Sterols in blood of normal and Smith-Lemli-Opitz subjects

Benfang Ruan, William K. Wilson, Jihai Pang, Nicolas Gerst, Frederick D. Pinkerton, James Tsai, Richard I. Kelley, Frank G. Whitby, Dianna M. Milewicz, James Garbern, George J. Schroepfer

Research output: Contribution to journalArticle

Abstract

Smith-Lemli-Opitz syndrome (SLOS) is a hereditary disorder in which a defective gene encoding 7-dehydrocholesterol reductase causes the accumulation of noncholesterol sterols, such as 7- and 8-dehydrocholesterol. Using rigorous analytical methods in conjunction with a large collection of authentic standards, we unequivocally identified numerous noncholesterol sterols in 6 normal and 17 SLOS blood samples. Plasma or erythrocytes were saponified under oxygen-free conditions, followed by multiple chromatographic separations. Individual sterols were identified and quantitated by high performance liquid chromatography (HPLC), Ag+-HPLC, gas chromatography (GC), GC-mass spectrometry, and nuclear magnetic resonance. As a percentage of total sterol content, the major C27 sterols observed in the SLOS blood samples were cholesterol (12-98%), 7-dehydrocholesterol (0.4-44%), 8-dehydrocholesterol (0.5-22%), and cholesta-5,7,9(11)-trien-3β-ol (0.02-5%), whereas the normal blood samples contained 8, Δ8(14), Δ5,8(14), Δ5,24, Δ6,8, Δ6,8(14), and Δ7,24 sterols. The results are consistent with the hypothesis that the Δ8(14) sterol is an intermediate of cholesterol synthesis and indicate the existence of undescribed aberrant pathways that may explain the formation of the Δ5,7,9(11) sterol. 19-Norcholesta-5,7,9-trien-3β-ol was absent in both SLOS and normal blood, although it was routinely observed as a GC artifact in fractions containing 8-dehydrocholesterol. The overall findings advance the understanding of SLOS and provide a methodological model for studying other metabolic disorders of cholesterol synthesis.

Original languageEnglish (US)
Pages (from-to)799-812
Number of pages14
JournalJournal of Lipid Research
Volume42
Issue number5
StatePublished - 2001

Fingerprint

Sterols
Smith-Lemli-Opitz Syndrome
Blood
Gas chromatography
Trientine
Cholesterol
High performance liquid chromatography
Gas Chromatography
High Pressure Liquid Chromatography
Gene encoding
Artifacts
Gas Chromatography-Mass Spectrometry
Mass spectrometry
Magnetic Resonance Spectroscopy
Erythrocytes
Nuclear magnetic resonance
Oxygen
Plasmas
Genes
cholesta-5,8-dien-3 beta-ol

Keywords

  • Ag-HPLC
  • Cholesta-5,8-dien-3β-ol
  • GC-MS
  • NMR
  • Noncholesterol sterols
  • SLOS

ASJC Scopus subject areas

  • Endocrinology

Cite this

Ruan, B., Wilson, W. K., Pang, J., Gerst, N., Pinkerton, F. D., Tsai, J., ... Schroepfer, G. J. (2001). Sterols in blood of normal and Smith-Lemli-Opitz subjects. Journal of Lipid Research, 42(5), 799-812.

Sterols in blood of normal and Smith-Lemli-Opitz subjects. / Ruan, Benfang; Wilson, William K.; Pang, Jihai; Gerst, Nicolas; Pinkerton, Frederick D.; Tsai, James; Kelley, Richard I.; Whitby, Frank G.; Milewicz, Dianna M.; Garbern, James; Schroepfer, George J.

In: Journal of Lipid Research, Vol. 42, No. 5, 2001, p. 799-812.

