scholarly journals Measurement of Homocysteine and Other Aminothiols in Plasma: Advantages of Using Tris(2-carboxyethyl)phosphine as Reductant Compared with Tri-n-butylphosphine

2001 ◽  
Vol 47 (10) ◽  
pp. 1821-1828 ◽  
Author(s):  
Jakub Krijt ◽  
Martina Vacková ◽  
Viktor Kožich
Keyword(s):  
Low Molecular Weight ◽  
Total Glutathione ◽  
Deming Regression ◽  
Mixed Disulfides ◽  
Total Homocysteine ◽  
The Mean ◽  
Effects Of Temperature ◽  
Temperature Time ◽  
Reliable Methods

Abstract Background: Aminothiols have been implicated in the pathogenesis of arteriosclerosis, and reliable methods are needed to determine their concentrations in body fluids. We present a comparison of two analytical methods and focus on the reduction of low-molecular weight and protein-mixed disulfides of homocysteine, cysteine, cysteinyl-glycine, and glutathione. Methods: The plasma total aminothiol profile was determined by HPLC with fluorescence detection after derivatization with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate. Disulfides and protein-bound aminothiols were reduced by either tri-n-butylphosphine (the TBP method) or tris(2-carboxyethyl)phosphine (the TCEP method); the effects of temperature, time of reduction, and concentration of reductants were evaluated. Results: The intraassay imprecision (CV) was <3% for all aminothiols using both methods. The interassay CVs for total cysteine (tCys), total cysteinyl-glycine (tCys-Gly), and total homocysteine (tHcy) were <4% and <8% for the TCEP and TBP methods, respectively, whereas for total glutathione (tGSH) the interassay CV was >12% for both methods. Deming regression and Bland–Altman difference plots showed positive biases for total aminothiol concentrations determined by the TCEP method relative to the TBP method. The mean proportional biases were 65%, 27%, 6%, and 60% for tCys, tCys-Gly, tHcy, and tGSH, respectively. The calculated concentrations of total aminothiols by the TCEP method were less influenced by changes in temperature and concentration of reducing agent or by calibrator matrix. Conclusions: The agreement between the TCEP and TBP methods was considerably lower for the determination of tCys, tCys-Gly, and tGSH than for tHcy. For total-aminothiol determination, the TCEP method yields better reproducibility and is more robust than the TBP method.

scholarly journals DETERMINATION OF THE COEFFICIENT OF HEAT TRANSFER DURING COOLING OF SILVER THERMO PROBES, TAKING INTO ACCOUNT THE EFFECTS OF TEMPERATURE-TIME DELAY

2017 ◽  
Vol 39 (5) ◽  
pp. 41-47 ◽  
Author(s):  
E. N. Zotov ◽  
A. A. Moskalenko A.A. ◽  
O. V. Rasumtseva ◽  
L. M. Protsenko
Keyword(s):  
Heat Transfer ◽  
Heat Flux Density ◽  
Cooling Capacity ◽  
Heat Transfer Process ◽  
Special Program ◽  
Nonstationary Heat Transfer ◽  
Density Surface ◽  
Effects Of Temperature ◽  
Temperature Time

Existing methods for determining the characteristics of the nonstationary heat transfer process (temperature field, heat transfer coefficient, heat flux density, surface temperature) are considered and analyzed when cooling silver spherical and cylindrical thermo-probes. New analytical solutions are proposed using a special program IQLab, which increase the accuracy of calculations when testing the cooling capacity of various liquids. The results of the calculations are compared with the experimental data.  

Characterization of 10 Biogenic Amines in Male Silkworm Moth by HPLC with Precolumn Derivatization

2020 ◽  
Vol 16 (5) ◽  
pp. 608-614
Author(s):  
Mingqin Fan ◽  
Yun Ai ◽  
Wenjie Zhao ◽  
Yanni Sun ◽  
Jianli Liu ◽  
...  
Keyword(s):  
Biogenic Amines ◽  
Biologically Active ◽  
Low Molecular Weight ◽  
Precolumn Derivatization ◽  
Mean Values ◽  
The Mean ◽  
Toxic Potential ◽  
First Time

