Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography

B. Bleiberg, Jacob J. Steinberg, S. D. Katz, J. Wexler, T. LeJemtel

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Assessment of lactate metabolism is of particular interest during exercise and in disease states such as diabetes, shock, and absorptive abnormalities of short-chain fatty acids by the colon. We describe an analytical method that introduces radio-active tracers and high-performance liquid chromatography (HPLC) to simultaneously analyze concentrations and specific activities (SAs) of plasma lactate. The HPLC conditions included separation on a reversed-phase column (octadecylsilane) and an isocratic buffer (30% acetonitrile in water). [3H]Acetate served as an internal standard. Lactate and acetate were extracted from plasma samples with diethyl ether following a pH adjustment to less than 1.0 and back-extracted into a hydrophilic phase with sodium carbonate (2 mM, pH > 10.0). Lactate is detected in the ultraviolet range (242 and 320 nm) by derivatization with α-bromoacetophenone. Control plasma samples were studied after an overnight fast for precision and analytical recovery. Calibration curves were linear in the range 0.18-6.0 mM (r=0.92). The precision was 3% and the analytical recovery was 87%. The detection limit of the method was 36 pmol. Determination of lactate metabolism was performed in a patient with chronic congestive heart failure who was administered primed-continuous l-[U-14C]lactate (10 μCi bolus and 0.3 μCi/min continuously) during a 60-min rest period. Mean arterial lactate concentration and SA were 1.69 ± 0.2 mM and 253.8 ± 22 dpm/μmol, respectively. Systemic lactate turnover was 25.65 μmol/kg per min. Lactic acid systemic turnover, organ uptake and release rates can be accurately determined by isocratic HPLC.

Original languageEnglish (US)
Pages (from-to)301-308
Number of pages8
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume568
Issue number2
DOIs
StatePublished - Aug 23 1991

Fingerprint

High performance liquid chromatography
Lactic Acid
High Pressure Liquid Chromatography
Plasmas
Metabolism
Acetates
Recovery
Radioactive tracers
Volatile Fatty Acids
Medical problems
Radio
Ether
Calibration
Limit of Detection
Shock
Buffers
Colon
Heart Failure
Exercise
Water

ASJC Scopus subject areas

  • Chemistry(all)
  • Clinical Biochemistry
  • Molecular Medicine

Cite this

Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography. / Bleiberg, B.; Steinberg, Jacob J.; Katz, S. D.; Wexler, J.; LeJemtel, T.

In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 568, No. 2, 23.08.1991, p. 301-308.

Research output: Contribution to journalArticle

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