scholarly journals Measuring Sulforaphane and Its Metabolites in Human Plasma: A High Throughput Method

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 829 ◽  
Author(s):  
Annie Langston-Cox ◽  
Dovile Anderson ◽  
Darren J. Creek ◽  
Kirsten Palmer ◽  
Euan M. Wallace ◽  
...  
Keyword(s):  
Human Plasma ◽  
High Throughput ◽  
Clinical Investigation ◽  
Healthy Human ◽  
Potential Health ◽  
Low Concentrations ◽  
High Throughput Method ◽  
Standard Calibration ◽  
Internal Standardization ◽  
Cost Efficient

(1) Background: There is increasing understanding of the potential health benefits of cruciferous vegetables. In particular sulforaphane (SFN), found in broccoli, and its metabolites sulforaphane-glutathione (SFN-GSH), sulforaphane-cysteine (SFN-Cys), sulforaphane cysteine-glycine (SFN-CG) and sulforaphane-N-acetyl-cysteine (SFN-NAC) have potent antioxidant effects that may offer therapeutic value. Clinical investigation of sulforaphane as a therapeutic antioxidant requires a sensitive and high throughput process for quantification of sulforaphane and metabolites; (2) Methods: We collected plasma samples from healthy human volunteers before and for eight hours after consumption of a commercial broccoli extract supplement rich in sulforaphane. A rapid and sensitive method for quantification of sulforaphane and its metabolites in human plasma using Liquid Chromatography–Mass Spectrometry (LC–MS) has been developed; (3) Results: The LC–MS analytical method was validated at concentrations ranging between 3.9 nM and 1000 nM for SFN-GSH, SFN-CG, SFN-Cys and SFN-NAC and between 7.8 nM and 1000 nM in human plasma for SFN. The method displayed good accuracy (1.85%–14.8% bias) and reproducibility (below 9.53 %RSD) including low concentrations 3.9 nM and 7.8 nM. Four SFN metabolites quantitation was achieved using external standard calibration and in SFN quantitation, SFN-d8 internal standardization was used. The reported method can accurately quantify sulforaphane and its metabolites at low concentrations in plasma; (4) Conclusions: We have established a time- and cost-efficient method of measuring sulforaphane and its metabolites in human plasma suitable for high throughput application to clinical trials.

scholarly journals Development of a High-Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1466
Author(s):  
Lisard Iglesias-Carres ◽  
Lauren A. Essenmacher ◽  
Kathryn C. Racine ◽  
Andrew P. Neilson
Keyword(s):  
Chlorogenic Acid ◽  
Gallic Acid ◽  
High Throughput ◽  
Anaerobic Fermentation ◽  
Inhibitory Effect ◽  
Healthy Human ◽  
High Throughput Method ◽  
Lyase Activity ◽  
Inhibitory Potential ◽  
Potential Maximum

Choline is metabolized by the gut microbiota into trimethylamine (TMA), the precursor of pro-atherosclerotic molecule trimethylamine N-oxide (TMAO). A reduction in TMA formation has shown cardioprotective effects, and some phytochemicals may reduce TMA formation. This study aimed to develop an optimized, high-throughput anaerobic fermentation methodology to study the inhibition of choline microbial metabolism into TMA by phenolic compounds with healthy human fecal starter. Optimal fermentation conditions were: 20% fecal slurry (1:10 in PBS), 100 µM choline, and 12 h fermentation. Additionally, 10 mM of 3,3-dimethyl-1-butanol (DMB) was defined as a positive TMA production inhibitor, achieving a ~50% reduction in TMA production. Gallic acid and chlorogenic acid reported higher TMA inhibitory potential (maximum of 80–90% TMA production inhibition), with IC50 around 5 mM. Neither DMB nor gallic acid or chlorogenic acid reduced TMA production through cytotoxic effects, indicating mechanisms such as altered TMA-lyase activity or expression.

