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Arzneimittelforschung 2011; 61(5): 301-311
DOI: 10.1055/s-0031-1296203
Analgesics · Anti-inflammatories · Antiphlogistics · Antirheumatic Drugs
Editio Cantor Verlag Aulendorf (Germany)

Simultaneous extraction of acetylsalicylic acid and salicylic acid from human plasma and simultaneous estimation by liquid chromatography and atmospheric pressure chemical ionization/tandem mass spectrometry detection

Ramakrishna Nirogi
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
,
Vishwottam Kandikere
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
,
Koteshwara Mudigonda
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
,
Devender Ajjala
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
,
Ramakrishna Suraneni
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
,
Parthasarathi Thoddi
1   Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Hyderabad, India
› Author Affiliations
Further Information

Publication History

Publication Date:
27 November 2011 (online)

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Abstract

A simple analytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in atmospheric chemical ionization mode (APCI) for the simultaneous estimation of acetylsalicylic acid (ASA, CAS 50-78-2) and its active metabolite salicylic acid (SA, CAS 69-72-7) in human plasma has been developed and validated. ASA and SA were analyzed simultaneously despite differences in plasma concentration ranges of ASA and SA after oral administration of ASA. In spite of having different chemical, ionization and chromatographic properties, ASA and SA were extracted simultaneously from the plasma sample using acetonitrile protein precipitation followed by liquid-liquid extraction. The analytes were separated on a reversed phase column with rapid gradient program using mobile phase consisting of ammonium acetate buffer and methanol. The structural analogue diclofenac was used as an internal standard. The multiple reaction monitoring (MRM) transitions m/z 179 → 137 for ASA, m/z 137 → 65 for SA and m/z 294 → 250 for IS were used. The assay exhibited a linear dynamic range of 0.02–10 μg/mL for ASA and 0.1–50 μg/mL for SA. The between-batch precision (%CV) ranged from 2.1 to 7.9% for ASA and from 0.2 to 5.2% for SA. The between-batch accuracy ranged from 95.4 to 96.7% for ASA and from 94.6 to 111.3% for SA. The validated method was successfully applied for the evaluation of pharmacokinetics of ASA after single oral administration of 650 mg test formulation versus two 325 mg reference formulations of ASA in human subjects.

Key words

Acetylsalicylic acid - Atmospheric chemical ionization mode - CAS 50-78-2 - CAS 69-72-7 - Salicylic acid - Tandem mass spectrometry
 

