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Method Abstract 374

MA 374 / ANALYSIS OF AMINO ACIDS IN OXIDIZED AND UNOXIDIZED FEED SAMPLES

Commission Regulation (EC) No 152-2009 published in Official Journal of European Union laid down the methods of sampling and analysis for the official control of feed. The Regulation describes methods of analysis to control the composition of feed materials and compound feed products. Establishing the Amino Acids profile is an important way to control quality and nutritional value of feeds. This regulation specifies HPLC with post-column derivatization with Ninhydrin reagent as the method of analysis for total and free amino acids. Pickering Laboratories developed the analytical method to comply with all the chromatographic requirements of Commission Regulation (EC) No 152-2009. The same method is used to analyze oxidized and unoxidized feed samples.

METHOD

Analytical conditions

Column: High-efficiency Sodium cation-exchange column, 4.0 x 150 mm, Catalog Number 1154150T
Guard: Cation-exchange GARD™, Catalog Number 1700-3102
Flow Rate: 0.4 mL/min
Mobile Phase: Na270, Na740, RG011. See method in Table 1
Injection Volume: 10 uL

 

Post-Column Conditions

Post-column System: Pinnacle PCX 
Reactor Volume: 0.5 mL
Reagent: Trione®
Column Temperature: See Method in Table 2
Reactor Temperature: 130 °C
Flow Rate: 0.25 mL/min
Detection: UV/VIS 570 nm for primary amino acids,440 nm for secondary amino acids

TABLE 1. HPLC PROGRAM
 

TIME, MIN   

NA270, %

NA740, % RG011, %

0

100 0 0

16

100 0 0

40

54 46 0

45

0 100 0

66

0 100 0
66.1 0 0 100
70 0 0 100
70.1 100 0 0
80 100 0 0

 
TABLE 2. COLUMN OVEN PROGRAM
 

TIME   

TEMP, ºC

0

55

32

55

33

65

41

65

42

55


 

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