Nationwide sampling in the U.S. of whole large eggs, to update values in the United States Department of
Agriculture (USDA) National Nutrient Database for Standard Reference (SR) (http://www.ars.usda.gov/
nutrientdata), was conducted in 2000–2001 and in 2010. Retail cartons of large eggs were obtained from
12 supermarket locations using statistical sampling plans based on market share and census data.
Cholesterol was analyzed at three laboratories using standard methods involving gas chromatography of
the saponified total lipid extract. Vitamin D3 and 25-OH-vitamin D3 (2010 samples only) were analyzed
by HPLC and UHPLC–MS/MS. Quality control materials were included to validate the accuracy and
precision of measurements. The mean cholesterol content decreased 51 mg/100 g (12%; p < 0.0001),
from 423 mg/100 g in 2000–2001 to 372 (range 344–405) in 2010. Over the same period, average
vitamin D3 increased by 60%, to 2.05 mg [80 IU]/100 g (range 0.97–12.1). Samples from 2010 contained
0.65 mg 25-OH-D3/100 g (range 0.43–1.32). The disparate vitamin D (and cholesterol) content of eggs
sampled from different locations may reflect industry efforts to modify poultry feed or supplements to
affect the nutrient profile of eggs. Cholesterol and vitamin D3 data from this work were included in SR
release 23, and support food consumption surveys, food and nutrition policy, and consumer education.