Novel Fast Chromatography-Tandem Mass Spectrometric Quantitative Approach for the Determination of Plant-Extracted Phytosterols and Tocopherols

Abstract

Phytosterols and tocopherols are commonly used in food and pharmaceutical industries for their health benefits. Current analysis methods rely on conventional liquid chromatography, using an analytical column, which can be tedious and time consuming. However, simple, and fast analytical methods can facilitate their qualitative and quantitative analysis. In this study, a fast chromatography-tandem mass spectrometric (FC-MS/MS) method was developed and validated for the quantitative analysis of phytosterols and tocopherols. Omitting chromatography by employing flow injection analysis—mass spectrometry (FIA-MS) failed in the quantification of target analytes due to analyte-to-analyte interferences from phytosterols. These interferences arise from their ambiguous MS fingerprints that would lead to false identification and inaccurate quantification. Therefore, a C18 guard column with a 1.9 µm particle size was employed for FC-MS/MS under isocratic elution using acetonitrile/methanol (99:1 v/v) at a flow rate of 600 µL/min. Analyte-to-analyte interferences were identified and eliminated. The false peaks could then be easily identified due to chromatographic separation. In addition, two internal standards were evaluated, namely cholestanol and deuterated cholesterol. Both internal standards contributed to the observed analyte-to-analyte interferences; however, adequate shift in the retention time for deuterated cholesterol eliminated its interferences and allowed for an accurate quantification. The method is fast (1.3 min) compared to published methods and can distinguish false peaks observed in FIA-MS. Seven analytes were quantified simultaneously, namely brassicasterol, campesterol, stigmasterol, β-sitosterol, α-tocopherol, δ-tocopherol, and γ-tocopherol. The method was successfully applied in the quantitative analysis of phytosterols and tocopherols present in the unsaponifiable matter of canola oil deodorizer distillate (CODD). β-sitosterol and γ-tocopherol were the most abundant phytosterols and tocopherols, respectively.

Liposomal Phytosterols as LDL-Cholesterol-Lowering Agents in Diet-Induced Hyperlipidemia

“…The resulting mixture was chilled for 1 h. During the chilling process, PS crystallized and was collected by vacuum filtration. The PS was composed of brassicasterol, campesterol, and β-sitosterol (Table ), and the purity of the extract was >90% as determined by our previously developed LC-MS/MS method . In brief, 1 mg/mL PS extract was prepared in chloroform and was analyzed in LC-MS/MS using freshly prepared calibration and quality control standards .…”

“…The PS was composed of brassicasterol, campesterol, and β-sitosterol (Table ), and the purity of the extract was >90% as determined by our previously developed LC-MS/MS method . In brief, 1 mg/mL PS extract was prepared in chloroform and was analyzed in LC-MS/MS using freshly prepared calibration and quality control standards . The development of the liposomal PS was reported in our published work, which showed a 90% entrapment efficacy. , In brief, the PS was formulated into liposomes composed of PC and tocopherols.…”

“…24 In brief, 1 mg/mL PS extract was prepared in chloroform and was analyzed in LC-MS/MS using freshly prepared calibration and quality control standards. 24 The development of the liposomal PS was reported in our published work, which showed a 90% entrapment efficacy. 25,26 In brief, the PS was formulated into liposomes composed of PC and tocopherols.…”

Advantages of Multiplexing Ability of the Orbitrap Mass Analyzer in the Multi-Mycotoxin Analysis

“…However, simultaneous introduction of numerous compounds into the ion-source results in abundant matrix effects leading to sensitivity loss [ 2 ]. Recently, dilute-and-shoot methods [ 3 ] and the use of a short or guard column in the FIA-MS method (i.e., fast chromatography) [ 4 ] have been developed to overcome this effect.…”

Investigating the Tocopherol Contents of Walnut Seed Oils Produced in Different European Countries Analyzed by HPLC-UV: A Comparative Study on the Basis of Geographical Origin

“…Several analytical protocols have been proposed in the literature for tocopherol analysis, employing high-pressure liquid chromatography coupled to diode array (HPLC-DAD) [ 27 , 28 , 29 ], UV–Vis [ 30 , 31 ], fluorescence (FLD) [ 32 ], or mass spectrometric (MS) detectors [ 33 , 34 , 35 , 36 ]. Undoubtedly, UV detectors allow a fast, reliable, and cost-effective analysis, especially when it comes to such a small number of compounds, where separation and identification can be achieved without much adversity.…”