6. AOACSPIFANMethods-2018Awards

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S CHIMPF ET AL . : J OURNAL OF AOAC I NTERNATIONAL V OL . 101, N O . 1, 2018 275

Figure 9. Chromatographic separation of carotenoids in an adult nutritional ready-to-feed nutritional (00729RF00).

Figure 10. Chromatographic separation of carotenoids in an adult nutritional ready-to-feed nutritional (00730RF00).

lutein curve; D 1

= first dilution volume in milliliters (10 mL = second dilution volume in milliliters

method, nutritionals are saponified and carotenoids are extracted with hexane and MtBE. A portion of the extract is concentrated and redissolved in 75 + 25 of 10% BHT in methanol – MtBE and analyzed with C30 chromatography and UV-Vis spectroscopy. During a single-laboratory validation of this method, most of the SPIFAN SMPRs were met. Repeatability for all three carotenoids ranged from 1.81 to 14.9%, and recoveries ranged from 89.3 to 109%. The method demonstrated good linearity for all three carotenoids and met LOQs. (1) Rodriguez-Amaya, D.B. (2001) A Guide to Carotenoid Analysis in Foods, OMNI Research , ILSI Human Nutrition Institute, Washington, DC (2) Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Gaithersburg, MD, Method 938.04 , http://www.eoma.aoac.org (3) Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Gaithersburg, MD, Method 2005.07 , http://www.eoma.aoac.org (4) Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Gaithersburg, MD, Method 2009.04 , http://www.eoma.aoac.org (5) Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Gaithersburg, MD, Method 2016.13 , http://www.eoma.aoac.org (6) Lombeida, W.O., Rubio, F., & Levy, L.W. (2016) J. AOAC Int. 99 , 1459 – 1469. doi:10.5740/jaoacint.16-0186 References

extraction solution); D 2

= total reconstitution weight in

(0.25 mL dilution solution); R TW

grams, if applicable; A 1

= aliquot of the first dilution pipetted; SW = = weight of powder reconstituted in

sample weight in grams; R PW

grams, if applicable; and 10 = conversion to 100 g. ( 3 ) cis- and trans- b -carotene and lycopene sample concentration. — C cis ; trans - BC ; Lyc = ð C DS × D 1 × D 2 × R TW Þ = ð A 1 × SW × R PW × 10 Þ where C cis,trans- BC,Lyc = cis- or trans - b -carotene or lycopene concentration in µg/100 g; C DS = cis- or trans - b -carotene or lycopene concentration extrapolated from the calibration curve in µg/L; D 1 = first dilution volume in milliliters (10 mL extraction solution); D 2 = second dilution volume in milliliters (0.25 mL dilution solution); R TW = total reconstitution weight in grams, if applicable; A 1 = aliquot of the first dilution pipetted; SW = sample weight in grams; R PW = weight of powder reconstituted in grams, if applicable; and 10 = conversion to 100 g. ( 4 ) Total lutein, b -carotene, or lycopene concentration (µg/100 g) = concentration of each cis isomer (µg/100 g) + concentration of trans isomer (µg/100 g).

Conclusions

A new reversed-phase HPLC method was developed to determine cis and trans isomers of lutein, b -carotene, and lycopene in infant, pediatric, and adult nutritionals. With this