prockop (Juva, K. and Prockop, D.J. (1966) Anal. Biochem. 15, 77–83), in which hydroxyproline is oxidized to pyrrole, and then extracted and purified by column chromatography before counting radioactivity.

Modulation of T4 5'-monodeiodination in rat anterior pituitary and liver homogenates by thyroid states and fasting.

In order to clarify the role of the pituitary conversion of L-thyroxine (T4) to 3, 5, 3'-L-triiodothyronine (T3) in regulating thyrotropin (TSH) secretion, the effect of altered thyroid states and fasting on intrapituitary T3 generation was investigated by a paperchromatographic procedure using the anterior pituitary homogenates. Hepatic T3 generation was also studied for comparison. The rate of pituitary and hepatic T3 generation in normal rats averaged 25.2 +/- 12.4 (mean +/- SE) fmoles T3/mg protein/min and 33.8 +/- 12.7 fmoles T3/mg protein/min, respectively. T4 treatment raised the hepatic T3 generation from T4 (46.7 +/- 3.1 fmoles T3/mg protein/min) and lowered the intrapituitary T3 generation (4.5 +/- 0.2 fmoles T3/mg protein/min). On the contrary, thyroidectomy slowed the hepatic T4 5'-deiodination (11.0 +/- 2.8 fmoles T3/mg protein/min), and accelerated the pituitary T4 5'-deiodination (64.3 +/- 1.4 fmoles T3/mg protein/min). In 48 h fasted rats, serum T4, T3 and TSH concentrations were all lower than those in fed rats, and both pituitary and hepatic T3 generations were also suppressed. Thus, altered thyroid states cause an opposite effect on pituitary and liver 5'-monodeiodination, whereas fasting causes similar changes. The findings suggest the existence of an autoregulatory mechanism for thyroid hormone activation within the target tissues.

Novel spray reagents for plant pigments

Paper chromatographic procedures involving the use of butanol∶acetic acid∶water (4∶1∶5 v/v) as solvent and 1 % aqueous uranyl acetate or uranyl nitrate solution as spray reagent, have been developed for detection of plant pigments when present in amounts of at least 5 μg each. Quinones, flavones and some of their O-glycosides having at least one hydroxyl group adjacent to carbonyl group, have been easily detected. The colour intensity has been found to increase in the case of compounds containing vicinal hydroxyl groups.

Radiochromatography of indium (111In) oxine

A paper chromatographic procedure is described for distinguishing complexed and uncomplexed indium in solutions resulting from the extraction of indium-111 with oxine (8-quinolinol).

Hyperresponsivitiy of spontaneously hypertensive rat to indirect measurement of blood pressure.

A chronic, indwelling, tail arterial cannula was implated in conscious undisturbed rats for measurement of blood pressure and heart rate and for obtaining blood samples. As an index of sympathetic activity, plasma levels of catecholamines in arterial blood of conscious animals were assayed by a radioenzymatic, paper-chromatographic procedure. Blood pressures of unrestrained spontaneously hypertensive (SHR) rats in their home cages (161 +/- 3/141 +/- 4 mmHg) were not different from those of pentobarbitol-anesthetized, hypertensive animals but were about 25 mmHg lower than awake animals during the restraint required for the tail-cuff procedure. Basal levels of plasma catecholamines in awake, undisturbed or in pentobarbital-anesthetized animals were similar in age-matched SHR and normotensive Wistar-Kyoto (WKY) rats. SHR rats were shown to have greater increase in plasma catecholamines than WKY rats during forced immobilization or restraint for indirect measurement of blood pressure.

A rapid and accurate procedure for assaying DNA polymerase, RNA polymerase, or ribosome dependent protein synthesis

A descending paper chromatographic procedure for separating trichloracetic acid soluble from precipitable material is described. Its use for assaying DNA, RNA, or protein synthesis from a variety of sources with various states of purity attests to its general applicability to reactions of interest to molecular biologists. Large numbers of analyses can be carried out in a short period of time without compromising accuracy or reliability. The use of this procedure for still more enzymes, those involved in modifying DNA, RNA, or proteins, is discussed.

