Measurement of Deuterium-Labeled Phylloquinone in Plasma by High-Performance Liquid Chromatography/Mass Spectrometry

Xueyan Fu†, James W. Peterson†, Mona Hdeib†, Sarah L. Booth*†, Michael A. Grusak‡, Alice H. Lichtenstein† and Gregory G. Dolnikowski†
Jean Mayer U.S. Department of Agriculture Human Nutrition Research Center on Aging at Tufts University, 711 Washington Street, Boston, Massachusetts 02111, and U.S. Department of Agriculture/Agricultural Research Service Children’s Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030
Anal. Chem., 2009, 81 (13), pp 5421–5425
DOI: 10.1021/ac900732w
Publication Date (Web): June 4, 2009
Copyright © 2009 American Chemical Society
* To whom correspondence should be addressed. Phone: 617-556-3231. Fax: 617-556-3149. E-mail: Sarah.booth@tufts.edu., †

Tufts University.

, ‡

Baylor College of Medicine.

Pharmacology

Abstract

Phylloquinone (vitamin K1) is a lipophilic compound present in plasma at low concentrations, which presents technical challenges for determining its bioavailability or metabolic fate using stable isotopes. We developed a method to simultaneously measure unlabeled and deuterium-labeled phylloquinone concentrations in plasma specimens using high-performance liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization (LC−APCI/MS). Phylloquinone was extracted from plasma using hexane, further purified by solid-phase extraction, and then quantified using high-performance liquid chromatography with an APCI/MS as a detector. Plotting the expected versus the measured amount of serial dilutions of either unlabeled or labeled phylloquinone gave correlation coefficients (R) of 0.999 for both compounds. The minimum detectable concentrations of unlabeled and labeled phylloquinone were 0.05 and 0.08 pmol/injection, respectively. Pooled plasma samples spiked with between 0.5 and 32 nmol phylloquinone/L gave average recoveries of 96.7% with 5.4% relative standard deviation (RSD) for unlabeled phylloquinone and 96.2% with 6.6% RSD for labeled phylloquinone. Plasma phylloquinone concentrations determined by LC-fluorescence and LC−APCI/MS methods from healthy subjects (n = 17) were not statistically different (P = 0.13). The LC−APCI/MS method is a sensitive technique for simultaneous determination of both unlabeled and labeled phylloquinone and can be applied to bioavailability studies.

View: Full Text HTML | Hi-Res PDF | PDF w/ Links

Citing Articles

View all 1 citing articles

Citation data is made available by participants in CrossRef's Cited-by Linking service. For a more comprehensive list of citations to this article, users are encouraged to perform a search in SciFinder.

This article has been cited by 1 ACS Journal articles (1 most recent appear below).

Tools

SciFinder Links

SciFinder subscribers: Click to sign in | Not a SciFinder subscriber? Learn more at www.cas.org

Explore by:

History

  • Published In Issue July 01, 2009
  • Article ASAPJune 04, 2009
  • Received: April 6, 2009
    Accepted: May 22, 2009

Recommend & Share

  • Share on ACS NetworkACS Network
  • Add to FacebookFacebook
  • Tweet ThisTweet This
  • Add to CiteULikeCiteULike
  • Add to NewsvineNewsvine
  • Digg ThisDigg This
  • Add to DeliciousDelicious

Related Content

Other ACS content by these authors: