Phytosterols HPLC

The chemical purity of these radiolabeled compounds is very high from most sources. If in question, the purity can be checked and restored by standard TLC and/or HPLC methods. 14C-labeled phytosterols are less available. 

Some of the tests involved relatively simple colour reactions such as the Baudouin reaction for sesame oil, and the Halphen test for cottonseed oil. In both cases a compound characteristic to an oil is used to determine the presence of the oil. Here again the test detected a component that today would be detected and quantified by gas chromatography (GC) or high performance liquid chromatography (HPLC). It was even possible to determine the presence of cholesterol or phytosterols, although, after separation, the identification as to which type was present depended on microscopic examination and fractional... 

Adulteration of fats and oils is an old problem. Many older tests involved determination of physical properties such as refractive index, melting point, and viscosity. However, color tests were later used for this purpose. Thus, Baudonin reaction for sesame oil and the Halpben test for cottonseed oil have been noted. In both cases, a compound characteristic to an oil determines the presence of the oil. However, today such detections and quantitations are carried out with GC and HPLC procedures. Thus, cholesterol and phytosterols may be determined by gas chromatography for fingerprinting purposes however, fatty acid analysis might also be used for higher levels of contamination (31). Detailed discussion of issues related to oil authentication and adulteration has taken place (11). 

Warner, K. and Mounts, T.L. 1990. Analysis of Tocopherols and Phytosterols in Vegetable Oils by HPLC with Evaporative Light-Scattering Detection. J. Am. Oil Chem. Soc. 67 827-831. 

Anacardium occidentale (Anacardiacae) Anacardic acid,alpha-linolenic acid, anacardol, antimony, cardol, europium, folacin, gingkol,leucine, glucuronic acid, glutamic acid, haftiium, serine, phytosterol, praline, vitamin c, kaempferol, terpenes, quercetin-glycoside Terpenes HPLC (59)... 

Likewise, although the quantification of phytosterols in VOO analysis is usually performed by GC with different detectors [80,85,86], recently published research has proposed new analytical methods for the precise determination of phytosterols in olive oil using HPLC-MS [22,87], and more recently, highly sensitive methods based on UHPLC-MS were applied in the determination of VOO phytosterols [88,89]. 

Furthermore, using UHPLC-MS in VOO phytosterols analysis led to a remarkable reduction in the analysis time (both in the sample preparation step and in the phytosterols separation). The separation was achieved in 5 min, time in which 14 phytosterol compounds were identified (Figure 9.8). Thereby, the analysis time using UHPLC-MS is four to six times faster than those of HPLC-MS methods [22,82,90], where the time required is between 25 and 30 min. The comparison is even more... 

Figure 1. HPLC-ELSD chromatogram of corn fiber oil. Insert shows the structure of sitostanyl-ferulate, the major ferulate-phytosterol ester in corn fiber oil (Norton, 1995). Abbreviations StE, phytosterol fatty acyl ester TAG, triacylglycerol, a-Toc, a-tocopherol FFA, free fatty acids CE, p-coumarate-phytosterol esters y-Toc, y tocopherol ST, free phytosterols 1,3-D, 1,2-diacylglycerols FE, ferulate-phytosterol esters 1,2-D, 1,2-diacylglycerols.

K Warner, TL Mounts. Analysis of tocopherols and phytosterols in vegetable oils by HPLC with evaporative hght-scattering detection. J Am Oil Chem Soc 67 827-831, 1990. 

Slavin, M. and L. (L) Yu. 2012. A single extraction and HPLC procedure for simultaneous analysis of phytosterols, tocopherols and lutein in soybeans. Food Chem. 135 2789-2795. 

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