Determination of carbonyl compounds in water by derivatization-solid-phase microextraction and gas chromatographic analysis

The solid-phase microextraction (SPME) technique was evaluated for the determination of 23 carbonyl compounds in water. The carbonyl compounds in water were derivatized with omicron-(2,3,4,5,6-pentafluorobenzyl)-hydroxylamine hydrochloride (PFBHA), extracted with SPME from liquid or headspace and analyzed by GC with electron capture detection (GC-ECD). The effects of agitation techniques and the addition of salt (NaCl) on extraction, the absorption-time and absorption-concentration profiles were examined. The precision of the SPME technique for the determination of carbonyl compounds was evaluated with spiked bidistilled water, ozonated drinking water, and rain water. The relative standard deviations obtained from different spiked water matrix were similar, and in the range of 5.7-21.1%. The precision can be further improved by using an internal standard. With 4 ml of water sample, the limits of detection for most of the tested carbonyl compounds using liquid or headspace SPME-GC-ECD were similar and in the range of 0.006-0.2 micrograms/l, except for glyoxal and methylglyoxal, which showed low sensitivity when using headspace SPME. In the analysis of an ozonated drinking water sample, the SPME techniques gave comparable results to those of the conventional liquid-liquid extraction method.     

Reactivity in tissue tryptophyl units in histofluorescence methods using carbonyl compounds

The contribution of tissue tryptophyl residues, with both amino and carboxyl groups linked to the peptide bonds, to visible fluorescence was studied following various histochemical methods. Tryptophan residues of chymotrypsinogen and trypsinogen exhibited visible fluorescence after (1) combined formaldehyde-HCl vapour, (2) combined formaldehyde and acetyl chloride vapour, and (3) glyoxylic acid vapour treatment.     

Comparative quantitative analysis of sucrose and related compounds using ion exchange and reverse phase chromatographic methods

Two analytical methods of sugar determination, namely ion exchange chromatography on an anionic resin coupled with electrochemical detection, and reverse phase chromatography on Nucleosil-NH2 resin equipped with a light scattering detector were tested and compared as regards their rapidity, sensitivity and accuracy with sucrose, fructose, glucose, raffinose, maltose, arabinose, fucose, rhamnose and xylose. Excellent resolution and highly reproducible results were obtained in both cases. Greater sensitivity up to the picomolar range was possible however only with ion exchange chromatography. Reverse phase chromatography was successfully applied to the time course of sucrose hydrolysis under chemical (acid) and enzymatic (invertase) conditions. The hydrolysis was monitored by determining sucrose degradation and the corresponding formation of glucose and fructose.     

Electron capture gas chromatographic detection of thromboxane B2

An electron capture gas chromatographic method is described for the detection of thromboxane B2. Thromboxane B2 is esterified with diazomethane, followed by treatment with pentafluorobenzylhydroxylamine hydrochloride and silylation with BSA. In pyridine, the free aldehyde form of the acetal ring is favored allowing rapid formation of a novel thromboxane B2 pentafluorobenzyloxime. The method has been applied to detect thromboxane B2 formation during aggregation of washed platelets. It must be emphasized that by ordinary analytical standards, the derivatization reproducibility from 50-375 nanograms is poor (+/- 11% - +/- 42%); however, the improved selectivity of the method and its ability to detect nanogram levels of thromboxane B2 make it a useful complement to commonly employed bioassay techniques.     

Comparison of four RNA extraction methods for the detection of porcine reproductive and respiratory syndrome virus by RT-PCR

We investigated the effect of repeated cold stress (RCS) on the capsaicin-evoked release of glutamate from the primary afferent fibers of the rat, and compared this with the effect of inoculation of complete Freund's adjuvant (adjuvant inoculation). The release of glutamate was measured using a fluorometric on-line continuous monitoring system in which the immobilized glutamate dehydrogenase column was connected to an in vitro superfusion system. In the presence of 0.3 microM tetrodotoxin, the application of 1 microM capsaicin to spinal dorsal horn slices evoked glutamate release (18.6 +/- 1.2 pmol mg(-1) protein, n = 11). In rats subjected to RCS (RCS rats), the release of glutamate evoked by 1 microM capsaicin was markedly increased to 272% (n = 6, P < 0.05) of the value for the control group, although the basal release was not significantly altered (n = 6, P > 0.05). Adjuvant inoculation produced a significant increase in the basal and capsaicin (1 microM) evoked release of glutamate to 141 and 344% (n = 6, P < 0.05) of the value for the control group, respectively. The present results suggest that the facilitated release of glutamate from capsaicin-sensitive primary afferent terminals in the spinal dorsal horn is, at least in part, involved in the hyperalgesia of RCS rats as well as the complete Freund's adjuvant-induced hyperalgesia.     

