Name: how maldi turns samples into ions
Uploaded: 2025-12-15T15:55:27.000Z
Duration: 1 min 8 s
Channel: JoVE

Matrix-assisted laser desorption ionization spectrometry or MALDI is an ionization technique employed to acquire accurate molecular mass data for polar biopolymers with molecular masses ranging from a few to a hundred thousand Daltons.
The sample preparation process involves mixing an analyte solution with a matrix material, such as 2,5-dihydroxybenzoic acid. This mixture is then deposited on a probe inserted into the mass spectrometer.
When the ultraviolet or infrared laser beam is focused on the surface, the matrix absorbs the laser energy, leading to desorption and ionization of sample molecules through a proton transfer from the matrix to the analyte molecule, generating protonated ions.
The newly formed ions are accelerated by an electric field and separated based on their mass-to-charge ratio in a time-of-flight mass analyzer.
The resulting mass spectrum can be analyzed to determine the molecular mass of the biomolecule under investigation.

matrix-geassisteerde laser desorptie ionisatie (maldi)

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-geassisteerde Laser Desorptie Ionisatie (MALDI) is een krachtige analytische techniek die wordt gebruikt in massaspectrometrie. Het maakt de identificatie en karakterisering van verschillende biomoleculen mogelijk, waaronder eiwitten, peptiden, nucle&#239;nezuren en koolhydraten. MALDI is een ionisatietechniek die veel wordt gebruikt in biologisch en medisch onderzoek, evenals in vakgebieden zoals farmacologie en biochemie.
De analyt van belang, een biomolecuul of een mengsel van biomoleculen, wordt gemengd met een geschikt matrixmateriaal. De matrix is &#8203;&#8203;meestal een organische verbinding, zoals een klein organisch zuur of een kristallijne substantie, die ionisatie vergemakkelijkt. Dit analyt-matrixmengsel wordt vervolgens op een doeloppervlak afgezet, in de massaspectrometer ingebracht en gedroogd. Tijdens het drogen vormt de matrix kleine kristallen die de analytmoleculen in hun structuur opnemen.
Zodra het monster is voorbereid, wordt een gepulseerde laser op het doeloppervlak gericht. De matrixmoleculen absorberen de laserstraalenergie, waardoor ze opwarmen en snel verdampen. Dit proces staat bekend als laserdesorptie. Terwijl de matrixmoleculen verdampen, dragen ze de ingebedde analytmoleculen. De verdampte analytmoleculen worden ge&#239;oniseerd via een proces dat protonoverdracht wordt genoemd. De protonoverdracht vindt plaats tussen de matrix en het analytmolecuul. Het matrixmolecuul doneert een proton aan het analytmolecuul, wat resulteert in de vorming van geprotoneerde ionen. De analytionen worden vervolgens versneld door een elektrisch veld en verder gescheiden op basis van hun massa-tot-ladingverhouding in een time-of-flight-massaanalysator.
Het massaspectrum verkregen uit MALDI-spectrometrie biedt waardevolle informatie over de massa en relatieve overvloed van de geanalyseerde biomoleculen.

Matrix-geassisteerde Laser Desorptie Ionisatie (MALDI)

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and ...

matrix-assisted-laser-desorption-ionization-maldi

Education

JoVE Core

Analytical Chemistry

Analytische chemie

Unit 1

Mass Spectrometry Fragmentation Methods

Fragmentatiemethoden in de massaspectrometrie

KEY TERMS AND CONCEPTS

KERNTERMEN EN CONCEPTEN

how maldi turns samples into ions

Matrix-assisted laser desorption ionization (MALDI) is a mass spectrometry technique that turns biomolecules into ions for analysis. It is used to identify and characterize proteins, peptides, nucleic acids, and carbohydrates. Because of this, it is common in biological and medical research, as well as pharmacology and biochemistry.

In MALDI, the analyte of interest is mixed with a matrix material. The matrix is usually an organic compound, such as a small organic acid or a crystalline substance, that helps ionization. The analyte-matrix mixture is placed on a target surface, inserted into the mass spectrometer, and dried. As the sample dries, the matrix forms small crystals that trap the analyte molecules inside them.

A pulsed laser is then aimed at the target surface. The matrix absorbs the laser energy, heats up, and vaporizes quickly. This step is called laser desorption. As the matrix leaves the surface, it carries the embedded analyte molecules with it.

The vaporized analyte molecules become ionized by proton transfer. In this step, the matrix donates a proton to the analyte molecule. This forms protonated ions, which can then be accelerated by an electric field. The ions are separated in a time-of-flight mass analyzer based on their mass-to-charge ratio, and the resulting mass spectrum shows the mass and relative abundance of the biomolecules studied.

