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Toward the Atomic-Level Mass Analysis of Biomolecules 337


Figure 1. Mass spectrum of valine.


Figure 3. Mass spectra of glutamine (above) and glutamic acid (below).


ions in the detection sequence. When the SWCNT zone is analyzed, most detected ions are singly and doubly charged carbon ions (Nishikawa et al., 2009). In the molecule zone, the number of the carbon ions is small and the number of other ions increases. Leucine and methionine are successfully mass analyzed, revealing stable clusters such as COOH and HCNH2 (Nishikawa et al., 2009). Example of mass spectra of valine and proline are shown


Figure 2. Mass spectrum of prolone.


hydrochloric acid (HCl) and 62mg of Lys-Lys (a-L-Lysyl- L-Lysine) was dissolved in 50mL of 0.1N HCl. A small ball (~10–20 µm in diameter) of tangled long


SWCNTs was attached to aWtip with silver paste. Then, the tip was dipped in the solution of sample molecules.


RESULTS


SAP analysis proceeds by field evaporating SWCNTs and molecules from the surface layers. Although the SWCNT and molecule zones are randomly mixed, the SWCNT zone can be separated from the molecules zone by plotting the


Table 1. The ratios of the number of atoms detected by analyzing valine and proline.


HC N


Valine Ratio of atoms


Proline Ratio of atoms


11 0 5 1 2


Ratio of counted number of atoms 9.08 4.98 1 2.74 9


5 1 2 Ratio of counted number of atoms 9.35 5.11 1 2.03


Ghitamine Ratio of atoms


in Figures 1 and 2. Clusters of strongly bound carboxy, HCNH2, and HCNH groups are detected. No ions corre- sponding to pyrrolidine were detected. The number of atoms forming all mass peaks are


counted and listed in Table 1. The ratios of the counted number of H, C, N, and O atoms are very close to the ratios of the atoms forming the molecules. This indicates that the SAP analysis is not only qualitative, but also quantitative. The structures of glutamine (NH2–CO–CH2–


CH2–CHNH2–COOH) and glutamic acid (HO–CO–CH2– CH2–CHNH2–COOH) are very similar. The only differences are the amino group of glutamine and the OH group of glutamic acid. The mass difference of them is 1 amu. Figure 3 shows the mass spectra of glutamine and glutamic acid, the mass peaks were very similar. An unexpected finding was that the carboxy group had an extra hydrogen atom. Only difference between the two mass spectra was an extra


Table 2. The ratios of the number of atoms detected by analyzing glutamine and glutamic acid.


HC N 10 9 O 5 2 3


Ratio of counted number of atoms 9.07 4.98 2 3.76 Glutamic acid Ratio of atoms


5 1 4 Ratio of counted number of atoms 9.13 5.06 1 3.54


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