Lunasin extract from wheat was identified using a SCIEX TripleTOF6600 mass spectrometer connected to a UPLC system (AB SCIEX, Framingham, MA, USA). The freeze-dried samples were dissolved in deionized water, ionized, and detected through time-of-flight mass spectrometers (TOFMS) scans and product ion scans. UPLC analysis was performed through an ACQUITY UPLC BEH Shield RP18 (1.7 µm, 2.1 mm × 100 mm, Waters Corp., Milford, MA, USA). The mobile phase consisted of Solution A (0.1% v/v formic acid in water) and Solution B (0.1% v/v formic acid in acetonitrile). The separation step was carried out as follows: 2 min 95% v/v A, 15 min 65% v/v A, 18 min 20% v/v A, 23 min 20% v/v A, 24 min 95% v/v A, and 30 min 95% v/v A.
Acquity uplc beh shield rp18
Manufactured by Waters Corporation
Sourced in United States
The ACQUITY UPLC BEH Shield RP18 is a reversed-phase liquid chromatography column. It is designed for high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC) applications. The column utilizes a bonded C18 stationary phase to separate a wide range of analytes based on their hydrophobic interactions.
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Lab products found in correlation
Acquity uplc h class sqd 2 system, by Waters Corporation (2 mentions)
Tripletof 6600, by AB Sciex (1 mentions)
Acquity ultra performance liquid chromatography, by Waters Corporation (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Autosampler injection and pump systems, by Waters Corporation (1 mentions)
Tb 04 ta, by Boeco (1 mentions)
Minipuls 3 peristaltic pump, by Gilson (1 mentions)
1290 infinity 2, by Waters Corporation (1 mentions)
8 protocols using acquity uplc beh shield rp18
1
Lunasin Identification in Wheat by UPLC-MS
2
UPLC-PDA Analysis of Purified Compounds
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The purified prescription was dissolved in 50% methanol/H2O and filtered through 0.22 μm nylon membrane microfilters (Shimadzu-GL, Japan). The chromatographic analysis was achieved using a Waters ACQUITY Ultraperformance Liquid chromatography (UPLC) with a photodiode detector (PDA) (Waters, Milford, MA, USA). Reversed-phase separation was performed on an ACQUITY UPLC BEH Shield RP18 (2.1 × 100 mm, 1.7 μm, Waters, Milford, MA, USA) column at 35°C. Mobile phases comprised (A) 0.2% formic acid in water and (B) acetonitrile. The sample was injected (2 μL injection volume) onto the column and eluted at a flow rate of 0.25 mL/min according to the following gradients: initial 5.0% B; 0.0–3.0 min/5.0–6.0% B; 3.0–14.0 min/6.0–7.0% B; 14.0–15.0 min/7.0–9.5% B; 15.0–15.5 min/9.5–10.0% B; 15.5–20.0 min/10.0% B; 20.0–20.5 min/10.0–11.0% B; 20.5–35.0 min/11.0% B; 35.0–36.0 min/11.0–11.5% B; 36.0–43.0 min/11.5% B; 43.0–57.0 min/11.5–16.0% B; 57.0–72.0 min/16.0–21.0% B; 72.0–78.0 min/21.0–24.0% B; 78.0–84.0 min/24.0–30% B; 84.0–90.0 min/30.0–38.0% B; 90.0–93.0 min/38.0–60.0% B; 93.0–94.0 min/60.0–100.0% B. Ultraviolet detection was set to 254 nm.
3
LC-ESI-MS Analysis of PEFA
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LC–ESI–MS analysis of PEFA was performed on a Waters Acquity UPLC H-Class/SQD II system (Waters Corp., Milford, MA, USA), equipped with an ACQUITY UPLC BEH Shield RP18 (100 mm × 2.1 mm, 1.7 μm) column. The chromatography separation was performed as described by Guerfali et al. [17 (link)]. The LC–MS instrument was operated in positive ion electrospray mode with an acquisition range of m/z 115–1700 with a scan rate of 0.5 spectra/s.