How MALDI Turns Samples Into Ions

mass spectrometry: long-chain alkane fragmentation

mass-spectrometry-long-chain-alkane-fragmentation

Mass Spectra of Linear Alkane Chains

Mass spectra of linear alkanes show a clear fragmentation pattern. The molecular ion often breaks at a carbon-carbon bond away from the end of the chain. This cleavage forms a stable carbocation and a stable radical, so the middle of the mass-to-charge plot usually has the strongest signals while the ends are weaker. As each bond breaks, a methyl group is lost and the main peaks in the spectrum appear 14 u apart. Smaller peaks near each main signal can also appear. These come from secondary...

mass spectrometry: branched alkane fragmentation

mass-spectrometry-branched-alkane-fragmentation

Branched Alkanes in Mass Spectra

Branched alkanes in mass spectrometry fragment quickly because branching can create stable carbocations. When cleavage happens at a branching point, a secondary or tertiary carbocation may form. That stability helps explain why the molecular ion peak is often very weak or missing compared with a linear alkane. The lesson uses 2-methylbutane, neopentane, and n-pentane to show how this pattern appears in mass spectra. 2-methylbutane and neopentane break apart to form secondary and tertiary...

mass spectrometry: cycloalkane fragmentation

mass-spectrometry-cycloalkane-fragmentation

Cycloalkane Ions and Mass Peaks

Cycloalkane ions in mass spectrometry often give strong peaks because the ring structure helps stabilize the molecular ion. Compared with linear or branched alkanes, this added stability changes how the molecule breaks apart. As a result, cycloalkanes can produce prominent signals in the mass spectrum. Cyclohexane is a clear example. Its molecular ion has an m/z of 84, and this peak is often stronger than the signal from hexane, a linear alkane. The closed ring helps hold the ion together,...

mass spectrometry: alkene fragmentation

Alkene Ion Fragmentation Patterns

Alkene ion fragmentation patterns in mass spectrometry begin when the molecule loses one electron from the unsaturated π bond. This ionization forms a stable molecular ion. The ion can then break apart by three main reaction pathways. The most common pathway is cleavage at the allylic position, which is the carbon next to the double bond. This produces an allylic carbocation that is stabilized by resonance. In terminal alkenes, this fragment appears at a mass-to-charge ratio of 41. In internal...

mass spectrometry: cycloalkene fragmentation

mass-spectrometry-cycloalkene-fragmentation

Cycloalkene Side-Chain Cleavage in Mass Spectra

Cycloalkenes can break apart in a mass spectrometer through retro-Diels–Alder fragmentation. In this process, two carbon-carbon bonds in the ring split, and ethene is released. The remaining fragment is a dienyl radical cation that is 28 u less than the molecular ion. This pathway is similar to the fragmentation of cycloalkanes that also releases ethene. The difference is the type of charged fragment that remains. Cycloalkanes form an alkyl species, while cycloalkenes form a dienyl species.

mass spectrometry: alkyne fragmentation

Alkyne Mass Spectra and Key Peaks

Alkyne mass spectra often show strong peaks from two main fragmentation paths. One path breaks the carbon-carbon bond between the alpha and beta carbons of the alkyne bond. This produces a 3-propynyl cation, also called a propargyl cation. In terminal alkynes, this is the only fragmentation path that gives the 3-propynyl cation. The unsubstituted 3-propynyl cation appears at a mass-to-charge ratio of 39. In internal alkynes, the 3-propynyl cation is substituted. For example, 2-pentyne forms a...

mass spectrometry: alcohol fragmentation

Alcohol Fragmentation Patterns in Mass Spectra

Alcohols fragment in mass spectrometry in predictable ways. When an alcohol, written as R-OH, ionizes, it can lose one non-bonded electron from the oxygen atom. That forms a molecular ion, but the molecular ion peak is often weak or may not appear because alcohols break apart quickly. Two main fragmentation routes are important for alcohols: β-cleavage and dehydration. In β-cleavage, the bond at the β-position next to the hydroxyl group breaks. This gives a resonance-stabilized cation and a...

mass spectrometry: aromatic compound fragmentation

mass-spectrometry-aromatic-compound-fragmentation

Mass Spectra of Benzene and Toluene

Mass spectra of aromatic compounds show a strong molecular ion peak. In ionization, the molecule becomes a molecular ion, which appears as a prominent peak in the spectrum. Benzene gives a molecular ion at m/z 78, and toluene gives one at m/z 92. Benzene’s molecular ion is especially stable. It does not fragment readily because breaking the aromatic benzene ring requires a large amount of energy. Alkyl-substituted benzenes behave differently in mass spectrometry. Toluene can lose a hydrogen...

