AOAC SMPR 2014.001 Table 1.
Method performance requirements:
trans-vitamin K1a
Analytical range 1–100b
Standard Method Performance Requirements for Limit of quantitation (LOQ) ≤1b
Vitamin K in Infant Formula and Adult/Pediatric
Nutritional Formula Repeatability (RSDr) 1–10 b
≤8%
>10b ≤5%
Recovery 90 to 110% of mean spiked
Intended Use: Reference Method for Dispute Resolution recovery over the range
of the assay
1 Applicability Reproducibility (RSDR) 1–10b ≤15%
Determination of trans-K1 and cis-K1 (phylloquinone) in all >10 b
≤10%
forms of infant, adult, and/or pediatric formula (powders, ready-to- a
Concentrations apply to (a) “ready-to-feed” liquids “as is”;
feed liquids, and liquid concentrates). (CAS 84-80-0) (b) reconstituted powders (25 g into 200 g of water); and (c) liquid
concentrates diluted 1:1 by weight.
2 Analytical Technique b
mcg/100 g reconstituted final product.
Any analytical technique that meets the following method
performance requirements is acceptable.
3 Definitions 2ʹ,3ʹ-trans-phylloquinone; phytonadione is a mixture of cis- and
Adult/pediatric formula.—Nutritionally complete, specially trans-phylloquinone). See Figure 1 for chemical structure of trans-
formulated food, consumed in liquid form, which may constitute vitamin K1.
the sole source of nourishment [AOAC Stakeholder Panel on Infant
4 Method Performance Requirements
Formula and Adult Nutritionals (SPIFAN); 2010], made from any
combination of milk, soy, rice, whey, hydrolyzed protein, starch, See Table 1.
and amino acids, with and without intact protein.
Infant formula.—Breast-milk substitute specially manufactured 5 System Suitability Tests and/or Analytical Quality Control
to satisfy, by itself, the nutritional requirements of infants during
the first months of life up to the introduction of appropriate Suitable methods will include blank check samples, and check
complementary feeding (Codex Standard 72-1981) made from any standards at the lowest point and midrange point of the analytical
combination of milk, soy, rice, whey, hydrolyzed protein, starch, range. Method should be able to accurately separate trans-
and amino acids, with and without intact protein.
Limit of detection (LOD).—The minimum concentration or mass phylloquinone from cis-phylloquinone.
of analyte that can be detected in a given matrix with no greater 6 Reference Material(s)
than 5% false-positive risk and 5% false-negative risk.
Limit of quantitation (LOQ).—The minimum concentration National Institute of Standards and Technology (NIST) Standard
or mass of analyte in a given matrix that can be reported as a Reference Material® (SRM) 1849a Infant/Adult Nutritional
quantitative result.
Repeatability.—Variation arising when all efforts are made Formula or equivalent. The SRM is a milk-based, hybrid infant/
to keep conditions constant by using the same instrument and adult nutritional powder prepared by a manufacturer of infant
operator, and repeating during a short time period. Expressed as the formula and adult nutritional products. A unit of SRM 1849a
repeatability standard deviation (SDr); or % repeatability relative
standard deviation (%RSDr). consists of 10 packets, each containing approximately 10 g of
Reproducibility.—The standard deviation or relative standard material. Certified value of NIST 1849a is 1.06 ± 0.17 mg/kg total
deviation calculated from among-laboratory data. Expressed vitamin K1 (cis + trans).
as the reproducibility relative standard deviation (SDR); or %
reproducibility relative standard deviation (%RSDR). Relevant reference material(s) with a known value of tran-
Recovery.—The fraction or percentage of spiked analyte that is phylloquine and cis-phylloquinone will be very helpful in the
recovered when the test sample is analyzed using the entire method. method verification but currently not available.
Vitamin K1.—CAS 84-80-0. 2-methyl-3-phytyl-1,4-
naphthoquinone (vitamin K1 produced in nature by plants is 7 Validation Guidance
Recommended level of validation: Official Methods of
AnalysisSM.
8 Maximum Time-to-Result
No maximum time.
Approved by AOAC Stakeholder Panel on Infant Formula and Adult
Figure 1. Chemical structure of trans-vitamin K1. Nutritionals (SPIFAN). Final Version Date: March 18, 2014.
© 2014 AOAC INTERNATIONAL