Background: Biogenic Amines (BAs) are biologically active nitrogenous organic compounds of low molecular weight, which are frequently found in a wide variety of foods, beverages and herbs due to their toxic potential in humans. Male Silkworm Moth (MSM), a Traditional Chinese Medicine (TCM), has been exploited and utilized as nutritious liquor based on its traditional effects in the Chinese community. Objective: The objective of this study was to develop an HPLC with Dns-Cl derivatization method for characterizing overall BAs in MSM and providing data for further evaluating its activities and safety profiles. Methods: The method has acceptable sensitivity, precision, accuracy, selectivity and recovery, and was successfully applied to the determination of the BAs contents in MSM for the first time. Results: In the analysis of 10 batches of MSM samples, serotonin and dopamine were not found in detectable concentrations in any samples, and the most abundant amine found was putrescine. The mean values of tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine determined in the samples were found to be 34.7 mg/ kg, 16.1 mg/ kg, 218.3 mg/ kg, 37.9 mg/ kg, 12.1 mg/ kg, 18.2 mg/ kg, 4.5mg/ kg, and 0.9 mg/ kg, respectively. Conclusion: The contents of BAs in 10 batches of MSM were below the maximum recommended limits, and MSM can be used safely.

Fully automated fluorescence assay for determining total homocysteine in plasma.

1989 ◽  
Vol 35 (9) ◽  
pp. 1921-1927 ◽  
Author(s):  
H Refsum ◽  
P M Ueland ◽  
A M Svardal
Keyword(s):  
Disulfide Bonds ◽  
Free Fraction ◽  
Column Switching ◽  
Plasma Homocysteine ◽  
Automated Assay ◽  
Mixed Disulfide ◽  
Cation Exchange Column ◽  
Mixed Disulfides ◽  
Total Homocysteine

Abstract Homocysteine exists in human plasma as various (mixed) disulfides. Most plasma homocysteine (about 70%) is protein bound, probably via a disulfide bond to albumin, whereas homocysteine-cysteine mixed disulfide is the predominating form in the free fraction. We here present a method for the determination of total homocysteine, which includes both fractions. Plasma was initially treated with sodium borohydride to reduce the disulfide bonds, and the liberated thiols were derivatized with monobromobimane. The derivatized sample, still containing the plasma proteins, was injected onto a strong cation-exchange column, from which the homocysteine derivative was directed by column switching into a cyclohexyl silica (CH) column. The homocysteine derivative was top-concentrated on the CH column, then rapidly eluted with a steep gradient of methanol. Both the derivatization procedure and chromatography were performed with a combined sample processor and sample injector from Gilson (Model 232-401). Within-run and between-run precision (CV) was less than 4%, and the detection limit of 0.2 pmol was sufficiently low for monitoring homocysteine in plasma. We verified the assay against two established manual methods for the determination of total homocysteine in plasma. This, the first fully automated assay for total plasma homocysteine, allows the unattended analysis of 70 samples per 24 h.

Formation and Binding of Histamine by Free Mast Cells of Rat Peritoneal Fluid

1956 ◽  
Vol 186 (2) ◽  
pp. 199-202 ◽  
Author(s):  
Richard W. Schayer
Keyword(s):  
Mast Cells ◽  
Peritoneal Fluid ◽  
Histidine Decarboxylase ◽  
Cell Suspensions ◽  
Stable Form ◽  
Effects Of Temperature ◽  
Temperature Time ◽  
Considerable Loss ◽  
Number Of Cells

Suspensions of the cells of rat peritoneal fluid have a marked ability to decarboxylate C14 l-histidine and bind the resulting histamine in stable form. The evidence is strong that this activity is due almost entirely to mast cells; these constitute about 4% of the total number of cells present. The effects of temperature, time, and ph on this process were found to be characteristic of enzymic reactions. Preincubation of cell suspensions with nonisotopic histamine had no effect on the formation and binding of C14 histamine; this is evidence that exogenous histamine cannot enter the bound condition in free mast cells. The effects of histidine analogs on binding is described. Disruption of the mast cells results in a considerable loss in histamine forming activity. However, a fairly stable, soluble histidine decarboxylase can be obtained from the disrupted cells. A new method for determination of histamine as the S35 dibenzenesulfonyl derivative is described.

