A validated HPLC method with electrochemical detection for simultaneous assay of 5-aminosalicylic acid and its metabolite in human plasma.
A high-performance liquid chromatographic method was developed, validated and applied to the simultaneous determination of 5-aminosalicylic acid (5-ASA) and its acetylated metabolite (acetyl-5-ASA) in human plasma. The method involves liquid-liquid extraction with methanol followed by isocratic reversed-phase chromatography on a Kromasil KR100 C(18) column with electrochemical detection. The recovery, selectivity, linearity, precision and accuracy of the method were evaluated from spiked human plasma samples. The effects of mobile phase composition, buffer concentration, mobile phase pH and concentration of organic modifiers on retention of 5-ASA, acetyl 5-ASA and internal standard were investigated. Limits' of detection were 5 ng/mL for 5-ASA and 10 ng/mL for acetyl-5-ASA, respectively. The method can be used for supporting therapeutical drug monitoring and pharmacokinetic studies.
Palumbo
Giancarlo
G
Dipartimento di Medicina Interna, Facoltà di Medicina, Università degli Studi di L'Aquila, Italy. gpalumbo@cc.univaq.it
Bacchi
Simona
S
Primavera
Luisa
L
Palumbo
Paola
P
Carlucci
Giuseppe
G
D000636:Aminosalicylic Acids; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood*; D001682:Biological Availability; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry; D006801:Humans; D019804:Mesalamine / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000636:Aminosalicylic Acids; D000894:Anti-Inflammatory Agents, Non-Steroidal; C505368:acetyl-5-aminosalicylic acid; D019804:Mesalamine