Page 40 - Clinical-Medical Solution for Clinical Chemistry

Determination of Unbound Urinary Amino Acids Incorporated
with Creatinine Normalization by LC-MS/MS Method with
CLAM-2000 Online Sample Pre-treatment

(x10,000)
5.0
Phe-d5 (A) 0.5 uM each analyte
CR
Ile
2.5 Pro Gly
Leu Cit
Trp Phe Val Ser-d3
Met Glu His Arg
Tyr Thr Asn Cys Lys
0.0
2.5 5.0 7.5 10.0 12.5 15.0 17.5 min
(x1,000,000)
(B) 50 uM each analyte CR Orn
2.0
Lys
Pro Cit Arg
1.0
Trp Ile Cys
Phe-d5Phe Leu Met Val Glu His
0.0 Tyr Thr Asp Ala Ser Ser-d3 Gly Asn
2.5 5.0 7.5 10.0 12.5 15.0 17.5 min

Figure 2: MRM (SIM for Gly) chromatograms of 22 AA and CRE in mixed standards with (5µL inj. volume).

Area Ratio (x10) Area Ratio (x10) Area Ratio
8 8 8
Phe Pro Ser
1.0 2.0 5.0

7 7 7
0.5 1.0 2.5

6 6 6
1 2 3 4 5 1 2 3 4 5 4 5
0.0 0.0 0.0
0.0 Conc. Ratio 0.0 Conc. Ratio 0.0 Conc. Ratio
Area Ratio (x10) Area Ratio (x10) Area Ratio (x10)
8 8 8
Asp Cys Gly (SIM
2.0 mode)
7 5.0 7 2.5 7
1.0

6 6
2 3 4 5 2 3 4 1 2 4 5 6
5
3
0.0 0.0 0.0
0.0 Conc. Ratio 0.0 Conc. Ratio 0.0 Conc. Ratio
Figure 3: Representative calibration curves of eight levels at from 0.1 µM to 100 µM on CLAM-LC-MS/MS.

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