The purpose of this analysis was to quantify oxalates present in plant leaves and animal feed. High-performance liquid chromatography analysis was performed on an Agilent 1260 Infinity Quaternary LC (Agilent Technologies) and Agilent 1100 Series (Agilent Technologies) based at two laboratories (Toxicology and Residue) for reproducibility purposes. The HPLC instrument from each laboratory comprised a binary pump, high-performance degasser, high-performance auto-sampler, column thermostat and a variable wavelength detector. The analytical column used was an Eclipse XDB-C18 4.6 mm ID × 250 mm (5 µm) 80Å. The chromatographic condition consisted of 20 mM sulphuric acid mobile phase, 1 mL/min flow rate for 10 min at a wavelength detection of 210 nm, ambient temperature and injection volume of 50 µL. The sample was prepared using previously reported methods (Ruan et al. 2013 (link)). A 1 mg/mL stock solution of sodium oxalate was prepared and appropriate volumes from the stock solution were further diluted to prepare standards of varying concentrations.
Agilent 1260 infinity quaternary lc
Manufactured by Agilent Technologies
Sourced in United States, Germany
The Agilent 1260 Infinity Quaternary LC is a high-performance liquid chromatography (HPLC) system. It features a quaternary solvent delivery system capable of delivering up to four different solvents simultaneously for advanced gradient capabilities. The system is designed to provide precise and reliable liquid chromatography analysis.
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Lab products found in correlation
Agilent 1100 series, by Agilent Technologies (1 mentions)
Sunfire c18 column, by Waters Corporation (1 mentions)
Hplc system, by Waters Corporation (1 mentions)
Zorbax stable bond c18, by Agilent Technologies (1 mentions)
Eclipse plus c18 column, by Agilent Technologies (1 mentions)
Ethyl acetate etoac, by Daejung Chemicals (1 mentions)
Kieselgel 60 f254, by Merck Group (1 mentions)
Kieselgel 60, by Merck Group (1 mentions)
Avance, by Bruker (1 mentions)
Agilent 6120 quadrupole, by Agilent Technologies (1 mentions)
8 protocols using agilent 1260 infinity quaternary lc
1
Quantifying Oxalates in Plants and Feeds
2
HPLC-MS Analysis of Plant Extracts
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HPLC analysis of extracts was carried out in a Waters HPLC system (Waters Corporation, Milford, MA) composed of a 1525 binary pump with a column oven, a 2707 autosampler, and a 2998 photodiode array detector (210–400 nm) using a Waters SunFire C18 column (5 μm, 4.6 × 150 mm). Plant extract, fractions, and single compounds were eluted by a linear gradient system using acetonitrile and water (0.1% HCOOH), ranging from 20% A to 90% acetonitrile (for 40 min) followed by an isocratic solvent 100% acetonitrile (10 min) at the flow rate of 1.0 mL/min. UV absorption was monitored under 300 nm. LC-ESI-MS experiments were performed on Agilent 6120 single quadruple MS hyphenated to Agilent 1260 Infinity quaternary LC (Agilent Technologies, Santa Clara, CA, USA) using a Thermo Acclaim Polar Advantage II (2.2 μm, 2.1 × 100 mm) in the positive mode, and developed by a gradient solvent mixture with acetonitrile and water (0.1% HCOOH), 20% to 100% acetonitrile for 25 min at 0.3 mL/min. The MS detection was carried out using electrospray ionization (ESI) with API source and MS spectra were obtained between m/z 100–1000 in a positive mode.
3
Triglyceride Species Analysis by HPLC
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After scaled-up synthesis and SPD, TAG species were analyzed by HPLC to further determine which enzyme worked best in producing MLM-type SL TAG species. TAG species determination was performed as described previously with modification 23) (link) . Tricaproin, tricaprylin, tricaprin, trilaurin, triolein and tristearin were used as standards. Their concentrations were at 1 mg/mL after dissolving in dichloromethane. Purified SL was prepared at 5 mg/mL concentration in dichloromethane. Agilent 1260 Infinity Quaternary LC (Agilent Technologies, Inc.) HPLC system equipped with a Sedex Model 85 ELSD (SEDERE SAS Inc., Alfortville, France) was used for TAG species analysis. Separation was achieved with a reverse phase Agilent Zorbax stable bond C18 (250 mm×4.6 mm, 5 µm) column. Column temperature was maintained at 50℃, flow rate at 0.5 mL/ min, injection volume was 5 µL. ELSD drift tube tempera-ture was set at 50℃, nebulizer gas pressure at 3.4 bar and gain at 10. 100％ methanol was used as mobile phase A, and mobile phase B was a mixture of acetone/isopropanol (1:1, v/v) . The following gradient system was employed: hold at 1％ TAG species were determined based on their equivalent carbon number (ECN) compared with the analyzed standards retention times in the chromatograph, and their relative mol％ was calculated based on their peak area.
