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It was also found that the mobile phase ratio was the most important factor in capsaicin separation.
Three factors including mobile phase ratio (ethanol:water), bed height and flow rate of mobile phase were analyzed.
The method remained unaffected, by small but deliberate variations, in the LC flow rate (± 10%) and mobile phase ratio (± 10%).
Robustness was studied by introducing small changes in the mobile phase ratio, flow rate, and detection wavelength of analysis.
Robustness of the method was checked by making slight changes in chromatographic conditions like mobile phase ratio, pH of buffer, flow rate.
Robustness of the proposed method was determined in two different ways, i.e., by making deliberate changes in the mobile phase ratio, flow rate, and detection wavelength of analysis.
Similar(51)
Water and methanol were used as a mobile phase, the ratio of water and methanol was 1 9 and the speed was 1.0 mL/min.
The parameters involved were: the pH of the phosphate buffer used in the mobile phase, the ratio between the components of the mobile phase, pH of the derivatization reaction mixture, and column temperature.
The evaluation was performed using a C18 column (150 mm × 4.6 mm i.d., 5 μm particle size) with a gradient flow rate of acetonitrile and diluted sulfuric acid (0.002 M) as mobile phase from ratio 15:85 to 80 20 (%v/v) at a flow rate of 2.0 ml/min at 265 nm.
The mobile phase composition ratio was reverted to the initial conditions and the column was allowed to re-equilibrate for 15 min for a total run time of 32 min.
TLC plates in Iodine chamber showed a significant results in 9 1, 8 2, 7 3 and 6 4 mobile phase solvent ratio (n-hexane : ethyl acetate) and found spots in TLC with Rf values 0.65, 0.69, 0.68, 0.67 and indicated different verities of compounds.
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