About Instrument and its applications-
Mass Spectrometry (MS) is the study of systems causing the formation of gaseous ions, with or without fragmentation, which are then characterised by their mass to charge ratios (m/z) and relative abundances. Matrix-Assisted Laser Desorption-Ionization (MALDI) mass spectrometry used as a powerful, highly versatile technology with a diverse range of applications. MALDI coupled to time-of-flight mass spectrometry (MALDI-TOF MS) is used to sequence proteins, map biomolecules in tissues, identify microorganisms, and analyze several thousand biochemical assays. The sample for analysis by MALDI-TOF MS is prepared by mixing or coating with a solution of an energy-absorbent matrix which entraps and co-crystallizes the sample when dried. The matrix is ionized with a laser beam, and transfers the charge to the analytes, generating singly charged ions from analytes in the sample that are then accelerated. Ions are separated from each other on the basis of their mass-to-charge ratio (m/z) before being detected and measured using the TOF mass analyzer. The high throughput and speed associated with MALDI-TOF MS has made this technology an important tool for large-scale proteomics, imaging studies, and other analyses.

General Information-
Make- Bruker
Model- UltrafleXtreme
Year of Installation- 2024

Specifications-
1) Resolution: up to 40,000
2) Mass Accuracy: 1.5 ppm
3) Source: The IR-laser based self-cleaning ion source
4) Laser: Smartbeam II
5) Laser frequency [Hz]: 2,000 (MS) / 1,000 (TOF/TOF)
6) Mass Range: up to 1,000,000
7) Software Suite: Flex Control for quick and easy data acquisition & Flex Analysis for automatic and interactive data analysis.

Sample Requirements-
Powder: 3-5mg
Liquid: 10-50µl (with suitable matrix if required)

General Instructions:
• Samples will not be analyzed unless complete information is provided.
• The user must bring samples to IIC before one day from the date of analysis.
• Choose matrix as per the molecular weight of the sample.
• Mix sample and matrix in ratio of 1:4 (v/v) and vortex well.
• TFA may be used depending on the sample
• 2 µL mixture will be loaded on the sample plate.
• Matrices for peptide and proteins are available in the laboratory for calibration purposes.
• Matrix for sample analysis to be supplied by users.

Sample Preparation Guidelines Samples:
1) Samples for MALDI-TOF analysis need to meet certain requirements for obtaining good spectra. The more careful you prepare samples the more likely a successful analysis will be.
2) Samples can be submitted in solid, liquid, or solution form. If you are submitting a sample in solution, you should indicate the solvent on the submission form that your sample is dissolved in and other solvents in which your sample is soluble.
3) Suitable solvents are ones that are volatile. For sample work up and purification fresh HPLC grade solutions should be use: water, ammonium hydrocarbonate, ammonium acetate, ammonium formiate,0.1% trifluoroacetic acid, 0.2% formic acid, methanol, acetonitrile, ethanol etc. Some samples can degrade if an acid or base is added to the sample. If you are submitting a sample that is sensitive to acid to base, please indicate so. Please supply the sample in a low-salt buffer (less than 20 mM).
4) Avoid the use of non-volatile agents like salts (NaCl, CaCl2, KH2PO4), detergents (Tween, Triton, SDS), chaotropic agents (Urea, Guanidinium salts) and non-volatile solvents like DMSO, DMF, or Glycerol. If you can’t avoid these agents, purify. Dialysis, ZipTips, and RP-HPLC are good purification methods. After purification, lyophilize if possible. Ion exchange beads may work well for salt removal. The range for many samples/preparations is not very large, therefore it is necessary to have a good estimate of the sample amount because the sample amount may need to be varied on the target. The total amount of sample needed for MALDI analysis depends on the sample type. The larger the molecular weight the more sample concentration is needed.
5) Give information like: structure, sequence, molecular weight, type of compound, biological activity, chemical reactivity, pH, sample amount/concentration, describe purification/isolation with focus on relative agents/solvents, known or suspected impurities, suitable solvents, hazardous properties: radioactivity, carcinogenicity, poison, or explosive.

Contact Person-
Dr. Jay Singh Meena
Technical Officer-II
Tel: +91-1332-285779/286170
E-mail: jsmeena.mic2018@iitr.ac.in