Simple method for the determination of 5-aminosalicylic and N-acetyl-5-aminosalicylic acid in rectal tissue biopsies.
We describe a sensitive high-performance liquid chromatographic method for the determination of 5-aminosalicylic acid and N-acetyl-5-aminosalicylic acid in rectal tissue biopsies. Samples were derivatised using propionic anhydride and proteins were precipitated with methanol. A Supelcosil ABZ column (150x4.6 mm I.D., 5 microm silica particles) was used with a mobile phase comprising 0.1 M acetic acid, acetonitrile and triethylamine (1600:114:6, v/v/v). Fluorescence detection was employed and detection limits were 0.2 ng/mg tissue at a signal-to-noise ratio of three (measured concentration: 5-aminosalicylic acid, 0.254 (0.228-0.286) ng/mg, C.V. 10.7%; N-acetyl-5-aminosalicylic acid, 0.18 (0.154-0.198) ng/mg, C.V 9.8%). This assay was validated for use with serum, urine and faecal samples for which it proved to be both precise and accurate (C.V.<10%, measured concentration within 10%).
Hussain
F N
FN
Department of Gastroenterology, Northern General Hospital, Sheffield, UK.
Ajjan
R A
RA
Moustafa
M
M
Anderson
J C
JC
Riley
S A
SA
D000636:Aminosalicylic Acids / Q000032:analysis* / Q000097:blood / Q000652:urine; D000812:Anhydrides; D001706:Biopsy; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003093:Colitis, Ulcerative / Q000188:drug therapy; D005243:Feces / Q000737:chemistry; D006801:Humans; D007202:Indicators and Reagents; D007413:Intestinal Mucosa / Q000737:chemistry; D007700:Kinetics; D019804:Mesalamine / Q000032:analysis* / Q000097:blood / Q000652:urine; D000432:Methanol; D011422:Propionates; D012007:Rectum / Q000737:chemistry*; D012680:Sensitivity and Specificity; D007507:Therapeutic Irrigation
D000636:Aminosalicylic Acids; D000812:Anhydrides; D007202:Indicators and Reagents; D011422:Propionates; D019804:Mesalamine; C028836:N-acetyl-5-aminosalicylic acid; C096126:propionic anhydride; D000432:Methanol