Amino-01 (July 2018)

Single Laboratory Validation Report for Total Amino Acids by UHPLC-UV in Infant Formulas and Adult Nutritionals

S A CA C is

×

=

s

s

×

is

Note: C s

is determined automatically when Empower is used to create the calibration curves.

:

Concentration of individual amino acid in the test sample solution in pmol/μL

C s A s C is A is

:

Peak area of individual amino acid in the test sample solution Concentration of internal standard injected in pmol/µL

:

:

Peak area of internal standard chromatogram

S :

Slope of the calibration curve

(all curves are forced through zero, equation: y=ax)

Calculate the mass fraction, w, of each amino acid, in milligram per 100 grams of product, using the following equation:

× × × d d V MW C w

×

=

1

2

s

AA

s

10

m

×

S

:

Molecular weight of individual amino acids in g/mol (see below)

MW AA

V s d 1 d 2 m s : : 10: : :

Volume of hydrolysis solution in mL (typically 5 mL) Dilution factor in the neutralization step (4) Dilution factor in the derivatization step (10)

mass of the test portion in mg

Combined factor to convert pg to mg (10 -9 ), mL to µL (10 3 ) and mg to 100 g (1/10 - 5 )

Note: Empower can be configured to calculate the mass fraction w

Molecular Weights ( MW AA

) of Amino Acids (g/mol)

Aspartic acid:

133.11 119.12 105.09 147.13 115.13

Methionine: Isoleucine:

149.21 131.18 131.18 181.19 165.19 146.19 155.16 174.20 125.15

Threonine:

Serine:

Leucine: Tyrosine:

Glutamic acid:

Proline: Glycine: Alanine: Cystine: Valine:

Phenylalanine:

75.07 89.10

Lysine:

Histidine: Arginine: Taurine:

240.30 117.15

Jaudzems, Guthrie, Lahrichi, and Fuerer – January 2018

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