Each 10-μl aliquot of sample was injected into a 2.1×100 mm (1.8 mm) ZORBAX SB-C18 column for subsequent rapid resolution liquid chromatography (6530 series) (both from Agilent Technologies, Inc.). A mixture of acetonitrile containing 0.1% formic acid (phase A) and water containing 0.1% formic acid (phase B) were used as the mobile phase for electron spray ionisation in positive mode (ESI+), while a mixture of acetonitrile (phase A) and water (phase B) was used as the mobile phase for ESI in negative mode (ESI−). The protocols for the linear mobile phase gradient were as follows: 95% A held for 1 min; decreased to 2% A by 10 min; held at 2% A until 13 min; increased to 95% A by 13.1 min; and held at 95% A until 20 min. The flow rate of the mobile phase was 0.3 ml/min at 40°C.
6530 series
Manufactured by Agilent Technologies
Sourced in Germany
The Agilent 6530 series is a high-performance liquid chromatography-quadrupole time-of-flight (LC-QTOF) mass spectrometry system. It is designed to provide accurate and precise mass measurements for the identification and characterization of unknown compounds.
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2 protocols using 6530 series
1
Rapid Liquid Chromatography for Metabolite Analysis
2
HPLC-QTOF-ESI Mass Spectrometry Analysis
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The analysis was completed using an HPLC system (1100 series, Agilent Technologies, Waldbronn, Germany) coupled to a quadrupole time-of-flight (QTOF) equipped with an orthogonal electrospray ionization source (ESI) with Jet Stream thermal focusing technology (6530 series, Agilent) and operating in positive or in negative ion modes (i.e. polarity-switch was not used, and the samples were analyzed twice). The HPLC system consisted of a degasser, a quaternary pump, an automatic injector, and a thermostatic column compartment. Agilent Mass Hunter Qualitative Analysis software (B.04.00) was used for MS control, data acquisition, and data analysis. A solution of compounds whose masses are known with great accuracy was continually infused to the system to allow constant mass correction for accurate mass calibration. Thus, during all analysis, two reference masses were used, m/z 121.0509 (C5H4N4) and m/z 922.0098 (C18H18O6N3P3F24) for positive ionization mode, and m/z 112.9856 (C2O2F3(NH4)) and m/z 1033.9881 (C18H18O6N3P3F24) for negative ionization mode.
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