Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Spectrophotometry”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 343 records · Page 19Linked to original sources

Hemoglobin determination in plasma or serum by first-derivative recording spectrophotometry. Evaluation of the procedure of Soloni, Cunningham, and Amazon.

The authors tested the plasma hemoglobin measurement procedure of Soloni, Cunningham, and Amazon (Am J Clin Pathol 1986;85:342-347), which uses recording derivative spectrophotometry. By this technique, the authors were able to measure plasma or serum hemoglobin down to a level of 10 mg/L (1 mg/dL). The method was found to be quantitative and not affected by bilirubin or lipemia. Methemoglobin did not interfere with the assay. The observed first derivative maxima and minima were reproducible but instrument dependent. The authors applied this technique to the measurement of serum hemoglobin in 100 healthy human subjects. They observed a geometric mean value of 68 mg/L (6.8 mg/dL) with the actual range 21-189 mg/L (2.1-18.9 mg/dL). The authors confirm that this technique is a simple, rapid, and reproducible method for determining plasma or serum hemoglobin.

Bilirubin↗

Cutaneous melanin density of Caucasians measured by spectrophotometry and risk of malignant melanoma, basal cell carcinoma, and squamous cell carcinoma of the skin.

Recent advances have enabled quite accurate estimation by spectrophotometry of the density of cutaneous melanin. The relation between skin cancers and this objective measure of skin phenotype is examined here. For this purpose, a population-based case-control study of subjects aged 20-59 years of northern European ancestry was conducted in Tasmania, Australia. Cases (n = 244) of cutaneous malignant melanoma during 1998-1999, and a sample of cases of basal cell carcinoma (n = 220) and squamous cell carcinoma (n = 195) of the skin were identified from cancer registrations. Controls (n = 483) were selected from a comprehensive population listing. Melanin at the upper inner arm was estimated from skin reflectance of light of 400 and 420 nm wavelengths. For melanoma, basal cell carcinoma, and squamous cell carcinoma, respectively, the odds ratios comparing the least with the highest of four melanin categories were 6.2 (95% confidence interval (CI): 2.3, 16.6), 6.3 (95% CI: 2.6, 15.1), and 4.2 (95% CI: 1.7, 10.8) for men and 1.9 (95% CI: 1.0, 3.7), 1.4 (95% CI: 0.7, 3.0), and 0.7 (95% CI: 0.3, 1.7) for women. The gender differences were not due to disparities in site of occurrence or (for melanoma) in thickness of the lesion. The authors conclude that, particularly for men, cutaneous melanin density at the upper inner arm is a strong predictor of risk of skin cancer.

Adult↗

Assessment of provitamin A determination by open column chromatography/visible absorption spectrophotometry.

Determination of provitamin A content by open column chromatography/visible absorption spectrophotometry is assessed using food samples of varying carotenoid composition. A general method consisting of extraction with acetone, transfer to petroleum ether, saponification (optional), concentration, separation on activated MgO:Hyflo Supercel column developed with 1 to 15% acetone in petroleum ether, and quantitation of individual provitamins spectrophotometrically demonstrates repeatability comparable with that of high-performance liquid chromatographic (HPLC) methods. Overnight saponification (10% methanolic KOH, ambient temperature) does not degrade the provitamins and is unnecessary for kale, tomato, and squash; however, it is required for good separation of papaya carotenoids due to the presence of carotenol esters. The current Association of Official Analytical Chemists (AOAC) method is found to be inappropriate because (1) the volume of extracting solvent is not adjusted to the type of sample; (2) the less active alpha- and gamma-carotene, alpha- and beta-cryptoxanthin, and 5,6-monoepoxy-beta-cryptoxanthin (50% active) are quantified as beta-carotene (100% active); (3) inactive carotenoids such as xi-carotene and zeinoxanthin are also quantified as beta-carotene.

