Quantification in MALDI-TOF mass spectrometry of modified polymers

Abstract

MALDI-TOF mass spectrometry quantification is hampered by the poor reproducibility of the signal intensity and by molecular-mass and compositional discrimination. The addition of a suitable compound as an internal standard increases reproducibility and allows a calibration curve to be constructed. The concept was also verified with synthetic polymers but no instructions for practical implementation were given [H. Chen, M. He, J. Pei, H. He, Anal. Chem. 75 (2003) 6531-6535.], even though synthetic polymers are generally non-uniform with respect to molecular mass and composition and access to the polymer of the same molecular mass distribution and composition as that of the quantified one is thus the exception rather than rule. On the other hand, relative quantification of polymers e.g., the content of the precursor polymer in a batch of a modified polymer, is usually sought. In this particular case, the pure precursor is usually available and the modified polymer can serve as an internal standard. However, the calibration curve still cannot be constructed and the use of the internal standard has to be combined with the method of standard addition in which the precursor polymer is added directly to the analyzed sample. The experiments with simulated modified polymers, mixtures of poly(ethylene glycol) (PEG) and poly(ethylene glycol) monomethyl ether (MPEG) of similar molecular-mass distribution, revealed a power dependence of the PEG/MPEG signal-intensity ratio (MS ratio) on the PEG/MPEG concentrations ratio in the mixture (gravimetric ratio). The result was obtained using standard procedures and instrumentation, which means that the basic assumption of the standard-addition method, i.e., the proportionality of the MS and gravimetric ratios, generally cannot be taken for granted. Therefore, the multi-point combined internal-standard standard-addition method was developed and experimentally verified for the quantification of the precursor in modified polymers. In this method, the two parameters of the power-type calibration curve - the proportionality constant and the exponent-are assumed. If the exponent strongly deviates from unity the minority component can be significantly underrepresented in the spectrum. Therefore, the absence of the precursor polymer signals in the MALDI-TOF mass spectrum of a modified polymer sample does not prove the absence of the precursor in the sample. Such a conclusion has to be corroborated by the standard-addition method.