Uplc system

Strigolactones were analyzed using an Acquity UPLC^TM System (Waters, Milford, MA, USA) coupled to a Xevo® TQ-XS tandem quadrupole mass spectrometer (Waters MS Technologies, Manchester, UK) with an electrospray interface. The separation was performed on an Acquity BEH C18 column (2.1 × 100 mm, 1.7 μm) at flow rate 0.45 mL/min and temperature 45 °C, with 12 min binary gradient elution as follows: 0–0.4 min (85% A), 0.4–5 min (60% A), 5–8 min (35% A), 8–8.7 min (35% A), 8.7–9.5 (5% A), column wash for 0.8 min (5% A) and final column equilibration for 3 min for initial conditions (85% A), where A = 15 mM formic acid/water and B = 15 mM formic acid/acetonitrile (v/v). The eluate was introduced in the ion source of the mass spectrometer and analysed using the following settings: ion source/desolvation temperature (120/550 °C), desolvation/cone gas (nitrogen) flow (1000/150 L/hr), collision gas (argon) flow 0.15 ml/min. Compounds were analyzed in multiple reaction monitoring mode (MRM) and quantified by diagnostic transitions of: 4-deoxyorobanchol (331 > 97), putative methoxy-5-deoxystrigol isomers (361 > 97) using optimized cone voltage (20–22 V) and collision energy (20 eV). The MassLynx^TM software, version 4.2 (Waters) was used to operate the instrument, acquire and process the data.

The phenolic composition of the Graciano extract (GE) was studied by ultra-high performance liquid chromatography and mass spectrometry using an ACQUITY UPLC^TM system with a diode array detector and coupled to a quadrupole time of flight mass spectrometer (Waters, Milford, MA, USA). The separation was carried out as described by Asensio-Regalado et al. [15 (link)] Briefly, the mobile phases were 0.1% (v/v) acetic acid in MeOH and 0.1% (v/v) acetic acid in water delivered at a flow rate of 0.35 mL/min; the volume injected was 5.0 µL; flavan-3-ols, hydroxycinnamic acids, and flavanols were detected at 280, 320, and 370 nm, respectively. Positive and negative ion modes mass spectra were recorded.

The thiols were separated by using an UPLC system (Waters, Milford, USA) equipped with a vacuum degasser, a binary solvent manager, and an autosampler. As stationary phase an analytical column (waters BEH C18, 100 × 2.1 mm, 1.7 μm) was used. Flow-rate was 0.3 mL/min and the composition of eluents was: solvent A (0.1% formic acid in water) and solvent B (0.1% formic acid in acetonitrile). The linear gradient for solvent B was as follows: 0 min, 15%; 13 min, 22%; 14 min, 30%; 18 min, 35%; 18.5 min, 100%; 21.5 min, 100%; and 22 min, 15%. The injection volume was set at 10 μL.
The derivatization Baijiu sample was analyzed by means of UPLC–MS/MS using precursor ion scan screening for compounds releasing the diagnostic ion (m/z X→143.5) (24 (link)). In the source, cone voltage of 23 V and collision energy of 20 eV was applied. The eleven derivatives were quantified in MRM mode by monitoring their corresponding precursor ion, product ion, cone voltage, and collision energy, respectively (Table 1). Each Baijiu sample was tested in three different sessions to obtain an average value.

For the UPLC-QToF-MS analysis, the powdered red ginseng samples were extracted in a manner similar to our previous studies using ultrasonic extraction [13 (link)]. In total, 300 mg of each powered red ginseng was weighed in a centrifuge tube (15 mL, PP-single use; BioLogix Group, Jinan, Shandong, China) and shaken vigorously after the addition of 6 mL of methanol. The extract was then placed in an ultrasonic cleaner (60 Hz; Wiseclean, Seoul, Korea) for 30 min. The solution was centrifuged (Legand Mach 1.6R; Thermo, Frankfurt, Germany) at a speed of 3000 rpm for 10 min, and an aliquot of the supernatant solution was filtered (0.2 µm; Acrodisk, Gelman Sciences, Ann Arbor, MI, USA) and injected into the UPLC system (Waters Co., Milford, MA, USA).

UPLC/MS analysis was performed on a UPLC system coupled with XEVO G2 Q-TOF mass spectrometer via an ESI source (Waters Corp., Milford, MA). For UPLC separation, 2 μL of sample solution was injected into an ACQUITY HSS T3 C₁₈ column (100 × 2.1 mm, 1.7 μm, Waters). The mobile phase consisted of ACN (A) and water containing 0.1% (v/v) formic acid (B). Linear gradient elution was applied (0–5 min, 5–30% A; 5–10 min, 30–40% A; 10–20 min, 40–65% A; 20–25 min, 65–90% A) at a flow rate of 0.4 mL/min. The column temperature was 45°C. For MS detection, accurate mass was maintained by the LockSpray interface of sulfadimethoxine (309.0658 [M-H]⁻). The operating parameters in negative ion mode were as follows: capillary voltage, 3.0 kV; cone voltage, 30 V; desolvation gas flow rate, 750 L/h; source temperature, 120°C; desolvation temperature, 350°C. MS data were acquired in centroid mode and processed by MassLynx software (Waters, version 4.1).

The identification of component peaks was carried out on an ultra‐performance liquid chromatography (UPLC) system (Waters, Milford, MA, USA) equipped with a photo‐diode array detector. Aliquots (2.0 μl) of test sample were then injected into an analytical column (BEH C₁₈, 2.1 × 100 mm, 1.7 μm, Waters) at a flow rate of 0.4 ml/min. The mobile phase consisted of water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B). The linear gradient was as follows: 0–1 min, 10% B; 1–5 min, 10–40% B; 5–9 min, 40–60% B; 9–11.2 min, 60–100% B; 11.2–13.2 min, 100% B; 13.2–15 min; return to 10% B. The quadrupole time‐of‐flight mass spectrometer (Q‐ToF Premier™, Waters) was operated in both positive‐ and negative‐ion mode under the following conditions: capillary voltage, 2.5 kV; cone voltage, 40 V; source temperature, 110°C; desolvation temperature, 350°C. A sprayer with a reference solution of leucine‐enkephalin ([M + H]⁺m/z 556.2771 and [M–H]⁻m/z 554.2615) was used as the lock mass.