Carotenoids↗

Estimation of perimortal percent carboxy-heme in nonstandard postmortem specimens using analysis of carbon monoxide by GC/MS and iron by flame atomic absorption spectrophotometry.

In decomposed, formalin-fixed, embalmed, exhumed, and some fire-dried cases in which normal blood is unavailable, the usual methods for determination of carboxyhemoglobin saturation frequently fail. To address these specimens, a method utilizing both gas chromatography/mass spectrometric (GC/MS) determination of carbon monoxide (CO) and flame atomic absorption spectrophotometry (FAAS) determination of iron (Fe), in the same specimen, was developed. The method is reported here, along with its application to seven pertinent forsensic death investigations. The CO analytical methodology involves acid liberation of the gas from the specimen aliquot in a headspace vial. After heating and equilibrating, a sample of the headspace vapor is injected into the GC/MS system with a gastight syringe. Quantitation is achieved by standard addition comparison utilizing the ideal gas law equation. Iron is quantified by FAAS analysis of the same aliquot used for the CO determination, following nitric acid digestion. The concentration is determined by comparison to a standard curve. A formula for determining the minimum percent carboxy-heme saturation was derived by using the ratio of the amount of CO to the amount of Fe in the aliquot analyzed. Tissue types analyzed include spleen, liver, muscle, dried blood, and unspecified decomposed tissue.

Calibration↗

Determination of hydroxocobalamin and cyanocobalamin by derivative spectrophotometry in cyanide poisoning.

Hydroxocobalamin (OHCo) and cyanocobalamin (CNCo) are determined directly in biological media, without extraction, by using first derivative spectrophotometry. We diluted 200 mL of plasma, urine, or standards with 1.8 mL of pH 6 buffer (boric acid, potassium dihydrogen orthophosphate, and potassium hydroxide). The first derivative spectra of the dilutions were plotted between 320 and 400 nm. At the exact zero-crossing point for hydroxocobalamin, the derivative values of cyanocobalamin concentration were determined. The same procedure was followed for hydroxocobalamin at the zero-crossing point for cyanocobalamin. The derivative values of the concentration curves are linear in the range 5-100 microM. The minimum detection limit is approximately 5 microM for hydroxocobalamin of cyanocobalamin on the determination of hydroxocobalamin or vice versa, although the spectra strongly overlap. The method is fast and simple to use, thus making it easy to assess the in vivo transformation of hydroxocobalamin into cyanocobalamin after the administration of high doses of hydrocobalamin in cyanide poisoning.

Cyanides↗

Determination of methemoglobin and total hemoglobin in toxicological studies by derivative spectrophotometry.

A method for the determination of methemoglobin in the presence of other hemoglobin subforms (i.e., oxy-, deoxy-, and carboxyhemoglobin) by use of derivative spectrophotometry is proposed. The method, which uses the first-derivative of the spectrum at 645 nm, is straightforward and expeditious, so it is of special interest to forensic toxicology laboratories as it allows the simultaneous determination of the methemoglobin saturation percentage and the hemoglobin concentration. This facilitates interpretation of the results and provides a better understanding of the significance of methemoglobin saturation in specific cases. Based on an analysis of interferences, the presence of other hemoglobin subforms or of endogenous components of plasma does not detract in any way from the performance of the method.

Forensic Medicine↗

A stationary cold-vapor technique for the determination of submicrogram amounts of mercury in biological tissues by flameless atomic absorption spectrophotometry.

A procedure is provided for the determination of mercury in animal tissue using a single unit stationary cold-vapor generator utilizing atomic absorption spectrophotometry. The entire reduction-absorption cell is fitted to the burner mount on the instrument. Sample digests are introduced into the sample compartment along with reductant, and background corrected absorbance is measured after three minutes of sample-reductant mixing. The method is sensitive to 2 ng Hg (0.50 ng/mL Hg at 0.0044 absorbance units), which corresponds to 25 ng Hg per gram wet tissue using the prescribed digestion procedure. Reproducibility and accuracy are comparable to conventional cold-vapor techniques and the fact that relatively small sample aliquots (4.0 mL) are used for analysis provides for multideterminations of Hg in a single tissue digest; this is not ordinarily possible using conventional cold-vapor methods. A total of 15 digested tissue sample can be analyzed per hour. The method is proposed as an alterative to presently used cold-vapor Hg techniques.

