Quantitation of hepatic ceramides was performed by multiple reaction monitoring (MRM) and/or parent ion scanning using a Waters UHPLC Acquity system coupled to a Waters Xevo TQMS with a Waters Acquity C18 BEH column (2.1 × 100 mm). The internal standard C17:0 ceramide was obtained from Sigma-Aldrich. Sample preparation and ceramide measurements were carried out as described previously (Jiang et al. 2015a (link)).
Acquity uhplc system
Manufactured by Waters Corporation
Sourced in United States, United Kingdom
The Acquity UHPLC system is a high-performance liquid chromatography (HPLC) instrument developed by Waters Corporation. It is designed for the separation, identification, and quantification of chemical compounds in complex mixtures. The Acquity UHPLC system utilizes ultra-high pressure technology to achieve rapid and efficient separation of analytes, providing improved resolution, sensitivity, and throughput compared to traditional HPLC methods.
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Lab products found in correlation
Triple tof 5600 system, by AB Sciex (16 mentions)
Masslynx v4, by Waters Corporation (5 mentions)
Acquity uplc beh c18 column, by Waters Corporation (4 mentions)
Xevo tq ms, by Waters Corporation (3 mentions)
Beh c18 column, by Waters Corporation (3 mentions)
Q tof micro mass spectrometer, by Waters Corporation (3 mentions)
C18 analytical column, by Waters Corporation (3 mentions)
Acquity beh c18, by Waters Corporation (2 mentions)
Progenesis qi data processing software, by Waters Corporation (2 mentions)
Acquity uhplc beh c18 column, by Waters Corporation (2 mentions)
Elambda dad, by Waters Corporation (2 mentions)
Xcalibur software, by Thermo Fisher Scientific (2 mentions)
Progenesis qi, by Waters Corporation (2 mentions)
Targetlynx, by Waters Corporation (2 mentions)
Triple quadrupole ms, by Waters Corporation (2 mentions)
Acquity hss t3 1.8 μm vanguard guard column, by Waters Corporation (2 mentions)
Acquity uplc hss t3 column, by Waters Corporation (2 mentions)
Xevo g2 qtof, by Waters Corporation (2 mentions)
Argon 99.995, by Linde (2 mentions)
Acquity beh c18 column, by Waters Corporation (1 mentions)
80 protocols using acquity uhplc system
1
Quantitative Hepatic Ceramide Analysis
2
Chromatographic Fingerprint Analysis of CS Powder
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Standard compounds were dissolved in 50% methanol. Dry CS powder was diluted with ultrapure water to a concentration of 10 mg/mL. Chromatogram fingerprints were analysed using an UHPLC Acquity system (Waters, Milford, MA), with a photodiode array detector (scanning from 200 to 400 nm) recording at 323 nm. The UHPLC conditions included a column ACQUITY BEH C18, 100 × 2.1 mm, 1.8 μm particle size (Waters, Milford, MA); column temperature, 35 °C; injection volume, 5 μL; mobile phase (A) 0.1% formic acid (B) acetonitrile; flow rate, 200 μL/min; gradient B, 5% (0 min), 5% (5 min), 20% (15 min), 28% (25 min), 60% B (30 min), 5% (32 min) and 5% (33 min).
3
UHPLC Analysis of Standard Compounds and FZHFZY
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Standard compounds and FZHFZY dry powder were dissolved in methanol containing 0.1% formic acid. Chromatogram fingerprints were collected with an UHPLC Acquity system (Waters, United States), which comprised a UHPLC pump and a photodiode array detector scanning from 200 to 400 nm and recording at 330 nm. The HPLC conditions were as follows: column: ACQUITY BEH C18, 100 × 2.1 mm, 1.8-μm particle size (Waters), column temperature, 40°C; mobile phase: (A) acetonitrile; (B) 0.1% formic acid; flow rate, 250 μl/min; injection volume, 3 μl; gradient, 8% A (0 min), 30% A (15 min), 55% A (17 min) and 60% A (20 min).
4
Trace Organic Contaminants Extraction
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Oasis Accell TM Plus QMA (500 mg, 6 mL) cartridges, in tandem with Oasis HLB (200mg, 6mL). After sample loading on SPE cartridges, for both solid and liquid samples, Oasis Accell TM Plus QMA cartridges were discarded and Oasis HLB were washed with HPLC water, dried under vacuum for 30 min and eluted with 4x2 mL of methanol. Extracts were evaporated under a gentle nitrogen stream and reconstituted with 1 mL methanol-water mixture (50:50, v/v), except for soil extracts, which were reconstituted in 0.5 mL methanol-water (50:50, v/v). Prior to instrumental analysis, extracts from solid manure and slurry samples were filtered through 0.22 µm PVDF syringe filters. PhACs were detected and quantified using an ultra-high-performanceliquid chromatography (UHPLC) Acquity system (Waters Corporation, MA, USA) coupled to a 5500 QTRAP hybrid quadrupole-linear ion trap tandem mass spectrometer (AB Sciex, Foster City, CA, USA). Detailed information about analytical methods, chromatographic conditions and performance parameters (e.g., extraction recoveries, instrumental detection limits, method detection and quantification limits and matrix effects) can be found in the supplementary material (Tables S5,S6, S7, S8 and S9).
