JASMI  Vol.3 No.3 A , November 2013
An Ion-Pair HPLC Method for Simultaneous Determination of Exogenous Phosphocreatine and Its Metabolite Creatine and Related ATP in Rabbit Plasma and RBC: Application to a Pharmacokinetic Study
Author(s) Li Lv, Heng Xi, Guozhu Han*
ABSTRACT


A specific, precise and accurate ion-pair HPLC-UV method has been developed and validated for simultaneous determination of phosphocreatine (PCr), and its metabolite creatine (Cr) as well as related ATP in plasma and red blood cell (RBC) of rabbits. After addition of TMP as IS, the samples were deproteinized with 6% PCA. The analytes were separated on a Kromasil C18 column using a tertiary gradient mobile phase composed of buffer A (0.2% KH2PO4 + 0.08% tetrabutyl ammonium hydrogen sulphate, pH 3.0), buffer B (buffer A adjusted to pH 7.5 with 1 mol/L NaOH) and MeOH. Detection wavelengths were set at 210 nm for PCr and Cr and 260 nm for ATP and TMP. Some blank samples were initially run for baseline subtraction. The linear detection responses were obtained for PCr concentration over a range of 10 - 7500 mg/ml (plasma) and 5 - 2500 mg/ml (RBC) and for both Cr and ATP concentrations of 10 - 1500 mg/ml (plasma) and 5 - 750 mg/ml (RBC) (r > 0.99). The QC samples of 3 analytes showed intra-day and inter-day precisions (RSD) of <10%, accuracy of 97% - 107%. The method was successfully used to simultaneously determine plasma and RBC concentrations of the 3 analytes and to study pharmacokinetics after iv administration of PCr to rabbits.



Cite this paper
L. Lv, H. Xi and G. Han, "An Ion-Pair HPLC Method for Simultaneous Determination of Exogenous Phosphocreatine and Its Metabolite Creatine and Related ATP in Rabbit Plasma and RBC: Application to a Pharmacokinetic Study," Journal of Analytical Sciences, Methods and Instrumentation, Vol. 3 No. 3, 2013, pp. 17-23. doi: 10.4236/jasmi.2013.33A003.
References
[1]   P. Korge, M. L. Silber and P. D. Gollnick, “Effect of Creatine Phosphate on the Contractile Activity in Acutely Failing Rat Heart,” Cardiologia, Vol. 43, No. 12, 1998, pp. 1345-1354.

[2]   V. A. Saks, I. V. Dzhaliashvili, E. A. Konorev, E. Strumia, “Molecular and Cellular Aspects of the Cardioprotective Mechanism of Phosphocreatine,” Biokhimiia, Vol. 57, No. 12, 1992, pp. 1763-1784.

[3]   V. G. Sharov, N. I. Afonskaya, M. Y. Ruda, N. M. Cherpachenko, EYa Pozin, R. A. Markosyan, II Shepeleva, M. B. Samarenko and V. A. Saks, “Protection of Ischemic Myocardium by Exogenous Phosphocreatine (Neoton): Pharmacokinetics of Phosphocreatine, Reduction of Infarct Size, Stabilization of Sarcolemma of Ischemic Cardiomyocytes, and Antithrombotic Action,” Biochemical Medicine and Metabolic Biology, Vol. 35, No. 1, 1986, pp. 101-114. http://dx.doi.org/10.1016/0885-4505(86)90064-2

[4]   E. Strumia, F. Pelliccia and G. D’Ambrosio, “Creatine Phosphate: Pharmacological and Clinical Perspectives,” Advances in Therapy, Vol. 29, No. 2, 2012, pp. 99-123. http://dx.doi.org/10.1007/s12325-011-0091-4

[5]   W. Martindale, “Martindale: The Extra Pharmacopocia,” 31st Edition, The Royal Pharmaceutical Society, 1996.

[6]   P. Korge, M. L. Silber and P. D. Gollnick, “Effect of Creatine Phosphate on the Contractile Activity in Acutely Failing Rat Heart,” Cardiologia, Vol. 43, No. 12, 1998, pp. 1345-1354.

[7]   P. Mastroroberto, L. Di Tommaso and M. Chello, “Creatine Phosphate Protection of the Ischemic Myocardium during Cardiac Surgery,” Current Therapeutic Research, Vol. 51, No. 1, 1992, pp. 37-45.

