Amino acids in the garlic powder samples were determined using an Agilent 1260 infinity quaternary liquid chromatograph (Hewlett Packard) with a multiple wave-length detector operating at 338 nm (excitation=340 nm). Separation was carried out with a Zorbax Eclipse AAA rapid resolution column (150×4.6 mm i.d., 5 μm particle size, Agilent Technologies). The mobile phase, comprising 40 mM Na2HPO4, pH 7.8 (solvent A) and ACN/MeOH/water 45:45:10 (v/v/v) (solvent B) was used with the following linear gradient profile: 0% B (0~1.9 min), 0~57% B (1.9~18.1 min), 57~100% B (18.1~18.8 min), 100% B (18.8~22.3 min), 100~0% B (22.3~23.2 min), and 0% B (23.2~26 min) was applied at a flow rate of 2.0 mL/min. The column was equilibrated for 5 min under initial conditions prior to injection of the next sample. The column temperature was 40°C. Determination of sample amino acid content was facilitated using pre-column derivatization with o-phthalaldehyde, and 0.5 μL portions were injected into the HPLC system. Data analysis was performed using the Chemstation software (Hewlett Packard).
Zorbax eclipse aaa rapid resolution column
Manufactured by Agilent Technologies
Sourced in United States
The Zorbax Eclipse AAA rapid resolution column is a high-performance liquid chromatography (HPLC) column designed for the analysis of amino acids. It features a small particle size and rapid separation capabilities, enabling efficient and reliable amino acid determinations.
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2 protocols using zorbax eclipse aaa rapid resolution column
1
Amino Acid Profiling of Garlic Powder
2
Quantifying Ergosterol in Fungi by HPLC
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Ergosterol concentrations were quantified by applying HPLC according to the protocol of Gulis and Bärlocher [27 (link)]. An Agilent 1260 Infinity Bioinert HPLC System equipped with an autosampler was used together with a Zorbax StableBond Aq Analytical Guard Column (4.6 × 12.5 mm; 5 µm particle size; 80 Å pore size, Agilent Technologies, Santa Clara, CA, USA) and a Zorbax Eclipse AAA Rapid resolution column (4.6 × 150 mm; 3.5 µm particle size, 80 Å pore size, Agilent Technologies). For detecting ergosterol, the wavelength of the UV detector was set at 282 nm. The mobile phase consisted of 100% methanol (HPLC grade, Sigma-Aldrich) with a flow rate of 0.8 mL/min. The injection volume was 400 µL and the samples were run for 20 min in total, including 5 min of purging at a column temperature of 25 °C. Ergosterol standards (HPLC grade, Sigma-Aldrich) were prepared in a range from 500 µM to 30 nM in methanol and run together in the same sequence batch with the fungal samples. Furthermore, the temperature of the autosampler was kept at 10 °C to protect them from potential degradation. Peak areas were used for calculating ergosterol concentrations in samples.
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