The effect of changing the pH of phosphate buffer solution was tested from 3.0 to 7.0.
For skin donors, skin samples, frozen in an 85% glycerol buffer, and skin storage solution were tested.
Local lesions typical of PMMoV infection were excised and crushed into 0.6 ml of inoculation buffer, and 350 µl of this solution were tested for the presence of PMMoV by RT-PCR using primers published by Hamada et al. [10], by electron microscopy, and by inoculation to one plant of Capsicum annuum var Yolo Wonder or Nicotiana benthamiana, two susceptible hosts for PMMoV.
Three different buffer solutions were prepared: pH 4 citrate buffer 0.1 M, pH 6 citrate buffer 0.1 M and PBS 1x (pH 7.4).
Both lipid and buffer solutions were fully degassed before loading into the appropriate cell.
Octanol and buffer solutions were presaturated.
For this reason, 0.01 M of CuCl2 solution, 7.5 mM of ethanol neocuproine solution, and 1.0 M of CH3COONH4 buffer solution were added to each test tube, respectively.
In order to investigate the effect of buffer components on the reaction, different buffer solutions of pH 8.5 were tested: Clark, Robinson, phosphate, borate and tris buffers.
Various ratios of acetonitrile and phosphate buffer solutions and different mobile phase pH values were tested using a C18 (150 × 4.5 mm, 3.5 μm) column, higher acetonitrile ratio resulted in shorter retention times of drugs Using this mobile phase ratio best results were obtained in terms of peak symmetry, selectivity and analysis time for drugs and the results are shown in Fig. 2.
To compensate for the removed 1.5 mL solution, 1.5 mL of fresh TRIS buffer solution was added to the original test solution.
