539.4.1 TubingFigure 9.5 shows the tubing and wiring arrangement used for calibration. Aninjector loop with a capacity of 1mL or more is required. However, the capacityof the injector loop does not need to be precise. When an appropriate loop isnot available, prepare a loop using tubing 2m or more in length with an internaldiameter of 0.8mm. Alternatively, prepare a loop using tubing 1.3m or more inlength with internal diameter of 1.0mm. Change to the “ ~ 10mL/min.” tubingconfiguration. (refer to section 8)PumpWater1mL loopInjectorInternal diameter of 0.25mm or lessIntegrator orrecorderRI-2031Port No.6Port No.3Figure 9.5 Tubing and Wiring Arrangement for Calibration9.4.2 Standard solution(1) Prepare approximately 1000mL of water. Sufficiently degas the water toprevent generation of bubbles during calibration.(2) Dissolve 106.6mg of saccharose in a 100mL volumetric flask using thewater prepared in (1) above. The refractive index of this aqueoussaccharose solution is 160x10 -6RIU greater than the refractive index of thewater.Note: Use the same water for the mobile phase and to dissovle thesaccharose. If different water is used, the correct refractive index maynot be obtained.
