Low Lactose ERP - Review Book

4. Does the method, as written, contain all appropriate precautions and warnings related to the method's reagents, components, instrumentation, or method steps that may be hazardous? If no, please suggest wording or option(s). 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 method description contains a Safety Precaution section, in which the potential safety hazards are highlighted.

Yes. (Please note that the equation it used to estimate the LOD is not common).

The SMPR specifies a reference material, NIST SRM 2383a, and Harmonization materials from National Milk Laboratories (UK) and from MUVA (Germany). The supporting data of this method shows results from four MUVA samples, the MUVA-ML-2308, 2309 (best used before: 06/2018, and the MUVA-ML-2310, 2311. The intermediate reproducibility from the determination of the four harmonization samples meet the SMPR requirement. However, the recovery was not evaluated. The reason given in the supporting document says that “the reference values of the materials were obtained from two different technologies, enzymatic assays, and HPLC, and none of them are reference methods for low lactose products”. I did a simple calculation to compare the HPLC values from the harmonization samples to the BIOMILK lactose results. We can see that for the high concentration (>100 mg/100g) lactose samples (-2308 and -2310 in Table 17), the recovery is within 10%, meet the SMPR. But for the lactose-free samples (- 2309 and -2311), the BIOMILK results are outside the SMPR limit on recovery. However, please note that even though the BIOMILK results are not within the SMPR limit on recovery for lactose-free samples, they are within the HPLC reference values’ uncertainty limits. In the supporting document, it contains a third-party validation report, in which the recovery was evaluated against a MUVA-ML-2311 UHT milk, which has a reference value of lactose 0.0066% (m/m), a recovery of about 68% was obtained. In that report, a conclusion was made that this level of bias was not significantly different from the reference value. However, this recovery is outside of the SMPR.