Physiological variations in plasma testosterone in normal adult male subjects measured by a competitive protein-binding technique

A method of estimating plasma testosterone utilizing a single paper chromatographic procedure and competitive protein-binding is described. The influence of some conditions of sample collection on plasma testosterone has been investigated. Delay in the separation of plasma from cells and the type of anticoagulant used were both without significant effect. There was an approx. two-fold variability in plasma testosterone concn. in samples withdrawn daily, weekly and monthly from four young, normal adult males.

Ergoline Alkaloidal Constituents of Hawaiian Baby Wood Rose, Argyreia nervosa (Burm. f.) Bojer

Chemical analysis showed that the seeds of Argyreia nervosa contain the highest percentage of indole alkaloid constituents (0.5-0.9%) of the genera of the Convolvulaceae thus far studied. A total of 19 indole alkaloids were identified by thin-layer and paper chromatographic procedures. Of these, lysergene, festu-clavine, setoclavine, isosetoclavine, agroclavine, elymoclavine, ergine, and isoergine were isolated by column chromatographic procedures and characterized by TLC and IR analyses. Penni-clavine, chanoclavine-I, chanoclavine-II, ergometrine, ergometrinine, lysergic acid α-hydroxyethylamide, isolysergol, racemic chanoclavine-II, molliclavine, lysergol, and isolysergic acid α-hydroxyethylamide were identified by TLC only. Of these, lysergene, setoclavine, isosetoclavine, chanoclavine-II, racemic chano-clavine-II, isolysergol, and molliclavine were identified for the first time in species of the Convolvulaceae. Ergine (0.136%) and isoergine (0.188%) were found in the highest concentration. In addition, 11 unidentified indole alkaloids were detected, these being found in very low concentration. Although the pericarp showed the same alkaloid pattern as the seeds, the concentration was much lower (0.0015%). No alkaloids could be delected in the vegetative tissues of nonflowering specimens.

METABOLISM OF THE TOXIC PRINCIPLE OF ASTRAGALUS MISER VAR. SEROTINUS IN RUMINANTS AND NON-RUMINANTS

Rats and mice fed timber milk vetch (TMV) developed symptoms within 36 hours and died approximately 24 hours later. The rapidity of the onset of symptoms depended on the amount of TMV consumed. The symptoms included a loss of equilibrium, staggered gait and a characteristic arching of the back. Extensive hemorrhages were seen in the gastric mucosa. There was a significant (P < 0.05) increase in the activity of serum glutamate oxaloacetate transaminase and serum isocitrate dehydrogenase of rats fed TMV. By paper chromatographic procedures it is shown that miserotoxin is hydrolyzed to glucose and 3-nitropropionic acid under the acidic conditions of the stomach in monogastric animals. In ruminants, miserotoxin is hydrolyzed to glucose and 3-nitro-1-propanol.

Determination of Arsenical Herbicide Residues in Plant Tissues

Paper chromatographic separation of hydroxydimethylarsine oxide (cacodylic acid), monosodium methanearsonate (MSMA), sodium arsenate, and sodium arsenite was achieved with the aid of four solvent systems. Aqueous extracts of plant tissues removed essentially all the arsenicals applied, but methanolic fractionation was required before the extracts could be analyzed by paper chromatographic procedures. A standard nitric-sulfuric acid digestion procedure was employed for arsenic analyses, but great care was taken to avoid sulfuric-acid-induced charring by first adding relatively large amounts of nitric acid to drive off chlorides present. Depending upon the amount of chloride present, substantial losses of arsenic as arsine chlorides were observed if the samples charred. Five minutes in fuming sulfuric acid to completely break the carbon-arsenic bonds was another critical requirement for the quantitative determination of arsenic from cacodylic acid and MSMA. The silver diethyldithiocarbamate colorimetric method was useful for detecting as little as 0.6 μg or as much as 20 μg of arsenic per sample.