Animals↗

Detection of benzoylecgonine in urine by means of UV spectrophotometry.

A simple and reliable method has been developed for the detection of benzoylecgonine in urine by UV spectrophotometry. Benzoylecgonine was isolated by liquid-liquid extraction and subsequently identified based on its hydrolysis by NaOH. The decrease in absorbance at 235 nm caused by this NaOH hydrolysis was used to identify benzoylecgonine even in the absence of its characteristic spectrum. None of the other drugs evaluated interfered with the method, which has a detection limit of 1.25 micrograms/mL.

Cocaine↗

An improved wet digestion procedure for the analysis of total arsenic in biological samples by direct hydride atomic absorption spectrophotometry.

The presence of dimethylarsinic acid (DMAA) in biological samples can cause an underestimation of total arsenic content when analyzed relative to an inorganic arsenic standard by direct hydride flame atomic absorption spectrophotometry. An acid digestion procedure is described that quantitatively recovers DMAA as well as monomethylarsonic acid, inorganic arsenic(III), and arsenic(V) from aqueous and biological samples. Methylated arsenicals are converted to inorganic arsenic by wet digestion with HNO3, H2SO4, and K2Cr2O7 and subsequently reduced to arsenic(III) with Nal. Arsine is generated with NaBH4 and converted to atomic arsenic following immediate introduction into a nitrogen-entrained air-hydrogen flame. This method produces a linear relationship to absorbance within a mass range of 50 to 300 ng arsenic/arsine reaction. A sensitivity of 2 ng arsenic and a detection limit of 7 ng arsenic/arsine reaction were also obtained. Recovery of DMAA from water, urine, feces, or whole blood ranged from 92 to 105% with a coefficient of variation of 5 to 10%.

Animals↗

A study of derivative spectrophotometry for the determination of carboxyhemoglobin in blood.

An application of derivative spectrophotometry to the determination of carboxyhemoglobin (HbCO) in blood was examined. The 1st, 2nd, 3rd, and 4th derivative spectra were recorded in the Soret region for HbCO, reduced hemoglobin (Hb), and their mixed solution. The 4th derivative spectrum had the highest sensitivity to the change of HbCO-Hb ratio in the solution. A maximum point of the 4th derivative curve was employed for the HbCO determination. Blood samples with varied HbCO concentrations were analyzed using this technique and the results were compared with those obtained by conventional spectrophotometric and gas chromatographic procedures. The derivative method was in satisfactory agreement with both procedures.

Carboxyhemoglobin↗

Deuteration kinetics of deoxyguanosine, deoxycytidine, and their polynucleotides by means of ultraviolet spectrophotometry.

The kinetics of the hydrogen-deuterium exchange reactions of deoxyguanosine (dG), deoxycytidine (dC), double-helical poly[d(G-C)] X poly[d(G-C], and double-helical poly(dG) X poly(dC) have been examined at 20 degrees C, pH 7.0, and in low-salt (0.15 M NaCl) medium by stopped-flow ultraviolet spectrophotometry, in the spectral region of 260 to 320 nm. The rate constant was found to be 78.9 s-1 for dG-NH, 2.2 s-1 for dG-NH2, 39.3 s-1 for dC-NH2, 2.4 s-1 (fast) and 0.94 s-1 (slow) for poly[d(G-C)] X poly[d(G-C)], and 2.2 s-1 (fast) and 0.92 s-1 (slow) for poly(dG) X poly(dC). From these values, the probability of base-pair opening of the G X C containing B-form double helix is estimated to be (3 +/- 1) X 10(-3). This is much greater than what is expected from an extrapolation of the van't Hoff plot at the helix-coil transition region, i.e. at about 110 degrees C. The mechanism of these base-pair openings at 20 degrees C (as well as the mechanism of base-pair reformation) is suggested to be totally different from those in the melting temperature range.