Gas chromatographic and gas chromatographic-mass spectrometric analysis of ampicillin

A method was developed for the quantitative gas chromatographic (GC) determination of ampicillin. The procedure requires silylation iwth hexamethyldisilazane, trimethylchlorosilane, trimethylsilylimidazole and N,O-bis(trimethylsilyl)acetamide in pyridine and subsequent GC on an OV-17 column, using 5 alpha-cholestane as an internal standard. The method was applied to the determination of ampicillin in some pharmaceutical products. The characteristics of the mass spectra and the derivatization GC of ampicillin are also discussed.     

Revision of methods for analysis of volatile fractions in food

The volatile part of both, food and drinks, had been object of important researching, because of the "aroma" is considered as a principal characteristic of food, which in many times, is the cause of acceptance or refusing by consumers. Therefore, it sets up an interesting sign for appraisal in quality of the nutritious products. This part of food is formed by a big number of volatile substances very different among them and in concentrations; and this is the fact which makes that investigation of aromatic composition of food becomes complicated. There are many analyzing methods developed on this way. All of them will have to take account of this great diversity in quality as well as in quantity. In this study, we try to realize a bibliographic review about the techniques most used until our days, in extraction and concentration of the essential parts of the aroma of food, pointing out, in each of them, their basis, advantages and objections, as well as their applicability to the analysis of the aroma of food.     

Comparison of screening methods in the detection of bladder cancer

PURPOSE: We prospectively evaluate and compare the sensitivity and specificity of urine cytology, BTA stat, NMP22, fibrin/fibrinogen degradation products (FDP), telomerase, chemiluminescent hemoglobin and hemoglobin dipstick to detect bladder cancer. MATERIALS AND METHODS: Single voided specimens were obtained from 57 patients with bladder cancer, and 139 without evidence of bladder malignancy on cystoscopy or a negative biopsy of indeterminate lesions. A cytology report was available for 125 patients and interpreted independently. BTA stat, NMP22 and FDP were analyzed according to manufacturer specifications. The telomerase assay was performed on cells collected from urine by centrifugation in preparation for polymerase chain reaction based amplification using the telomeric repeat amplification protocol assay. The chemiluminescent screening assay for hemoglobin in urine uses the pseudoperoxidase activity of hemoglobin on hydrogen peroxide and subsequent oxidation of 7-dimethylaminonaphthalene-1,2-dicarbonic acid hydrazide to generate chemiluminescence emission. Hemoglobin dipstick was interpreted as positive if the hemoglobin content in the urine was trace or greater. RESULTS: Overall sensitivity with urine cytology, BTA stat, NMP22, FDP, telomerase, chemiluminescent hemoglobin and the hemoglobin dipstick was 44, 74, 53, 52, 70, 67 and 47%, respectively. Specificity with cytology, telomerase and FDP was high (95, 99 and 91%, respectively) but BTA stat, NMP22 (optimized), chemiluminescent hemoglobin (optimized) and the hemoglobin dipstick demonstrated lower specificity of 73, 60, 63 and 84%, respectively. Stepwise logistic regression analysis revealed that for all tumors, and within each tumor grade and stage telomerase had the strongest association with bladder cancer among all tests (69% overall concordance). Telomerase was also positive in 91% of the patients (10 of 11) with carcinoma in situ. CONCLUSIONS: Urinary telomerase had the highest combination of sensitivity and specificity (70 and 99%, respectively) for bladder cancer screening in these patients. It was the strongest predictor with superior accuracy in patients with grade 1 and noninvasive tumors (pTa), and extremely useful in patients with carcinoma in situ. Telomerase appears to be promising and outperformed cytology, BTA stat, NMP22, FDP, chemiluminescent hemoglobin and hemoglobin dipstick in the prediction of bladder cancer.     

Comparison of C18-carboxypropylbetaine and glass bead DNA extraction methods for detection of Mycobacterium bovis in bovine milk samples and analysis of samples by PCR

The purpose of this prospective study was to compare two different milk preparation methods to assay for the presence of Mycobacterium bovis by PCR. Detection by a C18-carboxypropylbetaine (CB-18)-based sample processing method was compared to extraction of DNA from milk with glass beads. Samples from 17 skin test-positive cattle were analyzed. Following CB-18 processing and glass bead extraction, the sensitivity of IS6110-based PCR was 94.1 and 58.8%, respectively (P < 0.025). Because CB-18 processing will permit the proficient use of PCR for diagnosis and surveillance of bovine tuberculosis, it will contribute to the more efficient detection and control of tuberculosis.     

