Low Lactose 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. 5. Based on the supporting information, what are the pros/strengths of the method?

YES: There are.

YES: There are information demonstrating that the method system suitability tests and control as specified in the SMPR worked appropriately and expected.

Generally: The method is well described and substantively prepared. The project of the method is well integrated and includes a clear and concise description.

The developed method was evaluated following the definitions of AOAC SMPR 2018.009 with respect to values of LOD and LOQ, Linearity, Repeatability, Reproducibility, Recovery. Analytical data was collected for all compounds, and required / suitable matrices (such as food, infant formula, feed and pet food matrices) listed in AOAC SMPR 2018.009. In the APPENDIX A, Authors presented example chromatograms and potential interfering compounds. In the APPENDIX B, Authors wrote: The theoretical LOQ is 0.005%. MUVA ML-2311 Lactose-free UHT Milk has ~0.007% of lactose. Based on six replicate determinations of MUVA ML-2311 in a single batch, the standard deviation for lactose result was determined, and considered as the “noise level” in a true sample matrix. Therefore, LOQ can be calculated as ten times the noise.' ONLY: My additional recommends: Could Authors be evaluate more precisely matrix effect and influence of them on the results? Examples of chromatograms for some matrices have been omitted, or their suitable full results are not presented in the described method.

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

7. Any general comments about the method?

My additional remarks and recommendations to the method are as follows:

Page 12, Extraction Robustness Study – Special Spike Recovery Results, table: In my opinion Authors may add RSD values for lactose and maltose at three spiked levels (0.1%, 2%, 20%).

Page 12, Summary In summary, the XBridge BEH Amide column provides excellent separation of six common sugars that are found in food, infant formula, pet food, and feed matrices.' In my opinion Authors may add Rs values for all analytes.