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Page 1: MALDI micro MX RESOLUTION - waters.com · Matrix: The matrix was prepared and used in 2 stages, •10 mg/mL sinapinic acid in acetone (Matrix 1) •10 mg/mL sinapinic acid in 60:40

MALDI micro MX RESOLUTION

M. Palmer, E. Claude, M. SnelWaters Corporation, MS technologies Center, UK

INTRODUCTIONMass resolution is the ability to separate two adjacent masses. It defines the performance of an analyzer and is a measure of the extent to which adjacent ions can be distinguished from each other with less than a defined amount of overlap. For a single peak this can be calculated from the width of a peak at a defined height and is usually referred to as Full Width Half Maximum (FWHM). FWHM is the resolution measured with a 50% valley overlap between peaks of adjacent m/z.

Resolution can be calculated using the following equation:

where m is the observed m/z and ∆m is the measured width at half peak height. Two key advantages of increasing resolution are: • Increased ability to separate peaks which are close in m/z• Improved detection limits through the generation of narrower, taller peaks.It is often assumed that high resolution is always required for good mass accuracy, this has however been shown not to be the case1.

EXPERIMENTALThe resolving power of the Waters® Micromass®

MALDI micro MXTM is demonstrated in reflectron and linear mode, in positive ion and negative ion modes.

Resolution reflectron modeSample: Adrenocorticotropic hormone clip 18 – 39 (ACTH) at a concentration of 2 pmol/µL in 0.1%trifluoroacetic acid (TFA).Matrix: Purified α-cyano-4-hydroxy cinnamic acid matrix was obtained from Waters (186002331). Matrix was prepared at 2 mg/mL in 1:1 acetonitrile:aqueous 0.1% TFA.

Samples were mixed 1:1 with matrix and 1 µL was spotted directly onto the target plate and allowed to air dry prior to analysis.Data acquisition: Six separate acquisitions in positive and negative mode.

Resolution linear modeSample: Cytochrome C (equine) at a concentration of 2 pmol/µL in 0.1% TFA.Matrix: The matrix was prepared and used in 2 stages,•10 mg/mL sinapinic acid in acetone (Matrix 1)•10 mg/mL sinapinic acid in 60:40 aqueous 0.1% TFA:acetonitrile (Matrix 2).1 µL of Matrix 1 was applied to the target plate and allowed to air dry then Matrix 2 was mixed 1:1 with the sample and 1 µL was spotted on top of the Matrix 1thin film.Data acquisition: Six separate acquisitions in positive and negative mode.

RESULTSThe results obtained are summarised in Table 1. Examples for each mode are shown in Figures 2-5. The MALDI micro MX exceeded the specified resolution in every mode.

∆mmResolution =

Figure 1. Definition of FWHM for a mass spectral peak

Page 2: MALDI micro MX RESOLUTION - waters.com · Matrix: The matrix was prepared and used in 2 stages, •10 mg/mL sinapinic acid in acetone (Matrix 1) •10 mg/mL sinapinic acid in 60:40

Mode Polarity Average resolution (six acquisitions)

Instrument specification

Reflectron positive 14,158 10,000

Reflectron negative 11,354 10,000

Linear positive 1,082 1,000

Linear negative 1,124 1,000

Table 1. Average resolution over six acquisitions.

Figure 2. Example of resolution in reflectron mode (positive). Observed resolution ≈ 16,000.

Page 3: MALDI micro MX RESOLUTION - waters.com · Matrix: The matrix was prepared and used in 2 stages, •10 mg/mL sinapinic acid in acetone (Matrix 1) •10 mg/mL sinapinic acid in 60:40

Figure 2. Example of resolution in reflectron mode (negative). Observed resolution ≈ 13,000.

Figure 4. Example of resolution in linear mode (positive). Observed resolution ≈ 1350.

Page 4: MALDI micro MX RESOLUTION - waters.com · Matrix: The matrix was prepared and used in 2 stages, •10 mg/mL sinapinic acid in acetone (Matrix 1) •10 mg/mL sinapinic acid in 60:40

Figure 5. Example of resolution in linear mode (negative). Observed resolution ≈ 1150.

CONCLUSION• In this test the MALDI micro MX always surpassed the instrument specifications.• In reflectron, positive ion mode a resolution of ca 16,000 was measured, 60 % better than the specified value of 10,000

REFERENCE1. Tyler, A. N.; Clayton, E.; Green, B. N. Anal. Chem. 1996, 68, 3561-3569.

Waters, Micromass and MALDI micro MX are trademarks of Waters Corporation.All other trademarks are the property of their respective owners.©2005 Waters Corporation Produced in the U.S.A. April 2005 720001152EN AW-PDF


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