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PubMed1987

Purification and quantitative analysis of urinary prostanoids in human and in rat by packed and capillary gas chromatography-negative-ion chemical-ionization mass spectrometry.

We describe a method for the quantitative analysis of prostaglandin (PG) E2 and the major urinary metabolites PGI2 and thromboxane (Tx) A2 in human and in rat by combined gas chromatography and negative-ion chemical-ionization mass spectrometry. The procedure is based on the sequential use of small columns with distinct properties combined with a thin-layer chromatography step, for the extraction and the purification of urinary prostaglandins. The compounds are then analysed as their pentafluorobenzyl ester-O-methyloxime-trimethylsilyl ether derivatives, using either packed or capillary columns. Deuterated analogues are used as internal standards. The method was established by using tritiated prostaglandins covering the extremes of polarity in order to optimize the recovery of prostanoids as well as the quality of the chromatograms and spectra. The overall recovery was 24%. Standard curves were obtained by the same procedure and found to be reproducible, with a maximal day-to-day variation of +/- 5%. The relatively simple approach required for the sequential extraction and purification of prostaglandins on small columns of distinct properties, combined with the highly specific and highly sensitive method of detection, places this procedure among the most reliable method for measuring urinary prostanoids in both humans and animals. In addition, the procedure is faster than classical approaches and necessitates smaller amounts of samples and solvents.

6-Ketoprostaglandin F1 alpha↗
PubMed1983

Quantitative analysis of minaprine and some of its metabolites with application to kinetic studies in rats.

A simple and rapid high-performance liquid chromatographic method is described for the quantitative analysis of the psychotropic drug minaprine and three of its metabolites (M1, M3 and M11), including one as yet undetected metabolite (M11) known as a monoamine oxidase type A inhibitor in vitro. After selective extraction all four compounds were separated on a reversed-phase muBondapak C18 column using sodium acetate (0.03 M)-acetonitrile-methanol (88:7:5) (pH 3.3) as the mobile phase. The eluted compounds were detected with a UV detector at 254 nm. The sensitivity of the method is 0.02 microgram per millilitre of body fluid or per gram of tissue for M1 and M11 and 0.05 microgram per minaprine and M3. The method has been applied successfully to the determination of minaprine and the metabolites in plasma and brain and is compared here with an gas-liquid chromatographic method with an electron-capture detector previously developed for the detection of minaprine and M11. M11 was identified in rat urine by gas chromatography-mass spectrometry.

Animals↗
PubMed1977

A non-enzymic procdure for the quantitative analysis of (3-methoxy-4-sulphoxyphenyl)ethylene glycol(MHPG sulphate) in human urine using stable isotope dilution and gas chromatography-mass spectrometry.

A method is described for the quantitative analysis of (3-methoxy-4-sulphoxyphenyl)-ethylene glycol (MHPG sulphate) in human urine, based on selected ion monitoring gas chromatography--mass spectrometry and using a specifically deuterium-labelled analogue of MHPG sulphate as internal standard. The procedure involves extraction of the urine sample on Amberlite XAD-2, followed by isolation of MHPG sulphate by column chromatography on Sephadex LH-20. Cleavage of the sulphate conjugate and formation of the MHPG tris(trifluoroacetate) derivative are carried out in a one-step reaction, without recourse to enzymic hydrolysis.

Chromatography, Gas↗
PubMed1983

Quantitative analysis of aminophospholipids by high-performance liquid chromatography using succinimidyl 2-naphthoxyacetate as a fluorescent label.

A simple and rapid high-performance liquid chromatographic procedure for the quantitative analysis of ethanolamine- and serine-containing phospholipids in tissue is described. The technique involves reaction of lipid extracts with succinimidyl 2-naphthoxyacetate to give fluorescent derivatives of aminophospholipids. Reaction products are separated by a silica gel column with gradient elution. The eluate is monitored by fluorescence detection at 228 nm (excitation) and 342 nm (emission). Ethanolamine and serine plasmalogens can be measured indirectly by converting their derivatives into lysophosphatidylethanolamine and lysophosphatidylserine derivatives with exposure to hydrogen chloride fumes. The method is highly sensitive and selective.