Base Composition↗

Does the redox state of cytochrome aa3 reflect brain energy level during hypoxia? Simultaneous measurements by near infrared spectrophotometry and 31P nuclear magnetic resonance spectroscopy.

We studied cerebral oxygen metabolism during hypoxia to demonstrate whether the redox state of cytochrome aa3 (cyt.aa3), as measured by near infrared spectrophotometry (NIRS), reflects brain energy level. Rats (n = 6) subjected to hypoxia were simultaneously monitored by NIRS and 31P nuclear magnetic resonance spectroscopy (NMRS). Brain function was evaluated using the electroencephalogram (EEG). After a reduction of the fraction of inspired oxygen FIO2 from 0.21 to 0.15, we observed a significant increase in reduced cyt.aa3 (from 16.5% +/- 2.1% to 41.2% +/- 2.8%; P < 0.01), without significant changes in phosphocreatine (PCr) and beta-adenosine triphosphate (beta-ATP) levels. The PCr decreased significantly at a FIO2 of 0.10 (53.8% +/- 6.4% as compared with 97.7% +/- 10.9% at a FIO2 of 0.21; P < 0.05), and reached a minimum at a FIO2 of 0.04. beta-ATP did not change significantly at a FIO2 of 0.10 or 0.08. With a FIO2 of less than 0.08, cyt.aa3 was almost totally reduced. EEG activity slowed at a FIO2 of 0.08 and became isoelectric at 0.04. Significant correlations were found between the levels of cyt.aa3 and PCr (P < 0.001, r = 0.83) as well as between cyt.aa3 and beta-ATP (P < 0.001, r = 0.73) by using the overall values at FIO2 levels from 0 to 1.0. However, no significant correlations were observed among these variables when the FIO2 was less than 0.10. These findings suggest that the increase in reduced cyt.aa3 reflects brain energy depletion; however, the redox state of cyt.aa3 will not indicate brain energy depletion during extreme hypoxia because cyt.aa3 is reduced totally during hypoxia insufficient to deplete intracellular ATP.

Adenosine Triphosphate↗

Blood volume measurement at the bedside using ICG pulse spectrophotometry.

BACKGROUND: In the treatment of critically ill patients, blood volume (BV) measurement requires injection of some tracer substance and subsequent blood sampling to analyze the tracer concentration. To obviate both the sampling and laboratory analysis, techniques of pulse oximetry have been adapted to the noninvasive optical measurement in the patient's nose or finger of the arterial concentration of an injectable dye. METHODS: The authors report the clinical accuracy of a new noninvasive bedside BV measurement test that uses pulse spectrophotometry (the pulse method). The device detects pulsatile changes of tissue optical density of a nostril or a finger spanned by a probe emitting two infrared wavelengths (805 and 890 nm). After a peripheral or central intravenous injection of indocyanine green, the arterial dye concentration is continuously computed by reference to the previously measured blood hemoglobin concentration. Three types of tests of its accuracy are described here. RESULTS: In 10 healthy volunteers, the authors compared BV determined by the pulse method with an (131)I-labeled human serum albumin method. Three subject data sets were excluded because of motion artifact, a low signal:noise ratio, or both. For the other seven volunteers, the bias+/-SD of pulse spectrophotometric BV values were 0.20+/-0.24 l (or 4.2+/-4.9%) for the nose probe and 0.34+/-0.31 l (or 7.3+/-6.9%) for the finger probe, with a mean BV of 5 l. In 30 patients who underwent cardiac surgery, the pulse method was compared with a standard indocyanine green method using intermittent blood sampling. In three patients, the BV could not be determined by the pulse method because of motion artifact, low signal:noise ratio, or both. In 27 patients, the bias+/-SD of the BV by the pulse method was -0.23+/-0.37 l (-5.3+/-8.7%) for the nose and -0.25+/-0.5 l (-4.2+/-8.4%) for the finger. Patient BV ranged from 2.51 to 7.13 l (mean, 4.48 l). In 10 additional patients before cardiac surgery, BV was measured by the pulse method before and shortly after removal of 400 ml blood. The pulse method recorded a decrease of BV of 480+/-114 mL Three days after venesection, the mean BV was 117+/-159 ml less than the predonation control. CONCLUSIONS: In most patients, the pulse method provides bedside measurement of BV without blood sampling (except for hemoglobin determination), with an estimated error less than 10%. In 10-30% of tests the method failed because of motion distortion of the record during the 10-min data collection period or because of insufficient pulse amplitude in the test tissue.

