Parallel detection, quantification, and depth profiling of peptides with dynamic-secondary ion mass spectrometry (D-SIMS) ionized by C60(+)-Ar(+) co-sputtering.
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) using pulsed C(60)(+) primary ions is a promising technique for analyzing biological specimens with high surface sensitivities. With molecular secondary ions of high masses, multiple molecules can be identified simultaneously without prior separation or isotope labeling. Previous reports using the C(60)(+) primary ion have been based on static-SIMS, which makes depth profiling complicated. Therefore, a dynamic-SIMS technique is reported here. Mixed peptides in the cryoprotectant trehalose were used as a model for evaluating the parameters that lead to the parallel detection and quantification of biomaterials. Trehalose was mixed separately with different concentrations of peptides. The peptide secondary ion intensities (normalized with respect to those of trehalose) were directly proportional to their concentration in the matrix (0.01-2.5 mol%). Quantification curves for each peptide were generated by plotting the percentage of peptides in trehalose versus the normalized SIMS intensities. Using these curves, the parallel detection, identification, and quantification of multiple peptides was achieved. Low energy Ar(+) was used to co-sputter and ionize the peptide-doped trehalose sample to suppress the carbon deposition associated with C(60)(+) bombardment, which suppressed the ion intensities during the depth profiling. This co-sputtering technique yielded steadier molecular ion intensities than when using a single C(60)(+) beam. In other words, co-sputtering is suitable for the depth profiling of thick specimens. In addition, the smoother surface generated by co-sputtering yielded greater depth resolution than C(60)(+) sputtering. Furthermore, because C(60)(+) is responsible for generating the molecular ions, the dosage of the auxiliary Ar(+) does not significantly affect the quantification curves.
Department of Materials Science and Engineering, National Taiwan University, Taipei 106, Taiwan.
Chi-Jen
CJ
Chang
Hsun-Yun
HY
Chang
Yun-Wen
YW
You
Hua-Yang
HY
Liao
Yu-Ting
YT
Kuo
Wei-Lun
WL
Kao
Guo-Ji
GJ
Yen
Meng-Hung
MH
Tsai
Jing-Jong
JJ
Shyue
D000595:Amino Acid Sequence; D001128:Argon / Q000737:chemistry; D002138:Calibration; D037741:Fullerenes / Q000737:chemistry; D007477:Ions / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*
D037741:Fullerenes; D007477:Ions; D010455:Peptides; D001128:Argon; C069837:fullerene C60