Uplc xevo tqs micro

GlcCer and GlcSph were measured as previously described by a modification of the Bligh and Dyer method [27 (link),28 (link),29 (link),30 (link)]. Briefly, 25 µL of ¹³C₅-GlcSph (0.1 µM) were added to 50 µL of fly homogenate and lipids were extracted with methanol, chloroform, and ammonium formate buffer (100 mM ammonium formate buffer pH 3.1) (1:1:0.9; v/v/v) resulting in 2 phases. The upper phase, containing GlcSph, was dried under N₂ stream and further extracted with water/butanol (1:1; v/v) before being applied to the UPLC-MS. GlcCer (lower phase) was separated from neutral lipids (ceramides and phospholipids) by solid phase extraction using Bakerbond silica gel (SiOH) columns (J.T. Baker, VWR, Phillipsburg, New Jersey, USA). 10 µL of the internal standard C17-dh-Ceramide (20 µM) was added and the samples were deacylated in a microwave for 1 h with 500 µL of methanolic NaOH (0.1 M). Deacylated lipids were additionally extracted with water/butanol (1:1; v/v) before being applied to the UPLC-MS. Lipids were analyzed by reverse-phase liquid Chromatography using a Waters UPLC-Xevo-TQS micro and a BEH C18 column, 2.1 × 50 mm with 1.7 μm particle size (Waters Corps. Milford, MA USA). Data was processed with MassLynx 4.1 Software (Waters Corporation, Milford, MA, USA).

GlcCer and GlcSph content was evaluated in BM, spleen, and liver samples. Briefly, GlcCer and GlcSph were extracted by a modification of the Bligh and Dyer method using acidic buffer (100 mM ammonium formate buffer pH 3.1). Prior to extraction, 20 μL of the internal standard C17-dh-Ceramide (20 μM) and 20 μL 13C5-GlcSph (0.1 μM) were added to the homogenate. Briefly, lipids were extracted by adding methanol, chloroform, and ammonium formate buffer (1:1:0.9; v/v/v), which resulted in 2 phases. The upper phase was dried under N2 stream and further extracted with water/butanol (1:1; v/v) before being applied to the UPLC-MS. The lower phase was transferred to a Pyrex tube and deacylated in a microwave for 1 h with 500 μL of methanolic NaOH (0.1 M). Deacylated lipids were extracted with water/butanol (1:1; v/v) and applied to the UPLC-MS. Lipids were analyzed by reverse-phase liquid chromatography using a Waters UPLC-Xevo-TQS micro and a BEH C18 column, 2.1 × 50 mm with 1.7 μm particle size (Waters, USA). Data were processed with MassLynx 4.1 software (Waters Corporation, USA). The method has been described previously.³⁶ (link)

Pesticide extracts from celery and soil samples were analyzed using UPLC Xevo TQ-S micro (Waters, Milford, MA, USA). Chromatographic column: BEH C18 column 100 A (50 mm× 1.7 mm, particle size: 5 μm), Waters Corporation, USA; column temperature was 40 °C, and the liquid chromatography mobile phase and gradient elution conditions are shown in Table 1. Sampling volume: 10 uL; Ion source: ESI, scanning mode: positive and negative ion scanning; ion source temperature was 150 °C, desolventing temperature was 350 °C, N₂ gas flow rate was: 650 L/h, N₂ conical gas flow rate was 250 L/h, and the mobile phase flow rate was 0.20 mL/min. The mass spectrometry parameters for 6 target compounds, such as qualitative ion pair, quantitative ion pair, collision energy (Ce), and the conical voltage, are shown in Table S3.