LC–MS determination of bioactive molecules based upon stable isotope-coded derivatization method

Liquid chromatography (LC) coupled with mass spectrometry (MS) has been widely used for the analyses of various molecules in many research fields. The electrospray ionization of MS has contributed to the advancement of the LC–MS and LC–MS/MS methods. However, the detection sensitivity is not always sufficient in biological samples, in spite of the highly sensitive ionization method. To increase the sensitivity, chemical derivatization, providing ionization enhancement and avoiding the matrix effect, is effective for various functional groups in the target molecules. However, the accuracy and precision by the determination is sometimes insufficient, especially in complex matrices. In such a case, stable isotope-labeled analogs are often used as the internal standards for the determination of the analytes. When the target compound in samples is limited, a high accuracy and precision is usually obtained by the isotope dilution method. However, the use of individual isotope standards is very difficult for the analyses of multiple molecules in complex matrices. Instead of the use of an isotope analog of the analytes, the differential isotope labeling method based upon chemical derivatization (stable isotope-coded derivatization) (ICD) by both reagents possessing different isotopes is realized. The ICD technique utilizing mass-different isotope tags is known to be an efficient means for metabolite profiling analyses. Thus, the present paper reviews the ICD method reported in the past 10 years. The species of the ICD reagents, their features and the applications to biological specimens are also described in this review.

► Liquid chromatography (LC) coupled with mass spectrometry (MS) has been widely used for the analyses of various molecules in many research fields.
► Stable isotope-labeled analogs are often used as the internal standards for the determination of target compounds by LC–MS.
► However, the use of individual isotope standards is very difficult for the analyses of multiple molecules in complex matrices.
► In such case, stable isotope-coded derivatization utilizing mass-different isotope tags is realized.
► This paper reviews the stable isotope-coded derivatization methods and their applications to proteomics and metabolomics in the past 10 years.