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Summary
A UHPLC method for determination of lutein and β-carotene in infant formula and adult
nutritionals was validated using the SPIFAN sample kit, including both unfortified and
fortified samples. The data provided show that the method meets the requirements of the
SMPR.
Linearity
Linearity of the calibration solutions correlates to approximately 0.8-45 μg/100 g
as fed for samples prepared for the lowest sample concentrations. With dilutions
specified in the method, the range can be extended to approximately 2250 μg/100
as fed. This extends beyond the range of 1-1300 μg/100 g specified in the SMPR.
Linearity of the internal standard was also demonstrated.
LOD/LOQ
The LOD for lutein and β-carotene is 0.08 μg/100 g. The LOQ for both lutein and
β-carotene (0.27) meet that of ≤1 μg/100 g specified in the SMPR.
Specificity
All experiments showed separation of all-
trans
-lutein and β-carotene from their
major
cis
isomers, apocarotenal, α-carotene, lycopene, and zeaxanthin. Samples
spiked with all-
trans
-lutein and β-carotene showed no isomerization during
sample preparation.
Precision
For all measurements in the range of 1-100 μg/100 g, repeatability was ≤5.8%
RSD for lutein and ≤5.1% RSD for β-carotene. For measurements >100 μg/100 g,
repeatability was ≤1.1% RSD for lutein and ≤1.7% RSD for β-carotene. The
method meets the precision requirements of the SMPR.
Accuracy
The SMPR calls for 90-110% recovery from spiked samples, and all data meets
this requirement.
VALIDATION REPORT
FOR ERP USE ONLY
DO NOT DISTRIBUTE