OMB Meeting Book_9-11-14

Evaluation of Method Performance vs. SMPR requirements.

AOAC SMPR: 2011.005

2011.10 Determination of vitamin B12 in infant formula and adult nutritionals by HPLC

Method Reference # Method title:

Vitamin B12 is extracted from the sample using sodium acetate buffer (pH 4.5) and potassium cyanide at 105°C. Extracts are purifi ed and concentrated with C8 or C18 solid-phase extraction (SPE) cartridges and analyzed with size-exclusion and reversedphase chromatography. Determination of B12 is made by liquid chromatography with visible detection at 550 nm.

Principle of the method:

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

Method Performance

Please report

SMPR Requirement

Parameter

in units as stated in SMPR!

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. cyanocobalamin, aquocobalamin, hydroxycobalamin, methylcobalamin, and adenosylcobalamin

SPIFAN matrices

All SPIFAN matrices analyzed during SLV.

All required analytes are measured.

Method measures all required analytes.

0.01-5.0 mcg/100g reconstituted final product

0.25-2.5 mcg/100 mL range of standard curve > 0.025 mcg/100g reconstituted final product

Analytical Range.

vitamin B12

~0.025 mcg/100g reconstitued final product

 0.004 mcg/100g reconstituted final product

Limit of detection (LOD)

vitamin B12

~0.025 mcg/100g reconstitued final product

 0.01 mcg/100g reconstituted final product

Limit of detection (LOQ)

Spike recovery

90-110%

See Table 1 See Table 2 See Table 3

Accuracy/Recovery

Bias vs SRM

0.01 mcg/100g reconstituted final product :  15% 0.2, 0.5, 5.0 mcg/100g reconstituted final product :  15%

Repeatability (RSDr)

No SMRP requirements

Intermediate Reproducibility (RSDiR)

0.3,0.6,1.0,2.5,5.0 mcg/100g in reconstituted final product :  11% See Table 4

Reproducibility (RSDR)

Peak responses of precision injections of the high working standard must have an RSD of ≤ 2.0%. At each working standard concentration, the peak areas of standards injected at the beginning and end of a set of samples should not increase or decrease by more than 10% . A standard curve must have a correlation of at least 0.999 to be considered acceptable for sample calculations. For each set of samples, the control result must be within 3 standard deviations of the control means.

Adequate proof of performance via system suitability

Feedback from Users of the Method since being awarded First Action Official Methods Status

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Did Method Author Consider ERP’s Method Specific Recommendations (See web link to specific method comments):

Analytical equipment is commonly available in most labs.

Analytical equipment

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 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 1.0