Journal
ANALYTICAL BIOCHEMISTRY
Volume 291, Issue 2, Pages 279-289Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1006/abio.2001.5030
Keywords
PEITC; PEITC-NAC; PEITC-GSH; phenethyl isothiocyanate; pharmacokinetics; HPLC; plasma assay; isothiocyanates
Funding
- NCI NIH HHS [CA 46535, CA-166081-21, P01 CA046535] Funding Source: Medline
- NCRR NIH HHS [M01 RR000096, CRC-RR-00096] Funding Source: Medline
- NCPDCID CDC HHS [NCI-CN-55120] Funding Source: Medline
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Dietary and pharmacologic isothiocyanates (ITCs) may play a role in reducing the risk of certain cancers. The quantification of ITCs in humans is important both for epidemiological and pharmacokinetic studies. We describe a modification of an HPLC-based assay of urinary ITCs for use with human plasma The assay utilizes the cyclocondensation reaction of 1,2-benzenedithiol with ITCs present in human plasma, followed by a two-step hexane extraction and analysis by HPLC using W detection at 365 nm. The method shows linearity and reproducibility with human plasma over a range of 49-3003 nM phenethyl isothiocyanate (PEITC) (r(2) = 0.996 +/- 0.003). A similar degree of linearity was seen with two other biologically occurring conjugates of PEITC: PEITC-N-acetylcysteine (PEITC-NAC) and PEITC-glutathione (PEITC-GSH). The recovery of PEITC assessed on multiple days was 96.6 +/- 1.5% and was 100% for PEITC-GSH and PEITC-NAC. The reproducibility of the assay on multiday samplings showed a mean %CV of 6.5 +/- 0.3% for PEITC, 6.4 +/- 4.3 for PEITC-NAC and 12.3 +/- 3.9 for PEITC-GSH. In clinical studies, mean plasma ITC level of 413 +/- 193 nM PEITC equivalents was determined for a non-dietary-controlled group of 23 subjects. Multiday analysis data from pharmacokinetic plasma sets of 3 subjects taking a single dose of PEITC at 40 mg showed a good CV (range: 16-21%). The applicability of the methodology to pharmacokinetic studies of PEITC in humans is demonstrated. (C) 2001 Academic Press.
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