Helpful Info

Sample Analysis

There are several instrumental parameters that can utilized to provide optimal data acquisition. The descriptions below should be useful when completing the sample submission form to ensure that your analytical experiments are conducted in the best possible manner. Utilize the descriptions below to help you understand and make the best decisions about how you would like your samples to be analyzed.


Electrospray Ionization – ESI

ESI is an atmospheric pressure, soft ionization technique that utilizes high voltages to generate quasi-molecular ions, [M + H]+ or [M – H]-, from non-volatile, polar liquid-phase analytes. The ions generated are then desolvated over a defined region and introduced in the mass spectrometer inlet as distinct gas phase ions. Electrospray ionization can be used on small molecules ranging in molecular weights from 50-2000 Da. Multiple charging of peptides and proteins can be achieved to enable analysis of very large molecular weight species at lower molecular weights, and data can be transformed to enable parent ion molecular weight determination.

Atmospheric Pressure Chemical Ionization – APCI

In APCI, a sample solution flows through a heated tube where it is volatilized and sprayed into a corona discharge to generate a plasma. Inside the plasma, proton transfer reactions occur to generate charged analyte species. APCI produces [M+H]+ ions but the source parameters must be optimized to avoid significant fragmentation. Polar, semi-volatile samples are the best candidates for analysis by APCI. Note that APCI has the ability to ionize less polar compounds than that of ESI.

Atmospheric Pressure Photonionization – APPI

APPI is ideal for non-polar speciation, aromatics, thiophenes, furans, et cetera, of compounds. LC eluent or sample is directly infused into a nebulizing tube to aid in desolvation of the sample. The desolvated ions are then exposed to a Krypton UV lamp, 120 nm photos ~10ev, where ionization can occur through direct ionization or proton-transfer reactions to generate either [M+H]+or M+⋅. The charged species are then swept into the mass spectrometry inlet for mass analysis. APPI is a soft ionization technique but ionization often time requires the selection of a dopant to increase ionization efficiency and sensitivity.


Positive ion electrospray analysis is commonly used to generate positively charged molecular ions through protonation to yield [M+H]+, where M signifies the molecular compound, and H, the additional proton. The m/z value at which one detects a singly charged ion is therefore 1 Dalton, Da, higher in mass than the molecular weight. Basic chemical functionalities that are most frequently ionized in the positive mode include basic nitrogen, pyridinic, furans and inorganic cations.

Negative ion electrospray analysis commonly used to generate negatively charged molecular ions through deprotonation to yield [M-H]-, where M signifies the molecular compound, and H, the lost proton. The m/z value where one would detect a singly charged deprotonated molecular ion is 1 Da lower in mass than the molecular weight. Acidic chemical functionalities that are most frequently ionized include neutral nitrogen, pyrrole, carboxylic acids and inorganic anions.

In special instances, sample adduction may be required to produce efficient ionization. Adduct formation is carried out in the solution phase prior to sample introduction. In negative ion ESI, the ionization of polar, neutral molecules or very weakly acidic species that do not generate stable negative ions through deprotonation often times form adducts with chloride ions. The resultant ion of increased molecular weight, +35 in the case of chloride, but possessing a single negative charge. In positive mode ESI, the addition of a salt, in very small quantities, less than 1mM, can facilitate ionization through the formation of a positively charged adduct with sodium, lithium, or ammonium. The observed positive ions result in a molecular weight increase equivalent to that of the salt ion, +23 for sodium, et cetera, and typically carry a single positive charge.