一种简单的HPLC-UV方法监测肺动脉高压患者的治疗依从性。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2025-02-01 DOI:10.1016/j.jchromb.2024.124443

Paweł K. Kunicki , Maciej T. Grymm , Tomasz Pawiński , Daniel Szulczyk , Marcin Waligóra , Grzegorz Kopeć

{"title":"一种简单的HPLC-UV方法监测肺动脉高压患者的治疗依从性。","authors":"Paweł K. Kunicki , Maciej T. Grymm , Tomasz Pawiński , Daniel Szulczyk , Marcin Waligóra , Grzegorz Kopeć","doi":"10.1016/j.jchromb.2024.124443","DOIUrl":null,"url":null,"abstract":"<div><div>A considerable percentage of ineffective treatment in pulmonary arterial hypertension (PAH) may be related to subtherapeutic dosage or non-adherence. The aim of the study was to develop a simple analytical method suitable for plasma determination of selected drugs: riociguat (RIO), bosentan (BOS) and macitentan (MAC) administered to PAH patients. An isocratic HPLC-UV system (Spectra Physics – Shimadzu) with a manual injector (50 μL loop) was applied. Chromatographic analysis was performed using a Suplecosil LC-CN column (150 × 4.6 mm, 5 μm) protected with a Supelguard precolumn at room temperature. The separation was carried out using the mobile phase: CH<sub>3</sub>CN:H<sub>2</sub>O:0.5 M KH<sub>2</sub>PO<sub>4</sub>:85 % H<sub>3</sub>PO<sub>4</sub> (172:324.2:3.7:0.1, v/v) at a flow rate of 1.8 mL/min. Ethyl acetate (4 mL) was used for 10-min liquid–liquid extraction from 0.4 mL alkalized plasma sample. Detection was performed at λ = 245 nm chosen as a compromise between signal intensity and matrix interference. The analytes were eluted at retention times of 4.4 min (RIO), 5.4 min (BOS), 8.9 min (MAC) and 7.8 min for gallopamil (internal standard, GAL). The method was found linear and calibrated in the ranges: 5–1000 ng/mL for RIO, 10–2000 ng/mL for BOS and 20–2000 ng/mL for MAC, with r<sup>2</sup> of 0.9991 for RIO, 0.9983 for BOS, and 0.9949 for MAC, respectively. Within the given ranges, the method ensured reliable results with the required precision and accuracy: ≤15 % (≤20 % for LLOQ). There was no significant carryover effect. The method has been successfully used in pilot study on adherence in patients treated for PAH, enabling monitoring of RIO, BOS and MAC. Drug concentrations were assessed in samples taken before (C0) and 3 h after drug administration (C3). For RIO, BOS and MAC, the developed method was suitable for both C0 and C3 samples, allowing steady-state drug determination if used. The presented method can be recommended to laboratories equipped with basic HPLC apparatus as an attractive analytical tool for both TDM and adherence studies.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1252 ","pages":"Article 124443"},"PeriodicalIF":2.8000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A simple HPLC-UV method for monitoring therapeutic adherence in pulmonary arterial hypertension\",\"authors\":\"Paweł K. Kunicki , Maciej T. Grymm , Tomasz Pawiński , Daniel Szulczyk , Marcin Waligóra , Grzegorz Kopeć\",\"doi\":\"10.1016/j.jchromb.2024.124443\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>A considerable percentage of ineffective treatment in pulmonary arterial hypertension (PAH) may be related to subtherapeutic dosage or non-adherence. The aim of the study was to develop a simple analytical method suitable for plasma determination of selected drugs: riociguat (RIO), bosentan (BOS) and macitentan (MAC) administered to PAH patients. An isocratic HPLC-UV system (Spectra Physics – Shimadzu) with a manual injector (50 μL loop) was applied. Chromatographic analysis was performed using a Suplecosil LC-CN column (150 × 4.6 mm, 5 μm) protected with a Supelguard precolumn at room temperature. The separation was carried out using the mobile phase: CH<sub>3</sub>CN:H<sub>2</sub>O:0.5 M KH<sub>2</sub>PO<sub>4</sub>:85 % H<sub>3</sub>PO<sub>4</sub> (172:324.2:3.7:0.1, v/v) at a flow rate of 1.8 mL/min. Ethyl acetate (4 mL) was used for 10-min liquid–liquid extraction from 0.4 mL alkalized plasma sample. Detection was performed at λ = 245 nm chosen as a compromise between signal intensity and matrix interference. The analytes were eluted at retention times of 4.4 min (RIO), 5.4 min (BOS), 8.9 min (MAC) and 7.8 min for gallopamil (internal standard, GAL). The method was found linear and calibrated in the ranges: 5–1000 ng/mL for RIO, 10–2000 ng/mL for BOS and 20–2000 ng/mL for MAC, with r<sup>2</sup> of 0.9991 for RIO, 0.9983 for BOS, and 0.9949 for MAC, respectively. Within the given ranges, the method ensured reliable results with the required precision and accuracy: ≤15 % (≤20 % for LLOQ). There was no significant carryover effect. The method has been successfully used in pilot study on adherence in patients treated for PAH, enabling monitoring of RIO, BOS and MAC. Drug concentrations were assessed in samples taken before (C0) and 3 h after drug administration (C3). For RIO, BOS and MAC, the developed method was suitable for both C0 and C3 samples, allowing steady-state drug determination if used. The presented method can be recommended to laboratories equipped with basic HPLC apparatus as an attractive analytical tool for both TDM and adherence studies.</div></div>\",\"PeriodicalId\":348,\"journal\":{\"name\":\"Journal of Chromatography B\",\"volume\":\"1252 \",\"pages\":\"Article 124443\"},\"PeriodicalIF\":2.8000,\"publicationDate\":\"2025-02-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Chromatography B\",\"FirstCategoryId\":\"1\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1570023224004525\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Chromatography B","FirstCategoryId":"1","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1570023224004525","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}

