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 D3 2000 IU D3 E 400 IU E 6 1 K1 100 µg K2 50 µg UPC 2 E A D3 K1 K2 1
ACQUITY UPC 2 ACQUITY UPC 2 A A+D3 D3 4 10 ml E 10 ml K 8 30 1.0 μm K2 30 1.0 μm PDA ACQUITY UPC 2 BEH 3.0 100 mm, 1.7 μm ACQUITY UPC 2 HSS C 18 SB 3.0 100 mm, 1.8 μm ACQUITY UPC 2 HSS C 18 SB 2.1 150 mm, 1.8 μm Empower3 A A A 2 1.325 1.394 1 2.0 2.5 ABPR 2176 psi CO 2 / 0.2% 97:3 90:10 3 2 2 2 A A 1.394 trans-vitamin A palmitate 0.40 0.30 0.20 0.10 cis-vitamin A palmitate 1.325 0.00 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 1 A 2 UPC 2
A/B ABPR ACQUITY UPC 2 HSS C 18 SB, 3.0 100 mm, 1.8 μm 2.0 ml/min 97 : 3 90 : 10 3 A CO 2 / B 0.2 % UV 320 nm 500 600 nm 1 μl 2176 psi 50 2 A 0.70 0.609 1.392 1.450 0.65 0.60 0.55 0.50 0.45 0.40 0.35 0.30 Vitamin A acetate Vitamin A palmitate Retinol 0.25 0.20 0.15 0.10 0.05 0.578 1.323 0.00-0.05 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 2 A A UPC 2 3
A+D3 A A D3 A 2.626 2.851 3 D3 6.862 3 D3 1.00 2.851 trans-vitamin A palmitate 0.024 0.75 0.018 0.012 0.50 0.006 cholecalciferol 6.862 0.000 0.25 4.20 4.40 4.60 4.80 5.00 5.20 5.40 5.60 5.80 6.00 6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40 8.60 8.80 9.00 0.00 cis-vitamin A palmitate 2.626 cholecalciferol 6.862 0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 3 A+D3 4 UPC 2
CO 2 / 0.2% 99:1 90:10 10 3 A/B ABPR ACQUITY UPC 2 HSS C 18 SB, 2.1 150 mm, 1.8 μm 1.0 ml/min 99 : 1 90 : 10 10 A CO 2 / B 0.2% UV 263 nm 500 600 nm 1 μl 2176 psi 50 3 A D3 D3 D3 3 A+D3 D36.867 0.048 0.036 0.024 0.012 cholecalciferol 6.867 0.000 0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 4 D3. UPC 2 5
E E 4 (d-, d-, d-, d- ) 90 5 CO 2 / 0.2% 98:2 95 5 1.5 4 α-tocopherol 0.489 1.04 0.78 γ-tocopherol 0.52 0.763 δ-tocopherol 0.26 β-tocopherol 0.695 0.893 0.00 0.00 0.12 0.24 0.36 0.48 0.60 0.72 0.84 0.96 1.08 1.20 5 E A/B ABPR ACQUITY UPC 2 BEH, 3.0 100 mm, 1.7 μm 2.5 ml/min 98 : 2 95 : 5 1.5 A CO 2 / B UV 293 nm 500 600 nm 1 μl 1885 psi 50 4 E 6 UPC 2
K1 K1 6 CO 2 / / = 1:1 = 99 : 1 RS>2.0 7 UV 246 nm 2 K 5 0.0104 trans-phylloquinone 0.0078 2.226 0.0052 0.0026 cis-phylloquinone 2.062 0.0000 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 6 K1 ACQUITY UPC 2 HSS C 18 SB, 2.1 150 mm, 1.8 μm 1.5 ml/min A / B = 99 : 1 A/B A CO 2 / B / UV 248 nm 300 400 nm 2 μl ABPR 1885 psi 50 5 K1 UPC 2 7
0.0014 0.0012 2.064 Peak 1 246.4 trans-phylloquinone 0.0010 0.0008 0.0006 0.0004 0.0002 0.0000 317.6 393.6 0.009 2.227 Peak 2 246.4 0.008 cis-phylloquinone 0.007 0.006 0.005 0.004 0.003 0.002 324.7 0.001 0.000 391.2 210.00 220.00 230.00 240.00 250.00 260.00 270.00 280.00 290.00 300.00 310.00 320.00 330.00 340.00 350.00 360.00 370.00 380.00 390.00 nm 7 2.064 2.227K UV 8 UPC 2
K2 K2 MK MK-3 MK-14 CO 2 / = 95 : 5 8 1 MK-7 MK-7 6 0.008 menaquinone-7 (MK7) 0.006 1.388 0.004 0.002 0.000 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 8 K2 A/B ABPR ACQUITY UPC 2 HSS C 18 SB, 3.0 100 mm, 1.8 μm 3.0 ml/min 95% A 5% B A CO 2 / B UV 248 nm 500 600 nm 1 μl 1885 psi 50 6 K2 UPC 2 9
ACQUITY UPC 2 6 A E K1 www.waters.co.jp 140-0001 1-3-12 5 TEL 03-3471-7191 FAX 03-3471-7118 532-0011 5-14-10 11F TEL 06-6304-8888 FAX 06-6300-1734 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. 2012 09 720004394JA PDF