Page 35 - Pharmaceutical Solution for Pharma Analysis

Application No. C149
News

„ Analysis of Standard Solution
This system was employed to analyze a standard mixed With CR-I (–), D-Gln and L-Lys, L-Ile and L-allo-Ile, and L-Thr
13
solution using C 6-L-Phe as the internal standard (Fig. 2). and L-allo-Thr, which each have the same MRM transition, are
Approximately equal area ratios were obtained with CR-I (+) detected with the same peak and therefore cannot be
and CR-I (–) for the amino acids other than Gln, Lys, Ile, allo-Ile, separated.
Thr, and allo-Thr, and this confirmed that the system is capable However, separation measurement can be performed for
of separation measurement (Table 2). these amino acids by utilizing two types of columns. For
With CR-I (+), L-Gln and D-Lys, D-Ile and D-allo-Ile, and D-Thr example, while D-Thr and D-allo-Thr cannot be separated
and D-allo-Thr, which each have the same MRM transition, are with CR-I (+) and L-Thr and L-allo-Thr cannot be separated
detected with the same peak and therefore cannot be with CR-I (–), interchanging the column types allows for
separated. separation measurement (Fig. 3).

Table 2 Analysis Results of Standard Solution
CR-I (+) CR-I (−) Ratio of CR-I (+) CR-I (–) Ratio of
Ratio of Ratio of Area Ratio of Ratio of Area
RT RT RT RT
Area Area (+)/(−) Area Area (+)/(−)
D-Ala 1.394 0.728 3.894 0.751 0.97 D-Leu 1.107 2.019 3.178 2.400 0.84
L-Ala 3.908 0.565 1.389 0.632 0.89 L-Leu 3.179 2.929 1.105 3.364 0.87
D-Arg 0.973 3.999 1.506 3.239 1.23 D-Lys 2.181 4.621 with L-Gln 7.395 1.641 2.82
L-Arg 1.499 5.633 0.981 6.718 0.84 L-Lys 7.348 1.795 2.161 5.118 with D-Gln 0.35
D-Asn 1.255 1.018 2.036 1.030 0.99 D-Met 1.259 1.704 4.554 1.859 0.92
L-Asn 2.036 0.805 1.263 0.911 0.88 L-Met 4.556 0.938 1.25 1.060 0.89
D-Asp 1.253 0.742 2.039 0.863 0.86 D-Phe 1.101 1.568 2.087 1.974 0.79
L-Asp 2.036 0.72 1.259 0.775 0.93 L-Phe 2.089 2.175 1.106 2.280 0.95
D-Cys 1.183 0.405 2.307 0.458 0.89 DL-Pro 0.957 2.756 0.971 3.105 0.89
L-Cys 2.308 0.789 1.186 0.797 0.99 D-Ser 1.222 0.224 1.756 0.253 0.89
D-Gln 1.247 2.111 2.161 3.478 with L-Lys 0.61 L-Ser 1.758 0.307 1.226 0.301 1.02
L-Gln 2.183 4.947 with D-Lys 1.239 3.686 1.34 D-Thr 1.023 1.339 with D-allo-Thr 1.53 0.968 1.38
D-Glu 1.246 2.972 4.426 3.262 0.91 L-Thr 1.533 0.851 1.033 1.324 with L-allo-Thr 0.64
L-Glu 4.388 3.506 1.24 3.731 0.94 D-allo-Thr 1.023 1.339 with D-Thr 1.205 0.573 2.34
Gly 2.827 0.037 2.796 0.039 0.93 L-allo-Thr 1.197 0.480 1.033 1.397 with L-Thr 0.34
D-His 0.967 2.797 1.099 3.917 0.71 D-Trp 1.105 2.839 1.99 3.344 0.85
L-His 1.09 3.699 0.977 2.969 1.25 L-Trp 1.988 3.458 1.111 3.510 0.99
D-Ile 0.988 4.745 with D-allo-Ile 1.446 2.983 1.59 D-Tyr 1.103 1.203 2.016 1.560 0.77
L-Ile 1.44 2.325 0.998 4.408 with L-allo-Ile 0.53 L-Tyr 2.016 1.448 1.109 1.455 1.00
D-allo-Ile 0.988 4.745 with D-Ile 1.313 2.926 1.62 D-Val 0.999 1.826 1.337 2.052 0.89
L-allo-Ile 1.308 1.844 0.998 4.101 with L-Ile 0.45 L-Val 1.331 3.170 1.008 3.251 0.97

indicates amino acids that can be separated by one column but not the other.
CROWNPAK CR-I (+) CROWNPAK CR-I (–)
120.10>74.00 (+) 450000 120.10>74.00 (+)
600000
D-Thr + D-allo-Thr 400000 L-Thr + L-allo-Thr
500000 350000
D-Thr
300000
400000 D-allo-Thr
L-allo-Thr L-Thr 250000
300000
200000
200000 150000
100000
100000
50000
0 0
0.5 1.0 1.5 0.5 1.0 1.5

Analysis Result of D/L-Threonine and D/L-allo-Threonine

References [1] Nakano,Y., Konya,Y., Taniguchi,M., Fukusaki,E., Journal of Bioscience and Bioengineering, 123, 134-138 (2016)

The analysis method presented in this edition of Application News was developed by the Fukusaki Lab in the School/Graduate School of
Engineering at Osaka University.

First Edition: Apr. 2017
For Research Use Only. Not for use in diagnostic procedure.
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