Agilent 1100 series hplc system

An Agilent 1100 HPLC Series system was used (Agilent Technologies, Darmstadt, Germany), connected with an Agilent Ion Trap SL mass spectrometer (MS) supplied with an electrospray or APCI ion origin. The separation of allicin was carried out utilizing a Synergy Polar 100 × 2.0 mm i.d., 4 μm columns (Phenomenex, SUA). The mobile phase is composed of 100% ammonium acetate, 1 mM in water, isocratic elution, flow 0.6 mL/min. A silver nitrate solution 1 mM in water was added post column, with a flow of 10 μL/min. The MS operated in positive multiple reaction monitoring mode, using an electrospray ion source and nitrogen as nebulizing and dry gas. The nebulizer was set at 60 psi, the dry gas flow was 10 L/min at 355°C. The device was set to record the transition m/z (449 + 451) >m/z (269; 271; 287; 289), specific to allicin-silver complex. The retention time of allicin in the above described conditions was 0.9 min.[19 ]

An Agilent 1100 HPLC series system (Agilent, USA) was employed for the content determination of SB, TV and TG, and the chromatographic conditions were optimized on the basis of literature with a slightly adjustment [3] .
Compounds SB, TV and TG in the samples were confirmed by comparing with the retention time of the references. The standard working curve created by an external standard method was used to determine the content of SB, TV and TG in the sample [3] . The study revealed that SB, TV and TG had good linearity in the range of 19.6–313.6 μg/mL, 20.0–100.0 μg/mL, and 19.4–310.4 μg/mL, respectively. The regression curves of SB, TV and TG were y = 5.9186x-199.82 (r² = 0.9990), y = 26.107x-213.45 (r² = 0.9994) and y = 7.4797x + 415.55 (r² = 0.9996), respectively, where × was the concentration of the standard sample (μg/mL) and y was the peak area.

An Agilent 1100 HPLC Series system (Agilent, USA) equipped with a degasser, binary gradient pump, column thermostat, autosampler, and U.V. detector was used. The HPLC system was coupled with an Agilent 1100 mass spectrometer (LC-MS analysis Ion Trap V.L.). For the separation, a reverse-phase analytical column was employed (Zorbax SB-C18, 100 × 3.0 mm i.d., 3.5 μm particle); the work temperature was 48 °C. The detection of the compounds was performed in both U.V. and M.S. mode. The U.V. detector was set at 330 nm until 17.5 min; then, it was set at 370 nm. The MS system was operated using an electrospray ion source in negative and positive modes. The chromatographic data were processed using ChemStation and DataAnalysis software from Agilent, USA. The mobile phase was a binary gradient: methanol and acetic acid 0.1% (v/v). The elution started with a linear gradient, beginning with 5% methanol and ending at 42% methanol, for 35 minutes; then, 42% methanol was used for the next 3 minutes. The flow rate was 1 mL min⁻¹ and the injection volume was 5 μL. Compounds were qualitatively identified based on spectral matching of the signals and spectra of each polyphenol with the available library and literature.