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1 PREP Calibration Mix DIOS Low i. Introduction Pre-packaged PREP Calibration Mixtures eliminate the need to purchase and store large quantities of the component calibration reagents, simplifying sample preparation for TOF mass spectrometry. In addition to calibration, the mixtures are also useful for diagnostic tests and relative comparisons between different instruments. The calibration mixtures are packaged as lyophilized solids in glass vials. The vials are vacuum-sealed in foil pouches to ensure protection from light and air. Contents I. introduction II. Recommended Usage of PREP Calibration Mix DIOS Low III. Preparation of PREP Calibration Mix DIOS Low a. Procedure b. Notes IV. storage and stability V. Peak List for PREP Calibration Mix DIOS Low Prep Calibration Mix- DIOS Low 1

2 II. Recommended Usage of PREP Calibration Mix DIOS Low Waters PREP Calibration Mix DIOS Low is intended for calibration of a MALDI TOF MS instrument used for small molecule detection between m/z 1 and 1500 in reflectron mode. i The mix contains polyethylene glycol (PEG) 200, 600, and 1000 with sodium iodide, rubidium iodide and cesium iodide (NaI, RbI and CsI). This mixture has not been optimized for use in electrospray systems. The following spectrum of the PREP Calibration Mix DIOS Low was processed by baseline subtraction, smoothing, and centroiding in preparation for calibration. Data was obtained on a Waters MALDI micro MX. Figure 1. spectrum of PREP Calibration Mix DIOS Low obtained on a Waters MALDI TOF mass spectrometer (MALDI micro MX TM ). Data presented was background subtracted, smoothed and centered in preparation for calibration. i Waters provides DIOS-target Plates for small molecule detection (see part numbers on the next page). DIOS stands for desorption/ionization on silicon. III. Preparation of PREP Calibration Mix DIOS Low The following methods are provided as a general guideline for preparation on DIOStarget Plates. The described method only serves as a starting point. Depending on the application, one may consider using other solvents and/or methods. For optimum performance, solvents of the highest purity commercially available are recommended. The following solvents were found to be satisfactory. CH 3 CN: Acetonitrile (J. T. Baker Cat. No.: ) H 2 O: High purity (18 M) water 3. Vortex the vial for several seconds to ensure that the solid is completely dissolved. 4. Apply 1 μl of the mixture onto a clean DIOS-target Plate. 5. Dry at room temperature before LDI (laser desorption/ionization) analysis. 6. Note that the PREP Calibration Mix is relatively concentrated and tends to leave a visible, shiny residue on the surface. Adjust your instrument laser power so that the signal does not saturate the detector. b. Notes i. Optimization for specific applications is recommended. Other relative ratios and solvents may be used for reconstitution. a. Procedure 1. For a dilution solution, make a 1:1 mixture of CH 3 CN and H 2 O. i 2. Add at least 250 μl of the dilution solution ii to the glass vial containing the lyophilized calibration mixture. ii. In some cases further dilutions of this solution may yield better spectra depending on the instrument and substrate used. The solution for the calibration spectrum shown previously was made by taking 50 µl of the calibration made in #2 and adding 450 µl of dilution solvent mix. Prep Calibration Mix- DIOS Low 2

3 iv. Storage and Stability It is recommended that PREP Calibration Mixtures remain in the foil pouch (in the dark) at room temperature until use. Once reconstituted in appropriate solvents, the PREP Calibration Mix solution can be used for at least 1 year when stored in the dark at 4 C. v. Peak List for PREP Calibration Mix DIOS Low The following reference file contains the masses of selected low mass clusters formed by the mixture of sodium, rubidium, and cesium iodide as well as the sodium (and some rubidium) adducts of polyethylene glycol oligomers with chemical formula HO[C 2 H 4 O]nH + Na (where n = 1,2,3,...) between 200 and 1500 Da. es were calculated using the following values (where M (x) indicates the mass, M, of the chemical species, x, within the parentheses): M(C) = M(H) = M(N) = M(O) = M(Na) = M(K) = M(Rb) = M(I) = M(Cs) = Below m/z 150 the sodium, rubidium, and cesium ions provide the more dominant peaks. Above m/z 200 the sodiated PEG peaks provide the more dominant peaks. Table 1: Calculated masses of selected low mass clusters formed by the mixture of sodium, rubidium, and cesium iodide with polyethylene glycol (PEG) oligomers Identity Comment Na sometimes weak Rb - primary primary peak Rb - isotope Cs primary peak PEG + Na very weak PEG + Na weak PEG + Rb weak PEG + Na main PEG + Rb main PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + N a primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak Prep Calibration Mix- DIOS Low 3

4 Table 1. Calculated masses of selected low mass clusters formed by the mixture of sodium, rubidium, and cesium iodide with polyethylene glycol (PEG) oligomers (continued) Identity Comment PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb PEG + Na primary peak PEG + Rb Prep Calibration Mix- DIOS Low 4

5 Sales Offices: Austria and European Export (Central South Eastern Europe, CIS and Middle East) Australia Belgium Brazil Canada China CIS/Russia Czech Republic Denmark Finland France (33) Germany Hong Kong Hungary India and India Subcontinent Ireland Italy Japan (81) Korea (82) Mexico The Netherlands +31 (0) Norway Poland (48) Puerto Rico Singapore Spain Sweden Switzerland Taiwan United Kingdom Waters Corporation,, The Science of What s Possible, Waters, MALDI micro MX TM and PREP are trademarks of Waters Corporation. September Rev B VW-PDF Waters Corporation 34 Maple Street Milford, MA U.S.A. T: F: Prep Calibration Mix- DIOS Low 5

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