5
Metabolomic Analysis of Transcriptome Samples
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The same material used for transcriptome sequencing was used for metabolome sequencing, with six replicates per treatment. Shanghai Lumine Biotechnology Co. carried out the metabolite extraction and liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis. The metabolic profiles in ESI-positive and ESI-negative ion modes were analyzed using an AB SCIEX Triple TOF 5600 system (AB SCIEX, Framingham, MA, USA) and an ACQUITY UHPLC system (Waters Corporation, Milford, MA, USA). For both positive and negative ion modes (1.7 µm and 2.1 × 100 mm), an ACQUITY UPLC BEH C18 column was utilized. Progenesis QI data processing software (Waters Corporation, Milford, MA, USA) was used to process the metabolite data, and the identified metabolites were analyzed using custom databases for metabolite identification along with public databases (e.g., http://www.lipidmaps.org/ , accessed on 1 April 2019) and http://www.hmdb.ca/ (accessed on 1 April 2019) that included public databases for metabolite identification. The criteria used to establish differential metabolites were VIP value > 1 and p-value < 0.05 [33 (link)].
6
Metabolic Profiling of M. nipponense
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Metabolic profiling analysis was performed to select the differentially expressed metabolites (DEMs) in M. nipponense caused by the alkali treatment, which were determined by liquid chromatography–mass spectrometry (LC/MS) analysis [17 (link)]. The detailed procedures for the metabolic profiling analysis have been well described in a previous published paper [18 (link)]. The metabolic profiling was analyzed by an ACQUITY UHPLC system (Waters Corporation, Milford, CT, USA) and an AB SCIEX Triple TOF 5600 System (AB SCIEX, Framingham, MA, USA) in both ESI positive and ESI negative ion modes. The criterion of a seven-fold cross-validation was employed to ensure the robustness and predictive ability of the model, and permutation tests were employed to perform further validation.
7
Bile Acids and SCFA Quantification
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The analysis of bile acids and short chain fatty acids was performed as published previously.69 (link),70 (link) Cecal content was isolated, frozen in liquid nitrogen and stored at −80°C. To avoid bacterial metabolism, further steps were performed on dry ice, including cutting for weighting. Cold Methanol was added prior homogenization and centrifugation. The Acquity UHPLC system (Waters) and a UHPLC column (Acquity™ UPLC BEH™ C8, Waters) were used for UHPLC. Mass spectrometry was performed with the amaZon ETD Ion Trap (Bruker Daltonics GmbH) in negative ionization mod and the maXis (Bruker Daltonics GmbH) in positive electorspray ionization mode for SCFAs.
8
Optimized UHPLC-MS/MS Quantification of Anthelmintic Residues
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All analytes were chromatographically separated using an in-house method as previously described by Whelan et al., 2010 [30 (link)]. Here analytes were separated on a stainless steel HSS T3 (100 mm × 2.1 mm, 1.8 μm particle size) column on a Waters Acquity UHPLC system, with a binary gradient. Anthelmintic residues were detected by a Waters Quattro Premier XE triple quad mass spectrometer (Milford MA, USA) with an electrospray ionisation (ESI) interface, coupled to the LC. All analysis was performed using rapid polar switching using a modified version of the acquisition described by [30 (link)]. Dwell times, collision energies (CE) and collision voltages (CV) were further optimised from the original method, with the modified conditions shown in Supplementary Information Table S2 .
9
LC/MS Analysis of Metabolites
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LC/MS analysis was performed using an Acquity UHPLC system (Waters Corporation, Milford, USA) coupled with an AB Sciex Triple TOF 5600 System (AB Sciex, Framingham, MA). Data acquisition was conducted in full-scan mode (the m/z ranged from 70 to 1000) in combination with information-dependent acquisition mode. The parameters were set as follows: ion spray voltage, 5500 V (+) and 4500 V (−); ion source temperature, 550 °C (+) and 550 °C (−); collision energy, 10 eV (+) and −10 eV (−); curtain gas of 35 PSI; interface heater temperature, 550 °C (+) and 600 °C (−).
10
Melatonin extraction and ethylene production
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Melatonin was extracted according to our previous study18 (link). The samples were separated on a Waters Acquity UHPLC system (Milford, MA, USA) equipped with a BEH C18 column (Waters, 2.1 mm internal diameter ×50 mm length, and 1.7 µm particle size). Mass spectrometry (MS) analyses were performed using a QTof-Micro mass spectrometer. The detection parameters and conditions of the ultra-high-performance liquid chromatography (UHPLC)-MS analysis were set according to our previous study26 (link).
The ethylene production rate was measured as described in a previous study18 (link). ACC extraction and determination were performed according to the method described by Tucker et al.59 (link).
The ethylene production rate was measured as described in a previous study18 (link). ACC extraction and determination were performed according to the method described by Tucker et al.59 (link).
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