Supplementary Material Digestibility of vitamin K (phylloquinone and menaquinones) in food and supplements Marie Bagge Jensena*, Anja Pia Biltoft-Jensenb, Jette Jakobsena a Research Group for Bioactives - Analysis and Application, National Food Institute, Technical University of Denmark, Kemitorvet, 2800 Kgs. Lyngby, Denmark b Research Group for Nutrition, Sustainability and Health Promotion, National Food Institute, Technical University of Denmark, Kemitorvet, 2800 Kgs. Lyngby, Denmark Marie Bagge Jensen, [email protected]. Table of Contents SM 1…………………………………...…………………………………………………………..1 SM2………………………………………………………………………………………………..2 SM 1: Four fractional factorial (23-1) experiments performed in order to validate the INFOGEST 2.0 model for assessment of bioaccessibility of vitamin K vitamers. All samples were digested in triplicates (n=3) and analysed for vitamin K vitamers using the LC-ESI-MS/MS method (Jensen et al., 2021b). Factor Experiment 1 0.8 g Raw spinach (A)/ 0.8 g Frozen boiled spinach (a) 0.2 g Oil (B)/ No oil (b) Bile (C)/ 2 x bile (c) Bioaccessibility (%) Result Sample 1 A Sample 2 Sample Sample 3 4 p-value < A a a 0.05 B b B b p-value < 0.05 C c c C p-value < 0.05 D E F G Sample 1 A Sample 2 Sample Sample 3 4 p-value > A a a Factor Experiment 2 100 ng PK, MK-4, MK-7, MK-9 (A)/ 200 ng PK, MK-4, MK-7, MK-9 (a) 100 ng vitamin D3 (B)/ No vitamin D3 (b) Bile (C)/ 2 x bile (c) Result Result 0.05 B b B b p-value > 0.05 C c c C p-value > 0.05 D E F G Factor Experiment 3 0.8 g standard meal (A)/ 0.4 g standard meal (a) 0.1 g raw spinach (B)/ 0.2 g raw spinach (b) Bile (C)/ 2 x bile (c) Result Result Sample 1 A Sample 2 Sample Sample 3 4 p-value < A a a 0.05 B b B b p-value > 0.05 C c c C p-value > 0.05 D E F G Factor Experiment 4 0.8 g standard meal (A)/ 0.6 g standard meal (a) 500 ng PK, MK-4, MK-7, MK-9 (B)/ 1000 ng PK, MK-4, MK-7, MK-9 (b) 500 ng vitamin D3 (C)/ 1000 ng vitamin D3 (c) Result Result Sample 1 A Sample 2 Sample Sample 3 4 p-value < A a a 0.05 B b B b p-value > 0.05 C c c C p-value > 0.05 D E F G 1 SM2: Chromatogram of vitamin K vitamers and internal standards in a natto sample. The sample was analysed for PK, MK-4, MK-5, MK-6, MK-7, MK-9, MK-10, d7-PK, d7-MK-4, d7-MK-7 and d7-MK-9. Only the highest peaks are indicated by arrows and labels. 2
