Drug tolerability in assisted reproduction techniques: a longitudinal study

Journal of Chromatographic Science

2020

Ganirelix is a synthetic decapeptide linked with nine different amino acids. To understand the peptide amino acid sequence or primary structure, the first step is to determine the amino acid composition of the peptide which can be a determining factor for the peptide immunogenicity. Edman degradation is not a suitable analytical technique to identify amino acid sequence present in Ganirelix due to the absence of uncharged N-terminal amino group. To address this challenge, a pre-column derivatization method was developed with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent. In the present work, the Ganirelix active pharmaceutical ingredient present in the injectable formulation was isolated by fraction collection and further purified by flash chromatography. The amino acid composition of Ganirelix is assayed by carrying out acid hydrolysis with 6 mol L−1 hydrochloric acid solution containing 1% phenol at 100°C for 24 h and derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate reagent solution, followed by determination of individual amino acids by reverse-phase chromatography using a C18 column. High resolution was achieved for the nine amino acid mixture. The amino acid composition results of temperature-stressed Ganirelix generic product and reference listed drug are in good agreement with the theoretical molar ratio of label information.

Determination of Amino Acid Composition of Ganirelix Acetate in an Injectable Formulation by Pre-column Derivatization with 6-Aminoquinolyl-N-hydroxysuccinimidyl Carbamate

Journal of Chromatographic Science

2020

Establishment of validated stability indicating purity method based on the stress degradation behavior of gonadotropin-releasing hormone antagonist (ganirelix) in an injectable formulation using HPLC and LC-MS-QTOF

Eur J Mass Spectrom (Chichester)

2021

Stress study of a drug substance or pharmaceutical drug product provides a vision into degradation pathways and degradation products of the active pharmaceutical ingredient and helps in interpretation of the chemical structure of the degradation impurities. In the current study, Ganirelix active ingredient presented in the Orgalutran® was stressed with acidic and alkali hydrolysis, photolysis, thermal and oxidation conditions as per the guidelines of International Conference on Harmonization (ICH) Q1A (R2). Ganirelix was found to be labile under thermal and alkali hydrolytic stress conditions, while it was stable to acid hydrolytic, oxidative and photolytic stress. All degradation products were separated with a resolution > 1.5 on a C18 column (2.6 µm, 25 cm×4.6 mm) using a hydrophilic ion pair such as sodium perchlorate, at a concentration <0.04 M. In total, four major degradant impurities were found during stress study. These impurities were fractionated and desalted by flash chromatography for identification of chemical structures. LC-MS-QTOF analysis revealed that two degradation products are diastereomers of Ganirelix, one degradation product is a deamination compound and other degradation product result from the insertion of a new amino acid residue in the Ganirelix peptide sequence. The developed method is sensitive enough to quantify the related substances of Ganirelix at the 0.04% level with that of Ganirelix test concentration.

Applications of Fourier Transform Infrared and UV-Visible Spectroscopy for the demonstrating sameness of Ganirelix peptide in Liquid Injection Formulation

2022

RJPT

Ganirelix (or Ganirelixum) is a synthetic peptide molecule made up of covalently bonded 10 amino acids. The structural order of peptides determines their biological properties. The minor change in the structure of therapeutic peptides can have a significant impact on their biological behaviors. In this regard, the ganirelix drug product was sterilized at 121°C for 15 mins and further exposed to thermal and humidity at 25°C/60%RH and 40°C/75% RH for 6 months. The ganirelix was isolated using flash chromatography and freeze- dried. We applied Fourier transform infrared (FTIR) and Ultraviolet -Visible (UV-Vis) spectroscopic techniques to investigate temperature and humidity effects on the chemical structure of Ganirelix. FTIR scans were conducted in the wave number range of 4000 – 400cm-1 and UV scans were conducted in the wavelength range of 190 – 400nm. The attained IR stretching vibrations, maximum UV absorbance results, overlayed IR and UV-Vis spectra of Ganirelix in temperature stressed generic drug product is closely comparable to reference listed drug (RLD). In addition, it showed homology in the finger print region of the FTIR with those of the reference listed drug (RLD).