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Quantitative analysis of d-tubocurarine chloride in curare by column liquid chromatography.

An improved quantitative analysis of d-tubocurarine chloride in the plant extract curare is presented. Gradient high-performance liquid chromatography on a hydrophobic stationary phase was found to be very suitable for the analysis of quaternary ammonium bases such as the complex mixture of curare alkaloids. Owing to the residual free silanol groups on the modified silica surface, the curare alkaloids are eluted from a reversed-phase column only if an electrolyte is added to the mobile phase. In order to optimize the separation, the effects of pH, the nature of the cation in the buffer and the concentration of the buffer of the retention of the alkaloids were investigated. Using a tetramethylammonium phosphate buffer at pH 4 in a gradient of water-methanol, undesirable retardation effects on the reversed-phase column could be suppressed sufficiently. As a result, an accurate method for the determination of d-tubocurarine chloride in curare was obtained. The coefficient of variation of this analysis is only 1.3%.

Chromatography, High Pressure Liquid↗

[Quantitative analysis of (-)-epigallocatechin gallate in tea leaves by high-performance liquid chromatography].

The quantitative analysis of (-)-epigallocatechin gallate (EGCG) in tea (Camellia sinensis L.) was performed by high-performance liquid chromatography (HPLC) with a C-18 reversed-phase column. EGCG was then eluted within 20 min by using methanol-water-acetic acid (20:75:5 (v/v/v)) as an eluent. As an internal standard, tryptophan was used. The content of EGCG in five kinds of green tea (sencha, gyokuro, bancha, matsucha and oolong tea) and in a cup of those was determined by both the extraction method with 50% (v/v) methanol and the infusion method with water. The largest amount of EGCG was obtained from matsucha by the extraction method, or from sencha by the infusion method. Furthermore, EGCG contents in various parts of the tea plant were examined. The first leaf had the highest concentration of EGCG, and the concentration of EGCG decreased with the aging of the leaf.

Catechin↗

[Quantitative analysis of deacylgymnemic acid by high-performance liquid chromatography].

A method of the quantitative analysis was established for the determination of deacylgymnemic acid (DAGA) in the alkaline hydrolysate of the sample containing gymnemic acids which are ingredients of Gymnema sylvestre R. BR. leaves, by means of high-performance liquid chromatography. This method was used for comparing the contents of gymnemic acids in various samples. The amount of gymnemic acids analyzed as DAGA in 70% ethanol extract of dry leaves was about twice that in hot water extract. The commercial health-supplemental foods of five companies were investigated for the contents of gymnemic acids as DAGA and there were large differences from 38 to 251 mg in the dosage per day recommended by each company.

Chromatography, High Pressure Liquid↗

A simple quantitative analysis system for coronary cineangiograms using a personal computer.

We have developed a simple quantitative analysis system for coronary cineangiograms using a personal computer and a standard projector-video camera system. The selected frame of cinefilm was projected onto the target area of a CCD video camera. To decrease spatial fluctuations due to quantum noise, the digital data were smoothed spatially by applying a moving averaged filter and a median filter. Following the smoothing process, the digital data with gray level were transformed to binary data by quantifying the threshold property of their gray level. To minimize the geometric influence, mainly pincushion effect, a cinefilm of a 1 cm grid was placed against the input screen of the image intensifier to correct the distortion by using 3rd degree polynomials. The accuracy of the contour detection procedure was validated on the basis of phantom models filled with a contrast medium. Despite the potential influence of many radiographic variables on computerized diameter measurements, the overall accuracy was found to be 0.26-0.33 mm over a wide range of clinically relevant artery diameters and radiographic conditions. Using this system we demonstrated one method of examining the effects of acetylcholine and nitroglycerin on coronary artery diameter in adult humans.

Acetylcholine↗

[Quantitative analysis of color of the nasal mucosa].

