Agilent 1290 infinity 2 liquid

The protocol analysis described by Pérez-Nevado et al. [30 (link)] was used to calculate the acrylamide content in the coffee beverage studied. A quantity of 2 g of coffee was filtered through a 0.45 μm nylon syringe filter, and the liquid was placed into two Telos columns (PCX, 200 mg/3 mL and PRP, 60 mg/3 mL) to obtain the eluate. Concentrations ranged from 50 to 150 ng mL⁻¹ of acrylamide, and a standard addition method was used. Finally, an Agilent 1290 Infinity II liquid chromatograph (Agilent Technologies) coupled to an Agilent 6460 triple quadrupole mass spectrometer was used to obtain the concentration of this toxic substance. Helium was used as carrier gas with a flow rate of 0.8 mL min⁻¹. It was worked in isocratic mode with a flow of 0.25 mL min⁻¹, using eluent A (0.1% formic acid in Milli-Q-water) and eluent B (0.1% formic acid in methanol). The gas injector temperature was 340 °C, the nebulizer pressure was 40 psi, the sheath gas temperature was 400 °C with a flow rate of 12 L h⁻¹ and capillary voltages were +2.5 kV. The nozzle voltage was 300 V and dental EMV: 300.

The serum samples were defrosted in an ice water bath and then mixed in a vortex for 30 s. A total of 100 μL of aliquot of each individual sample was transferred to an Eppendorf tube. After the addition of equal volumes of extraction buffer (acetonitrile/methanol/water, 2/2/1, v/v/v), the samples were vortexed for 1 min. The samples were stored at −20°C for 2 h and then 100 μL of 0.2% acetic acid in acetonitrile was added. After centrifugation at 12,000 rpm for 15 min at 4°C, 150 μL of supernatant was collected and filtered. Samples were analyzed in Agilent 1290 Infinity II liquid chromatograph-6546 LC/Q-TOF, equipped with Agilent InfinityLab Poroshell 120 HILIC-Z (2.1 × 100 mm, 2.7 μm) column. Mobile phase A was 10 mM ammonium acetate in water, pH = 9, and the mobile phase B was 10 mM ammonium acetate solution: acetonitrile (1/9, v/v), pH = 9. The HPLC program was as follows: 0 min, 0%A; 11.5 min, 30%A; 12 min, 0% A; 20 min 0% A. The column temperature was 30°C, and the injection volume was 5 μL. The mass spectrometry detection was performed with both positive and negative electrospray ionization mode. The MS and MS/MS spectra were obtained in the range of m/z 50–750. The final optimized mass parameters were set as follows: nebulizer gas and heater gas at 40 psig, ion spray voltage at 3000 v, the turbo spray temperature at 300°C, and collision energy at 115 eV.