Development and validation of a stability-indicating analytical method for the quantitation of oxytocin in pharmaceutical dosage forms

In this work, the usefulness of capillary electrophoresis–electrospray ionization time-of-flight–mass spectrometry for the analysis of biopharmaceuticals was studied. Noncovalently bound capillary coatings consisting of Polybrene-poly(vinyl sulfonic acid) or Polybrene-dextran sulfate-Polybrene were used to minimize protein and peptide adsorption, and achieve good separation efficiencies. The potential of the capillary electrophoresis-mass spectrometry (CE-MS) system to characterize degradation products was investigated by analyzing samples of the drugs, recombinant human growth hormone (rhGH) and oxytocin, which had been subjected to prolonged storage, heat exposure, and/or different pH values. Modifications could be assigned based on accurate masses as obtained with time-of-flight–mass spectrometry (TOF-MS) and migration times with respect to the parent compound. For heat-exposed rhGH, oxidations, sulfonate formation, and deamidations were observed. Oxytocin showed strong deamidation (up to 40%) upon heat exposure at low pH, whereas at medium and high pH, mainly dimer (>10%) and trisulfide formation (6–7%) occurred. Recombinant human interferon-β-1a (rhIFN-β) was used to evaluate the capability of the CE-MS method to assess glycan heterogeneity of pharmaceutical proteins. Analysis of this N-glycosylated protein revealed a cluster of resolved peaks which appeared to be caused by at least ten glycoforms differing merely in sialic acid and hexose N-acetylhexosamine composition. Based on the relative peak area (assuming an equimolar response per glycoform), a quantitative profile could be derived with the disialytated biantennary glycoform as most abundant (52%). Such a profile may be useful for in-process and quality control of rhIFN-β batches. It is concluded that the separation power provided by combined capillary electrophoresis and TOF-MS allows discrimination of highly related protein species.

show abstract

“…Oxytocin purity is generally assessed using liquid chromatography (LC) with UV detection [46, 47], taking the method described in the Ph. Eur.…”

Section: Resultsmentioning

confidence: 99%

Capillary electrophoresis-mass spectrometry using noncovalently coated capillaries for the analysis of biopharmaceuticals

et al. 2011

View full text Add to dashboard Cite

show abstract

“…[20] The validation parameters addressed were specificity, precision, accuracy, linearity, and robustness. [21][22][23][24][25][26][27][28] …”

Section: Methods Development and Validationmentioning

confidence: 99%

Validated stability indicating HPLC method for estimation of theophylline from a novel microsphere formulation

Shidhaye¹,

Malke²,

Kadam³

2009

Asian J Pharm

View full text Add to dashboard Cite

A new, simple, specific, precise and robust isocratic reversed-phase (RP) stability-indicating high-performance liquid chromatographic (HPLC) method was developed and validated for determination of theophylline from a novel formulation. The liquid chromatographic separation was achieved isocratically using a mobile phase of acetonitrile: 50 mM sodium acetate buffer (15:85) adjusted to pH 6.5 using dilute hydrochloric acid. The analysis was carried out using Hi-Q-Sil C18 column [250 mm x 4.6 mm, 5 µm] at flow rate of 1 ml/min and the UV detection at 270 nm. The method was validated for accuracy, precision, linearity, range, selectivity, and robustness. The linearity of the proposed method was investigated in the range of 1 to 24 µg/ml (r 2 = 0.9995). The drug was subjected to oxidation, hydrolysis, heat, and photolysis to apply stress conditions. The method provided good peak parameters with retention time of 8.6 ± 0.3 min. Degradation products resulting from stress studies did not interfere with the detection of theophylline and the assay can thus be considered as stability-indicating.

show abstract

“…The solution was then placed in a refrigerator for approximately 24 hours or until all the PF-127 had dissolved and the solution was clear on visual inspection. The solutions were assayed for OT content using a previously validated HPLC method (26) before use, and the gels were set in molds of dimensions 23.6 × 11.0 × 8.0 mm (L × W × D) such that only one surface of the gel was exposed to the dissolution test medium. An appropriate amount of gel was weighed such that each dosage unit to be tested contained approximately 300 IU of OT, and the gels were set in a Gallenkamp drying cabinet (Weiss Gallenkamp, Loughborough, United Kingdom) for 30 min at 37 °C prior to in vitro release testing.…”

Section: Preparation Of the Dosage Formmentioning

confidence: 99%

“…The analytical method used for the in vitro assessment of OT dosage forms was previously reported (26). The system consisted of a Model P100 dual-piston solvent delivery module (Thermo Separation Products, San Jose, CA, USA), a Model AS100 autosampler (Thermo Separation Products, San Jose, CA, USA) fitted with a Rheodyne® Model 7010 injector (Rheodyne, Reno, Nevada, USA) and a fixed-volume 20-µL loop, and a Model 1725 GASTIGHT® …”

Section: Hplc Systemmentioning

confidence: 99%

The Comparison of In Vitro Release Methods for the Evaluation of Oxytocin Release from Pluronic® F127 Parenteral Formulations

Chaibva

Walker²

2007

Dissolution Technol.

Self Cite

View full text Add to dashboard Cite

The objective of these studies was to develop a discriminatory in vitro release test for assessing formulation factors that may affect oxytocin (OT) release during formulation development studies of a Pluronic® F127 OT in situ gel-forming parenteral dosage form. An appropriate release assessment method should be able to discriminate between the performance of different formulation compositions (1, 2), and this was the primary criterion used for selection of an appropriate test procedure during the test method development process. ANOVA and the difference (f 1 ) and similarity (f 2 ) factors were used to evaluate the discriminatory behavior of different test methods that were investigated in these studies. The in vitro release tests that were investigated included the use of USP Apparatus 1, 2, and 3; a dialysis bag in USP Apparatus 2; and a membrane-less diffusion method. It was concluded that the use of USP Apparatus 3 was best able to discriminate between OT release for the different formulations tested. USP Apparatus 3 was thus considered the most suitable in vitro release test apparatus for studying formulation factors affecting OT release during the development of a parenteral dosage form prepared using Pluronic® F127.

show abstract

Development and validation of a stability-indicating analytical method for the quantitation of oxytocin in pharmaceutical dosage forms

Cited by 17 publications

References 8 publications

Capillary electrophoresis-mass spectrometry using noncovalently coated capillaries for the analysis of biopharmaceuticals

Capillary electrophoresis-mass spectrometry using noncovalently coated capillaries for the analysis of biopharmaceuticals

Validated stability indicating HPLC method for estimation of theophylline from a novel microsphere formulation

The Comparison of In Vitro Release Methods for the Evaluation of Oxytocin Release from Pluronic® F127 Parenteral Formulations

Contact Info

Product

Resources

About