scholarly journals Microspectrophotometric Determination of Nonheme Iron in Maturing Erythroblasts and its Relationship to the Endocellular Hemoglobin Formation

Blood ◽  
1960 ◽  
Vol 16 (3) ◽  
pp. 1285-1297 ◽  
Author(s):  
C. A. SONDHAUS ◽  
BO THORELL

Abstract The strong near-UV absorption of inorganic iron makes possible the microspectrophotometric determination of nonheme intracellular iron as distinct from hemoglobin-bound iron. Erythroid cells of Salamandra Maculosa were therefore studied microspectrophotometrically in the near ultraviolet and visible region. Absorption curves from cytoplasmic areas in living and dried cells were obtained from 310 to 700 mµ., and two component analyses were made to determine the presence and amount of hemoglobin and nonhemoglobin absorption. Under the experimental conditions, intracellular hemoglobin was present largely as the carbon monoxide derivative, a stable form which provided a well defined standard to correct for nonspecific absorption and other spectral distortion due to the optical properties of single cells. Absorption curves from the youngest cells measured were found to differ from those of mature cells, the difference curve agreeing most closely with that of inorganic iron or ferritin in solution. In the youngest cells, nonheme iron appeared to exceed 6 per cent by weight of hemoglobin, or over 20 times molar hemoglobin concentration, dropping and remaining at about twice the hemoglobin molarity from the time hemoglobin synthesis was one-third complete until maturity. Allowing for the increase of total cell mass with growth, the data suggest that although young erythroblasts may already have enough Fe for subsequent Hb synthesis, they continue to incorporate excess Fe throughout maturation and may thus contain a reserve pool of nonheme iron.

2006 ◽  
Vol 31 (4) ◽  
pp. 31-38
Author(s):  
L. M. Lourenço ◽  
F. G. Martins ◽  
V. R. Balbo ◽  
A. C. Pimenta ◽  
J. R. M. Castro ◽  
...  

A sensitive and alternative method for the spectrophotometric determination of chromium(III) based on the formation of chromium(III)/azide complexes was established by investigating a new band in the ultraviolet region. The best experimental conditions for the analytical determination of this metallic ion were: ligand and perchloric acid analytical concentration = 493 and 12.0 mmol L-1, respectively; aqueous medium; T = 25.0 ºC; contact time = 1 hour. The maximum molar absorptivity coefficient occurred at 287 nm (average 1.481 ± 0.008 <FONT FACE=Symbol>´</FONT> 10(4) L mol-1 cm-1), leading to the determination of metal ion concentrations one hundred times lower than the ones formerly determined in the visible region. The system obeys Beer's Law and is suitable for chromium determination in the 0.702-2.81 mg L-1 concentration range (15-65% T, 1.00 cm-width quartz cells). Analytical applications of the current method were tested with a nutritional supplement containing chromium. Results were compared with those obtained with atomic absorption spectrometry.


1996 ◽  
Vol 76 (01) ◽  
pp. 005-008 ◽  
Author(s):  
Jean Claude Lormeau ◽  
Jean Pascal Herault ◽  
Jean Marc Herbert

SummaryWe examined the effect of the synthetic pentasaccharide representing the minimal binding site of heparin to antithrombin on the antithrombin-mediated inactivation of factor Vila bound to tissue factor. This effect was compared to the effect of unfractionated heparin. Using purified recombinant human coagulation factors and either a clotting or an amidolytic assay for the determination of the residual activity of factor Vila, we showed that the pentasaccharide was an efficient antithrombin-dependent inhibitor of the coagulant activity of tissue factor-factor Vila complex. In our experimental conditions, assuming a mean MW of 14,000 for heparin, the molar pseudo-first order rate constants for ATIII-mediated FVIIa inhibition by ATIII-binding heparin and by the synthetic pentasaccharide were found to be similar with respective values of 104,000 ± 10,500 min-1 and 112,000 ± 12,000 min-1 (mean ± s.e.m., n = 3)


1982 ◽  
Vol 47 (7) ◽  
pp. 1973-1978 ◽  
Author(s):  
Jiří Karhan ◽  
Zbyněk Ksandr ◽  
Jiřina Vlková ◽  
Věra Špatná

The determination of alcohols by 19F NMR spectroscopy making use of their reaction with hexafluoroacetone giving rise to hemiacetals was studied on butanols. The calibration curve method and the internal standard method were used and the results were mutually compared. The effects of some experimental conditions, viz. the sample preparation procedure, concentration, spectrometer setting, and electronic integration, were investigated; the conditions, particularly the concentrations, proved to have a statistically significant effect on the results of determination. For the internal standard method, the standard deviation was 0.061 in the concentration region 0.032-0.74 mol l-1. The method was applied to a determination of alcohols in the distillation residue from an oxo synthesis.


