Assessing the Photo-degradation of Biopharmaceutical Compounds by HPLC-MS (CAT#: STEM-B-0401-CJ)

1. Solvent Pumping and Sample Injection in HPLC.
2. Sample Detection and Identification in HPLC.
3. Sample Introduction: sample is converted into a gaseous phase (except with gaseous samples or samples that are thermally unstable) and is introduced through the inlet to the ionization chamber.
4. Ionization: gaseous sample is ionized to generate cations (in most cases but a few types of MS work with anions).
5. Separation: ions separate according to their mass/charge ratio by a mass analyzer.
6. Detection: a detector is used to determine the species and quantity of each ion.

Biopharmaceutica

In HPLC-MS, the two techniques (HPLC and MS) are connected by an interface that transfers the separated components from the liquid chromatograph column into the mass spectrometer ion source. An interface is needed since HPLC operates at high pressure and the MS system has a high vacuum.

HPLC includes a mobile phase and a stationary phase, both of which can be modified to suit the sample matrix and the desired properties to be determined. Usually, the mobile phase is adjusted to suit the sample, and the stationary phase is adjusted to work well with the mobile phase. The degree of compound separation is based on the compound’s affinity for the mobile phase.

HPLC methods can be divided into two main categories based on the properties of the stationary and mobile phases. A combination of polar stationary phase and non-polar mobile phase is called "normal-phase chromatography" and the opposite of that, the combination of non-polar stationery and polar mobile phase is called "reverse-phase chromatography".

• Sample: Peptides, Proteins, Vaccines, Virus-like particles, Antibody
• Equipment: Chemyx Fusion 100-X Syringe Pump, Pneumatic intensifier, Reciprocating pumps, Mass spectrometer

HPLC-MS is suitable for samples in liquid form.