Acquity beh shield rp18

Canolol content in oil was determined by the method of Yang et al. (2014) with a few modifications. Oil sample of 1.25 g was mixed with 1.5 ml of 80% methanol solution and 1.5 ml of hexane for three times. The extracts were combined and through a 0.22‐μm nylon filter. The analysis was carried out with ACQUITY ultra‐performance liquid chromatography (UPLC) (Waters) equipped with photodiode array (PDA) detector and Waters ACQUITY BEH Shield RP18 (100 × 2.1 mm, 1.7 μm, Waters). The column temperature was 30°C, and injection volume was 3 μl. Extracts were eluted with 2% acetic acid (w/w) (mobile phase A) and 100% methanol (mobile phase B) at the flow of 0.21 ml/min. The gradient elution was as follows: 5%–25% B (7.40 min), 25%–29% B (2.67 min), 29%–36% B (6.66 min), 36%–45% B (6.67 min), 45%–65% B (2 min), 45%–65% B (2 min), 65%−5% B (2 min), and 5% B (2.67 min). The detection wavelength was 270 nm.

The ceramide content was quantified according to a previously described method [16 (link)]. First, lipids were extracted from the plasma and the aorta. Twenty-five µl of sample was mixed with 150 µl of dichloromethane/methanol (1:2; v/v). Then, we added 100 µl of 1 M NaCl and 100 µl of dichloromethane. After centrifugation, 100 µl of the organic phase was transferred to a new tube. The organic phase was dried and dissolved it in 20 µl of 2% Igepal-CA630. Second, we detected the ceramide content. Two µl of lipid solution was mixed with 2 µl of ceramide substrate buffer and incubated at 37°C. After 1 h, this reaction solution was mixed with 56 µl of naphthalene-2,2-dicarboxaldehyde recreation buffer and incubated at 50°C for 10 min. Finally, the product was detected and quantified using the UPLC system. In the UPLC system, we used a reversed phase column (Acquity BEH Shield RP18, 2.1 × 50 mm, 1.7 µm, Waters, U.S.A.) to determine total ceramide.