Abstract Conclusions References  The intent of this presentation is to address the usage of nanomanipulation-coupled with nanospray mass spectrometry as applied to the extraction of small volume ink samples from questioned documents.  This technique will provide a non-destructive approach to the analysis of ink samples from questioned documents. Introduction  The process of ink dating is an important research area for ink analysis.  Major problems arise when analysis has to be done on an ancient document where preservation of its integrity has to be considered.  Iron gall ink were widely used between the Middle ages and early 20 th century. The ink is made mostly using tannins obtained form plant galls and Iron(II) sulphate. The ink contains enormous amount of iron(II) ions along with some significant Zn, Mn, Ca and Mg ions.  However, modern ink is made from organic pigments and dyes with little metal present.  The metal ions present in the iron gall ink can be extracted using chelating agents and analyzed in mass spectrometer. For organic ink, an appropriate solvent can be used for the extraction. Results/Discussion This novel technique will impact the forensic science community by providing a non destructive approach to analyzing ink samples from questioned documents with small sample volumes. This will be an improvement from past analytical methods such as HPLC and TLC where higher sample volumes were required along with more preparation time. This approach will provide a more direct and efficient way to do ink analysis.  Chen, Z., Sun, Q., Xi, Y, F., Owens, G. Speciation of metal-EDTA complexes by flow injection analysis with electrospray ion mass spectrometry and ion chromatography with inductively coupled plasma mass spectrometry. J. Sep. Sci. 2008,31,  Siegal J., Allison J., Mohr D., Dunn J. The use of laser desorption/ionization mass spectrometry in the analysis of inks in questioned documents. J. Siegel et al./ Talanta , Materials/Methods Nanomanipulation-Coupled to Nanospray Mass Spectrometry Applied to Ink and Document Analysis Ubisha Joshi, Dr. Teresa Golden and Dr. Guido F. Verbeck Department of Chemistry, University of North Texas, Denton,Texas  A very small drop of extracting solvent (approximately 2-3 µL) can be placed on the surface of the ink using a gel loader pipette tip.  The solvent is then allowed to sit for a predetermined amount of time to allow diffusion of ink.  The dissolved ink is then collected into the nanospray capillary and analyzed by nanospray mass spectrometry.  EDTA is used to chelate the metal ions from the Iron gall ink.  The extraction of red ink shows the peak at m/z 443. this is due to the presence of Rhodamine dye, found in red ink. Both Rhodamine 6B and Rhodamine 6G are isomers.  For the extraction of the historical document, the ink extraction shows a distinct peak at m/z which cannot be seen during the background extraction. The peak was reproducible but has not been identified yet.  The EDTA gives a de-protonated peak at m/z 291. The EDTA complex with iron gives a peak at m/z 344 from [EDTA-4H+Fe] -. However, EDTA-Mn complex gave a peak with highest intensity than with iron at m/z 343 form [EDTA-3H+Mn] -. Top: EDTA in ESI mode Bottom left: EDTA and Iron gall ink in ESI mode Bottom right : Extraction of Iron gall ink in NSI mode Fig 1:Extraction of red ink Fig 2: Background extraction from the historical document Fig 3: Ink extraction form the historical document Top: Nanomanipulator Left: Schematic diagram of Nanomanipulator  A nanospray capillary is filled with an appropriate solvent and mounted on a nanopositioner.  The capillary is then maneuvered very close to the surface of the document where the extraction is to be made. (a) Before extraction (b) Extraction Set-up (c) After Extraction b c a The historic document Ink extraction set up