separation of somatropin and its degradation products by high-performance liquid chromatography using a reversed-phase polymeric column
نویسندگان
چکیده
the accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. in this study, a gradient rp-hplc method is described for the separation of human growth hormone (hgh) variants as deamidated and oxidized forms. the methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1ml/min flow rate of a linear gradient of 0.1% v/v tfa/acetonitrile and tfa/water (ph= 2.0) mixture as the mobile phase. the overall run time of this method was 12 min and the average retention times were about 8.7 min for the native somatropin, 7.2 min for the deamidated form and 1.6 and 5.3 min for oxidized variants. the method was also validated in terms of selectivity, linearity, intra- and inter-day variations. in conclusion the method was found to have the potential for being applied as an initial and rapid evaluation method for assessing the quality and quantity of hgh during downstream processing, formulation and storage.
منابع مشابه
Separation of Somatropin and Its Degradation Products by High-Performance Liquid Chromatography Using a Reversed-Phase Polymeric Column
The accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. In this study, a gradient RP-HPLC method is described for the separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min flow rate of a linear gradient of 0.1% v/...
متن کاملSeparation of Somatropin and Its Degradation Products by High-Performance Liquid Chromatography Using a Reversed-Phase Polymeric Column
The accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. In this study, a gradient RP-HPLC method is described for the separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min flow rate of a linear gradient of 0.1% v/...
متن کاملSeparation of Somatropin and Its Degradation Products by High-Performance Liquid Chromatography Using a Reversed-Phase Polymeric Column
The accurate prediction of protein stability is one of the most challenging goals in protein formulation and delivery. In this study, a gradient RP-HPLC method is described for the separation of human growth hormone (hGH) variants as deamidated and oxidized forms. The methodology employed a polymeric poly (styrene-co-divinylbenzene) column and a 1mL/min flow rate of a linear gradient of 0.1% v/...
متن کاملSeparation of a complex mixture of heparin-derived oligosaccharides using reversed-phase high-performance liquid chromatography.
A reversed-phase ion-pairing high-performance liquid chromatography (RP-HPIPC) method for the separation of a complex mixture of heparin-derived oligosacchrides has been developed by a stepwise optimization of the mobile phase, in which the concentration of ion-pairing reagent, mobile phase pH, and acetonitrile concentration were varied. The resolution of more than 30 oligosaccharide components...
متن کاملSeparation of actinomycins and biosynthetic analogues by normal and reversed-phase high performance liquid chromatography.
Chromatographic procedures for the separation and identification of actinomycins have been reviewed1}. Only one report of HPLC has appeared2); this described reversed-phase separations of the C complex and of a biosynthetic mixture of actinomycin D (AMD)and congeners containing c/s-4-chloroproline in place of one or both prolines. In the present study a wider range of naturally occurring actino...
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عنوان ژورنال:
iranian journal of pharmaceutical researchجلد ۲۰۰۴، شماره ۱۰، صفحات ۲۰۹-۲۱۳
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