Low Lactose ERP - Review Book

III. Review of Supporting Information 1. Are the definitions specified in the SMPR used and applied appropriately in the supporting documentation (manuscripts, method studies, etc...)? If not, please explain the differences and if the method is impacted by the difference. 2. Is there information demonstrating that the method meets the SMPR Method Performance Requirements using the Reference Materials stated in the SMPR? If not, then specify what is missing and how this impacts demonstration of performance of the method.

Yes

The Reference Materials stated in the SPMR were most of them used, but:

-NIST 18492a Infant Formula: lactose level outside the scope of the SMPR. -NIST 1869 Infant Formula: the same level of lactose of NIST 2383a stated in the SMPR. Good agreement with the reference value but with only 4 replicates, the intermediate reproducibility (not calculated by the authors, the value was calculated by the reviewer) was higher than the stated in the SMPR. -MUVA CA-0904 Sodium Caseinate: only one value of this material. The recovery does not meet the SMPR. -MUVA 2310 UHT Low lactose milk: Good agreement with the reference value but with only 4 replicates, the intermediate reproducibility (not calculated by the authors, the value was calculated by the reviewer) was a little bit higher than the stated in the SMPR. -MUVA 2311 UHT Lactose free milk: Good data, 8 replicates were analyzed, good agreement with the reference value and the intermediate reproducibility meets the SMPR. There is not repeatability data. Regarding intermediate reproducibility data, it was calculated for a higher level than the stated in the SMPR (cheese spread 6,8% of lactose). The reviewer calculated some intermediate reproducibility data from the Reference Materials analyzed but data do not meet SMPR except for UHT Lactose Free Milk. Regarding recovery, it was only calculated for dry pet food sample. The recovery was calculated at 3 different levels but only one, the lowest level, must be considered. The other two are higher than the levels specified in the SMPR. The recovery for the level 1% meets the SMPR. Good agreement with some reference values of reference materials. Regarding interferences, not problems with other sugars (a lot of sugars tested) except from cellobiose. But it seems not to be an important interference for the matrices of the scope. Some authors mention that 25% ethanol is not enough for avoid enzyme activity, but the authors analyzed interference with beta galactosidase and the results were very good. Only sugars and enzymatic activity were tested, there are some potential interferences mentioned in the SMPR that were not considered. More data is needed.

3. Is there information demonstrating that the

method performs within the SMPR Method Performance REquirements table specifications for all analytes in the SMPR applicability statement? If not, please specify what is missing and whether or not the method's applicability should be modified.