Quantitative HPLC analysis was performed on an Agilent 1200 system equipped with a G1311A QuatPump, a G1322A degasser, a G1315D diode array detector, and a G1329A ALS with a 20 μL loop. Agilent Extend-C18 (250 mm × 4.6 mm i. d., 5 μm, Agilent) was used to analyze samples. The mobile phase consisted of water with 0.1% formic acid solution (A) - acetonitrile (B) in a linear gradient program as follows: 0–20 min, 10–30% B; 20–25 min, 30–45% B; 25–30 min, 45–95% B; 30–35 min, 95% B at a flow rate of 0.8 mL/min. Chromatograms were recorded at 280 nm. The injection volume was 20 μL.
Agilent extend c18
Manufactured by Agilent Technologies
Sourced in United States
The Agilent Extend-C18 is a reversed-phase liquid chromatography column designed for the separation and analysis of a wide range of compounds. It features a high-purity, spherical silica gel with a chemically bonded C18 stationary phase, providing a stable and versatile platform for various analytical applications.
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Lab products found in correlation
1200 system, by Agilent Technologies (1 mentions)
G1311a quatpump, by Agilent Technologies (1 mentions)
G1322a degasser, by Agilent Technologies (1 mentions)
G1329a als, by Agilent Technologies (1 mentions)
Tsq vantagetm, by Thermo Fisher Scientific (1 mentions)
Accela 1250, by Thermo Fisher Scientific (1 mentions)
Xcalibur software, by Thermo Fisher Scientific (1 mentions)
Chi raa art cellulose sc, by YMC (1 mentions)
Milli q quality water, by Merck Group (1 mentions)
3 adon, by Merck Group (1 mentions)
15 adon, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Uv 5300, by Hitachi (1 mentions)
Xevo tq s, by Waters Corporation (1 mentions)
Cell apoptosis and cycle kits, by BD (1 mentions)
J 810 circular dichroism spectropolarimeter, by Jasco (1 mentions)
600 mhz instrument, by Bruker (1 mentions)
Sephadex lh 20, by Cytiva (1 mentions)
J 810, by Jasco (1 mentions)
4 protocols using agilent extend c18
1
Quantitative HPLC Analysis of Compounds
2
Quantitative Analysis of Mycotoxins by LC-MS/MS
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LC–MS/MS analysis was performed as described by Yu et al. [15 (link)]. Mycotoxins were quantified by an Accela 1250 UPLC system (Thermo Fisher Scientific, San Jose, CA, USA) coupled to a TSQ VantageTM (Thermo Fisher Scientific, San Jose, CA, USA) triple-stage quadruple mass spectrometer. An Agilent Extend C18 chromatographic column (100 mm × 4.6 mm, 3.5 μm) was used at a flow rate of 0.35 mL/min at 30 °C and with a 10 μL injection volume. The mobile phase consisted of 5 mM ammonium acetate (A) and 100% methanol (B). The gradient was as follows: 0 min 15% B, 1 min 15% B, 6.5 min 90% B, 8.5 min 90% B, 9 min 15% B, and 12 min 15% B. Mass spectrometry analysis was carried out in both positive (ESI + 3.5 kV) and negative (ESI − 2.5 kV) ionization modes using selected reaction monitoring (SRM). For the MS/MS analysis, both the vaporizer and capillary temperatures were 300 °C, the sheath gas pressure was 50 psi, and the aux gas pressure was 5 psi. Raw data were analyzed using Xcalibur™ software (Thermo Fisher Scientific, San Jose, CA, USA, 2011).
The ratio of D3G and total DON was calculated based on the following formula:
The ratio of D3G and total DON was calculated based on the following formula:
3
Mycotoxin Analysis via LC-MS/MS
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The standard of DON, 3-ADON and 15-ADON were purchased from Sigma-Aldrich (St. Louis, MO, USA) and stored at −20 °C prior to use. Acetonitrile, isopropanol, hexane, methanol and ammonium acetate were purchased from Merck (Darmstadt, Germany). In addition, all organic solvents and acids were of analytical or HPLC grade. Milli-Q-quality water (Millipore, Billerica, MA, USA) was used throughout the experiments. The Agilent Extend-C18 (3.0 mm × 150 mm, 3.5 μm) column were obtained from Agilent Technologies Inc (Santa clara, CA, USA). The Waters UPLC BEH amidel (2.1 mm × 100 mm, 1.7 μm) column was purchased from Waters (Milford, MA, USA). The YMC -TRIART diol-HILIC (3.0 mm × 100 mm, 1.9 μm), YMC CHI-RAa ART Cellulose-sc (2.0 mm × 100 mm, 3 μm) and YMC CHIRAa ART Cellulose-sc (3.0 mm × 250 mm, 3 µm) columns were purchased from YMC (Kyoto, Japan).
4
Phytochemical Analysis and Bioactivity Evaluation
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Ultraviolet-visible (UV/Vis) spectra were acquired using a UV-Vis spectrophotometer, the UV-5300 (Hitachi, Tokyo, Japan). Infrared (IR) spectra were measured on a Bruke Vertex 80 (KBr disks) infrared spectrometer. Optical rotations were recorded in methanol (MeOH) solution on an AUTOPOL1 polarimeter at 28 °C (Rudolph, Wilmington, MA, USA). CD spectra were obtained on a J-810 circular dichroism spectropolarimeter (JASCO Corp., J-810, Tokyo, Japan). Electro-spray ionization mass spectrometry (ESIMS) and high-resolution electrospray ionization mass spectrometry (HRESIMS) analyses were measured on a Waters Xevo TQS and a Waters Xevo G2S Q-TOF LC/MS, respectively. NMR spectra were recorded on a Bruker 600 MHz instrument, and tetramethylsilane (TMS) was used as an internal standard. Column chromatography was performed on silica gel (200−300 mesh, Qingdao Marine Chemical Co., Ltd., Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, Uppsala, Sweden), respectively. HPLC analysis was performed on an Agilent 1260 with a C18 column (Agilent Extend-C18, 4.6 × 250 mm, 5 μm, 1 mL/min). HPLC separation was performed on Shimadzu Soviet production LC-16P with an ODS column (Silgreen C18AB, 10 × 250 mm, 5 μm, 180 Å, 3 mL/min). Cell apoptosis and cycle kits were purchased from BD Biosciences (San Jose, CA, USA).
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