Abstract
Octreotide is a synthetic cyclic octapeptide analogue of somatostatin-14. It is mainly administered for the treatment of acromegaly, severe diarrhea, and neuroendocrine neoplasias. In this work, a hydrophilic interaction liquid chromatography (HILIC) method with fluorescence (FL) detection was developed and validated for the quantitation of octreotide in solutions for injection. Chromatographic separation was performed on an XBridge®-HILIC analytical column under isocratic elution with a short chromatographic run time of less than 10 min. The mobile phase consisted of ammonium bicarbonate 8.6 mM (pH 8.1)/acetonitrile 35/65 (v/v). The high sensitivity and selectivity of the fluorescence detection, with the excitation wavelength (λexcitation) set at 280 nm and the emission wavelength set at (λemission) 330 nm, enabled a simple sample preparation procedure that included only dilution steps. The calibration curve showed good linearity with a correlation coefficient greater than 0.998. The method was successfully applied to the analysis of commercially available octreotide injection forms.
Highlights
The neuropeptide somatostatin is a cyclic tetradecapeptide originally isolated from ovine hypothalamus and has been shown to be distributed in the peripheral and central nervous system [1,2]
Reversed-phase eluent were formate (AMF)toand ammonium acetate (AMA), their concen-highperformance liquid chromatography (RP-high performance liquid chromatography (HPLC)) with UV detection was used to separate tration levels ranged from 5 to 35 mM in mobile phases that consisted of acetonitrile/ aquepoly(ethylene glycol)octreotide derivatives with various molecular weights [30] and ous buffer 70/30 v/v
As shown by the plots of the ionization fraction presented in Figure 1b, octreotide is a hydrophilic peptide
Summary
The neuropeptide somatostatin is a cyclic tetradecapeptide originally isolated from ovine hypothalamus and has been shown to be distributed in the peripheral and central nervous system [1,2]. Octreotide (SMS 201-995) is an eight amino acid peptide and the first synthesized analogue of somatostatin-14 [10,11] It is a cyclic octapeptide and the retention of the disulfide bond is essential for its biological activity (Figure 1a) [12]. Reversed-phase eluent were formate (AMF)toand ammonium acetate (AMA), their concen-highperformance liquid chromatography (RP-HPLC) with UV detection was used to separate tration levels ranged from 5 to 35 mM in mobile phases that consisted of acetonitrile/ aquepoly(ethylene glycol)octreotide derivatives with various molecular weights [30] and ous buffer 70/30 v/v. A hydrophilic interaction liquid chromatographic method with fluorescence (HILIC-FL) detection was developed and validated for the quantification of octreotide in formulations for injection. At the pH values tested, the residual silanols of the stationary phase were negatively charged
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