Improved detection of polycyclic aromatic compounds in complex mixtures by liquid chromatographic fractionation on poly(divinylbenzene) prior to gas chromatography-mass spectrometry. Application to the analysis of diesel particulates

The Japanese pufferfish Fugu rubripes has a 400 Mb genome with high gene density and minimal non-coding complexity, and is therefore an ideal vertebrate model for sequence comparison. The identification of regions of conserved synteny between Fugu and humans would greatly accelerate the mapping and ordering of genes. Fugu C9 was cloned and sequenced as a first step in an attempt to characterize the region in Fugu homologous to human chromosome 5p13. The 11 exons of the Fugu C9 gene share 33% identity with human C9 and span 2.9 kb of genomic DNA. By comparison, human C9 spans 90 kb, representing a 30-fold difference in size. We have also determined by cosmid sequence scanning that DOC-2, a tumour suppresser gene which also maps to human 5p13, lies 6-7 kb from C9 in a head-to-head or 5' to 5' orientation. These results demonstrate that the Fugu C9/DOC-2 locus is a region of conserved synteny. Sequence scanning of overlapping cosmids has identified two other genes, GAS-1 and FBP, both of which map to human chromosome 9q22, and lie adjacent to the Fugu C9/DOC-2 locus, indicating the boundary between two syntenic regions.     

Determination of acids and volatile compounds in red Txakoli wine by high-performance liquid chromatography and gas chromatography

The Txakoli is a wine from the north of Spain with acid characteristics and medium ethanol content. We report the comparative results obtained from red Txakoli elaborated with different grape varieties. We have used chromatographic methods for the characterization of this wine. The volatile compounds were analyzed the means of gas chromatography and the organic acid content was determined by a newly validated liquid chromatographic procedure. The aim of this study is to characterize the red Txakoli and to know the major differences between Hondarrabi Beltza grape variety and the other varieties. Hondarrabi Beltza grape variety is the one which was awarded the Generic Label. The major differences observed in the samples obtained from Hondarrabi Beltza variety are their higher contents in ethanol, propanol, ethyl acetate and tartaric and malic acids.     

The gas chromatographic analysis of the higher fatty acids of bile lipids

The method of performing of abdominoanal rectal resection with anal hemiresection for tumor, localized in inferoampullar rectal portion and spreading on anus.     

Q390高强抗震钢板残余应力检测方法对比及分析

利用盲孔法、X-Ray法、中子衍射法和超声法对Q390高强抗震板及U形弯曲件进行残余应力检测,并对这4种方法测试得到的应力结果进行分析研究。结果表明,以上4种检测方法均要考虑其特定的检测应用范围,X-Ray法针对钢板表面下微米级深度的微米级晶格尺度范围局部点应力进行检测,中子衍射法可在毫米级尺度局部区域进行应力检测,盲孔法反映的是因毫米尺度钻孔导致的钢板表面孔周边的应力变化,超声法则可以通过连续扫描检测整个钢板残余应力的宏观分布变化。因此,对于工业钢板,超声法测试残余应力更加科学、合理、准确。同时,通过U形试样建立稳定的应力应变场,利用超声设备使用不同参考态对Q390高强抗震板弯曲件及退火弯曲件进行测试,结果表明,测试参考态的选取对应力测试结果影响很大,材料各向异性的影响对应力测试结果不可忽视。     

Coulometric detection in high-performance liquid chromatographic analysis of cholesteryl ester hydroperoxides

From the secretion of neurotransmitters via synaptic vesicles to the expulsion of cellular waste via contractile vacuoles, exocytosis and its sequel, endocytosis, are being explored with a variety of new optical tools. Fluorescent markers, especially styryl dyes such as FM1-43 (which reversibly labels endosomal membranes), have been used to follow exo- and endocytic events in many cell types. Even though the development of new dyes is still largely empirical, some theoretical principles have emerged to guide future dye chemistry. Moreover, advances in optical imaging technology that augment conventional fluorescence microscopy are appearing. For example, interference reflection microscopy (which requires no flurophore) and total internal reflection microscopy have recently been used to observe single exocytic events at the contact point between a glass coverslip and the plasma membrane.     

Comparison of four methods for the detection of fecal streptococci in water

The objective of this study was to select an effective and fast method for the detection of socalled fecal streptococci in water by comparing a method according to the German drinking water standard, a membrane filtration method according to the ISO-draft standard 7899/2, the Chromocult Enterococcus Broth (Merck) and the Enterolert-System (IDEXX). The study was based on a collective of 297 water samples derived from different stages of water treatment and distribution, as well as from individual water supplies. The sensitivity, reliability, and selectivity of the single methods in relation to their practicability was evaluated. Concerning false positive and false negative results, the tests were proved by metabolic characterization of the isolated strains. The advantages and disadvantages of the methods resulting from the investigated criteria are discussed. The work is part of a comparative study within the scope of the DIN ad-hoc-committee "fecal streptococci".