BİLAL YILMAZ, YÜCEL KADIOĞLU, Kadem MERAL, FİKRET ÇELEBİ, YAVUZ ONGANER

Determination and pharmacokinetics of recombinant human growth hormone in rabbit plasma by spectrofluorometry method

Bu çalışmada, insan büyüme hormonunun (somatotropin) tavşan plazmasında tayini için yeni bir spektroflorometri yöntemi geliştirildi. Geçerlilik FDA’nın “Bioanalitik Yöntem Validasyonu” rehberinde belirtildiği koşullarda yapıldı. Kalibrasyon eğrileri 0.125-8.0 μg/mL derişim aralığında doğrusaldır. Plazmada somatotropin için gün içi ve günler arası kesinlik değerleri %13.5’den küçüktü ve doğruluk (bağıl hata) %12.0’dan daha iyiydi. Bütün örnekler için geri kazanım >%84.0 idi. Somatotropinin belirlenebilen (LOD) ve ölçülebilen (LOQ) en düşük değeri sırasıyla 0.075 and 0.10 μg/mL’dir. Tanımlanan spektroflorometri yöntemi tavşanlarda somatotropinin farmakokinetiğini incelemek için yeterli duyarlılığa ve özgüllüğe sahiptir ve klinik farmakokinetik çalışmalara da uyarlanabilir.

Anahtar Kelimeler:

kan plazması, tavşan, tespit etmek, metodoloji, farmakokinetik, somatotropin

Spektroflorometri yöntemi kullanarak tavşan plazmasında rekombinant insan büyüme hormonunun tayini ve farmakokinetiği

In this study, a new spectrofluorometry method was developed for the determination of human growth hormone (somatotropin) in rabbit plasma. The validation was assessed under a variety of conditions (FDA, Bioanalytical Method Validation Guidline). Calibration curves were linear between the concentration range of 0.125-8.0 μg/mL. Intra- and inter-day precision values for somatotropin in plasma were less than 13.5, and accuracy (relative error) was better than 12.0%. The recoveries for all samples were >84.0%. The limits of detection (LOD) and quantification (LOQ) of somatotropin were 0.075 and 0.10 μg/mL, respectively. The described spectrofluorometry method has adequate sensitivity and specificity to study pharmacokinetics of somatotropin in rabbits, and could be adapted also to clinical pharmacokinetic study.

Keywords:

blood plasma, rabbits, determination, methodology, pharmacokinetics, somatotropin,

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1. Cheng YH, Dyer AM, Jabbal-Gill I, Hinchcliffe M, Nankervis R, Smith A, Watts P: Intranasal delivery of recombinant human growth hormone (somatropin) in sheep using chitosan-based powder formulations. Eur J Pharm, 26, 9-15, 2005.
2. Ikehara M, Ohtsuka E, Tokunaga T, Taniyame Y, Iwai S, Kitano K, Miyamoto S, Ohgi T, Sakuragawa Y, Fujiyama K, Ikari T, Kobayashi M, Miyake T, Shibahara S, Ono A, Ueda T, Tanaka T, Baba H, Mıkı T, Sakurai A, Oishi T, Chisaka O, Matsubara K: Synthesis of a gene for human growth hormone and its expression in Escherichia coli. Proc Natl Acad Sci USA, 81, 5956-5960, 1984.
3. Chang CN, Key M, Bochner B, Heyneker H, Gray G: High-level secretion of human growth hormone by Escherichia coli. Gene, 55, 189-196, 1987.
4. Becker GW, Hsiung HM: Expression, secretion and folding of human growth hormone in Escherichia coli. FEBS Lett, 204, 145-150, 1986.
5. McNerney TM, Watson SK, Sim JH, Bridenbaugh RL: Separation of recombinant human growth hormone from Escherichia coli cell pellet extract by capillary zone electrophoresis. J Chromatogr, 744, 223-229, 1996.
6. Riggin RM, Shaar CJ, Dorulla GK, Lefeber DS, Miner DJ: High-performance size-exclusion chromatographic determination of the potency of biosynthetic human growth hormone products. J Chromatogr, 435, 307-318, 1988.
7. Gellerfors P, Eketorp G, Fhölenhag K, Pavlu B, Johansson S, Fryklund L: Characterisation of a secreted form of recombinant derived human growth hormone, expressed in Escherichia coli cells. J Pharm Biomed Anal, 7, 173-183, 1989.
8. Wu S, Hancock WS, Pavlu B, Gellerfors P: Application of high-performance hydrophobic-interaction chromatography to the characterization of recombinant DNA-derived human growth hormone. J Chromatogr, 500, 595-606, 1990.
9. Riggin RM, Dorulla GK, Miner DJ: A reversed-phase high-performance liquid chromatographic method for characterization of biosynthetic human growth hormone. Anal Biochem, 167, 199-209, 1987.
10. Teshima G, Canova-Davis E: Separation of oxidized human growth hormone variants by revesed-phase high-performance liquid chromatography. Effect of mobile phase pH and organic modifier. J Chromatogr, 625, 207-215, 1992.
11. Hepner F, Cszasar E, Roitinger E, Lubec G: Mass spectrometrical analysis of recombinant human growth hormone (Genotropin®) reveals amino acid substitutions in 2% of the expressed protein. Proteome Science, 3, 1-12, 2005.
12. Charman SA, Segrave AM, Edwards GA, Porter CJH: Systemic availability and lymphatic transport of human growth hormone administered by subcutaneous ınjection. J Pharm, 89, 168-177, 2000.
13. Vermehren C, Hansen HS, Thomsen MK: Time dependent effects of two absorption enhancers on the nasal absorption of growth hormone in rabbits. Int J Pharm, 128, 239-250, 1996.
14. Vahl N, Jensen SB, Rasmussen MH, Susgaard S, Jorgensen JOL, Christiansen JS, Rasmussen UF, Hilsted J: Bioavailability of recombinant human growth hormone in different concentrations and formulations. Pharmacol-Toxicol, 79, 144-149, 1996.
15. Katakam M, Ravis WR, Golden DL, Banga AK: Controlled release of human growth hormone following subcutaneous administration in dogs. Int J Pharm, 152, 53-58, 1997.
16. Mzoughi N, Dachraoui M, Villeneuve JP: Evaluation of aromatic hydrocarbons by spectrofluorometry in marine sediments and biological matrix: What reference should be considered. CR Chimie, 8, 97-102, 2005. 17. FDA guideline for submitting samples and analytical data for method validations, Rockville, MD, USA, February 1987. 18. Yilmaz B, Kadioglu Y: Determination of insulin in rabbit plasma using high-performance liquid chromatography. Kafkas Univ Vet Fak Derg, 18 (1): 31-36, 2012.