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Related Experiment Videos

Gas chromatography/mass spectrometry method to quantify blood hydroxycitrate concentration.

Y C Loe1, N Bergeron, N Rodriguez

  • 1Department of Nutritional Sciences, University of California at Berkeley, 119 Morgan Hall, Berkeley, California 94720-3104, USA.

Analytical Biochemistry
|April 26, 2001
PubMed
Summary

Hydroxycitrate (HCA), a weight loss supplement, is absorbed by humans but present in small plasma amounts. This study developed a new method to accurately measure HCA levels, aiding future research on its effectiveness.

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Area of Science:

  • Biochemistry
  • Pharmacokinetics
  • Analytical Chemistry

Background:

  • Hydroxycitrate (HCA) is a popular weight loss supplement that inhibits ATP-citrate lyase, a key enzyme in de novo lipogenesis (DNL).
  • Animal studies suggest HCA effectively inhibits DNL and promotes weight loss, but human studies show inconsistent results, possibly due to differing bioavailability.
  • Investigating HCA bioavailability in humans is crucial for understanding its efficacy as a weight loss aid.

Purpose of the Study:

  • To develop and validate a novel Gas Chromatography/Mass Spectrometry (GC/MS) method for quantifying hydroxycitrate (HCA) in human plasma.
  • To assess the acute absorption and plasma concentration of HCA in human subjects following oral ingestion.
  • To provide an accurate analytical tool for future pharmacokinetic and efficacy studies of HCA.

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Main Methods:

  • Development of a sensitive GC/MS assay utilizing [U-(13)C]citrate (CA*) as an internal standard for accurate quantification.
  • Derivatization of HCA and CA* with BSTFA + 10% TMCS, followed by analysis using positive chemical ionization (PCI/GC/MS).
  • Measurement of plasma HCA concentrations in four human subjects over 3.5 hours after ingesting a 2g dose.

Main Results:

  • The developed GC/MS method accurately measured trace amounts of HCA in human plasma with minimal sample preparation.
  • Plasma HCA concentrations ranged from 0.8 to 8.4 microg/ml at 30 minutes and 2 hours post-ingestion, respectively.
  • Demonstrated acute absorption of HCA, indicating it enters the bloodstream, albeit in low concentrations.

Conclusions:

  • Hydroxycitrate (HCA) is absorbed following oral administration in humans.
  • Acute HCA intake results in low concentrations in human plasma, potentially explaining inconsistent weight loss effects.
  • The validated GC/MS method provides a precise tool for measuring HCA, facilitating further research into its bioavailability and efficacy.