AOAC OMB Final Action Recommendation (December 2019)-2016.14
2016.14 (October 2019) – FOS-03 METHOD
FOR ERP USE ONLY DO NOT DISTRIBUTE
Figure 2016.14D. Example of calibration curves obtained on a CarboPac PA20 column, but without post- 359 column addition, resulting in reduced quantitative range. A: Glucose B: Fructose.
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Appendix A: Sucrase/β-amylase/pullulanase/maltase enzyme mixture check test 367
To check performance and suitability of sucrase enzyme mixture, equal aliquots (100 µL) of sucrose 368
solution (0.75 mg/mL) and fructooligosaccharide (FOS) ingredient solution (0.75 mg/mL) are mixed and 369
treated with the sucrase enzyme mixture (200 µL) to be tested. Water is added (100 µl) and the mixture 370
is incubated at 40 °C for 90 min (or the relevant conditions to be tested). After centrifugation (5 min at 371
10 000 x g ), an aliquot of the supernatant (100 µL) is transferred into a vial suitable for the autosampler 372
instrument, diluted with water (400 µL) and mixed. This solution is injected onto an HPAEC-PAD 373
instrument (conditions described below) and the chromatographic profile obtained is compared with the 374
one obtained from the same solution (sucrose/FOS) not treated with the enzyme mixture (200 µL of water 375
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instead of the enzyme mixture).
Note: It is highly recommended to perform this test with a FOS ingredient of low DP (average 3-5) which 377
has been produced by partial hydrolysis of inulin, as these seem to be the most sensitive to sucrase side 378
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activities or impurities.
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