  • References

  • 1 Julian DG, Chamberlain DA, Pocock SJ.. A comparison of aspirin and anticoagulation following thrombolysis for myocardial infarction (the AFTER study): a multicentre unblended randomized clinical trial. Br Med J. 1996; 313: 1429-31
    CrossrefPubMedGoogle Scholar
  • 2 Warner TD, Mitchell JA.. Cyclooxygenase-3 (COX-3): Filling in the gaps toward a COX continuum?. PNAS. 2002; 99: 13371-3
    CrossrefPubMedGoogle Scholar
  • 3 Lenz T, Wilson A.. Clinical pharmacokinetics of antiplatelet agents used in the secondary prevention of stroke. Clin Pharmacokinet. 2003; 42: 909-20
    CrossrefPubMedGoogle Scholar
  • 4 Awtry EH, Loscalzo J.. Aspirin. Circulation. 2000; 101: 1206-18
    CrossrefPubMedGoogle Scholar
  • 5 Bochner F, Lloyd JV.. Aspirin for myocardial infarction: clinical pharmacokinetic considerations. Clin Pharmacokinet. 1995; 28: 433-8
    CrossrefPubMedGoogle Scholar
  • 6 Trinder P.. Rapid determination of salicylate in biological fluids. Biochem J. 1954; 57: 301-3
    PubMedGoogle Scholar
  • 7 Bakar SK, Niazi S.. High performance chromatographic determination of aspirin and its metabolites in plasma and urine. J Pharm Sci. 1983; 72: 1020-3
    CrossrefPubMedGoogle Scholar
  • 8 Kees F, Jehnich D, Grobecker H. et al Simultaneous determination of acetylsalicylic acid and salicylic acid in human plasma by high-performance liquid chromatography. J Chro-matogrB. 1996; 677: 172-7
    PubMedGoogle Scholar
  • 9 McMahon GP, Kelly MT.. Determination of aspirin and salicylic acid in human plasma by column-switching liquid Chromatograph using on-line solid-phase extraction. Anal Chem. 1998; 70: 409-14
    CrossrefPubMedGoogle Scholar
  • 10 O’kruk RJ, Adams MA, Philip RB.. Rapid and sensitive determination of acetylsalicylic acid and its metabolites using reversed-phase high performance liquid chromatography. J Chromatogr B. 1984; 310: 343-52
    PubMedGoogle Scholar
  • 11 Mays DC, Sharp DE, Beach CA, Kershaw RA, Bianchine JR, Gerber N.. Improved method for the determination of aspirin and its metabolites in biological liquid by high performance liquid chromatography: application to human and animal studies. J Chromatogr B. 1984; 311: 301-9
    PubMedGoogle Scholar
  • 12 Peng GW, Gadalla MA, Smith V, Peng A, Ghiou WL.. Simple and rapid high pressure liquid chromatographic simultaneous determination of aspirin, salicylic acid, and slicylu-ric acid in plasma. J Pharm Sei. 1978; 67: 710-20
    CrossrefPubMedGoogle Scholar
  • 13 Shen J, Wanwimolruk S, Roberts MS, Clerk CR. A sensitive assay for aspirin and its metabolites using reversed-phase ion-pair high performance liquid chromatography. J Liq Chromatogr Rel Technol. 1990; 13: 751-61
    CrossrefPubMedGoogle Scholar
  • 14 Walter LJ, Biggs DF, Coutts RT. Simultaneous GLC estimation of salicylic acid and aspirin in plasma. J Pharm Sei. 1974; 63: 1754-8
    CrossrefPubMedGoogle Scholar
  • 15 Ranee MJ, Jordan BJ, Nichols JD. A simultaneous determination of acetylsalicylic acid, salicylic acid, and salicyla-mide in plasma. J Pharm Pharmacol. 1975; 27: 425-9
    CrossrefPubMedGoogle Scholar
  • 16 Hanse SH, Jensen ME, Bjorsdottir I. Assay of acetylsalicylic acid and three of its metabolites in human plasma and urine using non-aqueous capillary electrophoresis reversed electro osmotic flow. J Pharm Biomed Anal. 1998; 17: 1155-60
    CrossrefPubMedGoogle Scholar
  • 17 Bae SK, Seo KA, Jung EJ, Kim HS, Yeo CW, Shon JH, et al.. Determination of acetylsalicylic acid and its major metabolite salicylic acid, in human plasma using liquid chroma-tograph-tandem mass spectrometry: application to pharmacokinetics study of Astrix in Korean healthy volunteers. Biomed Chromatogr. 2008; 22: 590-5
    CrossrefPubMedGoogle Scholar
  • 18 Xu X, Koetzner L, Boulet J, Maselli H, Beyenhof J, Grover G. Rapid and sensitive determination of acetylsalicylic acid and salicylic acid in plasma using liquid chromatography-tandem mass spectrometry: application to pharmacokinetic study. Biomed Chromatogr. 2009; 23: 973-9
    CrossrefPubMedGoogle Scholar
