The water-soluble fraction of the cheeses was prepared according to the method of Kuchroo and Fox (1982) . Total FAAs from the water-soluble extracts were determined by a Biochrom series 30 Amino Acid Analyzer (Biochrom Ltd., Cambridge Science Park, UK) as described by Siragusa et al.(2007) (link) by reversed-phase high-performance liquid chromatography (HPLC, 1200 Series Agilent chromatograph, Agilent Technologies, USA) using a Waters Nova Pack C18 column (3.9 × 300 mm, Waters Corporation, USA) and UV detection (254 nm).
Agilent 1200 series chromatograph
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1200 series chromatograph is a high-performance liquid chromatography (HPLC) system designed for analytical applications. It features a modular design, allowing for the integration of various components such as pumps, autosamplers, and detectors to create a customized analytical solution. The Agilent 1200 series chromatograph is capable of delivering precise and reliable separation and detection of chemical compounds in complex samples.
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Lab products found in correlation
Nova pack c18 column, by Waters Corporation (1 mentions)
Rezex roa column, by Phenomenex (1 mentions)
Agilent zorbax sb c18, by Agilent Technologies (1 mentions)
Agilent 6460 triple quad lc ms, by Agilent Technologies (1 mentions)
Masshunter software, by Agilent Technologies (1 mentions)
Refractive index detector, by Agilent Technologies (1 mentions)
7 protocols using agilent 1200 series chromatograph
1
Quantification of Free Amino Acids in Cheese
2
Profiling Amino Acids in Fermented Sausages
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The content of free amino acids in the fermented sausages was determined according to Aristoy & Toldrá (1991) using norleucine (65.6 µg) as internal standard. Phenylthiocarbamyl amino acids derivatives were analysed by reversed-phase HPLC (1200 Series Agilent chromatograph; Agilent, Palo Alto, CA, USA) using a Waters Nova Pack C18 column (3.9 x 300mm) (Waters Corporation, Milford, USA) and ultraviolet detection (254nm) as described by Flores, Aristoy, Spanier & Toldrá (1997) . The concentration of reduced glutathione, cysteine, glutathione and cystine was determined as described by Marušić, Aristoy & Toldrá (2013) using an Agilent 1100 Series HPLC with fluorescence detection (λ ex = 330 nm, λ em = 376 nm). The analysis was performed in three sausages at each sampling time and each sausage sample was derivatized in triplicate. The results were expressed as mg/100 g sausage in dry matter.
3
Quantification of Lactic Acid in LAB and G. geotrichum Bioreactor
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Samples taken during bioreactor culture of LAB and G. geotrichum were diluted with distilled water and centrifuged for 10 min at 13,000× g using a Biofuge Primo R centrifuge (Heraeus, Hanau, Germany). The supernatant obtained was passed through 0.45 µm mixed cellulose ester syringe filters (Lab Logistics Group GmbH, Meckenheim, Germany). The obtained samples underwent HPLC analysis to identify and quantify the lactic acid, using an Agilent 1200 series chromatograph with an autosampler and refractive index detector (RID) (Agilent Technologies, Santa Clara, CA, USA). The apparatus was equipped with a REZEX-ROA column (300 mm × 7.8 mm, Phenomenex, Aschaffenburg, Germany). The samples (10 µL) were injected into the HPLC column using the splitless mode. Lactic acid was analyzed at 40 °C with 0.005 N H2SO4 as a mobile phase at a flow rate of 0.6 mL/min. Lactic acid was identified by comparison with a reference compound based on the retention index. Quantitation was performed with an external standard curve. The analysis was carried out on samples taken during each day of culture in the bioreactor in order to study changes in lactic acid concentration during its duration.
4
Fluvalinate HPLC Analysis Protocol
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Fluvalinate standard (Chem Service, https://www.chemservice.com/tau-fluvalinate-n-13263-100mg.html (accessed on 8 March 2022) and purified fluvalinate stock solution (1 g/L) were subjected to HPLC analysis by an Agilent 1200 Series chromatograph (Agilent, Santa Clara, CA, USA). Sample separations were performed on an Agilent ZORBAX SB-C 18 (Agilent, Santa Clara, CA, USA)5 μm, 4.6 mm × 250 mm reversed-phase analytical column. For chromatographic conditions, the elution was carried out with a fixed ratio mixture consisting of 2 solvents (A + B = 70 + 30, v/v). Solvent A was 100% acetonitrile and solvent B was 2% acetic acid in acetonitrile. The flow rate was 1.5 mL/min, the column was kept at room temperature, and detection was performed at a wavelength of 230 nm. The injection volume consisted of 10 μL. HPLC patterns of the fluvalinate standard and purified fluvalinate stock solution were compared at a concentration of 1 g/L. HPLC results indicated that the patterns of purified fluvalinate stock solution and fluvalinate standard were identical (Figure S1 ), suggesting that the purification method was workable and efficient (purity > 90%).
5
Capillary HPLC-ESI-MS/MS Analysis Protocol
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Capillary HPLC–ESI–MS/MS analyses were performed with an Agilent 1200 series chromatograph (Agilent Technology, Waldbronn, Germany) equipped with a binary capillary pump, degasser, autosampler, thermostatically controlled column compartment and capillary Zorbax SB C18 column coupled to an electrospray ionization triple quadrupole mass spectrometer (Agilent 6460 Triple Quad LC/MS, Agilent Technologies, Santa Clara, CA, USA). All the operations, data acquisition and analysis were processed by MassHunter Software (Agilent Technology, USA). Operational parameters are summarized in Table S1 (see Electronic Supplementary Material).
A Bandelin Sonorex model 1210 ultrasonic bath (Germany) and MPW model 350R centrifuge (MPW Warsaw, Poland) were used for extraction procedures. Sample mineralization was performed with a Speedwave®4 microwave digestion system (Berghof, Germany).
A Bandelin Sonorex model 1210 ultrasonic bath (Germany) and MPW model 350R centrifuge (MPW Warsaw, Poland) were used for extraction procedures. Sample mineralization was performed with a Speedwave®4 microwave digestion system (Berghof, Germany).
6
SCFA Quantification of Prebiotic Xylooligosaccharides
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The determination of SCFAs obtained in the in vitro fermentation of the prebiotic xylooligosaccharides was performed using an Agilent 1200 series chromatograph coupled to a refractive index detector (Agilent Technologies, Santa Clara, CA, USA). For this purpose, an Aminex HPX-87H column (Bio-Rad, Hercules, CA, USA) was employed at 50 °C using a mobile phase of 0.003 M sulfuric acid in isocratic mode at a flow rate of 0.6 mL/min.
7
Carbohydrate Analysis by HPLC-RI
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The analysis was performed on Agilent 1200 Series chromatograph (Agilent Technologies, Santa Clara, CA, USA) coupled with Zorbax Hi-PlexCa (7.7 × 300 mm) column (Agilent Technologies, Santa Clara, CA, USA) and RI detector. The elution was isocratic (30 min) at 0.6 mL/min, using demineralized water as the mobile phase. Column temperature was set at 80 °C, injection volume was 10 μL. RI detector was set at 40 °C and the signal recorded at 2 s response time. The peaks were identified by retention time comparison with known standards. Internal standard calibration curves using glycerol (50 μg/mL) were established for sucrose, glucose, fructose, xylitol and maltitol. The results are expressed as the means with the standard deviation.
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