Detection of Circulating Tissue Factor and Factor VII in a Normal Population

1996 ◽  
Vol 75 (05) ◽  
pp. 772-777 ◽  
Author(s):  
Sybille Albrecht ◽  
Matthias Kotzsch ◽  
Gabriele Siegert ◽  
Thomas Luther ◽  
Heinz Großmann ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Normal Population ◽  
Mean Value ◽  
Factor Vii ◽  
Significant Difference ◽  
Age Dependent ◽  
The Mean ◽  
Highly Correlated ◽  
And Gender

SummaryThe plasma tissue factor (TF) concentration was correlated to factor VII concentration (FVIIag) and factor VII activity (FVIIc) in 498 healthy volunteers ranging in age from 17 to 64 years. Immunoassays using monoclonal antibodies (mAbs) were developed for the determination of TF and FVIIag in plasma. The mAbs and the test systems were characterized. The mean value of the TF concentration was 172 ± 135 pg/ml. TF showed no age- and gender-related differences. For the total population, FVIIc, determined by a clotting test, was 110 ± 15% and the factor VIlag was 0.77 ± 0.19 μg/ml. FVII activity was significantly increased with age, whereas the concentration demonstrated no correlation to age in this population. FVII concentration is highly correlated with the activity as measured by clotting assay using rabbit thromboplastin. The ratio between FVIIc and FVIIag was not age-dependent, but demonstrated a significant difference between men and women. Between TF and FVII we could not detect a correlation.

Determination of Plasminogen in Clots and Thrombi

1966 ◽  
Vol 16 (01/02) ◽  
pp. 038-050 ◽  
Author(s):  
Ulla Hedner ◽  
Inga Marie Nilsson ◽  
B Robertson
Keyword(s):  
Mean Value ◽  
Main Mechanism ◽  
Clot Lysis ◽  
Post Mortem ◽  
Digestion Time ◽  
Normal Volunteers ◽  
Casein Solution ◽  
The Mean

SummaryThe plasminogen content was determined by a casein method in plasma and serum from 20 normal volunteers. The mean plasminogen content was found to be 10.1 ACU (the arbitrary caseinolytic unit defined in such a way that using a 3% casein solution and a digestion time of 20 min. at 37°C, 10 ACU gave an extinction of 0.300). No difference between serum and plasma regarding the plasminogen content was found.Plasminogen was determined in drained and drained plus washed clots prepared from 2 ml plasma. The highest values found in the drained clots were 0.9 ACU/clot and 0.2 ACU/clot in the drained plus washed clots.Plasminogen was also determined in drained and drained plus washed clots prepared from plasma with added purified plasminogen. The plasminogen was recovered in the washing fluid. According to these tests, then, purified added plasminogen is washed out of the clots.The plasminogen content of 20 thrombi obtained post mortem was also determined. The mean value was found to be 0.7 ACU/cm thrombus. Judging from our results, the “intrinsic clot lysis theory” is not the main mechanism of clot dissolution.

SERUM TRIIODOTHYRONINE DETERMINATION BY SATURATION ANALYSIS

1973 ◽  
Vol 72 (4) ◽  
pp. 714-726 ◽  
Author(s):  
A. Burger ◽  
B. Miller ◽  
C. Sakoloff ◽  
M. B. Vallotton
Keyword(s):  
Ionic Strength ◽  
Limit Of Detection ◽  
Improved Method ◽  
Accuracy Of Measurement ◽  
Tracer Amount ◽  
The Mean ◽  
Physiological Values ◽  
Hyperthyroid Patients ◽  
Solvent Blank

ABSTRACT An improved method for the determination of serum triiodothyronine (T3) has been developed. After addition of a tracer amount of the hormone, T3 was extracted from 1 ml serum under conditions of pH and ionic strength which favoured T3 extraction (89%) over thyroxine (T4) extraction (58%). Chromatography of the extracted material on Sephadex LH-20 separated T3 completely from residual T4. The T3 eluate was dried, then re-dissolved in 0.5 ml NaOH 0.04 n. To 0.2 ml duplicate aliquots, a standard amount of TBG was added for the competitive protein analysis. After one hour incubation at 4°C, separation of bound from free T3 was achieved on small Sephadex G-25 columns. Overall recovery was 67 ± 10.8% and correction for the loss was made. The solvent blank was 37 ± 27 (sd) ng/100 ml. Accuracy of measurement of known quantities of T3 added to serum was 98.4%. The coefficient of variation within the assay was 6.2% and between the assays it was 11.4%. The limit of detection (0.1 ng) corresponded to a concentration of 25 ng/100 ml. T4 added to serum did not interfere with T3 determination until high non-physiological values were reached. The mean ± sd serum T3 in 54 euthyroid subjects was 153 ± 58 ng/100 ml and in 24 hyperthyroid patients it was 428 ±186 ng/100 ml; 4 out of the 24 hyperthyroid values were within 2 sd of the mean euthyroid group. All the values found in the euthyroid group were well above the limit of detection of the method.