Author(s):  
Timothy Aljoscha Frede ◽  
Marlene Dietz ◽  
Norbert Kockmann

AbstractFast chemical process development is inevitably linked to an optimized determination of thermokinetic data of chemical reactions. A miniaturized flow calorimeter enables increased sensitivity when examining small amounts of reactants in a short time compared to traditional batch equipment. Therefore, a methodology to determine optimal reaction conditions for calorimetric measurement experiments was developed and is presented in this contribution. Within the methodology, short-cut calculations are supplemented by computational fluid dynamics (CFD) simulations for a better representation of the hydrodynamics within the microreactor. This approach leads to the effective design of experiments. Unfavourable experimental conditions for kinetics experiments are determined in advance and therefore, need not to be considered during design of experiments. The methodology is tested for an instantaneous acid-base reaction. Good agreement of simulations was obtained with experimental data. Thus, the prediction of the hydrodynamics is enabled and the first steps towards a digital twin of the calorimeter are performed. The flow rates proposed by the methodology are tested for the determination of reaction enthalpy and showed that reasonable experimental settings resulted. Graphical abstract A methodology is suggested to evaluate optimal reaction conditions for efficientacquisition of kinetic data. The experimental design space is limited by thestepwise determination of important time scales based on specified input data.


Materials ◽  
2021 ◽  
Vol 14 (2) ◽  
pp. 328
Author(s):  
Kamil Anasiewicz ◽  
Józef Kuczmaszewski

This article is an evaluation of the phenomena occurring in adhesive joints during curing and their consequences. Considering changes in the values of Young’s modulus distributed along the joint thickness, and potential changes in adhesive strength in the cured state, the use of a numerical model may make it possible to improve finite element simulation effects and bring their results closer to experimental data. The results of a tensile test of a double overlap adhesive joint sample, performed using an extensometer, are presented. This test allowed for the precise determination of the shear modulus G of the cured adhesive under experimental conditions. Then, on the basis of the research carried out so far, a numerical model was built, taking the differences observed in the properties of the joint material into account. The stress distribution in a three-zone adhesive joint was analyzed in comparison to the standard numerical model in which the adhesive in the joint was treated as isotropic. It is proposed that a joint model with three-zones, differing in the Young’s modulus values, is more accurate for mapping the experimental results.


2015 ◽  
Vol 4 (4) ◽  
pp. 378-381 ◽  
Author(s):  
Mannem Sri Varsha ◽  
N. Raghavendra Babu ◽  
Yenumula Padmavathi ◽  
P. Ravi Kumar

A new simple, sensitive and specific procedure has been developed for determination of tenofovir disoproxil fumarate in bulk and pharmaceutical dosage forms using MBTH reagent. The purpose of this analytical validation procedure is to validate it by laboratory experiments to prove that the method meets the minimum standards for laboratory use. 3-methyl-2-bezothiazoline hydrazone reacts with the secondary amine group of tenofovir in the presence of oxidizing agent, ferric chloride. The resulting apple green coloured chromogen when measured spectrophotometrically in visible region (i.e., 400-800nm) shows a maximum absorbance at 626.5nm. This method can be successfully applied for the determination of drug content in pharmaceutical formulations. The results of analysis have been validated statistically.DOI: http://dx.doi.org/10.3329/icpj.v4i4.22620 International Current Pharmaceutical Journal, March 2015, 4(4): 378-381 


2021 ◽  
pp. 1-9
Author(s):  
Etsuo Niki

Reactive oxygen and nitrogen species have been implicated in the onset and progression of various diseases and the role of antioxidants in the maintenance of health and prevention of diseases has received much attention. The action and effect of antioxidants have been studied extensively under different reaction conditions in multiple media. The antioxidant effects are determined by many factors. This review aims to discuss several important issues that should be considered for determination of experimental conditions and interpretation of experimental results in order to understand the beneficial effects and limit of antioxidants against detrimental oxidation of biological molecules. Emphasis was laid on cell culture experiments and effects of diversity of multiple oxidants on antioxidant efficacy.


Molecules ◽  
2021 ◽  
Vol 26 (11) ◽  
pp. 3101
Author(s):  
Mariana N. Oliveira ◽  
Oriana C. Gonçalves ◽  
Samir M. Ahmad ◽  
Jaderson K. Schneider ◽  
Laiza C. Krause ◽  
...  

