Writing sequence identification of seals and signatures in documents using ambient mass spectrometry imaging with...

Identifying the writing sequence of seals and signatures in documents is often performed and difficult to resolve in forensic determination. Morphological and physical-chemical analysis methods are often limited by the destructive nature of samples, a high signal response strength and specific materials. Mass spectrometry imaging (MSI) has been used as an alternative method because it can generate molecular images from many surfaces and produce rich chemical information. Herein, we developed a sequence identification method by coupling an air flow-assisted desorption electrospray ionization (AFADESI)-MSI system with a chemometric analysis, which can holistically and directly analyse document samples under ambient, moderate and selectable conditions and maintain the original appearance of the paper documents after sampling. By integrating principal component analysis (PCA) and the partial least squares discriminant analysis (PLS-DA), equivocal point analysis can be objectively performed, where knowing the components of the seal or signature is not necessary to identify the sequence. In total, 28 prepared samples with known sequences and two original blind test samples were analysed. One prepared sample was analysed in negative ionization mode, and other samples were inferred in positive ionization mode. All writing sequences were in accordance with the actual case. The writing sequence of the blind testing of the original samples was correctly identified. This study provided a convenient, objective and quasi-nondestructive method to investigate the sequence differences among equivocal document samples and is promising for providing an alternative method for the sequence identification of seals and signatures in questionable documents.