High-throughput determination of ultra-low concentrations of LAG078, a lipid modulator, in human plasma

2005 ◽  
Vol 37 (5) ◽  
pp. 1039-1048
Author(s):  
Tapan K. Majumdar ◽  
Ray Bakhtiar ◽  
Cindy Chen ◽  
Luis Ramos ◽  
Francis L.S. Tse
Keyword(s):  
Human Plasma ◽  
High Throughput ◽  
Low Concentrations

Simple, cost-efficient and high throughput method for separating single-wall carbon nanotubes with modified cotton

Carbon ◽  
2021 ◽  
Vol 178 ◽  
pp. 157-163
Author(s):  
Timur Khamidullin ◽  
Shamil Galyaltdinov ◽  
Alina Valimukhametova ◽  
Vasiliy Brusko ◽  
Artur Khannanov ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
High Throughput ◽  
Single Wall Carbon Nanotubes ◽  
Single Wall Carbon ◽  
High Throughput Method ◽  
Cost Efficient

scholarly journals A New, High-Throughput High-Performance Liquid Chromatographic/Mass Spectrometric Assay for Therapeutic Level Monitoring of Digoxin in Human Plasma

2009 ◽  
Vol 92 (5) ◽  
pp. 1390-1395 ◽  
Author(s):  
Laurian Vlase ◽  
Daniela-Saveta Popa ◽  
Dana Muntean ◽  
Dan Mihu ◽  
Sorin Leucuta
Keyword(s):  
Human Plasma ◽  
High Throughput ◽  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Selected Ion Monitoring ◽  
High Throughput Method ◽  
Instrument Analysis ◽  
Liquid Chromatographic ◽  
Level Monitoring ◽  
Chromatographic Mass

Abstract A new, high-throughput HPLC/MS assay for the quantification of digoxin in human plasma was developed and validated. The separation was performed on a Zorbax SB-C18 column under isocratic conditions using a 55 + 45 (v/v) mixture of methanol and 0.1 (v/v) formic acid in 10 M sodium acetate as the mobile phase at 45C with a flow rate of 1 mL/min. The detection of digoxin was performed in the selected ion monitoring mode (m/z 803.5). The human plasma samples (0.2 mL) were deproteinized with 7 perchloric acid in water, and aliquots of 20 L from supernatants obtained after centrifugation were directly injected into the chromatographic system. The method showed good linearity (r > 0.9926), precision (CV > 18.9), and accuracy (bias > 5.6) over the range of 0.550 ng/mL plasma. The lower LOQ was 0.5 ng/mL, and the recovery was between 92 and 106. The method is not expensive, it needs a minimum time for plasma sample preparation, and it has a run time of 2.3 min for instrument analysis (the retention time of digoxin was 1.9 min). The developed and validated high-throughput method is very simple, rapid, and efficient, with wide applications in clinical level monitoring, pharmacokinetics, and bioequivalence studies.

scholarly journals Development of a High Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation

2021 ◽  
Author(s):  
Lisard Iglesias-Carres ◽  
Lauren A. Essenmacher ◽  
Kathryn C. Racine ◽  
Andrew P. Neilson
Keyword(s):  
Chlorogenic Acid ◽  
Gallic Acid ◽  
High Throughput ◽  
Anaerobic Fermentation ◽  
Inhibitory Effect ◽  
Healthy Human ◽  
High Throughput Method ◽  
Lyase Activity ◽  
Inhibitory Potential ◽  
Potential Maximum

Choline is metabolized by the gut microbiota into trimethylamine (TMA), the precursor of pro-atherosclerotic molecule trimethylamine N-oxide (TMAO). Reduction of TMA formation has been shown to provide to cardioprotective effects, and some phytochemicals may produce such reduction. This study aimed to develop an optimized, high-throughput anaerobic fermentation methodology to study inhibition of choline microbial metabolism into TMA by phenolic compounds with healthy human fecal starter. Optimal fermentation conditions were: 20 % fecal slurry (1:10 in PBS), 100 M choline, and 12 h fermentation. Also, 10 mM of 3,3-dimethyl-1-butanol (DMB) was defined as a positive TMA production inhibitor, achieving a ~50 % reduction in TMA production. Gallic acid and chlorogenic acid reported higher TMA inhibitory potential (maximum of 80 -90 % in. TMA production inhibition), with IC50 around 5 mM. Nor DMB neither gallic acid and chlorogenic acid reduced TMA production through cytotoxic effects, indicating mechanisms such as altered TMA lyase activity or expression.

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