Quantum access

This was performed on a Thermo Fisher TSQ LC–MS/MS system consisted of an Accela Autosampler, an Accela pump and a Quantum Access triple quadrupole mass spectrometer (Thermo Fisher Scientific Co., San Jose, CA, USA). Data acquisition was performed with the Xcalibur software version 2.0.7, and data processing using the Thermo LCquan 2.5.6 data analysis program (Thermo Fischer). The chromatographic separation was achieved using an XTerra-MS C₁₈ column (150 mm × 2.1 mm i.d., 3.5 μm, Waters Corp., Milford, MA, USA). The two eluents were: (A) 5 mM HA–0.5% DEA in water, pH adjusted to 10 with acetic acid; and (B) 50% acetonitrile in water. The mobile phase consisted of linear gradient of A and B: 0–15 min, 100–80% A (v/v); 15–35 min, 80–70% A; 35–45 min, 70–45% A; 45–46 min, 45–0% A; 46–50 min, 0–0% A; 51–70 min, 100–100% A. The liquid flow-rate was set at 0.3 mL/min, and the column temperature was maintained at 35 °C. For all RN and dRN, the following optimized parameters were obtained. The sheath gas pressure reached 40 psi. The ionspray voltage was set at 3000 V for negative mode and 4000 V for positive mode, respectively and the temperature at 350 °C. The Auxiliary gas pressure was 15 psi. Quantification was performed using multiple reactions monitoring (MRM) as previously published18 (link).

This was performed on a Thermo Fisher TSQ LC–MS/MS system consisted of an Accela Autosampler, an Accela pump and a Quantum Access triple quadrupole mass spectrometer (Thermo Fisher Scientific Co., San Jose, CA, USA). Data acquisition was performed with the Xcalibur software version 2.0.7, and data processing using the Thermo LCquan 2.5.6 data analysis program (Thermo Fischer). The chromatographic separation was achieved using an XTerra-MS C₁₈ column (150 mm X 2.1 mm i.d., 3.5 μm, Waters Corp., Milford, MA, USA). The two eluents were: (A) 5 mM HA–0.5% DEA in water, pH adjusted to 10 with acetic acid; and (B) 50% acetonitrile in water. The mobile phase consisted of a linear gradient of A and B: 0–15 min, 100–80% A (v/v); 15–35 min, 80–70% A; 35–45 min, 70–45% A; 45–46 min, 45–0% A; 46–50 min, 0–0% A; 51–70 min, 100–100% A. The liquid flow-rate was set at 0.3 mL/min, and the column temperature was maintained at 35 °C. For all RN and dRN, the following optimized parameters were obtained. The sheath gas pressure reached 40 psi. The ionspray voltage was set at 3000 V for negative mode and 4000 V for positive mode at a temperature of 350 °C and auxiliary gas pressure of 15 psi. Quantification was performed using multiple reactions monitoring (MRM) as previously published18 (link).

This was performed on a Thermo Fisher TSQ LC-MS/MS system consisted of an Accela Autosampler, an Accela pump and a Quantum Access triple quadrupole mass spectrometer (Thermo Fisher Scientific Co., San Jose, CA, USA). Data acquisition was performed with the Xcalibur software version 2.0.7, and data processing using the Thermo LCquan 2.5.6 data analysis program (Thermo Fisher). The chromatographic separation was achieved using an XTerra-MS C18 column (150 mm × 2.1 mm i.d., 3.5 µm, Waters Corp., Milford, MA, USA). The two eluents were: (A) 5 mM HA–0.5% DEA in water, pH adjusted to 10 with acetic acid; and (B) 50% acetonitrile in water. The mobile phase consisted of a linear gradient of A and B: 0–15 min, 100%–80% A (v/v); 15–35 min, 80%–70% A; 35–45 min, 70%–45% A; 45–46 min, 45%–0% A; 46–50 min, 0%–0% A; 50–51 min, 0%–100% A; 51–70 min, 100–100 % A. The liquid flow-rate was set at 0.3 mL/min, and the column temperature was maintained at 35 °C. For all dRNs, the following optimized parameters were obtained. The sheath gas pressure reached 40 psi. The ion spray voltage was set at 3000 V for negative mode and 4000 V for positive mode at a temperature of 350 °C and auxiliary gas pressure of 15 psi. Quantification was performed using multiple reactions monitoring (MRM) as previously published [24 (link)].