Highly sensitive fluorescent method for the detection of cholesterol aldehydes formed by ozone and singlet molecular oxygen

Fernando V. Mansano, Rafaella M.A. Kazaoka, Graziella E. Ronsein, Fernanda M. Prado, Thiago Cardoso Genaro De Mattos, Miriam Uemi, Paolo Di Mascio, Sayuri Miyamoto

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Cholesterol oxidation gives rise to a mixture of oxidized products. Different types of products are generated according to the reactive species being involved. Recently, attention has been focused on two cholesterol aldehydes, 3β-hydroxy-5β-hydroxy-B-norcholestane-6β- carboxyaldehyde (1a) and 3β-hydroxy-5-oxo-5,6-secocholestan-6-al (1b). These aldehydes can be generated by ozone-, as well as by singlet molecular oxygen-mediated cholesterol oxidation. It has been suggested that 1b is preferentially formed by ozone and 1a is preferentially formed by singlet molecular oxygen. In this study we describe the use of 1-pyrenebutyric hydrazine (PBH) as a fluorescent probe for the detection of cholesterol aldehydes. The formation of the fluorescent adduct between 1a with PBH was confirmed by HPLC-MS/MS. The fluorescence spectra of PBH did not change upon binding to the aldehyde. Moreover, the derivatization was also effective in the absence of an acidified medium, which is critical to avoid the formation of cholesterol aldehydes through Hock cleavage of 5α-hydroperoxycholesterol. In conclusion, PBH can be used as an efficient fluorescent probe for the detection/quantification of cholesterol aldehydes in biological samples. Its analysis by HPLC coupled to a fluorescent detector provides a sensitive and specific way to quantify cholesterol aldehydes in the low femtomol range.

Original languageEnglish (US)
Pages (from-to)6775-6781
Number of pages7
JournalAnalytical Chemistry
Volume82
Issue number16
DOIs
StatePublished - Aug 15 2010

Fingerprint

hydrazine
Molecular oxygen
Ozone
Aldehydes
Cholesterol
Fluorescent Dyes
Oxidation
Fluorescence
Detectors

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Highly sensitive fluorescent method for the detection of cholesterol aldehydes formed by ozone and singlet molecular oxygen. / Mansano, Fernando V.; Kazaoka, Rafaella M.A.; Ronsein, Graziella E.; Prado, Fernanda M.; Cardoso Genaro De Mattos, Thiago; Uemi, Miriam; Mascio, Paolo Di; Miyamoto, Sayuri.

In: Analytical Chemistry, Vol. 82, No. 16, 15.08.2010, p. 6775-6781.

Research output: Contribution to journalArticle

Mansano, Fernando V. ; Kazaoka, Rafaella M.A. ; Ronsein, Graziella E. ; Prado, Fernanda M. ; Cardoso Genaro De Mattos, Thiago ; Uemi, Miriam ; Mascio, Paolo Di ; Miyamoto, Sayuri. / Highly sensitive fluorescent method for the detection of cholesterol aldehydes formed by ozone and singlet molecular oxygen. In: Analytical Chemistry. 2010 ; Vol. 82, No. 16. pp. 6775-6781.
@article{43c214cd064d4bfa82c86698452336ea,
title = "Highly sensitive fluorescent method for the detection of cholesterol aldehydes formed by ozone and singlet molecular oxygen",
abstract = "Cholesterol oxidation gives rise to a mixture of oxidized products. Different types of products are generated according to the reactive species being involved. Recently, attention has been focused on two cholesterol aldehydes, 3β-hydroxy-5β-hydroxy-B-norcholestane-6β- carboxyaldehyde (1a) and 3β-hydroxy-5-oxo-5,6-secocholestan-6-al (1b). These aldehydes can be generated by ozone-, as well as by singlet molecular oxygen-mediated cholesterol oxidation. It has been suggested that 1b is preferentially formed by ozone and 1a is preferentially formed by singlet molecular oxygen. In this study we describe the use of 1-pyrenebutyric hydrazine (PBH) as a fluorescent probe for the detection of cholesterol aldehydes. The formation of the fluorescent adduct between 1a with PBH was confirmed by HPLC-MS/MS. The fluorescence spectra of PBH did not change upon binding to the aldehyde. Moreover, the derivatization was also effective in the absence of an acidified medium, which is critical to avoid the formation of cholesterol aldehydes through Hock cleavage of 5α-hydroperoxycholesterol. In conclusion, PBH can be used as an efficient fluorescent probe for the detection/quantification of cholesterol aldehydes in biological samples. Its analysis by HPLC coupled to a fluorescent detector provides a sensitive and specific way to quantify cholesterol aldehydes in the low femtomol range.",
author = "Mansano, {Fernando V.} and Kazaoka, {Rafaella M.A.} and Ronsein, {Graziella E.} and Prado, {Fernanda M.} and {Cardoso Genaro De Mattos}, Thiago and Miriam Uemi and Mascio, {Paolo Di} and Sayuri Miyamoto",
year = "2010",
month = "8",
day = "15",
doi = "10.1021/ac1006427",
language = "English (US)",
volume = "82",
pages = "6775--6781",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "16",

