Amino-01 (July 2018)

Amino-01 (JULY 2018) 180124 AA SPIFAN SLV Appendix J

FOR ERP USE ONLY DO NOT DISTRIBUTE

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

ERP request: Include system suitability step System Suitability

A working standard at 5pmol/ mL was injected 5 times on a UPLC / PDA systems used for this study. Working at concentration of 5.0 pmol/mL, was injected 5 times and the SR calculated using both the Area and Area/IS response ratio. Per USP 30 <621>, S R < 2 %. Suitability for all 18 amino acids was less than 1%.

PDA1 Amino acid stand 5pmol Suit for 5 injections Response: # Suitability Summary Table Suitability Component Summary For Response # SampleNaHis Tau Ser

Pro

Lys

Tyr

xCys

Met

Val

NVa

Ile

Leu

Thr

Ala

Asp

Glu

Gly

Arg

1 Suit mix 2 Suit mix 3 Suit mix 4 Suit mix 5 Suit mix

0.7009 0.7117 0.6968 0.6794 0.6886 0.6944 0.6884 0.7092 0.7226 0.6546 0.8853 0.7135 0.7013 0.7139 0.6985 0.6802 0.6863 0.6933 0.6869 0.7095 0.7205 0.6518 0.8759 0.7121 0.7021 0.7119 0.6997 0.6819 0.6903 0.6941 0.6874 0.7087 0.7192 0.6542 0.8704 0.7125 0.7023 0.7125 0.6992 0.6821 0.689 0.6933 0.6882 0.7089 0.7186 0.6542 0.8687 0.7165 0.7015 0.7159 0.6955 0.6798 0.6874 0.6923 0.6883 0.709 0.7203 0.6533 0.8544 0.7124

0.6984 0.6711 81031.36 0.7041 0.6951 0.6967 0.6694 81212.37 0.7025 0.6965 0.6977 0.6692 81121.22 0.7014 0.6938 0.6997 0.6707 80914.57 0.7023 0.6939 0.6973 0.6691 81033.11 0.703 0.6953

Mean

1 0

1 0

1 0

1 0

1 0

1 0

1 0

1 0 0

1 0

1 0

1 0

1 0

1 0

1 81063

1 0

1 0

Std. Dev.

0

111 0.1

% RSD

0.1

0.2

0.3

0.2

0.2

0.1

0.1

0.2

0.2

1.3

0.3

0.2

0.1

0.1

0.2

Project NaReported by User: Greg Jaudzems (JAUDZEG) Report MeDate Printed: 5368 23.07.2015Report Method ID: 5368 10:54:04US/Eastern

Area: #

Suitability Summary Table SuitabilityComponent Summary For Area # SampleNaHis Tau Ser

Glu

Thr

Ala

Pro

Lys

Tyr

xCys

Met

Val

NVa

Ile

Leu

Arg

Gly

Asp

1 Suit mix 2 Suit mix 3 Suit mix 4 Suit mix 5 Suit mix

56797 57669 56461 55056 55797 56267 55785 57471 58553 53044 71739 57812 56952 57974 56730 55241 55732 56308 55789 57618 58511 52937 71131 57829 56953 57746 56759 55315 56001 56303 55759 57493 58346 53067 70609 57798 56829 57653 56572 55190 55751 56094 55686 57362 58148 52936 70290 57972 56841 58011 56358 55085 55700 56102 55773 57451 58364 52937 69231 57724 56874 57811 56576 55177 55796 56215 55758 57479 58384 52984 70600 57827

56592 54384 81031 57053 56324 56582 54365 81212 57051 56563 56601 54286 81121 56902 56284 56615 54271 80915 56828 56144 56507 54217 81033 56963 56343 56579 54305 81063 56959 56332

Mean

Std. Dev.

73

170 0.3

172 0.3

108 0.2

120 0.2

108 0.2

42

92

160 0.3

65

942 1.3

90

42

69

111 0.1

97

151 0.3

% RSD

0.1

0.1

0.2

0.1

0.2

0.1

0.1

0.2

xCys stand 5pmol Suit for 5 injections Response: # Suitability Summary Table Suitability Component Summary For Response # SampleNaHis Tau Ser

Glu

Thr

Ala

Pro

Lys

Tyr

xCys

Met

Val

NVa

Ile

Leu

Arg

Gly

Asp

1 Suit xCys 2 Suit xCys 3 Suit xCys 4 Suit xCys 5 Suit xCys

0.722 0.724

83168.61 82873.99 83148.13 82882.29 82966.21

0.72

0.722

0.72

Mean

1 0

83008

Std. Dev.

142 0.2

% RSD Area: #

0.2

Suitability Summary Table SuitabilityComponent Summary For Area # SampleNaHis Tau Ser

Arg Leu (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) (µV*sec) 1 Suit xCys 60012 83169 2 Suit xCys 59988 82874 3 Suit xCys 59882 83148 4 Suit xCys 59873 82882 5 Suit xCys 59775 82966 Mean 59906 83008 Std. Dev. 96 142 % RSD 0.2 0.2 Gly Asp Glu Thr Ala Pro Lys Tyr xCys Met Val NVa Ile

Jaudzems, Guthrie, Lahrichi, and Fuerer – January 2018

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