ACQUITY UPC 2 ACQUITY UPC 2 A A+D3 D ml E 10 ml K μm K μm PDA ACQUITY UPC 2 BEH mm, 1.7 μm ACQUITY UPC 2 HSS C 18 SB 3.

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1 UPC 2 Andrew Aubin Waters Corporation, Milford, MA, USA Waters ACQUITY UPC 2TM 6 tert- UltraPerformance Convergence Chromatography TM UPC 2 ACQUITY UPC 2 1 A A D3 E D3 K1 K2 UPC 2 A E K1 ACQUITY UPC 2 ACQUITY UPC 2 ACQUITY UPC 2 PDA Empowe TM 3 1 / A 10,000 IU A A+D3 10,000 IU A 2000 IU D3 D IU D3 E 400 IU E 6 1 K1 100 µg K2 50 µg UPC 2 E A D3 K1 K2 1

2 ACQUITY UPC 2 ACQUITY UPC 2 A A+D3 D ml E 10 ml K μm K μm PDA ACQUITY UPC 2 BEH mm, 1.7 μm ACQUITY UPC 2 HSS C 18 SB mm, 1.8 μm ACQUITY UPC 2 HSS C 18 SB mm, 1.8 μm Empower3 A A A ABPR 2176 psi CO 2 / 0.2% 97:3 90: A A trans-vitamin A palmitate cis-vitamin A palmitate A 2 UPC 2

3 A/B ABPR ACQUITY UPC 2 HSS C 18 SB, mm, 1.8 μm 2.0 ml/min 97 : 3 90 : 10 3 A CO 2 / B 0.2 % UV 320 nm nm 1 μl 2176 psi 50 2 A Vitamin A acetate Vitamin A palmitate Retinol A A UPC 2 3

4 A+D3 A A D3 A D D trans-vitamin A palmitate cholecalciferol cis-vitamin A palmitate cholecalciferol A+D3 4 UPC 2

5 CO 2 / 0.2% 99:1 90: A/B ABPR ACQUITY UPC 2 HSS C 18 SB, mm, 1.8 μm 1.0 ml/min 99 : 1 90 : A CO 2 / B 0.2% UV 263 nm nm 1 μl 2176 psi 50 3 A D3 D3 D3 3 A+D3 D cholecalciferol D3. UPC 2 5

6 E E 4 (d-, d-, d-, d- ) 90 5 CO 2 / 0.2% 98: α-tocopherol γ-tocopherol δ-tocopherol 0.26 β-tocopherol E A/B ABPR ACQUITY UPC 2 BEH, mm, 1.7 μm 2.5 ml/min 98 : 2 95 : A CO 2 / B UV 293 nm nm 1 μl 1885 psi 50 4 E 6 UPC 2

7 K1 K1 6 CO 2 / / = 1:1 = 99 : 1 RS>2.0 7 UV 246 nm 2 K trans-phylloquinone cis-phylloquinone K1 ACQUITY UPC 2 HSS C 18 SB, mm, 1.8 μm 1.5 ml/min A / B = 99 : 1 A/B A CO 2 / B / UV 248 nm nm 2 μl ABPR 1885 psi 50 5 K1 UPC 2 7

8 Peak trans-phylloquinone Peak cis-phylloquinone nm K UV 8 UPC 2

9 K2 K2 MK MK-3 MK-14 CO 2 / = 95 : MK-7 MK menaquinone-7 (MK7) K2 A/B ABPR ACQUITY UPC 2 HSS C 18 SB, mm, 1.8 μm 3.0 ml/min 95% A 5% B A CO 2 / B UV 248 nm nm 1 μl 1885 psi 50 6 K2 UPC 2 9

10 ACQUITY UPC 2 6 A E K TEL FAX F TEL FAX Waters Waters Corporation ACQUITY UPC 2 UltraPerformance Convergence Chromatography UPC 2 Empower The Science of What s Possible Waters Corporation 2012 Waters Corporation. Printed in Japan JA PDF

LC ACQUITY UPLC H-Class Bio TUV Sigma, mg/ml 0.01 N 276 nm μl 0.4 ml/min L g/l / 99% / 65/15/20 v/v/v 10% 10% 0.01 N ACQUITY

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