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Salbutamol identification in liver and urine by high-performance thin-layer chromatography and densitometry.

This paper describes a rapid method for the identification of salbutamol in liver and urine. Salbutamol is extracted from liver with an acid solution, purified on Baker columns and eluted with methanol. After derivatization, salbutamol is detected on HPTLC plates as a blue spot. Urine samples are directly purified on the C18 columns and then the same procedure is followed as for the liver samples. Using this screening method, salbutamol can be semi-quantitatively determined at the micrograms/kg level.

Albuterol↗

Long-term follow-up examination of experimental cataracts in rats by Scheimpflug photography and densitometry.

Development of experimental cataract can be objectively monitored in rats by application of Scheimpflug photography with the SL 45 Topcon camera and subsequent densitometric image analysis. The method has been used to study naphthalene cataract in Brown-Norway rats (BN-CPB), as well as diabetic cataract induced by streptozotocin injection in Sprague-Dawley rats. The values obtained by linear microdensitometric image analysis allowed precise characterization of the opacification with respect to size, topography, and time progress so that statistical evaluation of the efficacy of certain drugs in prevention or delay of experimental cataracts was possible.

Animals↗

Predicting fractures in women by using forearm bone densitometry.

In 1,076 women the forearm bone mineral content (BMC) had been measured with single photon gamma absorptiometry (SPA) 10-16 years ago. The incidence of fractures incurred in the ensuing years (1975-1985) was recorded. The BMC had been significantly less in those women who were to have the types of fractures that are related to bone fragility but only in women measured during the first post- menopausal decades. From age 50-69, diminished BMC at the distal forearm site was associated with a threefold increase of relative risk whereas at the proximal site there was about a sixfold increase in relative risk. This association was not evident in the population over 70 years of age because of a high frequency of fractures (mostly of the spine) in subjects with relatively high BMC values. Body weight and grip strength were significantly lower in the fracture group over 70 years of age.

Adult↗

Bone densitometry of excised vertebrae; anatomical relationships.

Bone mineral density (BMD) was determined in 32 excised vertebrae using three methods: (1) dual-energy quantitative computed tomography (QCT), (2) dual-photon absorptiometry (DPA) with 153-Gd in an anteriorposterior projection and (3) scanning slit X-ray absorptiometry (SSXA) in both AP and lateral projections. The QCT region-of-interest in the anterior vertebral body had a lower density than that of the total trabecular portion of the body, but was highly correlated to this larger region (r = 0.96; SEE = 8 mg/cm3). The anterior QCT region also correlated moderately with BMD from DPA (r = 0.77; SEE = 18 mg/cm3). Measurements of the vertebral body in lateral projection were less well correlated (r = 0.5-0.7) to QCT densities. Both the anterior QCT region (r = 0.81; SEE = 18 mg/cm3) and the BMD from DPA (r = 0.86; SEE = 16 mg/cm3) and the BMD from DPA (r = 0.86; SEE = 16 mg/cm3) were similarly predictive of density of the integral vertebral body. Differences among densitometric methods on the spine depend on the projection used and the region examined.

Absorptiometry, Photon↗

Soft-laser scanning densitometry performed on a 35-mm slide illustrating SDS-PAGE separation of adenovirion polypeptides.

Virion polypeptides (35 S), methionine-labeled and purified by CsCl gradient centrifugation, were separated by SDS-gel electrophoresis. Analysis of their band pattern was performed by scanning the images of the SDS-gels shown on a 35-mm slide. The densitometric tracings revealed the presence of 17 protein bands, although only 15 of them were visible to the naked eye. The high sensitivity and resolving capacities of the soft-laser scanning densitometer enabled us to detect trace amounts of protein bands separated in SDS-gels and to obtain a resolution compatible to that of electrophoresis. Fourfold electronic amplification of the densitometric tracings, produced by a computer, generated new information regarding the pattern of the electrophoregram. The facility to amplify peaks of importance is particularly advantageous when faint or overlapping protein bands revealed on a gel are assessed.

Adenoviridae↗

Monitoring of ANP secretion from single atrial myocytes using densitometry.

