Amino-01 (July 2018)

Amino‐01 (July 2018)  Method Evaluation Form (MEF)

Evaluation of Method Performance  vs. SMPR requirements. AOAC SMPR: 2014.013

 FOR ERP USE ONLY  DO NOT DISTRIBUTE

N/A

Method Reference #

Method title:

Single Laboratory Validation Report for Total Amino Acids by UHPLC‐UV in Infant Formulas and Adult Nutritionals

Principle of the method:

Separation by UHPLC or derivatized amino acids after acid hydrolysis

Weighting  factor for  parameter

Suitability Ranking (1‐ 3‐5)  (select from drop‐down  list, 5 = best)

Method Performance                Please  report in units as stated in SMPR!

SMPR Requirement

Parameter

Applicable to all forms of infant, adult, and/or pediatric  formula (powders, ready‐to‐feed liquids, and liquid  concentrates).  Any combination of milk, soy, rice, whey, hydrolyzed  protein, starch, and amino acids, with and without intact  protein. Determination of free and/or total proteinogenic L‐α‐amino  acids and taurine

 SPIFAN matrices

1

Yes

All analytes defined in the  applicability statement are  measured.

Method is for total amino acids only. Tryptophan is  not included in the scope of this method.

1

The method covers a narrower range (from about  2 to 820 mg/100 g rec.prod., this varies for each  analyte), but almost all actual measurements fall  within this range. No sample value below lower  concentration of the calibration curve. See SLV report and appendices for details.

0.4–2500 mg/100g reconstituted product

Analytical Range.

1

All Analytes

N/A

Limit of detection (LOD)

1

All Analytes

0.4 mg/100 g reconstituted product

2.5 mg/100 g reconstituted product

This LoQ is in line with the actual analyte  concentrations measured across the SPIFAN II kit. See the SLV report and appendices for details.

Limit of quantification (LOQ)

2

0.5–5.0: 88‐112% 5.0‐150: 90‐110%                                                                150‐ 2500: 93‐107%

From Table 1, average spike recovery is 99%. See Table 1 as well as SLV report and appendices  for detailed results. From Table 2, 14 out of 17 analytes are within the  reference range. See Table 2 as well as SLV report and appendices  for detailed results. From Table 3, the average repeativility is 1.1%. See Table 3 as well as SLV report and appendices  for detailed results.

2

Spike recovery  (%)

Accuracy/Recovery

3

Bias vs SRM

same

0.5–5.0: ≤7% 5.0‐150: ≤5%                                                                    150‐ 2500: ≤ 3 %

All Analytes

Repeatability (RSDr)

3

From Table 3, the average intermediate  reproducibility is 5.0%. See Table 3 as well as SLV report and appendices  for detailed results.

All Analytes

Not in the SMPR

Intermediate Reproducibility  (RSDiR)

1

0.4–5.0: ≤15% >5.0: ≤10%

All Analytes

Not done in the SLV

Reproducibility (RSDR)

1

ADDITIONAL EVALUATION PARAMETERS

Adequate proof of performance via system suitability

N/A

Did Method Author Consider ERP’s Method Specific  Recommendations  (See web link to specific method  comments): Feedback from Users of the Method since being awarded  First Action Official Methods  Status

N/A

N/A

Bias against established method 

Is there a bias Yes/No ? N/A

Analytical equipment is commonly available in most labs. Waters Acquity systems were used for this SLV but  other equipement could in principle be used No unique proprietary equipment/accessories are required.  While the method is based on Waters AccQ‐Tag  technologiy, we propose alternative reagents that can be used.

Analytical equipment

Proprietary equipment

Method does not require any special safety precautions e.g. personal protection from highly toxic solvents.  Hydrolysis in hot hydrocholoric acid, neutralization with NaOH, acetonitrile for separation

Laboratory safety

The SLV report details the results for each analyte/product combination, according to the specific  requirements determined by the level of the analyte in the sample.

Other Considerations

0

Overall Score

Decision by ERP

Recommendation of ERP  2 years after First Action Status

move to Final Action/repeal/remove/expand 2 year term

Notes: a  Concentrations apply to (1) "ready to feed liquids" "as‐is"; (2) reconstituted powders (25 g into 200 g water); and (3) liquid concentrates diluted 1:1 by weight.

b    Units

SPIFAN ERP Checklist v 1.6

27.06.2013