Determination of the Probability Distribution of the Mean Sound Level

2010 ◽  
Vol 35 (4) ◽  
pp. 543-550 ◽  
Author(s):  
Wojciech Batko ◽  
Bartosz Przysucha
Keyword(s):  
Probability Distribution ◽  
Probability Distribution Function ◽  
Mean Value ◽  
Sound Level ◽  
Function Form ◽  
Noise Levels ◽  
Logarithmic Mean ◽  
The Mean ◽  
Estimation Of Uncertainty

AbstractAssessment of several noise indicators are determined by the logarithmic mean <img src="/fulltext-image.asp?format=htmlnonpaginated&src=P42524002G141TV8_html\05_paper.gif" alt=""/>, from the sum of independent random resultsL1;L2; : : : ;Lnof the sound level, being under testing. The estimation of uncertainty of such averaging requires knowledge of probability distribution of the function form of their calculations. The developed solution, leading to the recurrent determination of the probability distribution function for the estimation of the mean value of noise levels and its variance, is shown in this paper.

Simultaneous Determination of Alogliptin, Linagliptin, Saxagliptin, and Sitagliptin in Bulk Drug and Formulation by UPLC Q-TOF-MS

2020 ◽  
Vol 17 (1) ◽  
pp. 95-105
Author(s):  
Ramji Rathod ◽  
Faraat Ali ◽  
Amrish Chandra ◽  
Robin Kumar ◽  
Meenakshi Dahiya ◽  
...  
Keyword(s):  
Internal Standard ◽  
Gradient Elution ◽  
Ultra Performance Liquid Chromatography ◽  
Pharmaceutical Dosage Forms ◽  
Sensitivity And Selectivity ◽  
The Mean ◽  
Bulk Drug ◽  
Positive Ion ◽  
Higher Sensitivity

Background: A simple and sensitive Ultra Performance Liquid Chromatography-Mass Spectrometry method was developed and validated to measure the concentrations of Alogliptin (ALO), Linagliptin (LIN), Saxagliptin (SAX), and Sitagliptin (SIT) using Pioglitazone (PIO) as an internal standard. Methods: Chromatographic separation of six gliptins was achieved on a C-18 column (100×2.1 mm, 2.7 μm) using a mobile phase consisting of formic acid in water, 0.1%v/v: acetonitrile in gradient elution. Electrospray ionization (ESI) source was operated in the positive ion mode. Targeted MS/MS mode on a QTOF MS was used to quantify the drug utilizing the transitions of 340.1(m/z), 473.2 (m/z), 316.2 (m/z), 408.1 (m/z), and 357.1 (m/z) for ALO, LIN, SAX, SIT and PIO respectively. Results: As per ICH Q2R1 guidelines, a detailed validation of the method was carried out and the standard curves were found to be linear over the concentration ranges of 1516.0-4548.1 ng mL-1, 519.8- 1559.4 ng mL-1, 1531.4-4594.3 ng mL-1and 1519.6-4558.8 ng mL-1 for ALO, LIN, SAX and SIT respectively. Precision and accuracy results were within the acceptable limits. The mean recovery was found to be 98.8 _ 0.76 % (GEM), 102.2 _ 1.59 % (LIN), 95.3 _ 2.74 % (SAX) and 99.2 _ 1.75 % (SIT) respectively. Conclusions: The optimized validated UPLC QTOF-MS/MS method offered the advantage of shorter analytical times and higher sensitivity and selectivity. The optimized method is suitable for application in quantitative analysis of pharmaceutical dosage forms for QC laboratory.

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