Sq detector 2 mass spectrometer

Optical rotations were obtained on a JASCO P-1020 (JASCO Corporation, Tokyo, Japan) digital polarimeter. UV spectra were recorded on Waters 2487 (Waters Corporation, Milford, MA, USA), while the ECD spectrum were recorded on JASCO J-815 spectropolarimeter (JASCO Corporation, Tokyo, Japan). ¹H NMR, ¹³C NMR, DEPT and 2D NMR spectra were recorded on an Agilent 500 MHz DD2 spectrometer (Agilent Technologies Inc., Santa Clara, CA, USA). HRESIMS and ESIMS spectra were obtained using a Thermo Scientific LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) on positive ionisation mode. Column chromatography (CC) was performed with silica gel (200-300 mesh, Qingdao Marine Chemical Inc., Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, San Francisco, CA, USA). MPLC was performed on a Bona-Agela CHEETAHTM HP100 (Beijing Agela Technologies Co., Ltd., Beijing, China). RP-HPLC was performed on an ODS column (HPLC (YMC-Pack ODS-A, 10 × 250 mm, 5 µm, 3 mL/min)) (YMC Co., Ltd., Kyoto, Japan). LC-MS was performed using an Acquity UPLC H-Class coupled to a SQ Detector 2 mass spectrometer using a BEH C₁₈ column (1.7 μm, 2.1 × 50 mm, 1mL/min) (Waters Corporation, Milford, MA, USA).

The ethanol extract of MF was prepared as described previously (Jin et al., 2017 (link)). TSN and Ohchinin (OCN) were purchased from ChemFaces (Wuhan, China) (purity >98%) and used as experimental compounds and standards for ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) (Akihisa et al., 2013 (link)). The components of MF were separated using by a several Waters ACQUITY UPLC H-Class modules (Waters Corporation, Milford, MA, USA) coupled to the SQ detector 2 mass spectrometer (Waters Corporation, Milford, MA, USA) with electrospray ionization (ESI). Chromatographic separation was achieved with a Waters Acquity UPLC BEH C₁₈ octadecylsilane column (2.1 mm × 100 mm, 1.7 µm) and mobile phase (80% B) using 10 mM ammonium acetate in water or acetonitrile. The MS conditions were as follows: ion spray voltage, 3.5 kV; capillary voltage, 20 V; capillary temperature, 350°C; and tube lens voltage, 40 V. Analysis for TSN and OCN was performed in negative and positive mode, respectively (Ong and Ong, 2007 (link)).

The probe was designed by the authors and the synthesis was custom made from Serinno Holdings Limited. This probe will be commercially available after this work is disclosed. ¹H NMR (500 MHz, CD₃CN) δ = 10.48 (1H, s), 10.44 (1H, s), 7.91 (1H, d, J = 7.8 Hz), 7.82 (1H, s), 7.68 (1H, d, J = 7.8 Hz), 6.86 (1H, br s), 6.59 (1H, br s). ¹³C NMR (100 MHz, CD₃CN) δ = 193.3, 192.9, 171.4, 171.1, 146.8, 137.0, 135.2, 134.7, 131.7, 131.7, 80.5, 71.4. Analytical reversed-phase high-performance liquid chromatography (RP-HPLC) separations involving a mobile phase of 0.1% trifluoroacetic acid (TFA) (v/v) in acetonitrile (Solvent A) and 0.1% TFA (v/v) in water (Solvent B) were performed on a Waters UPLC H-class system equipped with an ACQUITY UPLC photodiode array detector and a Waters SQ Detector 2 mass spectrometer using a Waters ACQUITY BEH C18 column (1.7 μm, 130 Å, 2.1 ×50 mm) at a flow rate of 0.4 mL/min. The calculated purity was >95%. ESI–MS (m/z): calculated for C₁₉H₂₂N₂O₄S₂ [M + H]⁺: 407.53; found [M + H]⁺: 407.26; [M+Na]⁺: 429.51; found [M+Na]⁺: 429.27; [M+Na+H₂O]⁺: 447.52; found [M+Na+H₂O]⁺: 447.30.

Specific rotations were obtained on a JASCO P-1020 digital polarimeter developed by JASCO Corporation, Tokyo, Japan. UV spectra were carried out on Waters 2487 developed by Waters Corporation, Milford, MA, USA. NMR spectra were recorded on Agilent 500 MHz DD2 spectrometers made by Agilent Technologies Inc., Santa Clara, CA, USA, using tetramethylsilane as an internal standard, and the chemical shifts were recorded in δ values. HRESIMS spectra were obtained on a LTQ Orbitrap XL mass spectrometer made by Thermo Fisher Scientific, Waltham, MA, USA. The compounds were purified by HPLC made by the Waters company, Milford, MA, USA, equipped with a 2998 PDA detector and a C18 column (YMC-Pack ODS-A, 10 × 250 mm, 5 µm, 3 mL/min). Medium-pressure preparative liquid chromatography (MPLC) was performed on a Bona-Agela CHEETAH HP100 made by Beijing Agela Technologies Co., Ltd., Beijing, China. Column chromatography (CC) was performed with silica gel (100−200 mesh, 200−300 mesh, Qingdao Marine Chemical Inc, Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, San Francisco, CA, USA), respectively. LC-MS was recorded in ESI mode on an Acquity UPLC H-Class connected to a SQ Detector 2 mass spectrometer using a BEH C18 column (1.7 µm, 2.1 × 50 mm, 1 mL per minute) constructed by Waters Corporation, Milford, CT, USA.