Lc 20ad uflc xr pumps

The HPLC system consisted of a SIL-20AC XR auto-sampler and LC-20AD UFLC XR pumps (Shimadzu, Tokyo, Japan). Samples were separated on a Gemini C18 chromatographic column (3 μM 110A, 100 x 2.0 mm) coupled with a Security Guard Cartridge (Gemini-NX C18 4.0 x 2.0 mm), both provided by Phenomenex (Torrance, CA, USA) and thermostatically controlled at 25°C. The mobile phases (MP) used for chromatographic separation were 0.1% CH₃COOH/bidistilled water (MP A) and 0.1% CH₃COOH/acetonitrile (MP B). The HPLC system was set up with a flow rate of 0.3 mL/min and the following linear gradient:
step 1: the initial condition of 95% MP A held for 1 min;
step 2: from 95% MP A to 30% over 5.5 min;
step 3: constant for 1.5 min;
step 4: from 30% MP A to 10% over 0.5 min;
step 5: kept constant for 1.5 min;
step 6: from 10% MP A to the initial condition over 1 min and reconditioning for 7 min.
The total run time was 18 min.

The HPLC system consisted of a SIL-20AC XR auto-sampler and LC-20AD UFLC XR pumps (Shimadzu, Tokyo, Japan). The chromatographic separation of the samples were conducted on a SunFire^™ C18 column (3.5 μM, 92 Å, 2,1 x 150 mm) coupled with a Security Guard Cartridge (SunFire^™ C18 2.1 x 10 mm), both provided by Waters (Milford, MA, USA) and thermostatically controlled at 30°C. The mobile phases (MP) were 0.1% HCOOH/bidistilled water (MPA) and 0.1% HCOOH/CH₃CN (MPB). In the presented method, the following linear gradient was used, with a flow rate of 0.2 mL/min: (step 1) from the initial condition of 60% MPA to 0% over 12 min; (step 2) kept constant for 2 min; (step 3) from 0% MPA to the initial condition over 1 min; (step 4) reconditioning for 6 min. The total run time was 21 min.

The HPLC system was a SIL-20AC XR auto-sampler and LC-20AD UFLC XR pumps (Shimadzu, Tokyo, Japan). The mobile phases (MP) consisted of ultrapure water with 0.1% HCOOH (phase A) and methanol/isopropanol (9:1, v/v) with 0.1% HCOOH (phase B). The chromatographic separation was obtained on the Luna Omega Polar C18 column (3 μM, 100 Å, 50 x 2.1 mm) coupled with a Security Guard Cartridge (Polar, C18, 4 x 2.0 mm), both provided by Phenomenex (Castel Maggiore (BO), Italy). The mass spectrometry system used for the detection was an API 4000 triple quadrupole (AB SCIEX, Massachusetts, USA) with a TurboIonSpray source operating in positive ion mode. To optimize source and compound dependent parameters, solutions of each analyte at the concentration of 100 ng/mL were used with a flow rate of 20 μL/min. Data were processed with Analyst 1.6.3 and the quantification of the peaks was done with MultiQuant 2.1 (software package AB SCIEX).