Determination of acetylsalicylic acid and salicylic acid in blood and plasma by HPLC

Abstract

A method is described for the determination of acetylsalicylic acid and salicylic acid in human whole blood and plasma which uses liquid chromatography with UV detection. The enzymatic degradation of acetylsalicyclic acid in blood and plasma is examined. Addition of potassium fluoride as enzyme inhibitor and deep freezing after plasma preparation allows storage of plasma for one week. A mixture of acetonitrile and diluted acetic acid as mobile phase give the best peak shape. Traces of iron lead to slight peak broadening; addition of EDTA results in excessive tailing. The detection limit for acetylsalicylic acid and salicylic acid is about 250 ng/ml for a 1.0 ml primary sample. The method is sensitive enough for the monitoring of both drugs in bioavailability studies.

Review of HPLC methods and HPLC methods with mass spectrometric detection for direct determination of aspirin with its metabolite(s) in various biological matrices

“…ASA (p K a 3.5) and other metabolites, most researchers preferred to use acidic substances, e.g. p ‐toluic acid (Amick and Mason ; Rumble et al ., ; Bakar and Niazi ; Gaspari and Locatelli, ), benzoic acid and its analogs (Lo and Bye ; Buskin et al ., ; Nieder and Jaeger ; Kees et al ., ), as candidates for IS. Other examples include the use of simvastatin (p K a 5.5; Bae et al ., ) and a structure analog of SA (Xu et al ., ) on a mass spectrometer.…”

“…dichloromethane (Harrison et al, 1980;Mays et al, 1984;Klimes et al, 1992) and chloroform (Lo and Bye, 1980). In addition to two methods discussed in beginning ethereal extraction approach was also followed by several groups (Wahlin-Boll et al, 1981;Nieder and Jaeger, 1983;Bakar and Niazi, 1983;Brandon et al, 1985;Siebert and Bochner, 1987;Shen et al, 1990;Bae et al, 2008). All of these methods were developed on a UV detector (229-280 nm) for plasma and at 305 nm for urine (Bakar and Niazi, 1983), except for the reports of Siebert and Bochner (1987) and Bae et al (2008).…”

“…Therefore blood specimens should be collected in chilled tubes containing esterase inhibitor. Nieder and Jaeger () published the results of extensive studies on ASA hydrolysis in biological matrices and concluded that enzymatic hydrolysis of ASA is drastically reduced by potassium fluoride; with additional cooling, the loss of ASA was less than 1% in 15 min in plasma. Further, no detectable loss was seen in 5 days for frozen plasma samples (at 1 °C) while only a 5–10% loss was observed after 4 weeks of storage.…”

Sensitive determination of aspirin and its metabolites in plasma by LC–UV using on-line solid-phase extraction with methylcellulose-immobilized anion-exchange restricted access media

“…Most of these methods utilized an off-line sample procedure involving solvent extraction and therefore required evaporation and reconstitution steps. Since evaporation leads to the loss of ASA and/or SA because of sublimation [24,32,35,[44][45][46][47], the use of such procedures generally results in reduced recovery of the compounds being analyzed.…”

Comparative Analytical Study for Determination of Acetylsalicylic Acid in Bulk and in Pharmaceutical Formulations

“…Several analytical methods were developed to prepared aspirin alone or with other drugs [15,16] including liquid chromatography, UV derivative spectrophotometric method [17].…”