AOAC SPIFAN ERP Meeting Book-March 16, 2016

VitK-01 MEF FOR ERP USE ONLY DO NOT DISTRIBUTE

Evaluation of Method Performance  vs. SMPR requirements.

AOAC SMPR: ………….. AOAC SMPR 2014.001

VITK‐01

Method title: Principle of the method: Method Reference #

A LC‐MS/MS Method for Vitamin K Analysis in Infant Formula and Adult Nutrition Samples

Fat in sample is hydrolyzed by lipase after adding internal std. Vitamin K is extracted by a solvent extraction. The reconstituted extract is analyzed by LC‐MS/MS for vitamin K.

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.

1

 SPIFAN matrices

The method is applicable to all the matrices listed  in the SMPR.

All analytes defined in the  applicability statement are  measured.

1

Analytical Range.

1

Analyte 1 Analyte 2 Analyte 3 Analyte 4 Analyte 5 Analyte 1 Analyte 2 Analyte 3 Analyte 4 Analyte 5

trans‐vitamin K1 = 0.5‐100 mcg/100g  cis‐vitamin K1 = 0.1‐100 mcg/100g  Total‐vitamin K1 = 0.6‐100 mcg/100g 

trans‐vitamin K1 = 1‐100 mcg/100g recostituted sample cis‐vitamin K1 = 1‐100 mcg/100g recostituted sample Total‐vitamin K1 = 1‐100 mcg/100g recostituted sample

Limit of detection (LOD)

1

trans‐vitamin K1 = <1 mcg/100g recostituted sample cis‐vitamin K1 = <1mcg/100g recostituted sample Total‐vitamin K1 = <1mcg/100g recostituted sample

trans‐vitamin K1 = 0.5 mcg/100g 

cis‐vitamin K1 = 0.1/100g 

Limit of quantification (LOQ)

2

Total‐vitamin K1 = 0.6mcg/100g 

2

Spike recovery  (%) Vitamin K1 (cis + trans) = 90‐100%

Vitamin K1 (cis + trans) = 97.8 + 5.2% (n=44)

Accuracy/Recovery

NIST SRM 1849a Vitamin K1 = 1.00 + 0.10 mg/Kg  (n=5)

3

Bias vs SRM NIST SRM 1849a Vitamin K1 = 1.06 + 0.17 mg/Kg

<8% RSD <5% RSD

Repeatability (RSDr)

3

7.90% 6.30%

1‐10 mcg/100g >10 mcg/100g

<8% RSD <5% RSD

Intermediate Reproducibility  (RSDiR)

1

Reproducibility (RSDR)

1

ADDITIONAL EVALUATION PARAMETERS

Blank check and lowest and midrange calibration

Adequate proof of performance via system suitability

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

Bias against established method 

Is there a bias Yes/No ?

Analytical equipment

Analytical equipment is commonly available in most labs.

Proprietary equipment

No unique proprietary equipment/accessories are required.

Method does not require any special safety precautions e.g. personal protection from highly toxic solvents.

Laboratory safety

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