Ltq orbitrap xl mass spectrometer system

Separation was carried out on a Welch Ultimate XB-C₁₈ HPLC column (250 × 4.6 mm, 5 µm) at 35 °C. The mobile phase system was composed of acetonitrile (A) and 0.1% formic acid aqueous solution (B). The gradient program was used as follows: 0–20 min, 95% B; 20–25 min, 95–80% B; 25–30 min, 80–70% B; 30–45 min, 70–95% B. The flow rate was set at 0.80 mL/min. Two microliters supernatant samples were injected into LTQ–Orbitrap XL mass spectrometer system (Thermo Scientific, Bremen, Germany) for analysis.
The operating conditions of mass spectrometry were as follows: electrospray ionization (ESI) source was employed, and positive and negative ion mode were selected; sheath gas flow, 40 A.U.; auxiliary gas flow, 20 and 10 A.U.; source voltage 3.8 kV; capillary temperature of 300 °C; capillary voltage 25 and −35 V; tube lens, 110 and −110 V. The samples were analyzed using FT full scan with mass in the m/z 100–1500 range. All the raw data were processed using Xcalibur 3.0 software¹⁹ (Thermo Fisher Scientific, Waltham, MA, USA).

HL Granule was dissolved in double distilled water, and ultrasound extracted for 30 min by methanol. The extraction was centrifuged by 12,000 rpm/min at 4°C for 10 min, and supernatant was injected into the instrument for components’ detection. Chromatographic separation was performed on Waters ACQUITY UPLC H-CLASS system (Waters, Milford, MA, United States) with a AQUITY UPLC HSS T3 column (2.1 × 100 mm, 1.8 μm, United States). The column compartment was maintained at 35°C. The mobile phase was consisted of solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile). Gradient conditions were as follows: 0 min, 10% B, 0–23 min, 10%-90% B, 23–24 min, 90% B, 24–26 min, 90%-10% B, 26–30 min, 10% B, at a flow rate of 0.2 ml/min.
Mass spectrometry data was acquired by LTQ ORBITRAP XL mass spectrometer system (Thermo Scientific, United States) both in the positive and negative ionization mode. The temperature of the heated capillary was 270°C and that of the ESI probe was 300°C. The flows of sheath gas and auxiliary gas were set to 40 and 10 units, respectively. The scan range of the Orbitrap analyzer was m/z 500–1800 and MSⁿ data were acquired by LTQ in data dependent acquisition mode. Both of the positive and negative adopted the same MS parameters.