Analytical HPLC Application

20070031
Antibiotics, Antiinflammatory in Meat - separation of
Flunixin, Tiamulin Hydrogen Fumarate, Toltrazuril,
Toltrazurilsulfon - using a Chromolith column with LC-MS
detection

Chromatography
Substance class :

Analgesics

Matrix :

Meat

Chromatographic conditions
Column :

Chromolith Performance RP 18-e 100x4,6 mm (Cat. No. 1.02129)

Precolumn :

Chromolith Guard CartridgeRP-18e 5x4,6mm (Cat. No. 151451)

Mobile Phase :

Mobile Phase TIA: Methanol (LiChrosolv Cat. No.1.06007)-0,1% acetic acid in

water (LiChrosolv Cat. No.1.15333) 55 45; FLU,TOL,TOLS: A: Acetonitrile
(LiChrosolv Cat. No.1.00030) B: 0,1 acetic acid in water (LiChrosolv Cat.
No.1.15333)

Gradient :

TIA: Isocratic; FLU,TOL,TOLS: 0min 65%B;4min 30%B; 7min 65%B;8min 65%B;

Flow rate :

TIA: 1,2mL/min; FLU,TOL,TOLS: 0-4min 1,2 mL/min; 4-7min 2mL/min; 7-8min

1,2mL/min

Detection :

MS

Temperature :

26C

Inject. volume :

TIA:20L;FLU,TOL,TOLS10L

Sample :

FLU: Flunixin; TIA:Tiamulin Hydrogen Fumarate; TOL: Toltrazuril; TOLS:

Toltrazurilsulfon

Sample preparation
dried extracts solved in 0,6mL 70% methanol, 30% 0,022M 1-heptane sulfonic acid sodium salt and 0,01M
di-sodium hydrogen phosphate; then 2 times washed with 0,5mL n-hexane (discard organic layer), filtered,
then injected

Merck KGaA, 64271 Darmstadt, Germany, Tel.

+49(0)6151 72-2440
EMD Millipore Corporation, 290 Concord Road,
Billerica, MA 01821, USA, +1-978-715-4321

2008-09-11

Page 1 of 2

Analytical HPLC Application

20070031
Figure

Source
Victor Hormazbel, Magne Yndestad, Oyvin OstensvikJournal of Liquid Chromatography & Related
TechnologiesVol. 26, No. 5,pp. 791-8012003

Ordering Information
Product

Catalog No.

Chromolith RP-18 endcapped 5-4.6 guard cartridges (3 pieces)

151451

Water for chromatography LiChrosolv

115333

Methanol gradient grade for liquid chromatography LiChrosolv Reag. Ph Eur

106007

Acetonitrile gradient grade for liquid chromatography LiChrosolv Reag. Ph Eur

100030

Chromolith Performance RP-18 endcapped 100-4.6 HPLC column

102129

Merck KGaA, 64271 Darmstadt, Germany, Tel.

+49(0)6151 72-2440
EMD Millipore Corporation, 290 Concord Road,
Billerica, MA 01821, USA, +1-978-715-4321

2008-09-11

Page 2 of 2