Principles of Mass Spectrometry

Mass spectrometry works by converting substances into charged particles (ions) that can be manipulated and analyzed. In Time of Flight (ToF) mass spectrometry, particles are first ionized to form 1+ ions, then accelerated to equal kinetic energy, and finally measured based on how long they take to travel a fixed distance.

The ionization stage can occur through different methods. Electron impact (or electron ionization) involves bombarding vaporized samples with high-energy electrons from a heated filament, knocking off electrons to create positive ions. This process can be represented as: X(g) → X⁺(g) + e⁻.

Once ionized, the particles are attracted toward a negatively charged electric plate that accelerates them into the next stage of the instrument. This technique works particularly well for elements and lighter molecules that can be easily vaporized.

💡 Think of electron impact like a snooker break - the high-energy electrons are the cue ball that strikes the sample molecules, knocking electrons away and leaving positively charged ions behind.

Ionization Methods

Electron impact ionization is ideal for analyzing elements and compounds with lower formula mass. When molecules are ionized this way, the resulting 1+ ion is called a molecular ion. For example, methane undergoes the reaction: CH₄(g) → CH₄⁺(g) + e⁻. Though not required for exams, it's worth knowing that these molecular ions often fragment into smaller pieces that also appear in the spectrum.

Electrospray ionization works differently and is perfect for larger molecules like proteins. The sample is dissolved in a volatile solvent (like water or methanol) and sprayed through a fine needle attached to a positive voltage. As the tiny droplets leave the needle, they gain a proton (H⁺) from the solvent, creating XH⁺ ions with a mass of M + 1.

The reaction can be shown as: X(g) + H⁺ → XH⁺(g). As the solvent evaporates, the charged particles are drawn toward a negative plate for acceleration. This is considered a "soft" ionization technique because it rarely causes the molecules to break apart.

💡 Electrospray ionization is like adding a backpack (a proton) to each molecule rather than breaking bits off them - that's why it's called a "soft" technique!

Acceleration and Kinetic Energy

After ionization, all positive ions are accelerated through an electric field until they reach the same kinetic energy. This is where the physics gets interesting! Kinetic energy is calculated using the formula: KE = ½mv².

Since all particles now have identical kinetic energy but different masses, their velocities must differ. Using the equation v = √$2KE/m$, we can see that lighter particles will travel faster than heavier ones. This difference in velocity is the key to separating ions based on their mass.

The ions then pass through a hole in the negatively charged plate into a flight tube. The time each ion takes to travel through this tube depends entirely on its mass—the lighter the ion, the faster it moves and the sooner it reaches the detector.

💡 Imagine runners with different weights all given the same amount of energy - the lighter runners will naturally move faster and reach the finish line first, just like lighter ions in the flight tube!

The Flight Tube

In the flight tube, ions travel at speeds determined by their mass. The relationship between flight time and mass is given by: t = d√$m/2KE$, where t is time, d is the length of the flight tube, m is particle mass, and KE is kinetic energy.

This equation reveals that flight time is proportional to the square root of mass. So if one ion is four times heavier than another, it will take twice as long to travel the same distance. This principle allows us to separate isotopes of the same element that differ by just one or two neutrons.

For example, the three isotopes of magnesium (²⁴Mg, ²⁵Mg, and ²⁶Mg) will separate in the flight tube, with ²⁴Mg reaching the detector first, followed by ²⁵Mg, and finally ²⁶Mg. All ions begin their journey together, but gradually separate based on their mass differences.

💡 The flight tube works like a race where everyone starts at the same time but finishes at different times based on their weight—no need for fancy separation equipment, just physics doing the work!

Detection and Mass Spectra

When ions reach the end of the flight tube, they hit a negatively charged detector plate. As the positive ions contact this plate, they gain electrons and generate a measurable electric current. The size of this current indicates the number of ions hitting the plate at a particular time.

A computer processes this data to create a mass spectrum, which plots the mass-to-charge ratio $m/z$ against relative abundance. Since most ions carry a 1+ charge, the m/z value effectively shows the mass of each ion. The spectrum appears as a series of peaks, with each peak representing a different ion.

The mass spectrum of magnesium, for example, shows three distinct peaks at m/z values of 24.0, 25.0, and 26.0, corresponding to its three isotopes. From this data, we can calculate the relative atomic mass using the formula: Ar = (sum of [abundance × mass for each isotope]) ÷ (total abundance)

For magnesium: Ar = $(79.0 × 24.0) + (10.0 × 25.0) + (11.0 × 26.0)$ ÷ (79.0 + 10.0 + 11.0) = 24.3

💡 Think of the mass spectrum as a population census that tells you not just who's present, but exactly how many of each type of particle exists in your sample!

Interpreting Mass Spectra

When analyzing molecular compounds, the peak with the highest m/z value usually represents the molecular ion, giving us the relative molecular mass of the compound. For example, in propane's mass spectrum (following electron impact ionization), the highest significant peak appears at m/z 44, indicating its molecular mass.

Small peaks at slightly higher m/z values $like m/z 45 for propane$ often indicate molecular ions containing heavier isotopes such as ¹³C or ²H. Peaks at lower m/z values typically represent fragments of the original molecule, though you won't need to analyze these for your exams.

Proteins analyzed by electrospray ionization show different patterns. Since they gain a proton during ionization (creating MH⁺ ions), their relative molecular mass is calculated by subtracting 1 from the m/z value of the main peak. In the example shown, the protein's MH⁺ peak appears at 521.1, meaning its actual relative molecular mass is 520.1.

💡 Reading a mass spectrum is like decoding a molecular fingerprint—the highest peak shows the identity of the molecule, while smaller peaks reveal its isotopic makeup!