Targeted Quantification of Analytes by HPLC-MS/MS

The HPLC-MSMS system was composed of a LC- Shimadzu prominence LC20AD (Shimadzu, Maryland), with an ultramino 3 um 150 x 2.1 mm 100A aminopropylsilaneRestek column (Restek, Bellafonte, PA) with a flow rate of 0.30 ml/min with degasser and auto-sampler SIL-20AC HT (Shimadzu, Maryland), and column oven CTO-20A (Shimadzu, MD) run at 40°C. Mobile phase A was HPLC grade water with 0.1% formic acid and phase B was acetonitrile with 0.1% formic acid. Elution was programmed to start at 70% phase B for 0.5 minutes, then fall to 5% B at 2 minutes, return to 70%B at 2.5 minutes and equilibrate for two minutes prior to the next sample injection. MS/MS-API 5500 (AB Sciex, US) with turbo-ion probe (ESI) operated at 600C. The transitions monitored described before are listed with the retention times in Table 1 [17 (link)].