Table 2.

The LCMS/MS conditions and instrument settings used to assess ADAC concentrations in rat perilymph.

Min	%B	Flow rate (mL/min)
0	20	0.5
1	20	0.5
5	30	0.5
6	90	0.7
6.5	90	0.7
7.5	20	0.5
10	20	0.5

Mobile Phase A: 4.5 mM ammonium formate pH 7.0.

Mobile Phase B: 100% acetonitrile.

Column: Phenomenex® Gemini 5 µm C18 110 Å, 150 × 3.00 mm S/N: 625787-3.

Injection volume: 20 µL.

Detection wavelength: 254 nm, bandwidth 20 nm.

Source parameters: gas temp: 275°C; gas flow: 10 L/min; nebulizer: 25 psi; sheath gas temp: 350°C; sheath gas flow: 12 L/min. Capillary voltage: 2750 V (positive) and 3500 V (negative); nozzle voltage: 2000 V.

Multiple reaction monitoring (MRM) transitions and compound-dependent parameters: precursor ion (m/z) Q1: 577.4; product ion (m/z) Q3: 445.4; collision energy: 20 V; fragmentation: 150 V; dwell time: 100 ms.

Run time: 10 min; retention time ~2.5 min.

Thermostat control: 5°C.