Microfluidic Analysis of Complex Samples with Minimal Sample Preparation Using Gradient Elution Moving Boundary Electrophoresis

2009 ◽  
Vol 81 (24) ◽  
pp. 10201-10207 ◽  
Author(s):  
Elizabeth A. Strychalski ◽  
Alyssa C. Henry ◽  
David Ross
Keyword(s):  
Sample Preparation ◽  
Moving Boundary ◽  
Gradient Elution ◽  
Complex Samples ◽  
Minimal Sample ◽  
Microfluidic Analysis

Expanding the Capabilities of Microfluidic Gradient Elution Moving Boundary Electrophoresis for Complex Samples

2011 ◽  
Vol 83 (16) ◽  
pp. 6316-6322 ◽  
Author(s):  
Elizabeth A. Strychalski ◽  
Alyssa C. Henry ◽  
David Ross
Keyword(s):  
Moving Boundary ◽  
Gradient Elution ◽  
Complex Samples

No lyse no wash flow cytometry for maximizing minimal sample preparation

Methods ◽  
2018 ◽  
Vol 134-135 ◽  
pp. 149-163 ◽  
Author(s):  
Jordi Petriz ◽  
Jolene A. Bradford ◽  
Michael D. Ward
Keyword(s):  
Flow Cytometry ◽  
Sample Preparation ◽  
Minimal Sample

scholarly journals The analysis of acetaminophen (paracetamol) and seven metabolites in rat, pig and human plasma by U(H)PLC–MS

Bioanalysis ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 485-500
Author(s):  
Rebecca Dargue ◽  
Isobelle Grant ◽  
Leanne C Nye ◽  
Andy Nicholls ◽  
Theo Dare ◽  
...  
Keyword(s):  
Stable Isotope ◽  
Sample Preparation ◽  
Oral Administration ◽  
Human Plasma ◽  
Wistar Rats ◽  
Rat Plasma ◽  
Mouse Serum ◽  
Internal Standards ◽  
Minimal Sample

A U(H)PLC–MS/MS method is described for the analysis of acetaminophen and its sulphate, glucuronide, glutathione, cysteinyl and N-acetylcysteinyl metabolites in plasma using stable isotope-labeled internal standards. P-Aminophenol glucuronide and 3-methoxyacetaminophen were monitored and semi-quantified using external standards. The assay takes 7.5 min/sample, requires only 5 μl of plasma and involves minimal sample preparation. The method was validated for rat plasma and cross validated for human and pig plasma and mouse serum. LOQ in plasma for these analytes were 0.44 μg/ml (APAP-C), 0.58 μg/ml (APAP-SG), 0.84 μg/ml (APAP-NAC), 2.75 μg/ml (APAP-S), 3.00 μg/ml (APAP-G) and 16 μg/ml (APAP). Application of the method is illustrated by the analysis of plasma following oral administration of APAP to male Han Wistar rats.

scholarly journals Blood targeted proteomics: centrifugal filter sample preparation vs dilute-and-shoot

2015 ◽  
Author(s):  
Tore Vehus ◽  
Ole Kristian Brandtzaeg ◽  
Elsa Lundanes ◽  
Steven Ray Wilson
Keyword(s):  
Sample Preparation ◽  
Beta Catenin ◽  
Targeted Proteomics ◽  
Liquid Chromatography Mass Spectrometry ◽  
Blood Samples ◽  
Minimal Sample ◽  
External Standard ◽  
Nano Liquid Chromatography ◽  
Potential Challenge ◽  
Considerable Complexity

Blood has a complex proteome with a huge span of protein abundances, and we are currently exploring sample preparation strategies addressing this potential challenge. We first evaluated the possibility to simply reduce complexity by fractionating blood samples (prior to targeted proteomics), using combinations of centrifugal filters that are promoted as having differing molar mass (M M ) selectivity. However, systematic fractionation was not possible, as the units had surprisingly unpredictable M M filtration profiles. Hence, labeling implying M M selectivity of centrifugal filters shouldn’t be taken literally. One filter combination (100+50K) however appeared to reduce human serum albumin (HSA) compared with much of the proteome (assessed using gel electrophoresis and staining). However, for our target protein beta-catenin, a “dilute-and-shoot” approach (without any attempt to remove high abundant proteins) was what enabled identification of beta-catenin in blood with nano liquid chromatography mass spectrometry (nano-LC-MS). Thus, minimal sample preparation can be an option in targeted blood proteomics. All blood samples prepared and analyzed (using ultrafast nano-LC-MS) were of considerable complexity, and we document the importance of using external standard spiking and minimum three MS/MS transitions to confirm the presence of target proteins.

Total reflection X-ray fluorescence based quantification of gold nanoparticles in cancer cells

2018 ◽  
Vol 33 (3) ◽  
pp. 395-403 ◽  
Author(s):  
Gabriella Mankovskii ◽  
Ana Pejović-Milić
Keyword(s):  
Gold Nanoparticles ◽  
Sample Preparation ◽  
Cancer Cells ◽  
Sample Volume ◽  
Total Reflection ◽  
X Ray ◽  
Minimal Sample

A small (5 μl) sample volume and minimal sample preparation steps are required to accurately quantify AuNP uptake in cancer cells.

scholarly journals Targeted Metabolic Profiling of Methionine Cycle Metabolites and Redox Thiol Pools in Mammalian Plasma, Cells and Urine

Metabolites ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 235 ◽  
Author(s):  
Sidney Behringer ◽  
Victoria Wingert ◽  
Victor Oria ◽  
Anke Schumann ◽  
Sarah Grünert ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Plasma Cells ◽  
Genetic Diseases ◽  
Simultaneous Detection ◽  
Embryonic Stem ◽  
Methionine Sulfoxide ◽  
Diagnostic Value ◽  
Control Analysis ◽  
Reference Ranges ◽  
Minimal Sample

The concentration of thiol and thioether metabolites in plasma has diagnostic value in genetic diseases of B-vitamin metabolism linked to methionine utilization. Among these, cysteine/cystine (Cys/CSSC) and glutathione/oxidized glutathione (GSH/GSSG) act as cellular redox buffers. A new LC-MS/MS method was developed for the simultaneous detection of cystathionine (Cysta), methionine (Met), methionine sulfoxide (MSO), creatinine and the reduced and oxidized pairs of homocysteine (Hcy/HSSH), cysteine (Cys/CSSC) and glutathione (GSH/GSSG). A one-step thiol-blocking protocol with minimal sample preparation was established to determine redox thiol pairs in plasma and cells. The concentrations of diagnostic biomarkers Hcy, Met, Cysta, and Cys in a cohort of healthy adults (n = 53) agreed with reference ranges and published values. Metabolite concentrations were also validated in commercial samples of human, mouse, rat and Beagle dog plasma and by the use of a standardized ERNDIM quality control. Analysis of fibroblasts, endothelial and epithelial cells, human embryonic stem cells, and cancer cell lines showed cell specificity for both the speciation and concentration of thiol and thioether metabolites. This LC-MS/MS platform permits the fast and simultaneous quantification of 10 thiol and thioether metabolites and creatinine using 40 µL plasma, urine or culture medium, or 500,000 cells. The sample preparation protocols are directly transferable to automated metabolomic platforms.

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