This work entailed the development, optimization, validation, and application of a novel analytical approach, using the bar adsorptive microextraction technique (BAμE), for the determination of the six most common tricyclic antidepressants (TCAs; amitriptyline, mianserin, trimipramine, imipramine, mirtazapine and dosulepin) in urine matrices. To achieve this goal, we employed, for the first time, new generation microextraction devices coated with convenient sorbent phases, polymers and novel activated carbons prepared from biomaterial waste, in combination with large-volume-injection gas chromatography-mass spectrometry operating in selected-ion monitoring mode (LVI-GC-MS(SIM)). Preliminary assays on sorbent coatings, showed that the polymeric phases present a much more effective performance, as the tested biosorbents exhibited low efficiency for application in microextraction techniques. By using BAμE coated with C18 polymer, under optimized experimental conditions, the detection limits achieved for the six TCAs ranged from 0.2 to 1.6 μg L−1 and, weighted linear regressions resulted in remarkable linearity (r2 > 0.9960) between 10.0 and 1000.0 μg L−1. The developed analytical methodology (BAμE(C18)/LVI-GC-MS(SIM)) provided suitable matrix effects (90.2–112.9%, RSD ≤ 13.9%), high recovery yields (92.3–111.5%, RSD ≤ 12.3%) and a remarkable overall process efficiency (ranging from 84.9% to 124.3%, RSD ≤ 13.9%). The developed and validated methodology was successfully applied for screening the six TCAs in real urine matrices. The proposed analytical methodology proved to be an eco-user-friendly approach to monitor trace levels of TCAs in complex urine matrices and an outstanding analytical alternative in comparison with other microextraction-based techniques.


1993 ◽  
Vol 48 (1) ◽  
pp. 58-67 ◽  
Author(s):  
Joseph Grobe ◽  
Duc Le Van ◽  
Gudrun Lange

The course of the reactions o f fluorophosphaalkenes F3CP = C (F)OR [R = Me (1), Et (2)] with methanol or ethanol strongly depends on the experimental conditions. Thus at 70 °C a mixture of the 2-phosphapropionic acid ester F3CP (H )CO2R [R = Me (3), Et (4)] and trifluoromethylphosphane H2PCF3 is formed [molar ratio: 3 or 4 /H2 CF3 ≈1/1]. If the precursors F3CP (H )CO2R [R = Me (3), Et) are used as starting materials, the reaction with ROH under the same conditions affords 3 and 4, respectively, (90 to 95% yield) with only traces of H2PCF 3. In the presence o f iPr2NH these precursors react with R′OH to give the novel trifluoromethylphosphaalkenes F3CP = C (OR )OR [R /R′: Me/Me (6); E t/E t (7); Me/Et (8)]. With Et2NH , 3 undergoes an addition/elimination process yielding the interesting push/pull system Et2N(F)C = P-CO2Me (5). 1 and 2 react with primary amines R′NH2 (R′= tBu, Me) with stereoselective formation of the fairly labile phosphaalkenes F3CP = C(OR)NHR′ [R /R′: Me/tBu (9), Et/tBu(10), Me/Me (11)] with trans-positions for CF3 and NHR′.The new compounds 3 -11 were characterized by spectroscopic investigations (1H , 19F, 31P, 13C NMR ; IR, MS) and determination of M+ or typical fragment ions [M+ -OR ] by high resolution mass spectrometry.


Blood ◽  
1981 ◽  
Vol 57 (6) ◽  
pp. 1065-1067 ◽  
Author(s):  
JA Strauchen ◽  
W Alston ◽  
J Anderson ◽  
Z Gustafson ◽  
LF Fajardo

Abstract Because we recently observed two patients with severe diabetic hyperglycemia and spuriously elevated electronically determined hematocrit and mean corpuscular volume (MCV), we investigated the effect of hyperglycemia on two popular automated hematology systems, the Coulter S and Ortho ELT-8. Marked hyperglycemia (blood glucose 800-- 2000 mg/dl) caused consistent overestimation of the electronically determined MCV compared to that derived from a simultaneous spun microhematocrit. The resultant overestimation and underestimation, respectively, of the derived values for hematocrit and mean corpuscular hemoglobin concentration may be clinically misleading. The mechanism of MCV elevation in hyperglycemia appears to be swelling of hyperosmolar glucose “loaded” erythrocytes when diluted into “isotonic” counting medium. This effect is readily circumvented by determination of a spun microhematocrit.


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