Assay for the simultaneous determination of acetaminophen–caffeine–butalbital in human serum using a monolithic column

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 36; no. 4; pp. 737 - 741
• Main Authors: Pistos, C., Stewart, J.T.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 19.11.2004; Elsevier Science
• Subjects: Acetaminophen - blood; Acetaminophen - chemistry; Acetaminophen–caffeine–butalbital; Analysis; Analytical, structural and metabolic biochemistry; Barbiturates - blood; Barbiturates - chemistry; Biological and medical sciences; Caffeine - blood; Caffeine - chemistry; Chromatography, High Pressure Liquid - methods; Drug Combinations; Fundamental and applied biological sciences. Psychology; General pharmacology; HPLC; Humans; Medical sciences; Monolithic column; Pharmacology. Drug treatments; Serum; Serum; Acetaminophen–caffeine–butalbital; HPLC; Monolithic column; Human; Acetaminophen-caffeine-butalbital; Biological fluid; CNS stimulant; Psychotropic; Antimigrainous agent; HPLC chromatography; Hypnotic; Respiratory analeptic; Butalbital; Paracetamol; Analgesic; Simultaneous measurement; Antipyretic; Xanthine derivatives; Sedative; Barbiturates; Caffeine
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2004.07.042

A fast and sensitive high performance liquid chromatography (HPLC) assay was developed on a C18 monolithic column for the simultaneous determination of acetaminophen–caffeine–butalbital in human serum. Serum samples were treated with a solid phase extraction procedure. The analytes were separated using a mobile phase of 95:5 (v/v) 0.1 M potassium phosphate monobasic (pH 2.41)–acetonitrile on the C18 monolithic column with detection at 220 nm. Benzoic acid was used as the internal standard (IS). The method was validated over the range of 1.25–100 μg/ml for each drug and found to be linear ( r > 0.995, n = 12) with RSD less than 8.3%. The method proved to be accurate (percent bias for all calibration samples varied from −14.6 to −1.3%) and precise (ranged from 2.9 to 13.4%). The mean percent absolute recoveries from serum were 89.7 ± 3.6 for acetaminophen, 95.5 ± 4.5 for caffeine, 99 ± 5.2 for butalbital and 83.4 ± 3.9% for the internal standard.