4
HPLC Analysis of Purine Metabolites
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Inosine, guanosine, hypoxanthine, guanine, xanthine, and uric acid were quantified by HPLC using the method described by Li et al. (2014) (link), with slight modifications. Analyses were carried out using a Zorbax SB-C18 (5 μm, 4.6 × 250 mM) column connected to an HPLC device (Agilent 1,260 Infinity Quaternary LC) with a diode array detector (Agilent Technologies, Waldbronn, Germany). Working solutions (1.3 mM) of all compounds were prepared in phosphate buffer (K3PO4, 100 mM, pH 7), cleaned and sterilized by passing the solution through a filter (0.22 μm pore size; Nalgene 176–0020 nylon), and degassed by sonication. The separation of the compounds was achieved by using an isocratic flow (0.5 mL/min) of methanol and 0.1% of acetic acid in Milli-Q water (3:97, v/v). The retention times at 245 nm were 5.00, 4.75, 2.46, 2.36, 2.24, and 2.09 min for guanosine, inosine, xanthine, hypoxanthine, guanine, and uric acid, respectively.
Compound quantification was carried out by developing standard curves built using the corresponding pure compounds (Sigma, Alcobendas, Madrid). The analyses were carried out in triplicate.
Compound quantification was carried out by developing standard curves built using the corresponding pure compounds (Sigma, Alcobendas, Madrid). The analyses were carried out in triplicate.
5
Quantification of Aromatic Compound Degradation
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The method for detection of anthranilate and catechol degradation was carried out as previously described [46 (link)]. Bacterial isolates were tested in biological triplicate and analyzed with two technical replicates. The compounds and degradation products were separated on an Agilent 1260 Infinity Quaternary LC (Agilent Technologies, Santa Clara, CA, USA) outfitted with an Eclipse Plus C18 column (4.6 mm ID × 250 mm, 5 µm particle size, Agilent Technologies) and using a flow rate of 1 mL/min and a 15 min gradient from 5–25% MeOH (0.1% “v/v” formic acid), monitoring 210 nm and 230 nm. Standard curves were generated for both anthranilate and catechol using known concentrations of each compound, prepared in triplicate.
6
Phytochemical Analysis of Termantia asiaticum
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1H NMR data were recorded at 600 MHz on a Bruker Avance Neo 600 (Brucker, Billerica, MA, USA) spectrometer in the Central Laboratory of Kangwon National University (Chuncheon, Korea). The Mass experiements were performed on a Waters Xevo G2 qTOF mass spectrometer (Waters MS Technologies, Manchester, UK) with the UPLC system through an electrospray ionization (ESI) interface for the MN study, as well as the exact mass analysis. Silica gel Kieselgel 60 (40–60 μm, 230–400 mesh, Art. 9385, Merck, Darmstadt, Land Hessen, Germany) and Diaion HP-20 (Mitsubishi Chemical Industries Ltd., Chiyoda-ku, Tokyo, Japan) were used for column chromatography. Thin layer chromatography (TLC) was performed to monitor the different fractions of T. asiaticum using a Kieselgel 60 F254 (Art. 5715, Merck, Darmstadt, land Hessen, Germany) and an RP-C18 F254 (Art. 15389, Merck, Darmstadt, Land Hessen, Germany). Semipreparative HPLC was performed on an Agilent 1260 Infinity Quaternary LC (Agilent, Santa, CA, USA) with an InspireTM 5 μm C18 (250 × 21.2 mm, Dikima, Foothill Ranch, CA, USA). Methanol (MeOH), n-hexane, ethyl acetate (EtOAc) and n-butanol (n-BuOH) were purchased from Daejung (Si-heung, Korea). The other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).
7
Synthesis and Characterization of DNP-PEG2-Acid
8
HPLC Analysis of Dietary Sugars
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The apple pomace extract and the supernatant collected after in vitro digestion of biscuits and bread were analyzed according to Englyst et al. (1999) . Sugar were separated by an HPLC (Agilent 1260
Infinity Quaternary LC, Agilent Technologies, Germany) equipped with an auto-injector (1260 ALS), a chromatographic column (Amino 100 A, 5 µm, 250 mm, 4.6 mm, SephaChrom, Rho, Italy), a temperature control system (1260 TCC) and a quaternary pump (1260 Quat Pump), generating a flow rate of 1 mL/min. The mobile phase was represented by water and acetonitrile and sucrose (0.05 g/mL) was diluted to 1:5, 1:10, 1:20 (v/v), added with L-(+)-arabinose (0.01 g/mL) as internal standard and used for calibration.
Infinity Quaternary LC, Agilent Technologies, Germany) equipped with an auto-injector (1260 ALS), a chromatographic column (Amino 100 A, 5 µm, 250 mm, 4.6 mm, SephaChrom, Rho, Italy), a temperature control system (1260 TCC) and a quaternary pump (1260 Quat Pump), generating a flow rate of 1 mL/min. The mobile phase was represented by water and acetonitrile and sucrose (0.05 g/mL) was diluted to 1:5, 1:10, 1:20 (v/v), added with L-(+)-arabinose (0.01 g/mL) as internal standard and used for calibration.
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