Lpg 3 00 pump

The HPLC system is consisted of Dionex system with LPG 3 × 00 pump, ACC-3000 auto sampler, column oven and DAD-3000(RS) diode array ultraviolet (UV)/visible spectroscopy detector. Separation was accomplished using a Shiseido C₁₈ column (4.6 mm I.D. × 250 mm, 5 μm pore size). Column temperature was maintained at 30°C. The mobile phase was composed of 0.1% trifluoroacetic acid aqueous solution (a) and acetonitrile (b) at a flow rate of 1.0 ml/min. The gradient flows were as follows: 5% B at 0-10 min, 5-17% B at 10-20 min, 17-30% B at 30-40 min, 30-80% B at 40-60 min. The injection volume was 20 μL. The wavelength of UV detection was set at 205 nm, 250 nm, 280 nm and 330 nm, respectively.

To identify the phycocyanin from water extracted S. maxima sample, extracted S. maxima and purchased C-phycocyanin were analyzed by HPLC-DAD. C-phycocyanin from Spirulina sp. was purchased from Sigma–Aldrich (St. Louis, MO, USA, Cat # P2172). The high-performance liquid chromatography (HPLC) (Dionex) equipped with an LPG 3 × 00 pump, an ACC-3000 autosampler, a DAD-3000 (RS) diode array ultraviolet (UV)/visible detector, and a column oven. Both samples were separated on a Jupiter C₅ column (5 μm, 300 Å, 4.6 mm × 250 mm) at 25°C. The mobile phase consisted of 20% (v/v) aqueous acetonitrile (ACN) solution containing 0.1% (v/v) trifluoroacetic acid (TFA) and all reagents were purchased from J. T. Baker as HPLC grade. The flow rate was 1.0 mL/min and the concentration of S. maxima sample was 100 ppm and phycocyanin was 1000 ppm. The injection volume of each sample was 20 μL. The output signal of the detector was recorded using a Dionex Chromeleon^™ Chromatography Data System. The UV wavelength was 580 nm and 640 nm, respectively, and the chromatograms were acquired at 580 nm.