Simple correction improving long-term reproducibility of HPLC-MS

Tóth, Eszter and Hevér, Helga and Ozohanics, Olivér and Telekes, András and Vékey, Károly and Drahos, László (2015) Simple correction improving long-term reproducibility of HPLC-MS. JOURNAL OF MASS SPECTROMETRY, 50. pp. 1-14. ISSN 1076-5174 (Submitted)

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Abstract

Chromatographic peak areas in long series of HPLC-MS experiments often vary, which decreases reproducibility and may cause bias in the results. It was found that the sensitivitiy of various components change differently; in our case variability is in the order of 20-40%; and it is most likely due to changing conditions in ESI ionization. The most often used peak area correction methods do not take this effect into account. The change in peak areas can be well described by a polynomial function; we found that a 4th order polynomial is most often suitable. We suggest a simple correction algorithm based on polynomial fitting. When the experiments were inherently well reproducible, this correction improved reproducibility from 12% to 3% (on average for various components). When random errors were larger, this improvement was less significant (15% to 12% in nano-ESI), but nevertheless essential in order to avoid possible bias in the results.

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