An automated method for the rapid chromatographic analysis of serum valine, alanine and glycine

An automated system is described for the analysis of serum valine, alanine and glycine for use in nutrition surveys. The method represents an improvement on the paper Chromatographic procedure for the measurement of the amino acid ratio. It has the advantage that the results are not expressed as ratios of amino acid concentrations but in absolute values.

The separation of chlorinated derivatives of hydroquinone using thin-layer and paper chromatography

Thin-layer and paper chromatographic procedures are described for the separation and identification of six chlorinated derivatives of hydroquinone. Adsorption thin-layer chromatography (silica gel) or reversed-phase paper chromatography (olive oil—stationary phase and diluted acetic acid—mobile phase) did not enable satisfactory separation of dichloro-derivatives. These compounds have been clearly separated on kieselguhr thin layers and paper impregnated with formamide using three solvent systems.

Paper chromatographic techniques for the determination of cephalothin and desacetylcephalothin in body fluids.

Paper chromatographic procedures may be used to detect cephalothin and its metabolite desacetylcephalothin in urine, plasma, synovial fluid, and cerebrospinal fluid. Protein-bound antibiotics are released from plasma, cerebrospinal fluid, and synovial fluid by dilution with an equal volume of dimethylformamide. Data are presented on the sample preparation, paper chromatographic system, and other specific techniques.

Labeled complexes in liver cystosol after administration of 4-14C-corticosterone and 4-14C-cortisol.

Previous work showed a distribution of labeled cortisol in rat liver from 30 sec to 40 min after intravenous administration. The ultracentrifuged supernatant fraction (cytosol), containing about 2'3 of the label in the liver, has now been separated into a sharp, Sephadexchromatographed peak containing most of the supernatant label. Various findings indicate that this material is associated in a complex; this may be of molecular weight 5000 or a little less, a complex that could possibly be composed of steroid, polynucleotide and polypeptide. Further column and paper chromatographic procedures separated this material into 2 bands, each labeled and possessing 260 mμ absorption characteristics. Studies with corticosterone gave similar results. (Endocrinology 87: 486, 1970)

真珠の黄色色素-II.

The pigments responsible for the coloration of the yellow nacre of pearl were investigated. Yellow pigments were extracted with HCl-MeOH, fractionated by paper chromatographic procedure, and analyzed on their constituents. The yellow pigments consist of four kinds of pigments which were classified into three types based on their spectral characteristics in the ultraviolet region and color reactions with various kinds of spray reagents for paper chromatography. Among them, major yellow pigment, after acid hydrolyzed, gave rise to many amino acids and the substance giving color reaction characteristic of sugars. From these results, it was deduced that conchiolin-like proteinous substance bearing some sugar derivatives as its constituent might be a major yellow pigment of the yellow pearl.

Paper Chromatographic Techniques for the Determination of Cephalothin and Desacetylcephalothin in Body Fluids

Paper chromatographic procedures may be used to detect cephalothin and its metabolite desacetylcephalothin in urine, plasma, synovial fluid, and cerebrospinal fluid. Protein-bound antibiotics are released from plasma, cerebrospinal fluid, and synovial fluid by dilution with an equal volume of dimethylformamide. Data are presented on the sample preparation, paper chromatographic system, and other specific techniques.

Persistent mild hyperphenylalaninemia in various ethnic groups in Israel.

CLASSICAL PHENYLKETONURIA (PKU) may be defined as a hereditary metabolic disorder due to phenylalanine hydroxylase deficiency, leading to persistently very elevated blood phenylalanine levels (above 20 mg/100 cc) and urinary excretion of phenylpyruvic acid. Recently introduced programs for mass screening of newborn for the detection of this disease revealed that hyperphenylalaninemia may represent a sign of several distinct disorders.1-4In the present communication, we report on the occurrence of a type named persistent mild hyperphenylalaninemia4in the Israeli population. The genetic nature of this type is discussed. Methods Screening for PKU in the newborn was performed by the method of Guthrie and Susi5usually at the age of 3 to 5 days. When a presumptive positive result was observed, additional samples of blood dried on filter paper were requested and examined by the Guthrie method as well as a paper chromatographic screening procedure for amino acids.6