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Special Issue “Chemical Speciation of Organic and Inorganic Components of Environmental and Biological Interest in Natural Fluids: Behaviour, Interaction and Sequestration”
Open AccessArticle

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

Department of Obstetrics and Gynaecology, Monash University, Monash Medical Centre, 246 Clayton Road, Clayton, VIC 3168, Australia
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia
Author to whom correspondence should be addressed.
Molecules 2020, 25(4), 829;
Received: 18 December 2019 / Revised: 6 February 2020 / Accepted: 7 February 2020 / Published: 13 February 2020
(This article belongs to the Section Analytical Chemistry)
(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. View Full-Text
Keywords: sulforaphane; Liquid Chromatography–Mass Spectrometry; pharmacokinetic sulforaphane; Liquid Chromatography–Mass Spectrometry; pharmacokinetic
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Langston-Cox, A.; Anderson, D.; Creek, D.J.; Palmer, K.; Wallace, E.M.; Marshall, S.A. Measuring Sulforaphane and Its Metabolites in Human Plasma: A High Throughput Method. Molecules 2020, 25, 829.

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