Adult↗

Plasma selenium concentration in healthy Japanese children and adults determined by flameless atomic absorption spectrophotometry.

This study showed a rapid and direct method for determining selenium concentration in plasma by flameless atomic absorption spectrophotometry, and differences in plasma selenium concentration in healthy children and adults. A direct method is possible, since selenium is heat stable in the presence of nickel. With this method, the recovery of selenium added to plasma was 100.3 +/- 5.7%, and the relative standard deviation in repeated determinations of pooled plasma selenium was 3.0% and 6.8%. The plasma selenium concentration in adults was 99.4 +/- 12.5 ng/ml, lower than reported concentrations from the United States and Canada, and higher than those from New Zealand. These variations may reflect dietary habits, bioavailability of selenium compounds in diet, racial difference, or different analytical methods. The mean concentration of plasma selenium at 1 to 6 months of age (51.0 +/- 13.1 ng/ml) was significantly lower than in adults (p less than 0.001); it increased gradually and steadily to the adult level with age. This age-related difference of plasma selenium level is similar to that reported previously.

Adolescent↗

Egg colour matching in an African cuckoo, as revealed by ultraviolet-visible reflectance spectrophotometry.

Despite major differences between human and avian colour vision, previous studies of cuckoo egg mimicry have used human colour vision (or standards based thereon) to assess colour matching. Using ultraviolet-visible reflectance spectrophotometry (300-700 nm), we measured museum collections of eggs of the red-chested cuckoo and its hosts. The first three principal components explained more than 99% of the variance in spectra, and measures of cuckoo host egg similarity derived from these transformations were compared with measures of cuckoo host egg similarity estimated by human observers unaware of the hypotheses we were testing. Monte Carlo methods were used to simulate laying of cuckoo eggs at random in nests. Results showed that host and cuckoo eggs were very highly matched for an ultraviolet versus greenness component, which was not detected by humans. Furthermore, whereas cuckoo and host were dissimilar in achromatic brightness, humans did not detect this difference. Our study thus reveals aspects of cuckoo-host egg colour matching which have hitherto not been described. These results suggest subtleties and complexities in the evolution of host-cuckoo egg mimicry that were not previously suspected. Our results also have the potential to explain the longstanding paradox that some host species accept cuckoo eggs that are non-mimetic to the human eye.

Animals↗

The role of ultraviolet-A reflectance and ultraviolet-A induced fluorescence in the appearance of budgerigar plumage: insights from spectrofluorometry and reflectance spectrophotometry.

