Ms 8060 mass spectrometer

The UHPLC-MS/MS system consisted of Shimadzu UHPLC-30AD and MS-8060 mass spectrometer (Kyoto, Japan). Selective quantification of compounds was performed using the multiple reaction monitoring (MRM) mode. Two compounds were separated by UHPLC using a solvent system consisting of 10 mM ammonium formate-methanol (solvent A; v/v) and water (solvent B) along with an ACQUITY UHPLC ^® BEH C18 (100 mm × 2.1 mm, 1.7 µm) column. The gradient elution program for UHPLC-MS/MS analysis was as follows: 0 min, 40% A; 2.5 min, 45% A; 3 min, 50% A; 3.5 min, 95% A; 4 min, 45% A; 8 min, 40% A. The injection volume was 1 µL, and the flow rate was 200 µL/min. The column temperature was held at 40 °C.

Analyses were performed on LS-MS/MS analyses with a Shimadzu LC-30AD system coupled with a Shimadzu MS-8060 mass spectrometer (Kyoto, Japan). ZORBAX SB-C18 column (50 mm × 2.1 mm × 1.8 µm) was purchased from Agilent (Santa Clara, CA, USA). CPA224S electronic analytical balance was purchased from Sartorius Company (Göttingen, Germany). The refrigerator was obtained from Siemens (Munich, Germany). The HITACHI CT6E centrifuge was purchased from Hitachi, Ltd. (Tokyo, Japan). The 80350-CN grinder was supplied by Hamilton Beach Electric Co., Ltd. (Shenzhen, China). A DFT-100A portable high-speed universal crusher was purchased from Linda Machinery Co., Ltd. (Wenling, China). The HQ-60 vortex mixer was purchased from North Tongzheng Biotechnology Co., Ltd. (Beijing, China). The DTC-27J ultrasonic cleaner was provided by Dingtai Biochemical Technology Equipment Manufacturing Co., Ltd. (Wuhan, China). SPE column (Mega BE-SI, 1 g, 6 mL) was purchased from Agilent (Santa Clara, CA, USA). Pipettes (10 µL, 200 µL, 1 mL, 5 mL) were purchased from Eppendorf Company (Hamburg, Germany). A 1 mL syringe was purchased from Wangguan Medical Devices Co., Ltd. (Wuhan, China). A 0.22 µm organic phase filter was purchased in Millipore Company (Burlington, MA, USA).