Extraction of Plasma Catecholamines and Metanephrines using SPE Prior to LC-MS/MS Analysis

by | Articles, Bioclinical Analysis

Biotage has produced an application note describing a mixed-mode weak cation exchange SPE protocol for extraction of three catecholamines and three metanephrine metabolites from plasma prior to LC-MS/MS detection.

The method described in this application achieves high reproducible recoveries for a number of common catecholamine analytes in plasma. The application demonstrates how EVOLUTE® EXPRESS SPE products dramatically improve flow characteristics and enhance sample preparation productivity. By truly eliminating the need for column conditioning and equilibration, samples can be prepared using a simple, fast load-wash-elute procedure.

Epinephrine, Norepinephrine, Dopamine, Metanephrine, Normetanephrine and 3-Methoxytyramine (Sigma Aldrich Chemical Co, Poole, UK).

Internal Standards
D6-Epinephrine, D6-Norepinephrine, D4-Dopamine, D3-Metanephrine, D3-Normetanephrine (Sigma Aldrich Chemical Co, Poole, UK).

The solid-phase extraction procedure, UHPLC and MS conditions are given in the application note.

Extraction recoveries were calculated at a concentration of 5 ng/mL. Data is summarized from the results of n ≥ 7 extracted samples compared to n ≥ 4 blank samples fortified with an amount of analytes equivalent to 100% after extraction. Linearity was determined between 0.04 and 1.28 ng/mL for norepinephrine and dopamine and between 0.02 and 1.28 ng/mL for epinephrine, normetanephrine, metanephrine and 3-methoxytyramine. Example calibration lines using a manual full extraction can be found within the application note for each component.

Published  Sep 27, 2017

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