Alltech 3300

HPLC analysis was conducted using a Varian 9010 instrument (Varian Inc., Palo Alto, CA, USA) equipped with an Alltech 3300 evaporative light scattering detector (ELSD) (BÜCHI Labortechnik, AG, Flawil, CH). A Luna C18 column (250 × 4.6 mm, 5 µm particle size) from Phenomenex (Torrance, CA, USA) was used for separating triglycerides and FAMEs. HPLC conditions were as follows: eluent A, MeOH; eluent B, CH₂Cl₂; gradient: 0–3 min (A-B/80:20), 3–18 min (A-B/30:70), 18–23 min (A-B/30:70); flow rate: 1 mL min⁻¹. ELSD was set to a probe temperature of 40 °C and a gain of 16, and the nebulizer nitrogen gas was adjusted to 1.5 L min⁻¹.
GC analyses were conducted using Shimadzu GC-17A (Shimadzu Corporation, Kyoto, Japan) equipped with a fused silica capillary column from J&W Scientific (INNOWAX, 30 m, 0.25 mm, 0.25 µm) (Agilent Technologies, Santa Clara, CA, USA); nitrogen was used as the carrier gas (flow rate of 1 mL min⁻¹). GC conditions were as follows: the injector and detector temperatures were set at 250 and 280 °C, respectively; the aliquot of the reaction mixture was injected and analyzed using the following temp. prog.: 60 °C for 2 min, 60–200 °C at 10 °C min⁻¹, 200–250 °C at 5 °C min⁻¹, 250 °C for 5 min. Identification of different FAMEs and, eventually, the presence of tert-butyl glycerol ethers, was achieved by referring to the chromatograms of standard compounds.

After evaporation of methanol, the extract obtained by SPE is taken up in 1 mL of chloroform then possibly diluted according to the phospholipid content and injected (injection volume of 20 µL) in a liquid chromatography apparatus connected to an ELSD (Evaporative Light Scattering Detector) according to the procedure detailed in the literature [55 (link)]. The column used is of type Lichrospher 100 diol (5µ) 150 × 3 mm. The oven is set at a temperature of 40 °C and the detector Alltech 3300 (Büchi, Postfach, Switzerland) is operated at a temperature of 35 °C with an air supply at 4 bars and a flow rate of 1.6 L/h. Two eluents are prepared and mixed with a concentration gradient over time (see Supplementary Materials). The different phospholipids are identified by comparing their retention times with those of commercial standards. Quantification is performed by external calibration.

HPLC analysis was conducted using a Varian 9010 instrument (Varian Inc., Palo Alto, CA, USA) equipped with an Alltech 3300 evaporative light scattering detector (ELSD) (BÜCHI Labortechnik, AG, Flawil, CH). A Luna C18 column (250 × 4.6 mm, 5 µm particle size) from Phenomenex (Torrance, CA, USA) was used for separating triglycerides and FAMEs. HPLC conditions were as follows: eluent A, MeOH; eluent B, CH₂Cl₂; gradient: 0–3 min (A-B/80:20), 3–18 min (A-B/30:70), 18–23 min (A-B/30:70); flow rate: 1 mL min⁻¹. ELSD was set to a probe temperature of 40 °C and a gain of 16, and the nebulizer nitrogen gas was adjusted to 1.5 L min⁻¹.
GC analyses were conducted using Shimadzu GC-17A (Shimadzu Corporation, Kyoto, Japan) equipped with a fused silica capillary column from J&W Scientific (INNOWAX, 30 m, 0.25 mm, 0.25 µm) (Agilent Technologies, Santa Clara, CA, USA); nitrogen was used as the carrier gas (flow rate of 1 mL min⁻¹). GC conditions were as follows: the injector and detector temperatures were set at 250 and 280 °C, respectively; the aliquot of the reaction mixture was injected and analyzed using the following temp. prog.: 60 °C for 2 min, 60–200 °C at 10 °C min⁻¹, 200–250 °C at 5 °C min⁻¹, 250 °C for 5 min. Identification of different FAMEs and, eventually, the presence of tert-butyl glycerol ethers, was achieved by referring to the chromatograms of standard compounds.