Comparative study for purity assessment of recombinant human growth hormone using mass balance approach and amino acid-based isotope dilution mass spectrometry.

[Display omitted] • Combination of AA-based IDMS and mass balance approach for hGH quantification. • Mass balance method quantifies impurities in hGH using independent techniques. • Accurate hGH purity obtained without calibration graphs in AA-based IDMS. Growth hormone plays an unreplaceable role in regulating human growth and promoting protein synthesis, etc. For this reason, accurate quantification of its purity has become a focus of attention in the pharmaceutical industry and medical research. Herein, the purity assessment of human growth hormone (hGH) was established on the basis of mass balance approach and amino acid (AA)-based isotope dilution mass spectrometry (IDMS). In the first method, hGH purity was quantified by the determination of all the impurities in a hGH study material. In the second method, hGH purity was assessed by quantifying hydrolyzed AAs, e.g. leucine (Leu), phenylalanine (Phe), and proline (Pro). To accurately quantify these AAs, the corresponding isotope-labeled AAs were used as internal standards using high-performance liquid chromatography- mass spectrometry (HPLC-MS). The results of the two methods for determining the purity of hGH were consistent, with the mass balance method giving a result of (0.352 ± 0.010) g/g and IDMS a result of (0.363 ± 0.028) g/g. The work in this study could give a benefit for protein quantification and provides a reference for the development of hGH certified reference materials. [ABSTRACT FROM AUTHOR]

Copyright of Microchemical Journal is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)