Validation of a method for the quantitation of ghrelin and unacylated ghrelin by HPLC.

[en] An HPLC/UV method was first optimized for the separation and quantitation of human acylated and unacylated (or des-acyl) ghrelin from aqueous solutions. This method was validated by an original approach using accuracy profiles based on tolerance intervals for the total error measurement. The concentration range that achieved adequate accuracy extended from 1.85 to 59.30microM and 1.93 to 61.60microM for acylated and unacylated ghrelin, respectively. Then, optimal temperature, pH and buffer for sample storage were determined. Unacylated ghrelin was found to be stable in all conditions tested. At 37 degrees C acylated ghrelin was stable at pH 4 but unstable at pH 7.4, the main degradation product was unacylated ghrelin. Finally, this validated HPLC/UV method was used to evaluate the binding of acylated and unacylated ghrelin to liposomes.

Disciplines :

Pharmacy, pharmacology & toxicology

Author, co-author :

Rozet, Eric ; Université de Liège - ULiège > Département de pharmacie > Chimie analytique

Staes, Edith

Ucakar, Bernard

Hubert, Philippe ; Université de Liège - ULiège > Département de pharmacie > Chimie analytique

Preat, Veronique

Language :

English

Title :

Validation of a method for the quantitation of ghrelin and unacylated ghrelin by HPLC.

Publication date :

2010

Journal title :

Journal of Pharmaceutical and Biomedical Analysis

ISSN :

0731-7085

eISSN :

1873-264X

Publisher :

Elsevier, Amsterdam, Netherlands

Volume :

Issue :

Pages :