[8]   G. Prabhakar, L. Vona-Davis, D. Murray, P. Lakhani and G. Murray, “Phosphocreatine Restores High-Energy Phosphates in Ischemic Myocardium: Implication for Off-Pump Cardiacre Vascularization,” Journal of the American College of Surgeons, Vol. 197, No. 5, 2003, pp. 786-791. http://dx.doi.org/10.1016/j.jamcollsurg.2003.05.001

[9]   MYa Ruda, M. B. Samarenko, N. I. Afonskaya and V. A. Saks, “Redtion of Ventricular Arrhythmias by Phosphocreatine (Neoton) in Patients with Acute Myocardial Infarction,” American Heart Journal, Vol. 116, No. 2, 1988, pp. 393-397. http://dx.doi.org/10.1016/0002-8703(88)90611-4

[10]   M. L. Semenovsky, V. I. Shumakov, V. G. Sharov, G. M. Mogilevsky, A. V. Asmolovsky, L. A. Makhotina and V. A. Saks, “Protection of Ischemic Myocardium by Exogenous Phosphocreatine. II. Clinical, Ultrastructural, and Biochemicalevaluations,” Journal of Thoracic and Cardiovascular Surgery, Vol. 94, No. 5, 1987, pp. 762-769.

[11]   E. Lorenzi, G. Piacenza, E. Strumia and R. Borgoglio, “Pharmacokinetics of Phosphocreatine Following Intravenous Administration in Humans and Effect on Blood Levels of ATP,” Cardiologia, Vol. 32, No. 9, 1987, pp. 1031-1034.

[12]   M. Balestrino, M. Lensman, M. Parodi, L. Perasso, R. Rebaudo, R. Melani, S. Polenov and A. Cupello, “Role of Creatine and Phosphocreatine in Neuronal Protection from Anoxic and Ischemic Damage,” Amino Acids, Vol. 23, No. 1-3, 2002, pp. 221-229. http://dx.doi.org/10.1007/s00726-001-0133-3

[13]   G. J. Brewer and T. W. Wallimann, “Protective Effect of the Energy Precursor Creatine against Toxicity of Glutamate and Beta-Amyloid in Rat Hippocampal Neurons,” Journal of Neurochemistry, Vol. 74, No. 5, 2000, pp. 1968-1978. http://dx.doi.org/10.1046/j.1471-4159.2000.0741968.x

[14]   M. A. Tarnopolsky and M. F. Beal, “Potential for Creatine and Other Therapies Targeting Cellular Energy Dysfunction in Neurological Disorders,” Annals of Neurology, Vol. 49, No. 5, 2001, pp. 561-574. http://dx.doi.org/10.1002/ana.1028.abs

[15]   T. Teerlink, M. Hennekes, J. Bussemaker and J. Groeneveld, “Simultanous Determination of Creatine Compoumds and Adenine Nucleotides in Myocardial Tissue by High-Performance Liquid Chromatography,” Analytical Biochemistry, Vol. 214, No. 1, 1993, pp. 278-283. http://dx.doi.org/10.1006/abio.1993.1488

[16]   A. Ally and G. Park, “Rapid Determination of Creatine, Phosphocreatine, Purine Bases and Nucleotides (ATP, ADP, AMP, GTP, GDP) Inheart Biopsies by Gradiention-Pair Reversed-Phase Liquid,” Journal of Chromatography, Vol. 575, No. 1, 1992, pp. 19-27.

[17]   A. M. Persky, M. Müller, H. Derendorf, M. Grant, G. A. Brazeau and G. Hochhaus, “Singleand Multiple-Dose Pharmacokinetics of Oral Creatine,” Journal of Clinical Pharmacology, Vol. 43, No. 1, 2003, pp. 29-37. http://dx.doi.org/10.1177/0091270002239703

[18]   M. Levine, C. Conry-Cantilena, Y. Wang, R. W. Welch, P. W. Washko, K. R. Dhariwal, J. B. Park, A. Lazarev, J. F. Graumlich, J. King and L. R. Cantilena, “Vitamin C Pharmacokinetics in Healthy Volunteers: Evidence for a Recommended Dietary Allowance,” Proceedings of the National Academy of Sciences USA, Vol. 93, No. 8, 1996, pp. 3704-3709. http://dx.doi.org/10.1073/pnas.93.8.3704

 
 
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