The color of the nasal mucosa was quantitatively studied. Chromaticity of the nasal mucosa was indicated by two parameters, x (red component in RGB) and y (green in RGB) values, measured with a chromameter (MINOLTA Co., CS-100). According to preliminary studies in normal subjects, the following conditions were thought to make the method suitable for measuring the color of the nasal mucosa. 1) The chromameter can be held in the hand. 2) Brightness of the nasal mucosa should be kept as constant as possible. 3) The inferior turbinate mucosa is easier to measure than the septal one. In the preliminary study there was no significant difference in chromaticity measurements among the examiners. Chromaticity of the inferior turbinate was compared among patients with perennial nasal allergy (PNA, n = 19), with cedar pollinosis (CP, n = 11), and normal subjects serving as a control (n = 15). The x values in the PNA and CP groups were significantly lower than those in the control group (p < 0.01), and the y values were significantly higher (p < 0.02). The results agreed with the macroscopic observations made by trained rhinologists. The x values increased when azelastin was administered in five of the PNA group. This increase was related to improvement of their subjective symptoms. Quantitative analysis of the nasal mucosal color could be useful for objective evaluation of the clinical course of allergic rhinitis.

Color↗

[The reflection spectrometer and quantitative analysis design].

The reflection spectrometer with optical fibre transmission was designed and used in quantitative analysis. The reflection spectrometry was applied to the determination of reducing sugar in samples based on the reduction reaction of the reducing sugar with Fehling's reagent. The reduction reaction was carried out in a micro-reaction cell, and the reddish colored Cu2O produced was settled at the bottom of the reaction cell. The reflectance R(infinity), which is directly proportional to the amount of Cu2O produced, was measured by the reflection spectrometer. The fundamental principle, effect factors, and experimental conditions of the method were discussed. Linear relationship was obtained in the range of 20-120 microg of reducing sugar. The special features of this method were its simplicity in operation, relatively high sensitivity of determination, and the use of small amount of reagent. Satisfactory results were obtained in its application to the analysis of wine and honey samples.

English Abstract↗

Quantitative analysis of individual bile acids by gas-liquid chromatography: an improved method.

The quantitative analysis of individual bile acids by gas-liquid chromatography has been improved by column oven temperature programming and by a new liquid phase, SP-2401. The method is fast; bile acids are well resolved; retention times are reproducible; detector responses are linear and sensitive to 0.1 mug: and there is little adsorption onto the liquid phase. The method has been successfully used for bile, and it has the potential for use on serum.

Animals↗

Serial and quantitative analysis of mixed hematopoietic chimerism by PCR in patients with acute leukemias allows the prediction of relapse after allogeneic BMT.

Within a prospective study we analyzed hematopoietic chimerism in serial peripheral blood samples taken from 55 patients with acute leukemias (ALL 21, AML 20, MDS 14) with a median age of 13.5 years at very short time intervals following allogeneic bone marrow transplantation (allo-BMT). The investigation was performed to determine the implications of mixed hematopoietic chimerism (MC) with regard to the clinical outcome in patients with acute leukemias after allo-BMT. Analysis of chimerism was performed by PCR of variable number of tandem repeat (VNTR) sequences with a maximum sensitivity of 0.8%. Thirteen male patients transplanted with the marrow of a female donor were also studied by amplification of a Y-chromosome-specific alphoid repeat (0.1-0.01% sensitivity). VNTR analysis in 55 patients revealed complete chimerism (CC) in 36 cases, MC in 18 follow-ups and autologous recovery in one patient. Quantitative analysis of MC identified 10/18 patients with increasing autologous patterns in whom 9/10 subsequently relapsed. The patient with autologous recovery relapsed as well. Eight of 18 patients with MC showed decreasing amounts of autologous DNA and became CC upon further follow-up. In contrast, only 7/36 patients with CC in the prior analysis of chimerism status relapsed. However, in 4/7 patients the interval between last CC confirmation and relapse was more than 4 months. In 2/7 patients autologous DNA was not detectable in peripheral blood but in bone marrow aspirates. One of these seven patients developed a fulminant relapse within 3 weeks. The probability of relapse-free survival for patients with CC is 0.67 (n = 36), for patients with decreasing MC 1.0 (n = 8) and for patients with increasing MC 0.1 (n = 10). In summary, the results demonstrate that serial and quantitative chimerism analysis at short time intervals by PCR provides a reliable and rapid screening method for the early detection of recurrence of underlying disease and is therefore a prognostic tool to identify patients at highest risk of relapse.

Acute Disease↗

Quantitative analysis of dimethyl titanocene by iodometric titration, gas chromatography and NMR.

