1 Agilent 12 Infinity 1 Siji Joseph Agilent Technologies, Inc. Bangalore, India 12 1 2 4 5 3 2 1 4 1 12 14 4 2 Agilent 12 Infinity 2 4 5 3 2.5 1. 1.5 2. 2.5 3. 2 (RP-HPLC) Agilent Poroshell EC-C1 Agilent 12 Infinity LC Agilent 12 Infinity (DAD) 2 D Agilent 12 Infinity LC (UHPLC)
2 (FDA) 1 1 A D E K 1 LC UV 15 HPLC 3.5 UHPLC Agilent 12 Infinity LC Agilent 12 Infinity (G1311B) Agilent 12 Infinity (G13E) Agilent 12 Infinity (G131C) Agilent 12 Infinity (G4212B) mm Max-Light Agilent 12 Infinity LC UHPLC Agilent 12 Infinity (G422 A) 1 µl Jet Weaver Agilent 12 Infinity (G422A) Agilent 12 Infinity (G131C) Agilent 12 Infinity (G4212A) 1 mm Max- Light : Agilent Poroshell 12 EC-C1 2.1 mm 5 mm 2. µm (5-2) Agilent Poroshell 12 EC-C1 3. x 15 mm 2. µm (35-32) : Agilent ChemStation B.4.2 HPLC Milli Q (Millipore Elix 1 Millipore ) Lab- Scan () (THF) Sigma ( ) Fluka ( ) (K3) (- ) ( A ) ( A ) -cis ( A ) K2 (D3) ( E ) K1 Aldrich ( ) 2
UHPLC 1 5 mg 25 ml K1 K2 D3 B 2. mg/ml (2, ppm) + 4. ºC 1 µl 2 ppm 1, µl B 2 ppm 5 pg/µl 1 ng/µl (1 ) 2 D 45 5 g D 2 1:1 1 ml D Agilent 12 Infinity LC : 45 ºC 45 ºC : 2 Hz Hz Agilent 12 Infinity LC : 21 24 2 32 35 3 nm 21 24 2 32 35 3 nm : mm 1 mm : 5 µl ( 5 ) : 5 ºC 5 ºC 1 µl ( 3 ) A : 5:5 : THF +.5 % 5:5 : THF +.5 % B : 5:25:5 : : THF +.35 % : 3 % B 3 5 % B 1 % B 15 1 % B 15.1 3 % B : 5 1 5:25:5 : : THF +.35 % 5 % B. 5 % B 1. 1 % B 3.4 1 % B 3.5 5 % B :. ml/min. ml/min 1 Agilent 12 Infinity LC Agilent 12 Infinity LC 1. ml 2 µl B HPLC / 25 ng D 1 4 ºC 4 ºC D 5 µl B (RT) (SD) (RSD) LOD LOQ D 3
5 g 1 25 ng D / UHPLC LOD LOQ RT RSD UHPLC Agilent Poroshell 12 EC-C1 (3. mm x 15 mm 2. µm) 13 1 2 21 nm ChemStation 12 4 DAD1A, Sig = 21 DAD1D, Sig = 32 1 2.55 > 1..2.3 > 2..3 4.3 2 3.1.2 5.55 2 4 1 12 14 1.34 1.55 1 15 cm Agilent Poroshell 12 EC-C1 3 A Sl RT 1 K3 25 2.5 2 - < 21.1 3 A 32.2 4 A 35.3 5 -cis A 3.55 K2 K2 24 1.34 11.55 12.3 D3 2 1.55 E < 22 11.55 K1 K1 24 12.3 2 4
(LOD) (LOQ) S/N 3 LOD S/N 1 LOQ LOD LOQ 3 ( D) LOD LOQ 2 LOQ 3 3 4.4 4.35 4.3 4.25 4.2 4.15 4.1 1.5 LOQ LOD DAD1 C, Sig = 2 LEVEL 5 PG_LOQ DAD1 C, Sig = 2 LEVEL 25 PG_LOD DAD1 C, Sig = 2, BLANK 1.25 1.5 1.5 11 11.25 11.5 2 ( D) LOD (25 pg) LOQ (5 pg) Sl LOD (ng) LOQ (ng) (ng) R 2 = 1.25.5.5 25. 1 y = 33.12 x +.431423 2 1.5 2.5 2.5 5. y =1.122 x +.1412 3.5 1.5 1.5 5. y =.13132 x + 1.22412 4.25.5.5 25. 1 y = 4.3534 x +.32 5.25.5.5 25.5 1 y = 42.135 x + 5.452 K2.25.5.5 5. 11 y = 15.2555 x 1.25143.25.5.5 5 1. 11 y = 1.24 x 5.24 5 25 25 5. 4 y =.413 x 22.22 K1.5.15.15 5. 1 y = 14.24 x 1.324 3 LOD LOQ 5
