Tcc 3000rc

An Ultimate 3000 system equipped with a pump (LPG-3400RS), an autosampler (WPS-3000RS), a column oven (TCC-3000RC), and a VWD-3400RS UV/Vis detector (all from Thermo Scientific, Waltham, MA, USA) was used for the chromatographic separation, and the UV-Vis detector was operated at 214 nm. Data were collected and processed by Chromeleon software 7.2 SR5 (Thermo Scientific, Waltham, MA, USA). Separation was carried out on a Hypersil GOLD C18 column (100 mm × 2.1 mm, 5 μm) (Thermo Scientific, Waltham, MA, USA) at 45 °C, with an injection volume of 10 μL. Mobile phase A was 20 mM formic acid containing 5% of methanol (v/v), and mobile phase B was methanol containing 5% of 20 mM formic acid (v/v). Mobile phase B was increased from 20% to 80% within 3.5 min at a flow rate of 0.8 mL min⁻¹; afterward, it was decreased to 20% within 0.1 min, and this condition was kept for 2.5 min for equilibration.

An Ultimate 3000 system equipped with a pump (LPG-3400RS), an autosampler (WPS-3000RS), a column oven (TCC-3000RC), and a VWD-3400RS UV/Vis detector (Thermo Scientific, USA) was used for the chromatographic separation. Data were collected and processed by Xcilabur 3.0 (Thermo Scientific, USA). Separation was carried out on a Thermo Accucore C18 (100 mm × 2.1 mm, 2.6 μm) (Thermo Scientific, USA) at 40 °C, with an injection volume of 5 μl. The mobile phases consisted of 0.1% formic acid (A) and 0.1% acetonitrile (B). The flow rate was 0.4 ml/min, and the gradient program was as follows: 5–95% B (0–2.5 min), 95% B (2.5–3.8 min), 95–5% B (3.8–4 min), and 5% B (4–5 min).
Ionization was achieved using electrospray ionization (ESI) in positive mode with selected reaction monitoring (SRM). Mass spectrometry was performed with an ESI source in the positive-ionization mode with a sheath gas of 50 Arb and aux gas of 15 Arb. The ion spray voltage was set at 4000 V and the source temperature was 350 °C. The parameters for the quantification selected reaction monitoring (SRM) transitions are presented in Table 1.Table 1

The parameters for the quantification selected reaction monitoring transitions.

Analytes	Polarity	Parent (m/z)	Product (m/z)	Collision energy (eV)	Tube lens (V)
			95.207	17	89
Cantharidin	+	197.085	123.139	14	89
			134.996	14	89