Determination of glutathione and glutathione disulfide in human whole blood using HPLC with coulometric detection: A comparison with fluorescence detection

2011 ◽  
Vol 76 (4) ◽  
pp. 277-294 ◽  
Author(s):  
Roman Kanďár ◽  
Pavla Žáková ◽  
Miroslava Marková ◽  
Halka Lotková ◽  
Otto Kučera ◽  
...  
Keyword(s):  
Detection Limit ◽  
Whole Blood ◽  
Trichloroacetic Acid ◽  
Signal To Noise Ratio ◽  
Glutathione Disulfide ◽  
Simple Method ◽  
Human Whole Blood ◽  
Coulometric Detection ◽  
Coefficients Of Variation

We describe a relatively simple method for the determination of glutathione (GSH) and glutathione disulfide (GSSG) in human whole blood. We have used an HPLC with coulometric electrochemical detection for the simultaneous measurement of GSH and GSSG. Diluted and filtered trichloroacetic acid extracts were injected directly into the HPLC system and were eluted isocratically on a Polaris 5u C18-A, 250 × 4.6 mm analytical column. Glutathione in samples extracted with trichloroacetic acid and diluted with 1.0 mMhydrochloric acid was stable at 4 °C for at least 8 h. The analytical performance of this method is satisfactory: the intra-assay and inter-assay coefficients of variation were below 10%. Quantitative recoveries from spiked whole blood samples were at intervals 91.6–97.6% for GSH and 85.0–104.4% for GSSG. The linear range is 5.0–2000.0 μmol/l, with a detection limit of 2.1 μmol/l (signal-to-noise ratio = 3) for GSH and 2.0–250.0 μmol/l, with a detection limit of 0.9 μmol/l for GSSG.

Novel LC–MS/MS method for the determination of selumetinib (AZD6244) in whole blood collected with volumetric absorptive microsampling

Bioanalysis ◽  
2020 ◽  
Vol 12 (13) ◽  
pp. 883-892
Author(s):  
Ramakrishna R Voggu ◽  
Theodore S Brus ◽  
Chineta T Barksdale ◽  
Paul Severin ◽  
Patricia Hansen ◽  
...  
Keyword(s):  
Whole Blood ◽  
Ammonium Hydroxide ◽  
Bioanalytical Method Validation ◽  
Simple Method ◽  
Human Whole Blood ◽  
Regulatory Guidance ◽  
Validation Parameters ◽  
Diagnostic Applications ◽  
Volumetric Absorptive Microsampling

Aim: A method has been developed and validated for quantitation of selumetinib in human whole blood collected using a Mitra™ volumetric absorptive microsampling device. This device is patient-friendly, affording less-invasive sampling with broad applicability to clinical and diagnostic applications – specifically in pediatric populations. Materials & methods: In this method, drug is extracted from the Mitra device via sonication in methanol: Ammonium hydroxide, then analyzed by LC–MS/MS. The linear range for selumetinib analysis is 2.00–2000 ng/ml. Results: All validation parameters met acceptance criteria established in agreement with current regulatory guidance for bioanalytical method validation. The stability of selumetinib in Mitra tips was established at both ambient and frozen conditions. Conclusion: A simple method has been developed and validated for determination of selumetinib from human whole blood, collected using volumetric absorptive microsampling and analyzed by LC–MS/MS.

Determination of Residual Acrylamide Monomer in Medical Polyacrylamide Hydrogel by HPLC-SPE

2005 ◽  
Vol 288-289 ◽  
pp. 405-408
Author(s):  
Zi Yi Wan ◽  
Ting Fei Xi ◽  
P. Zhao ◽  
Y. Sun ◽  
Z.G. Feng
Keyword(s):  
Detection Limit ◽  
Solid Phase ◽  
Polyacrylamide Hydrogel ◽  
Simple Method ◽  
Lower Detection Limit ◽  
Response Range ◽  
Coefficients Of Variation ◽  
Lower Detection ◽  
Linear Response Range