(RT) 5 ng (RT) (RSD) RSD.35 % RT RSD.4 % RT RSD 4 3 % 5 % 4 RT K2 K2 4 2 5432 = 1.24*Amt 5.24 : 1. 2 4 [ng] 3.5 5 ng () RSD.4.35.3.25.2.15.1.5. RT RSD RSD RT RSD K2 K1 4 5 ng RT RSD
Sl () RT ( 3. %) ( 5. %) 1 (.) 2 %.2 ml/min. ml/min 2 (5 µl) 2 % 5.1 µl 3 (5 5 5 1) 4. µl 1 % 4.5 (5.5 1) 5 ( 4 ) 5.5 (5 2.5 1) 5 ( 4 ) 4 (ACN 5 %) 5 % % (:15:5) % (:25:5) 5 (45 ºC) 5 % 4.2 ºC (21 24 2 32 35 3 nm) 42. ºC 3 nm (21 24 2 32 35 3 nm) (213 243 23 323 353 33 nm) 5 4 D D 3 25 ng/µl D D 4 % 2 % D 5 2 1 2 1 2 1 DAD1 C Sig = 2 STD 1.5 1 11 12 13 14 DAD1 C Sig = 2 1.52 1 11 12 13 14 DAD1 C Sig = 2 1.52 : 422.2 : 12. : 4.2 1 11 12 13 14 5
D KOH D.3 µg/1 g 1 µg/. µg/1 g 12. µg/ 1 12 1 4 2 2 1 Agilent 12 Infinity LC 2 3 4 5 2 1 4 1 12 14 Agilent 12 Infinity LC 2 4 5 3.5 1. 1.5 2. 2.5 3. 1. 2. 3. 4. 5. cis. K2... K1 UHPLC UHPLC UHPLC 15 HPLC % ( ) ( ) ( ) A LOD LOQ UHPLC LOD LOQ 5 D 1 µ 1 ppm RT RSD RSD 1.25 % RT RSD.12 % Agilent 12 infinity HPLC 12 Infinity LC UHPLC Sl LOD (ng) LOQ (ng) RT RSD UHPLC (ng) R 2 1.25.5.5 2.5 1 2.5 1 1 5. 3.25.5.5 2. 1 4.5.1.1 2 1. 5 K2.5.1.1 5. 1.5.1.1 5. 1.5 1 1 5. K1.1.25.25 5. 5 Agilent 12 Infinity LC UHPLC LOD LOQ =
54231 Agilent Poroshell 12 EC-C1 Agilent 12 Infinity LC 15 HPLC A D E K D % Agilent 12 Infinity LC 3 UHPLC Agilent 12 Infinity LC Agilent 12 Infinity LC 1 12 4 = 3.221*Amt -.4525 :. 2 4 [ng]..5 5 ng D ( ) 1 µl % 1.4 1.2 1...4.2 RSD (1 ng ) RSD RT RSD K2 K1 1 ppm UHPLC RT RSD 1 µl
1. Analysis of water-soluble vitamins from multivitamin tablets for nutrition labeling, Agilent Application Note, publication number: 5-5EN, 211. 2. Leo M.L. Nollet, Food analysis by HPLC, Second edition revised and expanded, Chapter:, Page: 33-342. 3. Duncan Thorburn Burns, Klaus Danzer, and Alan Townshend, Use of the terms recovery and apparent recovery in analytical procedures, Pure Appl. Chem., Vol. 4, No. 11, pp. 221 225, 22. 1
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www.agilent.com/chem/jp Agilent Technologies, Inc., 211 Published in Japan, September 1, 211 5-JAJP