4
Soybean Bioactives: Antioxidant and Anti-inflammatory Properties
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The typical soybean varieties were harvested in northeastern China in 2018, Institute of Crop Sciences, provided by the Chinese Academy of Agricultural Sciences. The lunasin standard was from the Beijing Genomics Institute (Beijing, China). RAW264.7 macrophages and the MDA-MB-231 breast-cancer cell line originated from the Institute of Bioscience, Chinese Academy of Sciences (Shanghai). Penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonicacid) diammonium salt (ABTS), 1,1-diphenyl-2-picrylhydrazyl radical (DPPH), fluorescein sodium, and lipopolysaccharide (LPS) were purchased from Baierdi Biotechnology Co., Ltd. Dulbecco’s modified Eagle’s medium (DMEM) was from Thermo Fisher Scientific (Beijing, China) and fetal bovine serum (FBS) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Mass spectrometer (SCIEX TripleTOF6600®) Nitrogen generator (SCIEX, Massachusetts, USA). ACQUITY UPLC ®BEH Shield RP18 (Waters Corp, Milford, MA, USA).
5
Mass Spectrometry Analysis of Samples
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Mass spectrometry analyses were performed on a Quattro Premier™ XE Micromass MS Technologies triple quadrupole mass spectrometer configured with a Z-Spray™ electrospray ionization source (ESI, Waters, Milford, USA). An Acquity™ Ultra-High-Performance LC system (Waters, Milford) equipped with an autosampler injection and pump systems (Waters, Milford, USA) was employed. The autosampler vial tray was maintained at 4 °C. The separation was accomplished using an ACQUITY UPLC® BEH Shield RP18 (Waters, Milford, USA) analytical column (100 × 2.1 mm i.d., 1.7 μm). During the sample pretreatment, an electronic microbalance with a readability of 0.1 mg (Ohaus, model UMX2, Switzerland), an ultrasonic bath (Testlab (model TB-04 TA, Buenos Aires, Argentina)), a centrifuge (U-320R-BOECO, Germany), and a Minipuls 3 peristaltic pump (Gilson (Villiers-Le-Bell, France)) were employed.
6
HPLC-ESI-MS Analysis of Melanin-A
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HPLC–ESI–MS analysis of MEL-A was performed as a previously described method (Wang et al. 2019 (link)) on a Waters Acquity UPLC H-Class/SQD II system (Waters Corp., Milford, MA, USA) with an ACQUITY UPLC BEH Shield RP18 (100 mm × 2.1 mm, 1.7 μm) column. The MS detection was operated in positive ion electrospray mode with an acquisition range of m/z 50–1000 and a scan rate of 0.5 spectra/s. The mass spectral ions were identified by the calculation of elemental composition according to available literature (Goossens et al. 2016 (link)).
7
Estrogenic Substance Analysis by LC-MS/MS
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In parallel to bioassays, extracts were investigated by LC‐MS/MS and analyzed for the target estrogenic substances E1, E2, EE2, E3, BPA, and 4‐tert‐octylphenol. We used a method that was previously validated by comparing estrogenicity data (n = 33 effluent and surface water samples) with that of two other laboratories (see Könemann et al., 2018 (link)). Chemical analysis was performed in negative mode with an electrospray ionization source on an Agilent G6495A triple quadrupole mass spectrometer coupled to an ultrahigh performance liquid chromatography (UHPLC) system for chromatographic separation (Agilent 1290 Infinity II, Waters Acquity UPLC BEH Shield RP18, 130 Å, 2.1 mm × 100 mm, 1.7 µm column [p/n 186002854] with a 5‐mm precolumn [p/n 186003977]). A methanol/water + 5 mM NH3 gradient was applied as described in detail in Könemann et al. (2018 (link)) and Simon et al. (2019 (link)).
8
Carminative Compound Identification and Pain Relief
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Carminative and treatment of pain and bronchitis [1] The separation was performed on a Waters ACQUITY UPLC® BEH Shield RP18 (100 mm ×2.1 mm; particle size 1.7 μm). Column and auto sampler temperatures were set at 30°C and 20°C, respectively. The mobile phase consisted of 0.1% formic acid in deionized water (A) and acetonitrile (B). The gradient condition was set as the following: The % B was linearly increased from 30% to 33% in 4 min, then to 40% in 6 min, finally to 100% and kept there for another 0.5 min, then linearly ramped down to 30% again in 0.5 min. The total gradient run time was 13 min. The flow rate was set at 300 μL/min. The injection volume was 2 μL for all standards and samples.
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