Research output: Contribution to journalArticle

Ruan, B, Wilson, WK, Pang, J, Gerst, N, Pinkerton, FD, Tsai, J, Kelley, RI, Whitby, FG, Milewicz, DM, Garbern, J & Schroepfer, GJ 2001, 'Sterols in blood of normal and Smith-Lemli-Opitz subjects', Journal of Lipid Research, vol. 42, no. 5, pp. 799-812.
Ruan B, Wilson WK, Pang J, Gerst N, Pinkerton FD, Tsai J et al. Sterols in blood of normal and Smith-Lemli-Opitz subjects. Journal of Lipid Research. 2001;42(5):799-812.
Ruan, Benfang ; Wilson, William K. ; Pang, Jihai ; Gerst, Nicolas ; Pinkerton, Frederick D. ; Tsai, James ; Kelley, Richard I. ; Whitby, Frank G. ; Milewicz, Dianna M. ; Garbern, James ; Schroepfer, George J. / Sterols in blood of normal and Smith-Lemli-Opitz subjects. In: Journal of Lipid Research. 2001 ; Vol. 42, No. 5. pp. 799-812.
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title = "Sterols in blood of normal and Smith-Lemli-Opitz subjects",
abstract = "Smith-Lemli-Opitz syndrome (SLOS) is a hereditary disorder in which a defective gene encoding 7-dehydrocholesterol reductase causes the accumulation of noncholesterol sterols, such as 7- and 8-dehydrocholesterol. Using rigorous analytical methods in conjunction with a large collection of authentic standards, we unequivocally identified numerous noncholesterol sterols in 6 normal and 17 SLOS blood samples. Plasma or erythrocytes were saponified under oxygen-free conditions, followed by multiple chromatographic separations. Individual sterols were identified and quantitated by high performance liquid chromatography (HPLC), Ag+-HPLC, gas chromatography (GC), GC-mass spectrometry, and nuclear magnetic resonance. As a percentage of total sterol content, the major C27 sterols observed in the SLOS blood samples were cholesterol (12-98{\%}), 7-dehydrocholesterol (0.4-44{\%}), 8-dehydrocholesterol (0.5-22{\%}), and cholesta-5,7,9(11)-trien-3β-ol (0.02-5{\%}), whereas the normal blood samples contained 8, Δ8(14), Δ5,8(14), Δ5,24, Δ6,8, Δ6,8(14), and Δ7,24 sterols. The results are consistent with the hypothesis that the Δ8(14) sterol is an intermediate of cholesterol synthesis and indicate the existence of undescribed aberrant pathways that may explain the formation of the Δ5,7,9(11) sterol. 19-Norcholesta-5,7,9-trien-3β-ol was absent in both SLOS and normal blood, although it was routinely observed as a GC artifact in fractions containing 8-dehydrocholesterol. The overall findings advance the understanding of SLOS and provide a methodological model for studying other metabolic disorders of cholesterol synthesis.",
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AU - Gerst, Nicolas

AU - Pinkerton, Frederick D.

AU - Tsai, James

AU - Kelley, Richard I.

AU - Whitby, Frank G.

AU - Milewicz, Dianna M.

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AU - Schroepfer, George J.

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N2 - Smith-Lemli-Opitz syndrome (SLOS) is a hereditary disorder in which a defective gene encoding 7-dehydrocholesterol reductase causes the accumulation of noncholesterol sterols, such as 7- and 8-dehydrocholesterol. Using rigorous analytical methods in conjunction with a large collection of authentic standards, we unequivocally identified numerous noncholesterol sterols in 6 normal and 17 SLOS blood samples. Plasma or erythrocytes were saponified under oxygen-free conditions, followed by multiple chromatographic separations. Individual sterols were identified and quantitated by high performance liquid chromatography (HPLC), Ag+-HPLC, gas chromatography (GC), GC-mass spectrometry, and nuclear magnetic resonance. As a percentage of total sterol content, the major C27 sterols observed in the SLOS blood samples were cholesterol (12-98%), 7-dehydrocholesterol (0.4-44%), 8-dehydrocholesterol (0.5-22%), and cholesta-5,7,9(11)-trien-3β-ol (0.02-5%), whereas the normal blood samples contained 8, Δ8(14), Δ5,8(14), Δ5,24, Δ6,8, Δ6,8(14), and Δ7,24 sterols. The results are consistent with the hypothesis that the Δ8(14) sterol is an intermediate of cholesterol synthesis and indicate the existence of undescribed aberrant pathways that may explain the formation of the Δ5,7,9(11) sterol. 19-Norcholesta-5,7,9-trien-3β-ol was absent in both SLOS and normal blood, although it was routinely observed as a GC artifact in fractions containing 8-dehydrocholesterol. The overall findings advance the understanding of SLOS and provide a methodological model for studying other metabolic disorders of cholesterol synthesis.

AB - Smith-Lemli-Opitz syndrome (SLOS) is a hereditary disorder in which a defective gene encoding 7-dehydrocholesterol reductase causes the accumulation of noncholesterol sterols, such as 7- and 8-dehydrocholesterol. Using rigorous analytical methods in conjunction with a large collection of authentic standards, we unequivocally identified numerous noncholesterol sterols in 6 normal and 17 SLOS blood samples. Plasma or erythrocytes were saponified under oxygen-free conditions, followed by multiple chromatographic separations. Individual sterols were identified and quantitated by high performance liquid chromatography (HPLC), Ag+-HPLC, gas chromatography (GC), GC-mass spectrometry, and nuclear magnetic resonance. As a percentage of total sterol content, the major C27 sterols observed in the SLOS blood samples were cholesterol (12-98%), 7-dehydrocholesterol (0.4-44%), 8-dehydrocholesterol (0.5-22%), and cholesta-5,7,9(11)-trien-3β-ol (0.02-5%), whereas the normal blood samples contained 8, Δ8(14), Δ5,8(14), Δ5,24, Δ6,8, Δ6,8(14), and Δ7,24 sterols. The results are consistent with the hypothesis that the Δ8(14) sterol is an intermediate of cholesterol synthesis and indicate the existence of undescribed aberrant pathways that may explain the formation of the Δ5,7,9(11) sterol. 19-Norcholesta-5,7,9-trien-3β-ol was absent in both SLOS and normal blood, although it was routinely observed as a GC artifact in fractions containing 8-dehydrocholesterol. The overall findings advance the understanding of SLOS and provide a methodological model for studying other metabolic disorders of cholesterol synthesis.

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