The polyacrylamide hydrogel (PAMG) has been used in cosmetology in China, Ukraine and Russia since 1990s. Because the monomer acrylamide(AM) used to produce PAMG has been implicated as a potential mutagen and reproductive toxicant[1,2], it is important to accurately determine the amount of residual AM monomer in the PAMG. In this study, a quick, practical and simple method to determine AM is presented with respect to the hydrogel. AM is analysed quantitatively by ODS-3 column with ultraviolet (UV) absorbance detector. AM is separated from interferential component with an aqueous solution of 0.9%NaCl (NS) adjusted at pH~3.7 using hydrochloric acid and then detected at a UV wavelength of 210 nm. The results show that ODS-3 is effective approach for quantifying AM concentrations in PAMG. This method has a lower detection limit of 0.003µg/ml and a linear response range of 0.003 and 0.9 µg/ml (depending on the range required for analysis). Precision studies give coefficients of variation of <3.2%(n=5) for 0.003µg/ml. The recoveries for this method are greater than 90%. When AM content in PAMG is lower than the detection limit of this method, SPE (solid phase extraction) could be used to concentrate AM. In the case, C18 cartridge is used. And the recoveries are about 70% for SPE when AM concentration is lower than ppb.

Antigen-specific T cell responses: Determination of their frequencies, homing properties, and effector functions in human whole blood

Methods ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 77-83 ◽  
Author(s):  
Tanja Breinig ◽  
Martina Sester ◽  
Urban Sester ◽  
Andreas Meyerhans
Keyword(s):  
T Cell ◽  
Whole Blood ◽  
T Cell Responses ◽  
Human Whole Blood ◽  
Effector Functions ◽  
Cell Responses

LC–APCI–MS–MS for the Determination of Celastrol in Human Whole Blood

2007 ◽  
Vol 66 (9-10) ◽  
pp. 735-739 ◽  
Author(s):  
Qing Xu ◽  
Mei Huang ◽  
Micong Jin ◽  
Qilong Ren
Keyword(s):  
Whole Blood ◽  
Human Whole Blood

scholarly journals FORENSIC CASES IN THE NORTH OF CHILE: DETERMINATION OF ANTIDEPRESSANT DRUGS IN HUMAN WHOLE BLOOD

2013 ◽  
Vol 58 (2) ◽  
pp. 1733-1736 ◽  
Author(s):  
FELIPE BRAVO ◽  
CARMEN ZAMBRA ◽  
KARINA VENEGAS ◽  
DAVID RIOS ◽  
PEDRO BUC CALDERON ◽  
...  
Keyword(s):  
Whole Blood ◽  
Antidepressant Drugs ◽  
Human Whole Blood ◽  
The North

Direct solid-phase enzymoimmunoassay of testosterone in saliva.

1989 ◽  
Vol 35 (10) ◽  
pp. 2044-2047 ◽  
Author(s):  
K Howard ◽  
M Kane ◽  
A Madden ◽  
J P Gosling ◽  
P F Fottrell
Keyword(s):  
Detection Limit ◽  
Analytical Procedure ◽  
Solid Phase ◽  
Medical Applications ◽  
Microtiter Plates ◽  
Large Numbers ◽  
Coefficients Of Variation ◽  
Serial Sampling ◽  
Direct Solid

Abstract This competitive, solid-phase enzymoimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of the saliva samples. Repeated assay of three control saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical applications, particularly for projects involving serial sampling and yielding large numbers of samples.

Simultaneous determination of methotrexate and its eight metabolites in human whole blood by capillary zone electrophoresis

2003 ◽  
Vol 1014 (1-2) ◽  
pp. 93-101 ◽  
Author(s):  
Chien-Yuan Kuo ◽  
Hsin-Lung Wu ◽  
Hwang-Shang Kou ◽  
Shyh-Shin Chiou ◽  
Deng-Chyang Wu ◽  
...  
Keyword(s):  
Simultaneous Determination ◽  
Capillary Zone Electrophoresis ◽  
Whole Blood ◽  
Human Whole Blood ◽  
Zone Electrophoresis ◽  
Capillary Zone

scholarly journals Determination of Total Interleukin-8 in Whole Blood after Cell Lysis

2000 ◽  
Vol 46 (9) ◽  
pp. 1387-1394 ◽  
Author(s):  
Jochen Reinsberg ◽  
Jörg Dembinski ◽  
Christoph Dorn ◽  
Daniela Behrendt ◽  
Peter Bartmann ◽  
...  
Keyword(s):  
Whole Blood ◽  
Healthy Subjects ◽  
Room Temperature ◽  
Sample Pretreatment ◽  
Cell Lysis ◽  
Interleukin 8 ◽  
Simple Method ◽  
Sample Handling ◽  
A Cell

