{"title":"Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Eight Related Substances in Milrinone API","authors":"Yujiang Pang, Xiaoyan Shi, Zhengzhao Li, Zengyu Pan, Kai Liu, Xianyong Zhu, Zhong Feng","doi":"10.1007/s10337-023-04307-7","DOIUrl":null,"url":null,"abstract":"<div><p>A simple, quick, and economical reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative determination of related substance in milrinone API was developed and validated. The method has shown adequate separation of milrinone and eight kinds of related substance. Chromatographic separation was achieved on an Agilent ZORBAX Rx-C8 (250 × 4.6 mm, 5 µm) column at wavelength of 220 nm, using a mobile phase acetonitrile: phosphate buffer (85:15) in an isocratic elution mode at a flow rate of 1.0 mL min<sup>−1</sup>. The method was validated in terms of system suitability, specificity, linearity, range, accuracy, intermediate precision, and robustness. A linear relationship between peak area and concentration of milrinone and its related substances was observed in a level of 10–300% (determination coefficient, <i>r</i><sup>2</sup> ≥ 0.998), respectively. The method showed acceptable levels of precision (%RSD ≤ 2), accuracy (> 96% recovery), robustness (< 10% content difference), and stability (> 96% recovery) over varied environment and laboratory conditions. The validation results suggested that the developed method is sensitive enough and repeatable and could be used for qualitative and quantitative assessment of eight kinds of related substance in milrinone. As a significant part of drug development process, stress testing is also performed to identify the degradation products and validate the stability-indicating power of our analytical methods, in which milrinone is subjected to acidic, base, oxidation, thermal and photolytic stress environment. To sum up, the proposed method is suitable for purpose in quality-control laboratories for quantitative analysis of the drugs individually, as it is simple and rapid with good precision and accuracy.</p></div>","PeriodicalId":518,"journal":{"name":"Chromatographia","volume":"87 2","pages":"105 - 115"},"PeriodicalIF":1.2000,"publicationDate":"2024-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chromatographia","FirstCategoryId":"92","ListUrlMain":"https://link.springer.com/article/10.1007/s10337-023-04307-7","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
A simple, quick, and economical reverse-phase high-performance liquid chromatography (RP-HPLC) method for the quantitative determination of related substance in milrinone API was developed and validated. The method has shown adequate separation of milrinone and eight kinds of related substance. Chromatographic separation was achieved on an Agilent ZORBAX Rx-C8 (250 × 4.6 mm, 5 µm) column at wavelength of 220 nm, using a mobile phase acetonitrile: phosphate buffer (85:15) in an isocratic elution mode at a flow rate of 1.0 mL min−1. The method was validated in terms of system suitability, specificity, linearity, range, accuracy, intermediate precision, and robustness. A linear relationship between peak area and concentration of milrinone and its related substances was observed in a level of 10–300% (determination coefficient, r2 ≥ 0.998), respectively. The method showed acceptable levels of precision (%RSD ≤ 2), accuracy (> 96% recovery), robustness (< 10% content difference), and stability (> 96% recovery) over varied environment and laboratory conditions. The validation results suggested that the developed method is sensitive enough and repeatable and could be used for qualitative and quantitative assessment of eight kinds of related substance in milrinone. As a significant part of drug development process, stress testing is also performed to identify the degradation products and validate the stability-indicating power of our analytical methods, in which milrinone is subjected to acidic, base, oxidation, thermal and photolytic stress environment. To sum up, the proposed method is suitable for purpose in quality-control laboratories for quantitative analysis of the drugs individually, as it is simple and rapid with good precision and accuracy.
期刊介绍：
Separation sciences, in all their various forms such as chromatography, field-flow fractionation, and electrophoresis, provide some of the most powerful techniques in analytical chemistry and are applied within a number of important application areas, including archaeology, biotechnology, clinical, environmental, food, medical, petroleum, pharmaceutical, polymer and biopolymer research. Beyond serving analytical purposes, separation techniques are also used for preparative and process-scale applications. The scope and power of separation sciences is significantly extended by combination with spectroscopic detection methods (e.g., laser-based approaches, nuclear-magnetic resonance, Raman, chemiluminescence) and particularly, mass spectrometry, to create hyphenated techniques. In addition to exciting new developments in chromatography, such as ultra high-pressure systems, multidimensional separations, and high-temperature approaches, there have also been great advances in hybrid methods combining chromatography and electro-based separations, especially on the micro- and nanoscale. Integrated biological procedures (e.g., enzymatic, immunological, receptor-based assays) can also be part of the overall analytical process.
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