Retinoid quantification by HPLC/MS(n)

Peter McCaffery, James Evans, Omanand Koul, Amy Volpert, Kevin Reid, M David Ullman

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.
Original languageEnglish
Pages (from-to)1143-9
Number of pages7
JournalJournal of Lipid Research
Volume43
Issue number7
Publication statusPublished - 2002

Fingerprint

Retinoids
Tretinoin
High Pressure Liquid Chromatography
2-Propanol
Hexanes
Mass spectrometry
Vertebrates
Teratogens
Column chromatography
Atmospheric Pressure
Mass spectrometers
Cytoplasmic and Nuclear Receptors
Tandem Mass Spectrometry
Vitamin A
Isomers
Atmospheric pressure
Ionization
Chromatography
Transcription Factors
Cells

Keywords

  • Animals
  • Chromatography, High Pressure Liquid
  • Isomerism
  • Liver
  • Male
  • Mass Spectrometry
  • Rats
  • Rats, Sprague-Dawley
  • Reproducibility of Results
  • Retina
  • Retinoids
  • Sensitivity and Specificity
  • Tretinoin

Cite this

McCaffery, P., Evans, J., Koul, O., Volpert, A., Reid, K., & Ullman, M. D. (2002). Retinoid quantification by HPLC/MS(n). Journal of Lipid Research, 43(7), 1143-9.

Retinoid quantification by HPLC/MS(n). / McCaffery, Peter; Evans, James; Koul, Omanand; Volpert, Amy; Reid, Kevin; Ullman, M David.

In: Journal of Lipid Research, Vol. 43, No. 7, 2002, p. 1143-9.

Research output: Contribution to journalArticle

McCaffery, P, Evans, J, Koul, O, Volpert, A, Reid, K & Ullman, MD 2002, 'Retinoid quantification by HPLC/MS(n)', Journal of Lipid Research, vol. 43, no. 7, pp. 1143-9.
McCaffery P, Evans J, Koul O, Volpert A, Reid K, Ullman MD. Retinoid quantification by HPLC/MS(n). Journal of Lipid Research. 2002;43(7):1143-9.
McCaffery, Peter ; Evans, James ; Koul, Omanand ; Volpert, Amy ; Reid, Kevin ; Ullman, M David. / Retinoid quantification by HPLC/MS(n). In: Journal of Lipid Research. 2002 ; Vol. 43, No. 7. pp. 1143-9.
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AU - Ullman, M David

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N2 - Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

AB - Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

KW - Animals

KW - Chromatography, High Pressure Liquid

KW - Isomerism

KW - Liver

KW - Male

KW - Mass Spectrometry

KW - Rats

KW - Rats, Sprague-Dawley

KW - Reproducibility of Results

KW - Retina

KW - Retinoids

KW - Sensitivity and Specificity

KW - Tretinoin

M3 - Article

VL - 43

SP - 1143

EP - 1149

JO - Journal of Lipid Research

JF - Journal of Lipid Research

SN - 0022-2275

IS - 7

ER -