Library

Method Abstract 393

MA 393 / ANALYSIS OF AMINO ACIDS SUPPLEMENTS AND SPORTS DRINKS


Amino acids are building blocks of proteins and essential amino acids need to be obtain from diet as they cannot be synthesized by our
bodies. Amino acids supplements are popular among athletes and bodybuilders and are sold in the form of pills, powders and drinks.
They usually also contain vitamins, electrolytes, herbs, flavorings and other additives.

Analysis of these supplements is important to confirm the label claims and to maintain industry standards.

Amino Acids Analysis with post-column derivatization is a very sensitive, reproducible and rugged method and it has been a preferred
approach for laboratories running biological samples, protein, peptides and foods analysis. Pickering Laboratories Inc. products for
Amino Acid Analysis include post-column derivatization instruments, columns, eluants, reagents and standards. All products are
designed to work together to deliver optimum results for any chosen sample.

The amino acids present in the product determine if Sodium or Lithium columns and buffers need to be used. Lithium columns have
longer run time but have higher selectivity and are able to separate more amino acids than Sodium columns. Pickering Laboratories
will help you to identify the best method for your Amino Acid Analysis.


Sample Preparation

Grind 10 pills to fine powder before taking a sample for analysis. Mix all the powdered supplements thoroughly. Sample size depends
on concentrations of amino acids present.

To finely ground sample (0.05 g – 0.2 g) add 100 mL of Na220. Mix well and sonicate for 10 min. Filter through 0.45 um filter
and inject.
 

METHOD

Analytical Conditions

Column System: High-efficiency Sodium cation-exchange column,
      4.6 x 110 mm, Catalog Number 1154110T
Flow rate: 0.6 mL/min
Mobile Phase: See method in Table 1

Post-column Conditions

Post-Column System: Pinnacle PCX
Reactor Volume: 0.5 mL
Reactor Temperature: 130 ºC
Column Temperature: See method in Table 2
Flow Rate: 0.3 mL/min
Injection Volume: 10-50 uL


 

 

TABLE 1. HPLC PROGRAM FOR COLMN 1154110T

TIME

NA315%

NA425%

NA640%

RG011%

0.0

100

0

0

4.0

100

0

0

15.0

0

100 

0

0

16.0

0

100

0

31.0

0

100

0

31.1 0 0 0 100
33.0 0 0 0 100
33.1 100 0 0 0
40.0 100 0 0 0

 

TABLE 2. COLUMN OVEN PROGRAM

TIME

TEMP ºC

0

46

4

46

9

70

32

70

33

46

393-a
393-b
 
393-c
 
393-d
 
 

METHOD FOR LITHIUM COLUMN

Analytical Conditions

Column: High-efficiency Lithium cation-exchange column,
     4.6 x 75 mm, Catalog Number 0354675T
Flow rate: .55 mL/min
Mobile Phase: See method in Table 3
Injection Volume: 50 uL

Post-column Conditions

Post-Column System: Pinnacle PCX
Reactor Volume: 0.5 mL
Reagent: Trione®
Reactor Temperature: 130 ºC
Column Temperature: See method in Table 4
Flow Rate: 0.3 mL/min
Detection: UV/VIS 570 nm for primary amino acids,
     440 nm for secondary amino acids


 

  

 

TABLE 3. HPLC PROGRAM FOR COLMN 0354675T

TIME

NA315%

NA425%

NA640%

RG011%

0

100

0

0

15

100

0

0

35

40

60 

0

0

38

0

100 

0

0

43

0

100 

0

0

43.1 0 0 100 0
57.0 0 0 100 0
57.1 0 0 0 100
60.0 0 0 0 100
60.1 100 0 0 0
72 100 0 0 0

 

TABLE 2. COLUMN OVEN PROGRAM

TIME

TEMP ºC

0

34

6

34

30

45

32

70

59

70

60

34

393-e

 

393-f