High efficiency Amino Acid Analysis meets EP compliance requirements
Andrew D. Schaefer, Senior Scientist/Group Leader, Bio/Pharmaceutical Raw Materials
The European Pharmacopeia 8.0 Amino Acid Analysis 2.2.56, Method 1 – Post-Column Ninhydrin Derivatization, sets the foundation for new compliance requirements for testing of amino acids. Previous iterations of the compendia procedure utilized thin layer chromatography for separation of components coupled with ninhydrin exposure, to visually estimate amino acids. This technique only allowed for qualitative determination of amino acids, as the methodology had limited sensitivity.
The transition of the analytical technique to a high-pressure liquid chromatograph, equipped with an ion-exchange column, post-column derivatization system, and UV/Vis detector has allowed for more sensitive detection of Ninydrin Positive Substances. However the EP based methods employed are cumbersome and inefficient, often requiring 1 to 2 hour run times per injection. This creates lengthy analyses and translates into excessive turnaround times and potential costly delays in manufacturing.
Eurofins Lancaster Laboratories has worked extensively on establishing a compliant platform method which cuts analytical run times in half, while providing selectivity and sensitivity for a wide range of amino acids. This was accomplished by employing higher efficiency ion-exchange columns for separation of amino acids along with a pH and temperature gradient. Eurofins Lancaster Laboratories has significant investment in hardware and has two fully validated HPLC/derivatization systems in operation, dedicated for this analysis.
With sample analysis time greatly decreased, the platform method provides shorter overall TAT in testing, allowing for efficient and controlled scheduling for biopharmaceutical manufacturing. The platform method can also be easily modified and subsequently validated for any monographed amino acid.
Eurofins Lancaster Laboratories has validated several amino acid based materials against our established platform. The validation elements exceed the requirements of the individual monographs and the compendia. Having all method development work complete, turnaround times for establishment of any new amino acid material testing are short. With Eurofins Lancaster Laboratories’ vast capacity and extensive capabilities, we can ensure that critical manufacturing timelines are met for our customers.