Animals↗
PubMed1999

Quantitative analysis of CD1a (T6) positive Langerhans cells in human tonsil epithelium.

Fifty-one human palatine tonsils of both sexes and 4-54 years of age were studied for quantitative analysis of Langerhans cells in the epithelium using CD1a (T6), which is a specific immunological marker for Langerhans cells and indeterminate cells. Cryo-sections were stained using the avidin biotin peroxidase method. Using light microscopy, CD1a-positive dark brown cells with dendritic processes were identified as Langerhans cells, which were located in the epithelium, subepithelial tissue, follicles and interfollicular areas. The Langerhans cells were counted only in the tonsil epithelium per zone of 1.1 mm length of basement membrane. For each biopsy, 25 such zones were studied. The mean number (SEM) of Langerhans cells per zone of tonsil epithelium was found to be 37 (+/- 0.5). In the male, it was 36 (+/- 0.7) but in the female, it was 38 (+/- 0.2). In different age subgroups, the mean number (SEM) varied between 40 (+/- 1.7) and 14 (+/- 1.1). In the age subgroups of 11-15, 16-20, and 21-25 years, the mean number showed significant sex differences. Since the 11-15, 16-20, and 21-25 age subgroups in the female showed an increased number of Langerhans cells, it is concluded that the immunological role of the palatine tonsils is increased during puberty and adolescence. In the female, there was a negative correlation (r = -0.196, p < 0.01) between age subgroups and mean numbers, but in the male there was no correlation (r = 0.008). Overall, in all the 51 biopsies together, there was a negative correlation (r = -0.017, p < 0.01) and significant (p < 0.001) sex and age differences.

Adolescent↗
PubMed2006

Quantitative analysis of ochratoxin A in wine and beer using solid phase extraction and high performance liquid chromatography-fluorescence detection.

A reliable and accurate method is described for the quantitative analysis of ochratoxin A (OTA) in wine and beer. The method involves the use of disposable non-polar polymeric and aminopropyl solid-phase extraction cartridges to isolate the mycotoxin from alcoholic beverages. Extracts were subsequently analysed using reverse-phase high-performance liquid chromatography-fluorescence detection with post column ammoniation to improve the limit of detection. The precision of the method determined at three levels in both wine and beer was less than 5% (RSD). Standard addition studies in both wine and beer showed that the recovery of OTA varied between 90 and 106% over a concentration range of 0.016-1.284 microg l-1. The detection and quantification limits were shown to be better than 0.004 (S/N = 3) and 0.016 microg l-1 (S/N = 10) for both beer and wine.

Beer↗
PubMed2005

RP-HPLC method for the quantitative analysis of naturally occurring flavonoids in leaves of Blumea balsamifera DC.

A selective and sensitive reversed-phase (RP) high-performance liquid chromatographic method is developed for the quantitative analysis of five naturally occurring flavonoids of Blumea balsamifera DC, namely dihydroquercetin-7,4'-dimethyl ether (DQDE), blumeatin (BL), quercetin (QN), 5,7,3',5'-tetrahydroxyflavanone (THFE), and dihydroquercetin-4'-methyl ether (DQME). These compounds have been isolated using various chromatographic methods. The five compounds are completely separated within 35 min using an RP C18, Nucleosil column and with an isocratic methanol-0.5% phosphoric acid (50:50, v/v) mobile phase at the flow rate of 0.9 mL/min. The separation of the compounds is monitored at 285 nm using UV detection. Identifications of specific flavonoids are made by comparing their retention times with those of the standards. Reproducibility of the method is good, with coefficients of variation of 1.48% for DQME, 2.25% for THFE, 2.31% for QN, 2.23% for DQDE, and 1.51% for BL. The average recoveries of pure flavonoids upon addition to lyophilized powder and subsequent extraction are 99.8% for DQME, 99.9% for THFE, 100.0% for BL, 100.6% for DQDE, and 97.4% for QN.