633-9

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 14 January 2010

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Bibliography

Banga, A.K., (2006) Therapeutic Peptides and Proteins Formulation, Processing, and Delivery Systems. 2nd ed., , CRC/Taylor & Francis, Boca Raton, FL
Kojima, M., Hosoda, H., Date, Y., Nakazato, M., Matsuo, H., Kangawa, K., Ghrelin is a growth-hormone-releasing acylated peptide from stomach (1999) Nature, 402, pp. 656-660
Kojima, M., Kangawa, K., Ghrelin: structure and function (2005) Physiol. Rev., 85, pp. 495-522
Soares, J.B., Leite-Moreira, A.F., Ghrelin, des-acyl ghrelin and obestatin: three pieces of the same puzzle (2008) Peptides, 29, pp. 1255-1270
Akamizu, T., Kangawa, K., Translational research on the clinical applications of ghrelin (2006) Endocr. J., 53, pp. 585-591
Hosoda, H., Doi, K., Nagaya, N., Okumura, H., Nakagawa, E., Enomoto, M., Ono, F., Kangawa, K., Optimum collection and storage conditions for ghrelin measurements: octanoyl modification of ghrelin is rapidly hydrolyzed to desacyl ghrelin in blood samples (2004) Clin. Chem., 50, pp. 1077-1080
De Vriese, C., Gregoire, F., Lema-Kisoka, R., Waelbroeck, M., Robberecht, P., Delporte, C., Ghrelin degradation by serum and tissue homogenates: identification of the cleavage sites (2004) Endocrinology, 145, pp. 4997-5005
De Vriese, C., Hacquebard, M., Gregoire, F., Carpentier, Y., Delporte, C., Ghrelin interacts with human plasma lipoproteins (2007) Endocrinology, 148, pp. 2355-2362
Yoshimoto, A., Mori, K., Sugawara, A., Mukoyama, M., Yahata, K., Suganami, T., Takaya, K., Nakao, K., Plasma ghrelin and desacyl ghrelin concentrations in renal failure (2002) J. Am. Soc. Nephrol., 13, pp. 2748-2752
Akamizu, T., Shinomiya, T., Irako, T., Fukunaga, M., Nakai, Y., Nakai, Y., Kangawa, K., Separate measurement of plasma levels of acylated and desacyl ghrelin in healthy subjects using a new direct ELISA assay (2005) J. Clin. Endocrinol. Metab., 90, pp. 6-9
Paik, K.H., Choe, Y.H., Park, W.H., Oh, Y.J., Kim, A.H., Chu, S.H., Kim, S.W., Jin, D.K., Suppression of acylated ghrelin during oral glucose tolerance test is correlated with whole-body insulin sensitivity in children with Prader-Willi syndrome (2006) J. Clin. Endocrinol. Metab., 91, pp. 1876-1881
Broom, D.R., Batterham, R.L., King, J.A., Stensel, D.J., Influence of resistance and aerobic exercise on hunger, circulating levels of acylated ghrelin, and peptide YY in healthy males (2009) Am. J. Physiol Regul. Integr. Comp. Physiol., 296, pp. R29-R35
Rauh, M., Groschl, M., Rascher, W., Simultaneous quantification of ghrelin and desacyl-ghrelin by liquid chromatography-tandem mass spectrometry in plasma, serum, and cell supernatants (2007) Clin. Chem., 53, pp. 902-910
Beaumont, N.J., Skinner, V.O., Tan, T.M., Ramesh, B.S., Byrne, D.J., MacColl, G.S., Keen, J.N., Srai, K.S., Ghrelin can bind to a species of high density lipoprotein associated with paraoxonase (2003) J. Biol. Chem., 278, pp. 8877-8880
Bednarek, M.A., Feighner, S.D., Pong, S.S., McKee, K.K., Hreniuk, D.L., Silva, M.V., Warren, V.A., Heck, J.V., Structure-function studies on the new growth hormone-releasing peptide, ghrelin: minimal sequence of ghrelin necessary for activation of growth hormone secretagogue receptor 1a (2000) J. Med. Chem., 43, pp. 4370-4376
Makino, T., Matsumoto, M., Suzuki, Y., Kitajima, Y., Yamamoto, K., Kuramoto, M., Minamitake, Y., Yabuta, M., Semisynthesis of human ghrelin: condensation of a Boc-protected recombinant peptide with a synthetic O-acylated fragment (2005) Biopolymers, 79, pp. 238-247
De Vriese, C., Gregoire, F., De, N.P., Robberecht, P., Delporte, C., Ghrelin is produced by the human erythroleukemic HEL cell line and involved in an autocrine pathway leading to cell proliferation (2005) Endocrinology, 146, pp. 1514-1522
Pemberton, C., Wimalasena, P., Yandle, T., Soule, S., Richards, M., C-terminal pro-ghrelin peptides are present in the human circulation (2003) Biochem. Biophys. Res. Commun., 310, pp. 567-573
Ishimaru, M., Yoshizawa-Kumagaye, K., Kubo, S., Kitani, T., Chino, N., Kangawa, K., Kimura, T., Stability of the O-octanoyl group of rat ghrelin during chemical synthesis: counter-ion-dependent alteration of an ester bond breakage (2003) Lett. Pept. Sci., 10, pp. 41-50
Date, Y., Kojima, M., Hosoda, H., Sawaguchi, A., Mondal, M.S., Suganuma, T., Matsukura, S., Nakazato, M., Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesized in a distinct endocrine cell type in the gastrointestinal tracts of rats and humans (2000) Endocrinology, 141, pp. 4255-4261
Enomoto, M., Nagaya, N., Uematsu, M., Okumura, H., Nakagawa, E., Ono, F., Hosoda, H., Kangawa, K., Cardiovascular and hormonal effects of subcutaneous administration of ghrelin, a novel growth hormone-releasing peptide, in healthy humans (2003) Clin. Sci. (Lond.), 105, pp. 431-435
Bossola, M., Tazza, L., Luciani, G., Mechanisms and treatment of anorexia in end-stage renal disease patients on hemodialysis (2009) J. Ren. Nutr., 19, pp. 2-9
Mathot, F., Schanck, A., Van, B.F., Arien, A., Noppe, M., Brewster, M., Préat, V., Passive diffusion of polymeric surfactants across lipid bilayers (2007) J. Control. Release, 120, pp. 79-87
Bartlett, G.R., Phosphorus assay in column chromatography (1959) J. Biol. Chem., 234, pp. 466-468
Hubert, P., Nguyen-Huu, J.J., Boulanger, B., Chapuzet, E., Chiap, P., Cohen, N., Compagnon, P.A., Valat, L., Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal. Part I (2004) J. Pharm. Biomed. Anal., 36, pp. 579-586
Hubert, P., Nguyen-Huu, J.J., Boulanger, B., Chapuzet, E., Cohen, N., Compagnon, P.A., Dewe, W., Rozet, E., Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal. Part III (2007) J. Pharm. Biomed. Anal., 45, pp. 82-96
Hubert, P., Nguyen-Huu, J.J., Boulanger, B., Chapuzet, E., Chiap, P., Cohen, N., Compagnon, P.A., Rozet, E., Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal. Part II (2007) J. Pharm. Biomed. Anal., 45, pp. 70-81
International Conference on Harmonization (ICH) of technical requirements for registration of Pharmaceutical for human use, topic Q2(R1): Validation of analytical procedures: text and methodology, Geneva, 2005
Rozet, E., Ceccato, A., Hubert, C., Ziemons, E., Oprean, R., Rudaz, S., Boulanger, B., Hubert, P., Analysis of recent pharmaceutical regulatory documents on analytical method validation (2007) J. Chromatogr. A, 1158, pp. 111-125
Viswanathan, C.T., Bansal, S., Booth, B., DeStefano, A.J., Rose, M.J., Sailstad, J., Shah, V.P., Weiner, R., Workshop/conference report-quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays (2007) Aaps J., 9, pp. E30-E42
Boulanger, B., Dewe, W., Gilbert, A., Govaerts, B., Maumy-Bertrand, M., Risk management for analytical methods based on the total error concept: conciliating the objectives of the pre-study and in-study validation phases (2007) Chemom. Intell. Lab. Syst., 86, pp. 198-207
(1994) Part 1 to 6, , ISO 5725 Application of the statistics-accuracy (trueness and precision) of the results and methods of measurement, International Organization for Standardization ISO
Castanho, M.A., Fernandes, M.X., Lipid membrane-induced optimization for ligand-receptor docking: recent tools and insights for the "membrane catalysis" model (2006) Eur. Biophys. J., 35, pp. 92-103
Seelig, J., Thermodynamics of lipid-peptide interactions (2004) Biochim. Biophys. Acta, 1666, pp. 40-50