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Quantitative Determination of Sitosterol in the Presence of Cholesterol

IP.com Disclosure Number: IPCOM000085950D
Original Publication Date: 1976-Jun-01
Included in the Prior Art Database: 2005-Mar-03
Document File: 2 page(s) / 37K

Publishing Venue

IBM

Related People

Slocum, SH: AUTHOR [+2]

Abstract

The determination of the presence of sitosterol in blood may be valuable in the diagnosis of breast cancer, but interference from cholesterol makes this determination difficult because sitosterol is a compound with a molecular structure very similar to that of cholesterol (Fig. 1).

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Quantitative Determination of Sitosterol in the Presence of Cholesterol

The determination of the presence of sitosterol in blood may be valuable in the diagnosis of breast cancer, but interference from cholesterol makes this determination difficult because sitosterol is a compound with a molecular structure very similar to that of cholesterol (Fig. 1).

In detecting the presence of sitosterol in blood, it is necessary to use a technique capable of resolution high enough to distinguish between the two compounds. High-pressure liquid chromatography (HPLC) has this high- resolving power, but at the concentrations at which they are present in blood, sitosterol and cholesterol cannot be detected by the UV detectors generally used for this technique.

Treating these two compounds with the enzyme cholesterol oxidase yields products -- sitostenone and cholestenone -- which can be detected easily by the HPLC UV detector, since the products absorb UV light very strongly (Fig. 2).

As shown in Fig. 3, it is possible to separate cholestenone from sitostenone using a 1/4'' x 12'' mu Bondapak C(18) column (Waters Associates).

Gas chromatography (GC) can also be used to separate cholestenone and sitostenone [1], but considerably more effort must go into sample preparation when GC is used in place of HPLC. This difference is a significant advantage for HPLC in this analysis.

In summary, it is proposed that cholesterol oxidase be used to convert cholesterol and sitosterol in blood s...