July 2019 Sugar ERP - Review Book

2. Does the method contain system suitability tests or controls as specified by the SMPR? If not, please indicate if there is a need for such tests or controls and which ones. 3. Is there information demonstrating that the method system suitability tests and controls as specified in the SMPR worked appropriately and as expected? If no, please specify. 4. Based on the supporting information, is the method written clearly and concisely? If no, please specify the needed revisions.

Yes, SRM NIST 3233 Breakfast cereal; CRM BCR-644 Artificial foodstuff; SRM NIST 1849a infant formula drink are included in the study. However, no numbers are given as performance characteristics for these controls.

An overview is given of the performance of the reference materials. However, it seems that the summary in Table 11 does not meet SMPR for NIST3233 for Glu, Fruc and Malt. The other reference materials meet the SMPR for the analytes tested in the range they are present in the sample. Galactose is not included, glucose and maltose that were included via NIST3233 do not meet SMPR, so data is limited. Fructose has high RSD and a recovery at the low end in NIST3233, whereas it is more in line in BCR-644. What does it mean? The method is clearly written, the procedure is clear and apparatus & materials, reagents are very well documented. The results & discussion section is limited. A lot of data is given, but it needs extra explanation what is observed and how it fits the SMPR. Some small remarks: - Check 10’0000 mg/kL number in section (D.a). Is it correct? - Table 1: pyramid sector 2 is missing from the list, while in text it says all are covered - Check E.a (page 5), same number as above 10’0000 mg/kL - Section F, sample prep: No need to cut samples in advance of extraction? - Section H.c page 8: Should you start with a blank? - Section J calibration: 5% deviation of the retention time is a lot! Effect of interferents! This needs explanation. - Table 52: Missing numbers for Mean recovery and %RSD - Lack of a conclusion, summary - There is no internal standard used. It is unknown what is the effect of the precipitation/filtration of the samples on the final concentration of the sugars. An internal standard could indicate potential losses during these steps. - Needs demonstration that extraction procedure is effective for other matrices than the CRMs tested - Method is only valid from 0.01-44% - Needs more elucidation on safety aspects - Needs clarity around interferents and effect on quantification of samples. Could be an option to test with different column types/different suppliers for robustness of the study. A combination with SUG-003 could be an option? The method seems promising, however based on the current data it is difficult to judge whether it meets the SMPR. I am personally wondering whether it could be combined with SUG-003, since they look quite similar but make use of different type of equipment (which could also be the case when a method continues in MLT). - The materials and methods section is well documented - The method procedure is clearly written - Many matrices included - All sugars from SMPR are included

5. Based on the supporting information, what are the pros/strengths of the method? 6. Based on the supporting information, what are the cons/weaknesses of the method?

7. Any general comments about the method?