International descriptive and interventional survey for oxycholesterol determination by gas- and liquid-chromatographic methods.

Increasing numbers of laboratories develop new methods based on gas-liquid and high-performance liquid chromatography to determine serum concentrations of oxygenated cholesterol metabolites such as 7α-, 24(S)-, and 27-hydroxycholesterol. We initiated a first international descriptive oxycholesterol (OCS) survey in 2013 and a second interventional survey 2014 in order to compare levels of OCS reported by different laboratories and to define possible sources of analytical errors. In 2013 a set of two lyophilized serum pools (A and B) was sent to nine laboratories in different countries for OCS measurement utilizing their own standard stock solutions. In 2014 eleven laboratories were requested to determine OCS concentrations in lyophilized pooled sera (C and D) utilizing the same provided standard stock solutions of OCS. The participating laboratories submitted results obtained after capillary gas-liquid chromatography-mass selective detection with either epicoprostanol or deuterium labelled sterols as internal standards and high-performance liquid chromatography with mass selective detection and deuterated OCS as internal standard. Each participant received a clear overview of the results in form of Youden-Plots and basic statistical evaluation in its used unit. The coefficients of variation of the concentrations obtained by all laboratories using their individual methods were 58.5-73.3% (survey 1), 56.8-60.3% (survey 2); 36.2-35.8% (survey 1), 56.6-59.8, (survey 2); 61.1-197.7% (survey 1), 47.2-74.2% (survey 2) for 24(S)-, 27-, and 7α-hydroxycholesterol, respectively. We are surprised by the very great differences between the laboratories, even under conditions when the same standards were used. The values of OCS's must be evaluated in relation to the analytical technique used, the efficiency of the ample separation and the nature of the internal standard used. Quantification of the calibration solution and inappropriate internal standards could be identified as major causes for the high variance in the reported results from the different laboratories. A harmonisation of analytical standard methods is highly needed.

Cholesterol and stigmasterol within a sunflower oil matrix: Thermal degradation and oxysterols formation.

The characteristics of the lipid matrix surrounding sterols exert a great influence in their thermal oxidation process. The objective of this work was to assess the oxidation susceptibility of equal amounts of cholesterol and stigmasterol within a sunflower oil lipid matrix (ratio 1:1:200) during heating (180°C, 0-180min). Remaining percentage of sterols was determined and seven sterol oxidation products (SOPs) were analysed for each type of sterol along the heating treatment. Evolution of the fatty acid profile and vitamin E content of the oil was also studied. Overall oxidation status of the model system was assessed by means of Peroxides Value (PV) and TBARS. PV remained constant from 30min onwards and TBARS continued increasing along the whole heating treatment. Degradation of both cholesterol and stigmasterol fitted a first order curve (R(2)=0.937 and 0.883, respectively), with very similar degradation constants (0.004min(-1) and 0.005min(-1), respectively). However, higher concentrations of oxidation products were found from cholesterol (79µg/mg) than from stigmasterol (53µg/mg) at the end of the heating treatment. Profile of individual oxidation products was similar for both sterols, except for the fact that no 25-hydroxystigmasterol was detected. 7α-Hydroxy and 7-keto-derivatives were the most abundant SOPs at the end of the treatment. PUFA and vitamin E suffered a significant degradation along the process, which was correlated to sterols oxidation.

Stability of avocado oil during heating: comparative study to olive oil.

The stability of the saponifiable and unsaponifiable fractions of avocado oil, under a drastic heating treatment, was studied and compared to that of olive oil. Avocado and olive oil were characterised and compared at time 0h and after different times of heating process (180°C). PUFA/SFA (0.61 at t=0) and ω-6/ω-3 (14.05 at t=0) were higher in avocado oil than in olive oil during the whole experiment. Avocado oil was richer than olive oil in total phytosterols at time 0h (339.64; 228.27mg/100g) and at 9h (270.44; 210.30mg/100g) of heating. TBARs was higher in olive oil after 3h, reaching the maximum values in both oils at 6h of heating treatment. Vitamin E was higher in olive oil (35.52 vs. 24.5mg/100g) and it disappeared earlier in avocado oil (at 4 vs. 5h). The stability of avocado oil was similar to that of olive oil.