Fluorescence has so far been found in 52 parrot species when illuminated with ultraviolet-A (UVA) 'black' lamps, and two attempts have been made to determine whether such fluorescence plays any role in sexual signalling. However, the contribution of the reflectance versus fluorescence to the total radiance from feathers, even in the most studied species to date (budgerigars), is unclear. Nor has the plumage of this study species been systematically assessed to determine the distribution of fluorescent patches. We therefore used spectrofluorometry to determine which areas of budgerigars fluoresce and the excitation and emission spectra involved; this is the first time that such a technique has been applied to avian plumage. We found that both the yellow crown and (normally hidden) white downy chest feathers exhibit strong UVA-induced fluorescence, with peak emissions at 527 nm and 436 nm, respectively. Conversely, the bright-green chest and dark-blue tail feathers do not fluoresce. When comparing reflectance spectra (400-700 nm) from the yellow crown using illuminants with a proportion of UVA comparable to daylight, and illuminants with all UVA removed, no measurable difference resulting from fluorescence was found. This suggests that under normal daylight the contribution of fluorescence to radiance is probably trivial. Furthermore, these spectra revealed that males had fluorescent crowns with substantially higher reflectance than those of females, in both the UV waveband and at longer wavelengths. Reflectance spectrophotometry was also performed on a number of live wild-type male budgerigars to investigate the chromatic contrast between the different plumage areas. This showed that many plumage regions are highly UV-reflective. Overall our results suggest that rapid surveys using UVA black lamps may overestimate the contribution of fluorescence to plumage coloration, and that any signalling role of fluorescence emissions, at least from the yellow crown of budgerigars, may not be as important as previously thought.

Animals↗

Cryptic gentes revealed in pallid cuckoos Cuculus pallidus using reflectance spectrophotometry.

Many cuckoo species lay eggs that match those of their hosts, which can significantly reduce rejection of their eggs by the host species. However, egg mimicry is problematic for generalist cuckoos that parasitize several host species with different egg types. Some generalist cuckoos have overcome this problem by evolving several host-specific races (gentes), each with its own, host-specific egg type. It is unknown how generalist cuckoos lacking gentes are able to avoid egg rejection by hosts. Here we use reflectance spectrophotometry (300-700 nm) on museum egg collections to test for host-specific egg types in an Australian generalist cuckoo reported to have a single egg type. We show that the colour of pallid cuckoo (Cuculus pallidus) eggs differed between four host species, and that their eggs closely mimicked the eggs of the host they parasitized. These results reveal that pallid cuckoos have host-specific egg types that have not been detected by human observation, and indicate that gentes could be more common than previously realized.

Adaptation, Physiological↗

Redox poise and oxygenation of cytochrome bd in the diazotroph Azotobacter vinelandii assessed in vivo using diode-array reflectance spectrophotometry.

A ferrous oxygenated form of cytochrome d is characteristic of all cytochrome bd-type oxidases so far examined, but its participation in enzyme turnover is unclear. It is relatively stable, occurs in aerated cell suspensions and predominates during enzyme preparation. In this study, diode-array reflectance spectrophotometry was used to assess the redox poise and oxygenation of cytochrome bd in vivo, in the aerobic diazotroph Azotobacter vinelandii. Mutants either lacking or overproducing the cytochrome bd oxidase were used to confirm the reliability of the optical configuration. Changes in absorbance attributed to cytochromes b, c and d were followed as the O2 supply was altered either in suspensions of harvested cells or during steady-state growth. In washed cell suspensions, three states of cytochrome d, which differed in absorbance characteristics, were seen: (1) an oxygenated form that absorbs at 650 nm, (2) a form which has little absorbance at either 650 or 630 nm and (3) the reduced form that absorbs at 630 nm. The transition between states 2 and 3, but not 1 and 2, correlated with the changes in the redox states of cytochromes b595 and b560. The dissolved O2 concentration at which this transition occurred coincided approximately with the apparent O2 affinity for the oxidase in vivo (approx. 5 microM). During steady-state growth, the cytochromes were partially reduced and the oxygenated form of cytochrome d was undetected. These in situ measurements support the view that an oxygenated form of cytochrome d (absorbing at 650 nm) in the one-electron-reduced cytochrome bd-type oxidase does not take part in enzyme turnover.

Azotobacter vinelandii↗