}

TY - JOUR

T1 - Highly sensitive fluorescent method for the detection of cholesterol aldehydes formed by ozone and singlet molecular oxygen

AU - Mansano, Fernando V.

AU - Kazaoka, Rafaella M.A.

AU - Ronsein, Graziella E.

AU - Prado, Fernanda M.

AU - Cardoso Genaro De Mattos, Thiago

AU - Uemi, Miriam

AU - Mascio, Paolo Di

AU - Miyamoto, Sayuri

PY - 2010/8/15

Y1 - 2010/8/15

N2 - Cholesterol oxidation gives rise to a mixture of oxidized products. Different types of products are generated according to the reactive species being involved. Recently, attention has been focused on two cholesterol aldehydes, 3β-hydroxy-5β-hydroxy-B-norcholestane-6β- carboxyaldehyde (1a) and 3β-hydroxy-5-oxo-5,6-secocholestan-6-al (1b). These aldehydes can be generated by ozone-, as well as by singlet molecular oxygen-mediated cholesterol oxidation. It has been suggested that 1b is preferentially formed by ozone and 1a is preferentially formed by singlet molecular oxygen. In this study we describe the use of 1-pyrenebutyric hydrazine (PBH) as a fluorescent probe for the detection of cholesterol aldehydes. The formation of the fluorescent adduct between 1a with PBH was confirmed by HPLC-MS/MS. The fluorescence spectra of PBH did not change upon binding to the aldehyde. Moreover, the derivatization was also effective in the absence of an acidified medium, which is critical to avoid the formation of cholesterol aldehydes through Hock cleavage of 5α-hydroperoxycholesterol. In conclusion, PBH can be used as an efficient fluorescent probe for the detection/quantification of cholesterol aldehydes in biological samples. Its analysis by HPLC coupled to a fluorescent detector provides a sensitive and specific way to quantify cholesterol aldehydes in the low femtomol range.

AB - Cholesterol oxidation gives rise to a mixture of oxidized products. Different types of products are generated according to the reactive species being involved. Recently, attention has been focused on two cholesterol aldehydes, 3β-hydroxy-5β-hydroxy-B-norcholestane-6β- carboxyaldehyde (1a) and 3β-hydroxy-5-oxo-5,6-secocholestan-6-al (1b). These aldehydes can be generated by ozone-, as well as by singlet molecular oxygen-mediated cholesterol oxidation. It has been suggested that 1b is preferentially formed by ozone and 1a is preferentially formed by singlet molecular oxygen. In this study we describe the use of 1-pyrenebutyric hydrazine (PBH) as a fluorescent probe for the detection of cholesterol aldehydes. The formation of the fluorescent adduct between 1a with PBH was confirmed by HPLC-MS/MS. The fluorescence spectra of PBH did not change upon binding to the aldehyde. Moreover, the derivatization was also effective in the absence of an acidified medium, which is critical to avoid the formation of cholesterol aldehydes through Hock cleavage of 5α-hydroperoxycholesterol. In conclusion, PBH can be used as an efficient fluorescent probe for the detection/quantification of cholesterol aldehydes in biological samples. Its analysis by HPLC coupled to a fluorescent detector provides a sensitive and specific way to quantify cholesterol aldehydes in the low femtomol range.

UR - http://www.scopus.com/inward/record.url?scp=77955644918&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955644918&partnerID=8YFLogxK

U2 - 10.1021/ac1006427

DO - 10.1021/ac1006427

M3 - Article

VL - 82

SP - 6775

EP - 6781

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 16

ER -