[ Care and Use Manual ]

MassPREP Calibration Mix – DIOS Low Mass

i. Introduction Contents

Pre-packaged MassPREP™ Calibration Mixtures eliminate the need to purchase I. introduction and store large quantities of the component calibration reagents, simplifying sample preparation for TOF mass spectrometry. In addition to calibration, the II. recommended Usage of MassPREP Calibration Mix – DIOS Low Mass mixtures are also useful for diagnostic tests and relative comparisons between different instruments. The calibration mixtures are packaged as lyophilized III. Preparation of MassPREP Calibration Mix – DIOS Low solids in glass vials. The vials are vacuum-sealed in foil pouches to ensure Mass protection from light and air. a. Procedure b. Notes

IV. storage and stability

V. Peak List for MassPREP Calibration Mix – DIOS Low Mass

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II. Recommended Usage of MassPREP Calibration Mix – DIOS Low Mass

Waters MassPREP Calibration Mix – DIOS Low Mass 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 and cesium iodide (NaI, RbI and CsI). This mixture has not been optimized for use in electrospray systems. The following spectrum of the MassPREP Calibration Mix – DIOS Low Mass was processed by baseline subtraction, smoothing, and centroiding in preparation for calibration. Data was obtained on a Waters MALDI micro MX.

Figure 1. Mass spectrum of MassPREP Calibration Mix – DIOS Low Mass obtained on a Waters MALDI TOF mass spectrometer (MALDI micro MXTM). Data presented was background subtracted, smoothed and centered in preparation for calibration.

i Waters provides DIOS-target Plates for small molecule detection (see part 3. Vortex the vial for several seconds to ensure that the solid is numbers on the next page). DIOS stands for desorption/ionization on silicon. completely dissolved.

4. Apply 1 μL of the mixture onto a clean DIOS-target Plate. III. Preparation of MassPREP Calibration Mix – DIOS Low Mass 5. Dry at room temperature before LDI (laser desorption/ionization) analysis. The following methods are provided as a general guideline for preparation on DIOStarget Plates. The described method only serves as a starting point. 6. Note that the MassPREP Calibration Mix is relatively concentrated Depending on the application, one may consider using other solvents and/or and tends to leave a visible, shiny residue on the surface. Adjust methods. For optimum performance, solvents of the highest purity commer- your instrument laser power so that the signal does not saturate cially available are recommended. The following solvents were found to be the detector. satisfactory. b. Notes 3 CH CN: Acetonitrile (J. T. Baker Cat. No.: 9017-03) i. Optimization for specific applications is recommended. Other 2 H O: High purity (18 M) water relative ratios and solvents may be used for reconstitution. a. Procedure ii. In some cases further dilutions of this solution may yield better i spectra depending on the instrument and substrate used. The 1. For a dilution solution, make a 1:1 mixture of CH3CN and H2O. solution for the calibration spectrum shown previously was made ii 2. Add at least 250 μL of the dilution solution to the glass vial by taking 50 µL of the calibration made in #2 and adding 450 µL containing the lyophilized calibration mixture. of dilution solvent mix.

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iv. Storage and Stability Table 1: Calculated masses of selected low mass clusters formed by the mixture of sodium, rubidium, and cesium iodide with polyethylene glycol It is recommended that MassPREP Calibration Mixtures remain in the foil pouch (PEG) oligomers (in the dark) at room temperature until use. Once reconstituted in appropriate solvents, the MassPREP Calibration Mix solution can be used for at least 1 year Mass Identity Comment when stored in the dark at 4 °C. 22.989768 Na sometimes weak 84.911792 Rb - primary primary peak v. Peak List for MassPREP Calibration Mix – DIOS 86.909185 Rb - isotope Low Mass 132.905429 Cs primary peak The following reference file contains the masses of selected low mass clusters 217.1051919 PEG + Na very weak formed by the mixture of sodium, rubidium, and cesium iodide as well as the 261.1314067 PEG + Na weak sodium (and some rubidium) adducts of polyethylene glycol oligomers with 279.0272159 PEG + Rb weak 305.1576215 PEG + Na main HO[C2H4O]nH + Na (where n = 1,2,3,...) between 200 and 1500 Da. Masses were calculated using the following values (where M (x) 323.0534307 PEG + Rb main indicates the mass, M, of the chemical species, x, within the parentheses): 349.1838362 PEG + Na primary peak 367.0796455 PEG + Rb M(C) = 12.000000000 393.210051 PEG + Na primary peak M(H) = 1.007825040 411.1058602 PEG + Rb M(N) = 14.003074000 437.2362658 PEG + Na primary peak M(O) = 15.994914630 455.132075 PEG + Rb M(Na) = 22.989768 481.2624806 PEG + Na primary peak M(K) = 38.9637069 499.1582898 PEG + Rb M(Rb) = 84.911792 525.2886954 PEG + Na primary peak M(I) = 126.90446 543.1845046 PEG + Rb M(Cs) = 132.905429 569.3149102 PEG + Na primary peak 587.2107194 PEG + Rb Below m/z 150 the sodium, rubidium, and cesium ions provide the more 613.341125 PEG + Na primary peak dominant peaks. Above m/z 200 the sodiated PEG peaks provide the more 631.2369342 PEG + Rb dominant peaks. 657.3673398 PEG + Na primary peak 675.263149 PEG + Rb 701.3935546 PEG + N a primary peak 719.2893638 PEG + Rb 745.4197694 PEG + Na primary peak 763.3155786 PEG + Rb 789.4459841 PEG + Na primary peak 807.3417934 PEG + Rb 833.4721989 PEG + Na primary peak 851.3680081 PEG + Rb 877.4984137 PEG + Na primary peak 895.3942229 PEG + Rb 921.5246285 PEG + Na primary peak

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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)

Mass Identity Comment 939.4204377 PEG + Rb 965.5508433 PEG + Na primary peak 983.4466525 PEG + Rb 1009.577058 PEG + Na primary peak 1027.472867 PEG + Rb 1053.603273 PEG + Na primary peak 1071.499082 PEG + Rb 1097.629488 PEG + Na primary peak 1115.525297 PEG + Rb 1141.655702 PEG + Na primary peak 1159.551512 PEG + Rb 1185.681917 PEG + Na primary peak 1203.577726 PEG + Rb 1229.708132 PEG + Na primary peak 1247.603941 PEG + Rb 1273.734347 PEG + Na primary peak 1291.630156 PEG + Rb 1317.760562 PEG + Na primary peak 1335.656371 PEG + Rb 1361.786776 PEG + Na primary peak 1379.682586 PEG + Rb 1405.812991 PEG + Na primary peak 1423.7088 PEG + Rb 1449.839206 PEG + Na primary peak 1467.735015 PEG + Rb 1493.865421 PEG + Na primary peak 1511.76123 PEG + Rb

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©2007 Waters Corporation, , The Science of What’s Possible, Waters Corporation Waters, MALDI micro MXTM and MassPREP are trademarks of 34 Maple Street Waters Corporation. Milford, MA 01757 U.S.A. T: 1 508 478 2000 September 2007 715001478 Rev B VW-PDF F: 1 508 872 1990 www.waters.com

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