  • 19 Gangl ET, Annan MM, Spooner N, Vouros P. Reduction of signal suppression effects in ESI-MS using a nanosplitting device. Anal Chem. 2001; 73: 5635-44
    CrossrefPubMedGoogle Scholar
  • 20 Matuszewski BK, Constanzer ML, Chavez-Eng CM. Matrix effect in quantitative LC/MS/MS analyses of biological fluids: a method for determination of finasteride in human plasma at pictogram per milliliter concentrations. Anal Chem. 1998; 70: 882-9
    CrossrefPubMedGoogle Scholar
  • 21 Borts DJ, Bowers LD.. Direct measurement of urinary testosterone and epitestosterone conjugates using high-performance liquid chromatography/tandem mass spectrometry. J Mass Spectrom. 2000; 35: 50-61
    CrossrefPubMedGoogle Scholar
  • 22 King R, Bonfiglio R, Femandez-Metzler C, Miller-Stein C, Olah T. Mechanistic investigation of ionization suppression in electrospray ionization. J Am Soc Mass Spectrom. 2000; 11: 942-50
    CrossrefPubMedGoogle Scholar
  • 23 Klimes J, Sochor J, Zahradnicek M, Sedlacek J. Simultaneous high performance liquid chromatographic determination of salicylates in whole blood plasma and isolated erythrocytes. J Chromatogr. 1992; 584: 221-8
    CrossrefPubMedGoogle Scholar
  • 24 Gaspari F, Locatelli M. Determination of aspirin and salicylic acid in uremic patients’ plasma using reversed-phase high performance liquid chromatography. Ther Drug Monit. 1987; 9: 243-
    CrossrefPubMedGoogle Scholar
  • 25 Brandon RA, Eadie MJ, Smith MT. A sensitive liquid chromatographic assay for plasma aspirin and salicylate concentrations after low doses of aspirin. Ther Drug Monit. 1985; 7: 216-21
    CrossrefPubMedGoogle Scholar
  • 26 Nieder M. Jaeger H. J High Resolut Chromatogr Chromatogr Commun. 1983; 6: 655-60
    CrossrefPubMedGoogle Scholar
  • 27 Buskin JN, Upton RA, Williams RL. Improved liquid chromatography of aspirin, salicylate, and salicyluric acid in plasma, with a modification for determining aspirin metabolites in urine. Clin Chem. 1982; 28: 1200-
    PubMedGoogle Scholar
  • 28 Lo LY, Bye A.. Specific and sensitive method for the determination of aspirin and salicylic acid in plasma using reversed-phase high performance liquid chromatography. J Chromatogr. 1980; 181: 473-7
    CrossrefPubMedGoogle Scholar
  • 29 Harrison LI, Funk ML, Ober RE. High pressure liquid chromatographic determination of salicylsalicylic acid, aspirin and salicylic acid in human plasma and urine. J Pharm Sci. 1980; 69: 1268-71
    CrossrefPubMedGoogle Scholar
  • 30 Liu JH, Smith PC. Direct analysis of salicylic acid, salicyl acyl glucuronide, salicyluric acid and gentisic acid in human plasma and urine by high performance liquid chromatography. J Chromatogr B. 1996; 675: 61-70
    PubMedGoogle Scholar
  • 31 Krivosikova Z, Spustova V, Dzurik R. A highly sensitive HPLC method for the simultaneous determination of acetylsalicylic, salicylic and salicyluric acids in biological fluids: pharmacokinetic, metabolic and monitoring implications. Methods Find Exp Clin Pharmacol. 1996; 18: 527-32
    PubMedGoogle Scholar
  • 32 Coudray C, Mangournet C, Bouhadjeb S, Faure H, Favier A. Rapid high performance liquid chromatographic assay for salicylic acid in plasma without solvent extraction. J Chromatogr Sci. 1996; 34: 166-73
    PubMedGoogle Scholar
  • 33 Rumble RH, Roberts MS, Wanwimolruk S. Determination of aspirin and its major metabolites in plasma by high performance liquid chromatography without solvent extraction. J Chromatogr. 1981; 225: 252-60
    CrossrefPubMedGoogle Scholar
  • 34 Cham BE, Ross-Lee L, Bochner F, Imhoff DM. Measurement and pharmacokinetics of acetylsalicylic acid by a novel high performance liquid chromatographic assay. Ther Drug Monit. 1980; 2: 365-72
    PubMedGoogle Scholar
  • 35 Reilly IAG, Fitzgerald GA.. Aspirin in cardiovascular disease. Drugs. 1988; 35: 154-76
    PubMedGoogle Scholar
  • 36 Sorense S. Improved measurement of acetullsalicylic acid and esterase in serum. Clin Chem. 1983; 29: 491-4
    PubMedGoogle Scholar
  • 37 James GT. Inactivation of the protease inhibitor phenyl-methylsulfonyl fluoride in buffers. Anal Biochem. 1978; 86: 574-9
    CrossrefPubMedGoogle Scholar