mass spectrometry: amine fragmentation

Amines in Mass Spectra: Nitrogen Rule

Amines can be identified in mass spectrometry by their fragmentation patterns and by the nitrogen rule. The molecular ions of amines often break apart through β-cleavage, which cuts a carbon-carbon bond next to the nitrogen. This process forms an alkyl radical and a resonance-stabilized nitrogen-containing cation. The nitrogen rule helps predict the mass of the molecular ion. A compound with one nitrogen atom, or any odd number of nitrogen atoms, gives a molecular ion with an odd molecular...

mass spectrometry: alkyl halide fragmentation

mass-spectrometry-alkyl-halide-fragmentation

Mass Spectra of Alkyl Halides

Mass spectra of alkyl halides show clear isotope patterns from the halogen atom. Chlorine has two common isotopes, 35Cl and 37Cl, in a 3:1 ratio. Bromine has two common isotopes, 79Br and 81Br, in a 1:1 ratio. These isotope ratios create two molecular ion peaks, [M] and [M + 2], with relative heights that match the halide abundance. This pattern is easy to see in examples such as 2-chloropropane and 1-bromopropane. 2-Chloropropane gives peaks in a 3:1 ratio. 1-Bromopropane gives peaks in a 1:1...

mass spectrometry: aldehyde and ketone fragmentation

mass-spectrometry-aldehyde-and-ketone-fragmentation

Carbonyl Fragment Patterns in Mass Spectra

Aliphatic aldehydes and ketones break apart in mass spectrometry through a few key carbonyl fragment patterns. The main pathways are alpha-cleavage, inductive cleavage, and the McLafferty rearrangement. These patterns help chemists read the mass spectrum and spot a carbonyl group. Alpha-cleavage is a common break next to the carbonyl group. The bond beside the carbonyl splits, and this forms an alkyl radical and an acylium cation. The acylium cation is often seen in the spectrum because it is...

mass spectrometry: carboxylic acid, ester, and amide fragmentation

mass-spectrometry-carboxylic-acid-ester-and-amide-fragmentation

Mass Spectra of Carboxylic Compounds

Mass spectra of carboxylic acids, esters, and amides often show two key fragmentation routes: β-cleavage and McLafferty rearrangement. In β-cleavage, the carbon-carbon bond at the β-position next to the carboxylic group breaks. This produces a neutral radical and a cation. Long-chain compounds can also fragment by McLafferty rearrangement if they have a hydrogen on the γ-carbon. The γ-carbon is the third carbon away from the carboxylic group. This rearrangement gives a radical cation and a...

chemical ionization (ci) mass spectrometry

Using Reagent Gas in CI Mass Spectrometry

Chemical ionization mass spectrometry uses a reagent gas to help detect molecules that break apart too quickly in electron impact ionization. It is a milder ionization method that can extend the life of the ionized analyte, which makes molecular identification easier. In chemical ionization, the sample is mixed with an excess reagent gas. This setup makes the electron impact occur mainly on the reagent gas instead of on the analyte. The charged species formed from the reagent then protonates...

electrospray ionization (esi) mass spectrometry

electrospray-ionization-esi-mass-spectrometry

How ESI Turns Biomolecules Into Ions

Electrospray ionization, or ESI, is a mass spectrometry method used to detect biomolecules that are too large or nonvolatile for conventional electron impact ionization. These molecules may break down before they can ionize or evaporate. ESI helps prevent rapid fragmentation and lets scientists record a mass signal for the whole biomolecule. ESI uses electrical energy to move ions from the liquid phase into the gas phase. The analyte biomolecule is mixed with an ionic liquid, which is a...

matrix-assisted laser desorption ionization (maldi)

Matrix-assisted laser desorption ionization (MALDI) is a mass spectrometry technique that turns biomolecules into ions for analysis. It is used to identify and characterize proteins, peptides, nucleic acids, and carbohydrates. Because of this, it is common in biological and medical research, as well as pharmacology and biochemistry. In MALDI, the analyte of interest is mixed with a matrix material. The matrix is usually an organic compound, such as a small organic acid or a crystalline...

key terms and concepts

Learn how MALDI uses a matrix, laser desorption, and proton transfer to create ions for mass spectrometry analysis of biomolecules.

How MALDI Turns Samples Into Ions | JoVE K12

analytical chemistry

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How MALDI Turns Samples Into Ions

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