In this study we report the use of an automated iodometric titration method and a novel gas chromatography (GC) method for the quantitative analysis of dimethyl titanocene (DMT), a key raw material in drug synthesis. Both approaches are based on the reaction of DMT in toluene or tetrahydrofuran solutions with iodine. In the case of iodometric titration, excess iodine is titrated with a standardized aqueous sodium thiosulfate solution to a potentiometric end-point for the determination of DMT concentration. Alternatively, GC is employed to measure the concentration of iodomethane, a product of the reaction between DMT and iodine, in order to determine the concentration of DMT in the solution. Excellent agreement between iodometric titration, GC and NMR results using several DMT samples confirms the accuracy of the two methods and strongly supports the use of either method as a replacement to the expensive NMR for quantitative DMT analysis. The relatively few sources of error associated with the two methods, their ubiquitous nature and ease of application in routine analysis make them the analytical methods of choice, among all. Both methods have been validated according to ICH requirements. The use of iodometric titration method for DMT analysis is demonstrated with a couple of applications.

Chromatography, Gas↗

Quantitative analysis of two opioid peptides in plasma by liquid chromatography-electrospray ionization tandem mass spectrometry.

Quantitative analysis of two opioid peptides, DSLET [(D-Ser2)Leu-enkephalin-Thr6] and Met-enkephalin-Arg-Gly-Leu, was performed using microbore liquid chromatography interfaced to electrospray ionization tandem mass spectrometry. Validation of the methodology was demonstrated for each peptide in plasma. Quantitative analyses were performed through the use of a deuterium labelled peptide analog as an internal standard. Linearity was observed for the analysis of DSLET (5-1000 ng/ml) and Met-enkephalin-Arg-Gly-Leu (1-1000 ng/ml) in plasma with a limit of detection of 0.25 ng/ml for Met-enkephalin-Arg-Gly-Leu and 1.0 ng/ml for DSLET. In general, the observed concentrations showed good reproducibility with coefficients of variation of within 15%. In the concentration range studied, only 0.5 ml of plasma was required for optimal detection of Met-enkephalin-Arg-Gly-Leu and 0.25 ml for DSLET. Application of this method was demonstrated by studying the disposition of DSLET in a rat. DSLET administered to a rat exhibited a short half-life and a high clearance value.

Animals↗

Simplified micro-method for the quantitative analysis of putrescine, spermidine and spermine in urine.

A simplified micro-method for the quantitative analysis of urinary polyamines is described. After acid hydrolysis of urine, the polyamines are converted to fluorescent 1-dimethylaminonaphthalene-5-sulfonyl (Dns; dansyl) derivatives and separated by means of thin-layer chromatography. Dns-NH2, which has been reported to interfere with the determination of putrescine, is well separated from di-Dns-putrescine. Putrescine, spermidine and spermine are quantitated by in situ scanning of their fluorescent spots on the chromatogram. The present method is both sensitive and reproducible. It eliminates a number of time-consuming steps and thus reduces preparative losses. Yet an adequate chromatographic resolution is obtained. Representative polyamine analyses of urine from normal volunteers and from cancer patients are reported. Elevated levels occur in the urines of pregnant women and of patients with various types of cancer.

Chromatography, Thin Layer↗

Qualitative and quantitative analysis of uroliths in dogs: definitive determination of chemical type.

Effective treatment and prevention of urolithiasis depends on accurate determination of the chemical nature of the uroliths. A widely used qualitative chemical procedure was compared with quantitative crystallographic analysis of 272 canine uroliths. Agreement between the 2 methods was 78%. Qualitative analysis failed to detect 62% of calcium-containing uroliths and 83% of carbonate apatite uroliths. Qualitative analysis gave false-positive results for urates in 55% of cystine uroliths. Mixed uroliths comprising 6% of the total could not be classified without quantitative analysis. Silicate, cystine, and urate uroliths generally were of pure composition. Crystallographic analysis indicated the following distribution of major types: struvite, 69%; calcium oxalate, 10%; urate, 7%; silicate, 3.5%; cystine, 3.2%; calcium phosphate, 1%; and mixed, 6%. Among dogs with struvite uroliths, 66% had positive results of bacterial culturing from the urinary bladder. Six breeds (Miniature Schnauzer, Welsh Corgi, Lhasa Apso, Yorkshire Terrier, Pekingese, and Pug) had a significantly higher risk for urolithiasis, compared with other breeds. The German Shepherd Dog had a significantly lowered risk, compared with other breeds. Two breeds had significant relationship to a specific type of urolith: Miniature Schnauzer for oxalate, and Dalmatian for urate (P less than 0.001). It was concluded that quantitative analysis, using crystallography, was superior for the detection of calcium oxalate, carbonate apatite, cystine, urate, and mixed uroliths.