Asteraceae↗
PubMed1989

Quantitative analysis of the retinal ganglion cell layer and optic nerve of the barn owl Tyto alba.

The visual capacity of the common barn owl (Tyto alba) was studied by quantitative analysis of the retina and optic nerve. Cell counts in the ganglion cell layer of the whole-mounted retina revealed a temporal area centralis with peak cell density of 12,500 cells/mm2 and a horizontal streak of high cell density extending from the area centralis into the nasal retina. Integration of the ganglion cell density map gave an estimated total of 1.4 million cells for the ganglion cell layer. Electron microscopy of a single, complete section of the optic nerve revealed a bimodal fiber diameter spectrum (modes at 0.3 and 0.9 microns; bin width = 0.2 microns), with diameters ranging from 0.15 microns (unmyelinated) to 6.05 microns (myelinated, sheath included). The total axon count for the optic nerve was estimated from sample counts to be about 680,000 axons (25% unmyelinated). Therefore, roughly half of the cells in the retinal ganglion cell layer do not send axons into the optic nerve. With certain assumptions, the data predict a visual spatial acuity for barn owls on the order of 8 cycles/degree, a value similar to the known behaviorally measured acuities of masked owls (10 cycles/degree) and domestic cats (6 cycles/degree).

Animals↗
PubMed2006

Quantitative analysis of cytochrome C released from rice mitochondria using the adsorptive polarographic wave of guanidine modified Co(II)-cytochrome C complex.

A new, simple and sensitive method for the quantitative analysis of cytochrome C (Cyt C) based on the reduction wave of guanidine modified Co(II)-Cyt C complex at about -1.74 V (vs. SCE) by single sweep polarography in the solution containing 8 x 10(-6) mol L(-1) CoCl2, 0.04 mol L(-1) guanidine hydrochloride, 0.2 mol L(-1) NaOH and 0.5% Na2SO3. The peak height is linearly proportional to the concentration of Cyt C in the range of 0.005 approximately 1.500 mg L(-1) (correlation coefficient 0.999). Common amino acids, saccharide, organic acid and metal ions of appropriate concentrations have no interference on the Cyt C determination. The released Cyt C in the process of mitochondrial permeability transition of Hong-Lian cytoplasmic male sterile line of rice has been measured by the method, and the result is satisfactory.

Amino Acids↗
PubMed1991

[Prediction of one second forced expiratory volume after lung resection using multiple view lung perfusion images and "regions of interest" quantitative analysis].

In 17 patients mostly with bronchial carcinoma undergoing pneumonectomy (N = 2) or lobectomy (N = 15) multiple view lung perfusion images with 99mTc-MAA and "regions of interest" quantitative analysis of functional distributions to each lung or lobe (MV-ROI) were used for predicting postoperative FEV1. For comparison, other two methods were also conducted, i.e. the split function study by lung perfusion images combined with subtraction of segmental function (SF+SS) and the simple subtraction of segmental function (SSS). The correlation between predicted postoperative FEV1 and actual FEV1 measured 3-6 months after lung resection was better by MV-ROI method than by the other two. In all patients the correlation coefficient (r) was 0.957, 0.923 and 0.887 by three methods respectively; in lobectomy patients the (r) was 0.948, 0.907 and 0.869 by three methods respectively (all P less than 0.001). The mean error (liters), mean percent error and median of percent error between predicted and observed FEV1 in MV-ROI were less than in SF+SS or SSS. The difference between error of MV-ROI and error of SF+SS or SSS was significant, while the difference between error of SF+SS and error of SSS was insignificant. The number of cases having the error greater than 10% was 3,5,8 in all patients and 3,5,7 in lobectomy patients by three methods respectively. There was a tendency of overestimation in MV-ROI, which was attributed to the old age of our patients whose pulmonary functions recovered slowly and to the short period of the follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Quantitative analysis of DNA aberrations amplified by competitive polymerase chain reaction using capillary electrophoresis.