引用次数: 0

摘要

相当大比例的肺动脉高压（PAH）无效治疗可能与亚治疗剂量或不依从性有关。本研究的目的是建立一种简单的分析方法，适用于PAH患者血浆中所选药物的测定：瑞西奎特（里约热内卢）、波生坦（BOS）和马西坦（MAC）。采用手动进样（50 μL回路）等密度HPLC-UV体系（Spectra Physics - Shimadzu）。色谱分析采用Supelguard预柱保护的Suplecosil LC-CN柱（150 × 4.6 mm, 5 μm），室温下进行。采用流动相CH3CN:H2O:0.5 M kh2po4: 85% H3PO4 （172:324.2:3.7:0.1, v/v）进行分离，流速为1.8 mL/min。0.4 mL碱化血浆样品，用乙酸乙酯（4ml）液液萃取10min。在λ = 245 nm处进行检测，选择信号强度和矩阵干扰之间的折衷。洗脱时间为4.4 min（里约热内卢）、5.4 min （BOS）、8.9 min （MAC）和7.8 min （GAL）。该方法在里约热内卢的5-1000 ng/mL、BOS的10-2000 ng/mL和MAC的20-2000 ng/mL范围内呈线性关系，里约热内卢、BOS和MAC的r2分别为0.9991、0.9983和0.9949。在给定的范围内，该方法保证了可靠的结果，所需的精密度和准确度：≤15% （LLOQ≤20%）。没有明显的结转效应。该方法已成功用于PAH治疗患者依从性的试点研究，可以监测里约热内卢，BOS和MAC。在给药前（C0）和给药后3小时（C3）的样品中评估药物浓度。对于里约热内卢、BOS和MAC，该方法适用于C0和C3样品，使用该方法可以进行稳态药物测定。该方法可推荐给配备基本HPLC设备的实验室，作为TDM和粘附性研究的有吸引力的分析工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

A simple HPLC-UV method for monitoring therapeutic adherence in pulmonary arterial hypertension

A considerable percentage of ineffective treatment in pulmonary arterial hypertension (PAH) may be related to subtherapeutic dosage or non-adherence. The aim of the study was to develop a simple analytical method suitable for plasma determination of selected drugs: riociguat (RIO), bosentan (BOS) and macitentan (MAC) administered to PAH patients. An isocratic HPLC-UV system (Spectra Physics – Shimadzu) with a manual injector (50 μL loop) was applied. Chromatographic analysis was performed using a Suplecosil LC-CN column (150 × 4.6 mm, 5 μm) protected with a Supelguard precolumn at room temperature. The separation was carried out using the mobile phase: CH₃CN:H₂O:0.5 M KH₂PO₄:85 % H₃PO₄ (172:324.2:3.7:0.1, v/v) at a flow rate of 1.8 mL/min. Ethyl acetate (4 mL) was used for 10-min liquid–liquid extraction from 0.4 mL alkalized plasma sample. Detection was performed at λ = 245 nm chosen as a compromise between signal intensity and matrix interference. The analytes were eluted at retention times of 4.4 min (RIO), 5.4 min (BOS), 8.9 min (MAC) and 7.8 min for gallopamil (internal standard, GAL). The method was found linear and calibrated in the ranges: 5–1000 ng/mL for RIO, 10–2000 ng/mL for BOS and 20–2000 ng/mL for MAC, with r² of 0.9991 for RIO, 0.9983 for BOS, and 0.9949 for MAC, respectively. Within the given ranges, the method ensured reliable results with the required precision and accuracy: ≤15 % (≤20 % for LLOQ). There was no significant carryover effect. The method has been successfully used in pilot study on adherence in patients treated for PAH, enabling monitoring of RIO, BOS and MAC. Drug concentrations were assessed in samples taken before (C0) and 3 h after drug administration (C3). For RIO, BOS and MAC, the developed method was suitable for both C0 and C3 samples, allowing steady-state drug determination if used. The presented method can be recommended to laboratories equipped with basic HPLC apparatus as an attractive analytical tool for both TDM and adherence studies.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.