ANALYSIS OF PHYSIOLOGICAL AMINO ACIDS WITH THE MASSTRAK ™ AMINO ACID ANALYSIS SOLUTION Paula Hong, Thomas E. Wheat and Diane M. Diehl Following the benchtop pre-column derivatization of the analytes, separation and detection are achieved with a reversed-phase UPLC ® column and TUV detector, respectively. The analysis is complete in less than 32 minutes and allows for the identifica- tion and quantification of 42 common amino acids and related compounds in a single run. Full profile amino acid analyses are conducted using a defined analytical method. Electronic reports are generated using pre-defined software templates. These may be customized to suit specific reporting needs. INTRODUCTION The analysis of free amino acids in physiological fluids is an important tool in the study of physiological processes. Changes in the free amino acid concentrations can reflect altered states of metabolic pathways. These changes may be due to the influence of nutrition, environmental factors or genetic disturbances of metabolism. Expanding research in this field creates a demand for more amino acid analysis. A more robust method can offer the higher throughput that is required with this increasing sample load. Improved throughput is a function of both reducing run time and improving method robustness. Current ion exchange methods are time consuming and frequently require lengthy method adjust- ment. Waters has developed a new solution for research use only. The Waters MassTrak ™ AAA Solution combines a well-character- ized amino acid derivatization, QC tested columns and eluents, a pre-configured UltraPerfomance LC ® method, standards, and compliance-ready software. Figure 1. MassTrak AAA Solution Standard Chromatogram 250 µM PSer AMQ HyPro NH3 His PEA Asn 3MH Tau 1MH Ser Gln Carn Arg Gly Ans EA Asp Sar Glu Cit B-Ala Thr Ala GABA AADA Pro BAIB Hyl1 Hyl2 AABA Cy st Orn Cys Lys Tyr Deriv Peak Met Val Nva Ile aIle Leu HCys Phe Trp AU 0 .03 .06 .09 .12 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 min FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
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A NA LYSIS O F P H YSIO LOGI C A L AM INO AC I DS W IT H T H E MA S S T R A K ™ AM INO AC I D A NA LYSIS SO LU T IO N
Paula Hong, T homas E. Wheat and Diane M. Diehl
Following the benchtop pre-column derivatization of the analytes,
separation and detection are achieved with a reversed-phase
UPLC® column and TUV detector, respectively. The analysis is
complete in less than 32 minutes and allows for the identifica-
tion and quantification of 42 common amino acids and related
compounds in a single run. Full profile amino acid analyses are
conducted using a defined analytical method. Electronic reports
are generated using pre-defined software templates. These may be
customized to suit specific reporting needs.
INT RODUCT ION
The analysis of free amino acids in physiological fluids is an
important tool in the study of physiological processes. Changes
in the free amino acid concentrations can reflect altered states of
metabolic pathways. These changes may be due to the influence
of nutrition, environmental factors or genetic disturbances of
metabolism. Expanding research in this field creates a demand
for more amino acid analysis. A more robust method can offer
the higher throughput that is required with this increasing sample
load.
Improved throughput is a function of both reducing run time and
improving method robustness. Current ion exchange methods are
time consuming and frequently require lengthy method adjust-
ment. Waters has developed a new solution for research use only.
The Waters MassTrak™ AAA Solution combines a well-character-
ized amino acid derivatization, QC tested columns and eluents,
a pre-configured UltraPerfomance LC® method, standards, and
compliance-ready software.
Figure 1. MassTrak AAA Solution Standard Chromatogram 250 µM
PSer
AM
Q
HyP
roN
H3
His
PEA
Asn
3MH
Tau
1MH
Ser
Gln
Car
nA
rgG
ly Ans
EA Asp
Sar Glu C
it B-A
laTh
r Ala GA
BA AA
DA
Pro
BAIB
Hyl
1
Hyl
2
AA
BACy
st
Orn
Cys
Lys
Tyr
Deriv
Pea
kM
et
Val Nva
IleaI
le Leu
HC
ys
Phe
Trp
AU
0
.03
.06
.09
.12
4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 min
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
EX PERIMENTAL
Sample Preparation
Sample preparation utilizes the MassTrak AAA derivatization kit
and Total Recovery vials. The amino acids are derivatized with