Animals↗

Chemical characterization of the aroma of Grenache rosé wines: aroma extract dilution analysis, quantitative determination, and sensory reconstitution studies.

The aroma of a Grenache rosé wine from Calatayud (Zaragoza, Spain) has been elucidated following a strategy consisting of an aroma extract dilution analysis (AEDA), followed by the quantitative analysis of the main odorants and the determination of odor activities values (OAVs) and, finally, by a series of reconstitution and omission tests with synthetic aroma models. Thirty-eight aroma compounds were found in the AEDA study, 35 of which were identified. Twenty-one compounds were at concentrations higher than their corresponding odor thresholds. An aroma model prepared by mixing the 24 compounds with OAV > 0.5 in a synthetic wine showed a high qualitative similarity with the aroma of the rosé wine. The addition of compounds with OAV < 0.5 did not improve the model, whereas the aroma of a model containing only odorants with OAV > 10 was very different from that of the wine. Omission tests revealed that the most important odorant of this Grenache rosé wine was 3-mercapto-1-hexanol, with a deep impact on the wine fruity and citric notes. The synergic action of Furaneol and homofuraneol also had an important impact on wine aroma, particularly in its fruity and caramel notes. The omission of beta-damascenone, which had the second highest OAV, caused only a slight decrease on the intensity of the aroma model. Still weaker was the sensory effect caused by the omission of 10 other compounds, such as fatty acids and their ethyl esters, isoamyl acetate, and higher alcohols.

Chromatography, Gas↗

Quantitative analysis of dipyridamole-thallium images for the detection of coronary artery disease.

To determine if the detection of coronary artery disease by dipyridamole-thallium imaging is improved by quantitative versus qualitative analysis, and combining quantitative variables, 80 patients with chest pain (53 with and 27 without coronary artery disease) who underwent cardiac catheterization were studied. Segmental thallium initial uptake, linear clearance, monoexponential clearance and redistribution were measured from early, intermediate and delayed images acquired in three projections. Normal values were determined from 13 other clinically normal subjects. When five segments per view were used for quantitative analysis, sensitivity and specificity were 87 and 63%, respectively, for uptake, 77 and 67% for linear clearance, 60 and 60% for monoexponential clearance and 62 and 56% for redistribution. Of the four variables, uptake and linear clearance were the most sensitive (p less than 0.01) and specificity did not differ significantly. Using three segments per view, the specificity of uptake increased (p less than 0.05) to 78% without a significant change in sensitivity (85%). With this approach, sensitivity and specificity did not differ from those of qualitative analysis (85 and 78%, respectively). Stepwise logistic regression analysis demonstrated that the best quantitative thallium correlate of the presence of coronary artery disease was a combination variable of "either abnormal uptake or abnormal linear clearance, or both." Using five segments per view, the model's specificity (85%) was greater than that of uptake alone (p less than 0.02), with similar sensitivity (92%). Using three segments per view, the model's specificity (93%) was greater than that of uptake alone (p less than 0.05) and of qualitative analysis (p less than 0.05), with similar sensitivity (85%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Quantitative analysis of mRNA as a marker for viability of Mycobacterium tuberculosis].