We compared the use of capillary electrophoresis (CE) in a polymer network with the use of slab gel electrophoresis for the quantitative analysis of polymerase chain reaction (PCR)-amplified DNA samples. We quantified residual lymphoma cells carrying a translocation between chromosomes 14 and 18, in consecutive patient peripheral blood samples that were amplified by competitive PCR. For CE analysis we used a 4% linear polyacrylamide network. Results show that the calculated number of translocations in patient samples using both analyses were comparable. We conclude that CE is a sensitive, non-radioactive, fast and accurate method for quantitation of competitive PCR products.

Autoradiography↗
PubMed1988

Quantitative analysis of 6-keto-prostaglandin F1 alpha using immunoaffinity purification and gas chromatography-mass spectrometry.

This paper describes an immunoaffinity purification technique for 6-keto-prostaglandin F1 alpha (6KPGF1 alpha) prior to quantitative analysis by high-resolution gas chromatography-negative-ion chemical ionization mass spectrometry (HRGC-NICIMS). Polyclonal antibodies to 6KPGF1 alpha were partially purified using Staphylococcus aureus Protein A immobilized on Sepharose CL-4B. This partially purified fraction was covalently bound to silica gel using N-hydroxysuccinimidyl-functionalized silica. Columns constructed using this gel quantitatively bound 6KPGF1 alpha which could be eluted quantitatively with acetonitrile-water (19:1). Binding capacity was reconstituted by washing with 0.01 M phosphate buffer (pH 7.4). Human urinary and canine plasma 6KPGF1 alpha was sufficiently purified using these columns that HRGC-NICIMS analysis of the methoxime-pentafluorobenzyl-tris-trimethylsilyl derivative was interference-free.

6-Ketoprostaglandin F1 alpha↗
PubMed2005

Quantitative analysis of regional airways obstruction using dynamic hyperpolarized 3He MRI-preliminary results in children with cystic fibrosis.

PURPOSE: To investigate regional airways obstruction in patients with cystic fibrosis (CF) with quantitative analysis of dynamic hyperpolarized (HP) (3)He MRI. MATERIALS AND METHODS: Dynamic radial projection MRI of HP (3)He gas was used to study respiratory dynamics in a group of eight children with CF. Signal kinetics in a total of seven regions of interest (ROIs; three in each lung, and one in the trachea) were compared with the results of spirometric pulmonary function tests (PFTs). The tracheal signal intensity was used as a form of "input function" to normalize for input flow effects. RESULTS: A pattern of low flow rate in the upper lobes was observed. When the flow measurements from the peripheral ROIs were averaged to obtain an index of flow in the peripheral lung, a good correlation was found (P = 3.74 x 10(-5)) with the forced expired volume in one second (FEV1). CONCLUSION: These results suggest that a quantitative measurement of localized airways obstruction in the early stages of CF may be obtained from dynamic (3)He MRI by using the slope of the signal rise as a measure of air flow into the peripheral lung. This study also demonstrates that children can cooperate well with the (3)He MRI technique.

Adolescent↗
PubMed1998

Application of artificial neural networks for quantitative analysis of image data in chest radiographs for detection of interstitial lung disease.

The authors have developed an automated computeraided diagnostic (CAD) scheme by using artificial neural networks (ANNs) on quantitative analysis of image data. Three separate ANNs were applied for detection of interstitial disease on digitized chest images. The first ANN was trained with horizontal profiles in regions of interest (ROIs) selected from normal and abnormal chest radiographs for distinguishing between normal and abnormal patterns. For training and testing of the second ANN, the vertical output patterns obtained from the 1st ANN were used for each ROI. The output value of the second ANN was used to distinguish between normal and abnormal ROIs with interstitial infiltrates. If the ratio of the number of abnormal ROIs to the total number of all ROIs in a chest image was greater than a specified threshold level, the image was classified as abnormal. In addition, the third ANN was applied to distinguish between normal and abnormal chest images. The combination of the rule-based method and the third ANN also was applied to the classification between normal and abnormal chest images. The performance of the ANNs was evaluated by means of receiver operating characteristic (ROC) analysis. The average Az value (area under the ROC curve) for distinguishing between normal and abnormal cases was 0.976 +/- 0.012 for 100 chest radiographs that were not used in training of ANNs. The results indicate that the ANN trained with image data can learn some statistical properties associated with interstitial infiltrates in chest radiographs.