Abstract Background: It has been shown that a high percentage of interleukin-8 (IL-8) in blood is cell associated. Recently, a simple method for determination of cell-associated IL-8 in whole blood after cell lysis has been described. The purpose of this study was to evaluate this method, to examine the influence of preanalytic sample handling, and to establish the concentration range of total IL-8 and its relation to age and sex in healthy subjects. Methods: Total IL-8 content of whole blood was determined after lysing blood cells with Milenia® cell lysis solution. IL-8 in the resulting blood lysate was measured with the IMMULITE® IL-8 immunoassay. Results: When freshly drawn blood was stored up to 48 h on ice, no significant changes in total IL-8 were measured in the subsequently prepared lysate, whereas with storage at room temperature, total IL-8 increased after 3 h from 94 ± 13 ng/L to 114 ± 16 ng/L (n = 10). In lysate stored for 48 h at 4 °C, marginal changes of the IL-8 concentration were noted, with storage at room temperature, only 76% ± 5% (n = 12) of initial concentration was recovered. From lysate frozen at −20 and −80 °C, respectively, 84% ± 4% and 93% ± 2% of initial IL-8 was recovered after 70 days (n = 10). IL-8 was measured with comparable precision in plasma (CV, 3.2–4.2%) and blood lysate (CV, 3.7–4.1%). When plasma was diluted with cell lysis solution, a slightly overestimated recovery (125% ± 3%) was observed; for lysate specimens with a cell lysis solution content ≥75%, the recovery after dilution was 98% ± 2%. In lysate prepared from 12 blood samples with exogenous IL-8 added, IL-8 recovery was 104% ± 2% (recovery from plasma &lt;35%). The median total IL-8 in blood lysates from 103 healthy subjects (22–61 years) was 83 ng/L of blood (2.5–97.5 percentile range, 49–202 ng/L of blood). In females but not in males, total IL-8 increased significantly with advancing age (P &lt;0.002). We found grossly increased total IL-8 in six pregnant women with amniotic infection syndrome. Conclusions: The evaluated method allows the assessment of total IL-8 in blood with good performance when appropriate conditions of sample pretreatment are considered. The values in healthy volunteers all were above the detection limit of the IL-8 assay; therefore, slight changes of total IL-8 could be noted. Thus, the present method is a suitable tool to study the diagnostic relevance of total IL-8 in blood.

scholarly journals Simultaneous Determination of Some Phenothiazine Derivatives in Human Blood by Headspace Solid-Phase Microextraction and Gas Chromatography with Nitrogen-Phosphorus Detection

2008 ◽  
Vol 91 (6) ◽  
pp. 1354-1362 ◽  
Author(s):  
Natsuko Shinmen ◽  
Xiao-Pen Lee ◽  
Takeshi Kumazawa ◽  
Chika Hasegawa ◽  
Yasuhiro Ishiwata ◽  
...  
Keyword(s):  
Gas Chromatography ◽  
Whole Blood ◽  
Solid Phase Microextraction ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Regression Equations ◽  
Phenothiazine Derivatives ◽  
Headspace Solid Phase Microextraction ◽  
Coefficients Of Variation

Abstract Chlorpromazine, levomepromazine, promazine, triflupromazine, and trimeprazine were simultaneously determined in human whole blood and plasma by combining headspace solid-phase microextraction and gas chromatography with nitrogenphosphorus detection. Extraction efficiency for the phenothiazine derivatives was 0.0130.117 for both sample types. Regression equations were linear [correlation coefficient (r) 0.99510.9999] within the range 2.5200 ng/0.5 mL for triflupromazine and trimeprazine, and 6.3200 ng/0.5 mL for chlorpromazine, levomepromazine, and promazine. The limit of detection for each compound was 0.23.9 ng/0.5 mL whole blood and plasma. Intraday and interday coefficients of variation for all phenothiazines in both human samples were commonly &lt;15 and 20, respectively. We also report the determination of levomepromazine in human plasma after oral administration.

Sign in / Sign up

Export Citation Format

Share Document