HPLC separation and quantitation of cholesterol oxidation products with flame ionization detection

Maerker, G.; Nungesser, Edwin; Zulak, I. M.

doi:10.1021/jf00079a016

Cited by 24 publications

(10 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The best resolution was achieved by using a linear gradient of isopropanol (from 2 to 5%) and heptane at a flow rate of 0.6 mL/min. Our results were similar to those of Maerker et al [24], who reported that with isocratic solvent systems the full resolution over the entire polarity range of sterol oxidation products was not achieved. With isocratic elution (99:1, 98:2 and 97:3 heptane/isopropanol), 7-ketostigmasterol and 7b-OOH-stigmasterol coelute, complicating the quantitation of 7-ketostigmasterol.…”

Section: Applicability Of the Hplc Methods For Simultaneously Analyzinsupporting

confidence: 92%

Formation and decomposition of stigmasterol hydroperoxides and secondary oxidation products during thermo‐oxidation

Kemmo

Soupas

Lampi

et al. 2005

Eur. J. Lipid Sci. Technol.

View full text Add to dashboard Cite

The oxidation mechanisms of stigmasterol at 100 and 180 7C were investigated by using the HPLC-UV-FL method. An overall picture of the oxidation status was achieved with a single HPLC analysis, enabling us to monitor the formation and decomposition of both primary and secondary oxidation products. The oxidation behavior of stigmasterol was different at the two temperatures. At 180 7C, the amounts of hydroperoxides increased sharply during the first 10 min and then began to decrease. At 100 7C, the amounts of hydroperoxides increased over the entire experimental period. At 180 7C, all major secondary oxidation products, except 7-ketostigmasterol, reached a plateau after 40 min of oxidation, while at 100 7C their amounts increased constantly. The same oxidation products were formed at both temperatures, but their distribution differed. At 180 7C, the formation of free radicals at position 7 was more favorable than formation of radicals at position 25. The situation was the opposite at 100 7C; radicals formed more easily at the tertiary position 25. At 180 7C, 7-ketostigmasterol was dominant after 40 min of oxidation, whereas at 100 7C it was the main oxidation product over the entire experiment.

show abstract

Section: Applicability Of the Hplc Methods For Simultaneously Analyzinsupporting

confidence: 92%

Formation and decomposition of stigmasterol hydroperoxides and secondary oxidation products during thermo‐oxidation

Kemmo

Soupas

Lampi

et al. 2005

Eur. J. Lipid Sci. Technol.

View full text Add to dashboard Cite

show abstract

“…One of the problems encountered in these studies is the method of analysis; capillary gas chromatography (GC) (Park and Addis, 1985;van de Bovenkamp et al, 1988;Pie et al, 1990) and high-performance liquid chromatography (Tsai and Hudson, 1981;Maerker et al, 1988;Csallany et al, 1989) have been adopted, but it is generally difficult to resolve individual oxidized cholesterols satisfactory. Previous studies (Bascoul et al, 1986;Ryan et al, 1981;Yan and White, 1990) on the autoxidation of cholesterol by heating with fat such as lard and tallow detected 7a-hydroxycholesterol, 7~-hydroxycholesterol, 5a-epoxycholesterol, 5~-epoxycholesterol, cholestanetriol, and 7-ketocholesterol.…”

Section: Introductionmentioning

confidence: 99%

Oxidation of cholesterol by heating

Osada¹,

Kodama²,

Yamada³

et al. 1993

J. Agric. Food Chem.

View full text Add to dashboard Cite

Oxidation of pure cholesterol during heating in an air oven at high temperature was studied. Cholesterol was virtually stable during heating at 100 OC for 24 h but was unstable at temperatures above 120 OC. In the heated cholesterol preparations, a number of oxidized derivatives were detected by a combination of thin-layer chromatography and capillary gas chromatography-mass spectrometry. Major oxidized sterols were 7a-hydroxycho1estero1, 7,3-hydroxycholestero1, 5a-epoxycho1estero1, 5,3-epoxycholesterol, cholestanetriol, and 7-ketocholesterol. Various oxidized cholesterol derivatives were produced during heating above 120 "C within a relatively short time (1 h). The composition of the oxidized products differed depending on temperature and time of heating. When cholesterol was heated at 150 OC, the production of oxidized cholesterol was maximum, and 7-ketocholesterol was the most predominant oxidized product. Heating a t 120 OC also produced oxidized cholesterol to some extent, whereas only marginal amounts of oxidized cholesterols were produced a t 100 "C and at 200 "C cholesterol was almost decomposed in a short time.