Atrial myocytes secrete atrial natriuretic peptide (ANP) in response to mechanical stretch and can serve as a challenging model for studying stretch-secretion coupling. We have developed a technique for monitoring ANP secretion from single atrial myocytes, using neutral red and a CCD video camera. Atrial-specific granules (ASGs) containing ANP were stained with neutral red. The cells were illuminated with monochromatic light (550 mm) and the grey value monitored within the region of interest (ROI) surrounding the region in which ASGs were densely located. Assuming that neutral red is evenly distributed in ASGs, the change in optical density (OD) was considered to represent the total amount of secretion. Under control, non-stimulated conditions, the OD decreased spontaneously (19.7+/-1.4%/10 min, n=14). Direct mechanical stretch (cell length increased by 20%) with two micropipettes or hypotonic swelling (200 mOsm) accelerated the decrease in OD significantly (48.7+/-7.4%/10 min; n=3, 47.2+/-2.4%/10 min; n=7, respectively). In conclusion, this method allows monitoring of ANP secretion with a relatively high time resolution while mechanical stress is applied. Furthermore, patch-clamp or intracellular perfusion techniques can be combined with the present technique for studying cellular mechanisms of stretch-secretion coupling.

Animals↗

Determination of the phospholipid/lipophilic compounds ratio in liposomes by thin-layer chromatography scanning densitometry.

The determination of the ratio of phospholipid/lipophilic compounds in liposomes was achieved after thin-layer chromatography (TLC) by measuring the spot intensities of dipalmitoyl phosphatidylcholine and the lipophilic compound. The liposome components under study were separated on one TLC plate, developed in two steps, and detected after charring the plate with specific visualization reagents. The method shows good reproducibility and provides a simple way to quantify the level of lipophilic compound incorporated in the liposome bilayer.

Chromatography, Gas↗

Edge detection versus densitometry in the quantitative assessment of stenosis phantoms: an in vivo comparison in porcine coronary arteries.

The aim of this study was the in vivo validation and comparison of the geometric and densitometric technique of a computer-assisted automatic quantitative angiographic system (CAAS system). In six Landrace Yorkshire pigs (45 to 55 kg), precision-drilled phantoms with a circular lumen of 0.5, 0.7, 1.0, 1.4, and 1.9 mm were percutaneously introduced into the left anterior descending or left circumflex coronary artery. Twenty-eight coronary angiograms obtained with the phantom in a wedged intracoronary position could be quantitatively analyzed. Minimal lumen diameter, minimal cross-sectional area, percent diameter stenosis, and cross-sectional area stenosis were automatically measured with both the geometric and densitometric technique and were compared with the known phantom dimensions. When minimal lumen diameter was measured using the geometric approach, a nonsignificant underestimation of the phantom size was observed, with a mean difference of -0.06 +/- 0.14 mm. The larger mean difference observed with videodensitometry (-0.11 +/- 0.20 mm) was the result of the failure of the technique to differentiate the low lumen videodensities of two phantoms of smaller size (0.5 and 0.7 mm) from a dense background. Percent cross-sectional area stenosis measured with the two techniques showed a good correlation with the corresponding phantom measurements (mean difference between percent cross-sectional area stenosis calculated from the quantitative angiographic measurements and the corresponding phantom dimensions was equal to 2 +/- 6% for both techniques, correlation coefficient = 0.93 with both techniques, SEE = 5% with the geometric technique and 6% with the densitometric approach).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Densitometry as an aid in amniotic fluid gel acetylcholinesterase analysis.

We have performed gel acetylcholinesterase analysis blindly on 1,300 consecutive amniotic fluid samples sent to our laboratory for alpha-fetoprotein measurement. Faint acetylcholinesterase bands were identified in nine of 1,300 samples, and all of these were associated with pregnancies free of major fetal malformations. Dense acetylcholinesterase bands were identified in four samples, and all were associated with major fetal malformations. Densitometric analysis of acetylcholinesterase bands provided additional objectivity in classifying band density.

Acetylcholinesterase↗