OBJECTIVE: To study whether quantitative analysis of M. tuberculosis mRNA could be used to assess bacterial viability. METHODS: The levels of M. tuberculosis mRNA were compared 24, 48, and 72 hours after M. tuberculosis H(37)R(V) was treated with no drug, 1.0 micro g/ml and 10.0 micro g/ml rifampicin, 0.2 micro g/ml and 1.0 micro g/ml isoniazid, 2 micro g/ml and 4 micro g/ml ethambutol, 2 micro g/ml and 4 micro g/ml streptomycin, 1.25 micro g/ml and 5.00 micro g/ml ofloxacin. The levels of mRNA were determined by quantitative PCR. RESULTS: After exposure to isoniazid, rifampicin, and streptomycin for 24 h, the level of M. tuberculosis H(37)R(V) was reduced markedly. Similarly, after exposure to isoniazid, streptomycin, ethambutol, and ofloxacin for 72 h, the level of M. tuberculosis H(37)R(V) was identical, but higher than that after exposure to rifampicin. Exposure of M. tuberculosis H(37)R(V) to rifampicin for 24 h reduced the levels of 85B mRNA to 0.02% of the control; exposure to other drugs for 24 h reduced the levels to 1% approximately 10% of the control, and for 72 h to less than 1% of the control. CONCLUSIONS: The levels of mRNA can show the difference between M. tuberculosis H(37)R(V) exposed and unexposed to drugs, and are consistent with colony forming unit. mRNA could be used as a marker for assessing the viability of M. tuberculosis.

Biomarkers↗

Quantitative analysis of neuronal damage induced by tri-ortho-cresyl phosphate in Wistar rats.

A quantitative analysis of neuronal damage was performed on the fasciculus gracilis (FG) of the cervical spinal cord in male Wistar rats that received orally a single dose of tri-ortho-cresyl phosphate (TOCP) at 1500 mg/kg. FG tissues were sampled at 1, 2, and 3 weeks after treatment and examined histopathologically. Wallerian degeneration of myelinated nerve fibers was observed in FG at 2 weeks. Morphological changes were most evident at 3 weeks after treatment and the number of fibers was reduced. Ultrastructurally, axonal swelling due to the accumulation of cytoplasmic contents was observed near the node of Ranvier in the affected animals, indicating paranodal degeneration. Axonal atrophy and swelling in organophosphorus-induced delayed neuropathy (OPIDN) were evaluated quantitatively using a computer-assisted image analyzer. Morphometric examinations on semi-thin sections and frozen sections stained with Nauta's method were demonstrated to be useful for objective evaluation of OPIDN in the rat.

Animals↗

Quantitative analysis of two-dimensional gel electrophoresis protein patterns: a method for studying genetic relationships among Globodera pallida populations.

A method based in two-dimensional protein gel electrophoresis has been developed in order to improve the analysis of genetic relationships among populations of Globodera. It has been used to estimate genetic divergence among nine Globodera pallida nematode populations. Sixty-one anonymous polypeptide spots were resolved using silver-stained high-resolution 2D gels and they were quantified in each population to establish genetic variation among G. pallida populations. The results of this analysis were compared with those obtained after a study of allelic frequency variation, which was carried out using seven previously described loci. Genetic distances among populations were calculated by means of both studies, the quantitative analysis and the allelic frequency variation, and phylogenetic trees were constructed for each type of analysis. A correlation analysis between the two distance matrices was carried out and a bootstrap analysis was performed to determine the strength of the clusters obtained with each method. The results obtained support the idea that quantitative protein analysis can be successfully applied to phylogenetic analysis of G. pallida populations.

Animals↗

[Quantitative analysis of 123I-metaiodobenzylguanidine myocardial scintigraphy by myocardial uptake using a phantom].

To evaluate the quantitative analysis of 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy, total injected dose measured by first pass (FP) method (TFP) was compared with that measured by phantom method using an acrylic phantom in 45 patients with cardiac disease. Heart per mediastinum ratio (H/M) was compared to myocardial uptake calculated with TFP. The total injected dose measured using the phantom in which the syringe was set in depth of 3.5 cm (Tpham) was correlated with TFP (r = 0.73, p = 0.0001). When Tpham was corrected by body weight (c-Tpham), c-Tpham showed better correlation with TFP. MU calculated by TFP (MU-FP) was well correlated with MU by c-Tpham (MU-pham) (r = 0.94, p = 0.001). These results indicate that phantom method is sufficient to substitute for FP method. Though H/M was correlated with MU-FP (p < 0.001), the interpatient variation was relatively large. Then the analysis by H/M is insufficient to substitute for the myocardial uptake. It is though to be enough to use the phantom method on daily routine work, since this method is accurate and easy to quantitate the myocardial uptake of MIBG taking a short time.

3-Iodobenzylguanidine↗