Capillary Electrophoresis–Mass Spectrometry with Multisegment Injection and In-Capillary Preconcentration for High-Throughput and Sensitive Determination of Therapeutic Decapeptide Triptorelin in Pharmaceutical and Biological Matrices

Abstract

A capillary electrophoresis–tandem mass spectrometry method with a multisegment injection and an in-capillary field-enhanced sample stacking for determination of therapeutic peptide triptorelin in pharmaceutical and biological matrices was developed. The CE separation conditions were optimized in order to obtain maximal separation efficiency, analytical signal intensity and stability, and minimal adsorption of the analyzed peptide onto the capillary wall (1 M formic acid – HFo, pH 1.88). The implementation of the field-enhanced sample injection into CE improved the value of limit of detection 50 times while the multisegment injection increased the sample throughput three times in comparison to a conventional CE approach. The proposed method was characterized by favorable performance parameters, such as linearity (r2 ≥ 0.99), limit of detection (5 ng mL−1 in water matrix, 25 ng mL−1 in plasma matrix), precision (relative standard deviation, 1.5–9.4% for intraday and 2.3–11.9% for interday reproducibility), or accuracy (relative errors in the range of 80–109%). The FDA-validated method was successfully applied to the analysis of triptorelin in the commercial drug Diphereline® 0.1 mg (powder for injection) and in spiked human plasma samples. Favorable performance parameters along with proven application potentialities indicate the usefulness of the proposed method for its routine use in drug quality control laboratories and for clinical analysis, such as determination of triptorelin levels in plasma (for pharmacokinetic study).

Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

“…CZE-MS hyphenation has been mainly applied for the analysis of related process impurities, degradation products, and various PTMs, including glycosylation. Piešťanský et al recently developed a CZE-ESI-QQQ system for the analysis of immunogenic synthetic peptide in a conjugate with BSA as a carrier [224] and for quantitation of therapeutic peptide, triptorelin [225]. In the first work, an effective non-enzymatic release step of the peptide from the final peptide conjugate based on acid hydrolysis (2% formic acid) was successfully tested and implemented for simple, rapid, and robust quantification of immunogens in modern immunotherapeutics [224].…”

“…In the first work, an effective non-enzymatic release step of the peptide from the final peptide conjugate based on acid hydrolysis (2% formic acid) was successfully tested and implemented for simple, rapid, and robust quantification of immunogens in modern immunotherapeutics [224]. In the second work, an FDA-validated CZE-ESI-QQQ system with a multisegment injection and in-capillary FESI preconcentration for determination of triptorelin improved the limit of detection 50 times (5 ng/ml) and increased the sample throughput three times in comparison to a conventional CE approach [225]. To better characterize the heterogeneity of human chorionic gonadotropin (hCG), its analysis at the intact level by different electrophoretic techniques was undertaken for the first time by Camperi et al [226].…”

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

“…This approach has been used not only to decrease the analysis time of creatinine (by approximately 70%) [404,405] but also to inject multiple serum filtrate samples for metabolomic studies, including those of women in the last trimester of their pregnancy [406]. Other applications of this approach have been also recently described [407][408][409][410], noting that current analytical constraints of the approach include poor concentration sensitivity, potential loss in isobar/isomer resolution, sample carryover, its restriction to ionic metabolites [403], as well as the need to perform extensive method optimization to improve the compatibility with NACE protocols [411].…”

Capillary electrophoresis in the analysis of therapeutic peptides—A review

“…The already mentioned CZE-QQQ method, which uses MSI and FESI sample injection techniques, was applied not only in the analysis of a commercial drug containing triptorelin but also in the determination of spiked human plasma samples [240]. A simple sample pretreatment consisted of protein precipitation with ACN + 0.1% HFo.…”