Humans↗
PubMed2000

Approaches towards the quantitative analysis of peptides and proteins by reversed-phase high-performance liquid chromatography in the absence of a pure reference sample.

A reversed-phase HPLC protocol for the quantitative analysis of peptides and proteins is presented. It is applicable to purified samples and potentially to crude biological extracts. The key feature is that an analytically pure reference sample of the analyte is not required because the extinction coefficient for the UV absorbance at 280 nm can be accurately estimated from the amino acid sequence. The concentration of a protein can therefore be calculated from the peak area relative to an internal standard. Sources of error and limitations of the method are systematically considered. Tryptophan containing peptides gave closer agreement to expected values than those with only tyrosine. It was found that analogous, previously used methods could not be directly applied to lower wavelengths.

Chromatography, High Pressure Liquid↗
PubMed1978

Quantitative analysis of 6,11-dihydro-11-oxo-dibenz[b,e] oxepin-2-acetic acid (isoxepac) in plasma and urine by gas--liquid chromatography.

A method is described for the quantitative analysis of 6,11-dihydro-11-oxo-dibenz[b,e]-oxepin-2-acetic acid (isoxepac) in plasma and urine. Isoxepac and internal standard was analysed by gas-liquid chromatography using a flame ionization detector. The method is accurate and precise over the range 0.1--30 microgram/ml. The method has been applied to the analysis of plasma and urine from both healthy volunteers and patients receiving therapeutic oral doses of isoxepac.

Acetates↗
PubMed1992

Quantitative analysis of CT brain images: a statistical model incorporating partial volume and beam hardening effects.

The purpose of this study was to construct and evaluate a statistical model for the quantitative analysis of computed tomographic brain images. Data were derived from standard sections in 34 normal studies. A model representing the intracranial pure tissue and partial volume areas, with allowance for beam hardening, was developed. The average percentage error in estimation of areas, derived from phantom tests using the model, was 28.47%. We conclude that our model is not sufficiently accurate to be of clinical use, even though allowance was made for partial volume and beam hardening effects.

Adult↗
PubMed2005

Determination of the percentage of homopolymer component in Ziegler/Natta catalyst linear low-density polyethylene resins using high-temperature cell Fourier transform infrared and partial least squares quantitative analysis technique.

A new method for the determination of the percentage of homopolymer component, using high-temperature cell Fourier transform infrared (FT-IR) by partial least squares (PLS) quantitative analysis technique, was developed and applied to Ziegler Natta linear low-density polyethylene (LLDPE). The method is based on the IR spectrum changes between the 730 cm(-1) band and 720 cm(-1) band at the temperature of 110 degrees C, which is near the melting point of the polyethylene. The HD % (the percentage of high-density component, i.e., the percentage of homopolymer component) results obtained by CTREF (CRYSTAF in TREF mode) technique are used as the input data together with the respective FT-IR spectra for PLS analyses to establish a calibration curve. The PLS quality is characterized by a correlation coefficient of 0.997 (cross-validation) using four factors and a root mean square error of calibration (RMSEC) of 0.772. The HD% of the unknown can then be predicted by the PLS software from the unknown FT-IR spectrum. A control resin was tested seven times by CTREF and FT-IR. The HD% of the control resin was 28.59+/-0.88% by CTREF and 29.05+/-2.37% by FT-IR. It was found that the method was applicable for the same comonomer type of LLDPE within a melt index range and density.

Algorithms↗