show abstract

“…20α-Hydroxycholesterol 20αOC a 13 6-Ketocholestanol 6KetoC a 14 19-Hydroxycholesterol 19HOC a 15 5α-Cholestan-3,6-dione 5C36D c rivative standard. Figure 1 shows UV absorbance for cholesterol and eight available species of oxidized cholesterol derivative.…”

Section: Resultsmentioning

confidence: 99%

“…In contrast to GC analysis of oxidized cholesterol derivatives, the HPLC analysis condition of oxidized cholesterol derivatives is not completely established. Several studies measured the levels of oxidized cholesterol derivatives by HPLC with ultraviolet (UV) (5)(6)(7)(8), refractive index (RI) (9), evaporative light-scattering detection (ELSD) (10)(11)(12), and flame-ionization detector (FID) (13), using various adsorbent columns. Many of these analytical methods leave unresolved problems concerning sensitivity, baseline instability, and in application to samples derived from foods and biological specimen; however, Caboni et al (12) showed superior ELSD detection of oxidized cholesterol derivatives using cyano-bonded normal-phase column within 30 min.…”

mentioning

confidence: 99%

Rapid analysis of oxidized cholesterol derivatives by high‐performance liquid chromatography combined with diode‐array ultraviolet and evaporative laser light‐scattering detection

Osada

Ravandi

Kuksis

1999

J Americ Oil Chem Soc

View full text Add to dashboard Cite

Extensive evidence of the deleterious biological effects of oxidized 5-cholesten-3β-ol (cholesterol) derivatives has led to great interest in their detection. We observed that known oxidized cholesterol derivatives can be rapidly quantitated by combining reversed-phase high-performance liquid chromatography (HPLC) with ultraviolet (UV) absorption and evaporative laser light-scattering (ELSD) detection. Using a 20 × 0.46 cm C18 HPLC column and methanol/acetonitrile (60:40, vol/vol) as the mobile phase at 1.0 mL/min, 10 species of oxidized cholesterol derivative were measured by UV (205, 234, and 280 nm) while 5-cholestan-5α,6α-epoxy-3β-ol (5α-epoxycholesterol), 5-cholestan-5β,6β-epoxy-3β-ol (5β-epoxycholesterol), and 5-cholestan-3β,5α,6β-triol (cholestanetriol) were detected by only ELSD. The minimal limits of detection ranged from 100 to 500 ng depending on sterol and detector. The response was linear in the range 0-1000 or 0-2000 ng depending on detector. These oxidized cholesterol derivatives were also identified by HPLC/mass spectrometry analysis combined with UV detector.Heated tallow contained cholestanetriol, 5-cholesten-3β,7α-diol (7α-hydroxycholesterol), 5-cholesten-3β,7β-diol (7β-hydroxycholesterol), 5-cholesten-3β-ol-7-one (7-ketocholesterol), 5α-and 5β-epoxycholesterols under the developed analysis condition. Photooxidized cholesterol had cholestanetriol, 7α-and 7β-hydroxycholesterols and 3,5-cholestadien-7-one. On the other hand, 7α-and 7β-hydroxycholesterols, 7-ketocholesterol, 5α-and 5β-epoxycholesterols and 3,5-cholestadien-7-one were observed in copper-oxidized low-density lipoprotein. Thus, this developed HPLC analysis method could be applied to identification of oxidized cholesterol derivatives in food and biological specimen.

show abstract

HPLC separation and quantitation of cholesterol oxidation products with flame ionization detection

Cited by 24 publications

References 19 publications

Formation and decomposition of stigmasterol hydroperoxides and secondary oxidation products during thermo‐oxidation

Formation and decomposition of stigmasterol hydroperoxides and secondary oxidation products during thermo‐oxidation

Oxidation of cholesterol by heating

Rapid analysis of oxidized cholesterol derivatives by high‐performance liquid chromatography combined with diode‐array ultraviolet and evaporative laser light‐scattering detection

Contact Info

Product

Resources

About