Phuong Linh Ta, Lucia Bernat-Just, Adrián M. Alambiaga-Caravaca, Vicent Rodilla, Alicia López-Castellano, Francisco Bosch-Morell
{"title":"眼给药环糊精配合物中α-硫辛酸的高效液相色谱（HPLC-UV）定量方法优化与验证","authors":"Phuong Linh Ta, Lucia Bernat-Just, Adrián M. Alambiaga-Caravaca, Vicent Rodilla, Alicia López-Castellano, Francisco Bosch-Morell","doi":"10.1002/bmc.70133","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <p>Alpha-lipoic acid (ALA) is a natural antioxidant with therapeutic potential, but no method currently quantifies ALA in ocular structures. This study utilizes beta-cyclodextrins (β-CD) and (2-hydroxypropyl)-β-cyclodextrins (HP-β-CD) to enhance ALA solubility and stability in 1:1 M ratio. This study aims to optimize and validate a method for quantifying ALA in CD complexes using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) and to perform a diffusion study to evaluate ALA's permeability in ocular tissue. The HPLC method employed a C18 column (150 × 4.6 mm, 5 μm particle size) with an isocratic mobile phase of acetonitrile and water (50:50, v/v) at pH 4.0, a flow rate of 1 mL/min, detected ALA after 4.8 min at a wavelength of 334 nm. Validation of the method was conducted according to the ICH guideline M10 on bioanalytical method validation. The stability study revealed that better stability was obtained for ALA in β-CD inclusion in comparison with HP-β-CD. The <i>ex vivo</i> study using Franz diffusion cells was performed to confirm that ALA was able to permeate through the cornea and sclera via ocular administration. This accurate and precise HPLC-UV method paves the way for future research into ALA's potential to alleviate ocular conditions.</p>\n </section>\n \n <section>\n \n <h3> Summary</h3>\n \n <div>\n \n <ul>\n \n \n <li>This study optimized and validated a precise and accurate HPLC-UV method for quantifying ALA in CD complexes, following ICH guideline M10 on bioanalytical method validation.</li>\n \n \n <li>Stability studies revealed that β-CD complexes provided better thermal and solubility stability for ALA compared to HP-β-CD complexes.</li>\n \n \n <li>The <i>ex vivo</i> diffusion study confirmed that ALA successfully permeated the cornea and sclera via ocular administration.</li>\n \n \n <li>The validated method lays the groundwork for future research on ALA's potential to alleviate complications associated with ocular diseases.</li>\n </ul>\n </div>\n </section>\n </div>","PeriodicalId":8861,"journal":{"name":"Biomedical Chromatography","volume":"39 7","pages":""},"PeriodicalIF":1.8000,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bmc.70133","citationCount":"0","resultStr":"{\"title\":\"Optimization and Validation of a High-Performance Liquid Chromatography (HPLC-UV) Method for Quantification of α-Lipoic Acid in Cyclodextrins Complex for Ocular Delivery\",\"authors\":\"Phuong Linh Ta, Lucia Bernat-Just, Adrián M. Alambiaga-Caravaca, Vicent Rodilla, Alicia López-Castellano, Francisco Bosch-Morell\",\"doi\":\"10.1002/bmc.70133\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <p>Alpha-lipoic acid (ALA) is a natural antioxidant with therapeutic potential, but no method currently quantifies ALA in ocular structures. This study utilizes beta-cyclodextrins (β-CD) and (2-hydroxypropyl)-β-cyclodextrins (HP-β-CD) to enhance ALA solubility and stability in 1:1 M ratio. This study aims to optimize and validate a method for quantifying ALA in CD complexes using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) and to perform a diffusion study to evaluate ALA's permeability in ocular tissue. The HPLC method employed a C18 column (150 × 4.6 mm, 5 μm particle size) with an isocratic mobile phase of acetonitrile and water (50:50, v/v) at pH 4.0, a flow rate of 1 mL/min, detected ALA after 4.8 min at a wavelength of 334 nm. Validation of the method was conducted according to the ICH guideline M10 on bioanalytical method validation. The stability study revealed that better stability was obtained for ALA in β-CD inclusion in comparison with HP-β-CD. The <i>ex vivo</i> study using Franz diffusion cells was performed to confirm that ALA was able to permeate through the cornea and sclera via ocular administration. This accurate and precise HPLC-UV method paves the way for future research into ALA's potential to alleviate ocular conditions.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Summary</h3>\\n \\n <div>\\n \\n <ul>\\n \\n \\n <li>This study optimized and validated a precise and accurate HPLC-UV method for quantifying ALA in CD complexes, following ICH guideline M10 on bioanalytical method validation.</li>\\n \\n \\n <li>Stability studies revealed that β-CD complexes provided better thermal and solubility stability for ALA compared to HP-β-CD complexes.</li>\\n \\n \\n <li>The <i>ex vivo</i> diffusion study confirmed that ALA successfully permeated the cornea and sclera via ocular administration.</li>\\n \\n \\n <li>The validated method lays the groundwork for future research on ALA's potential to alleviate complications associated with ocular diseases.</li>\\n </ul>\\n </div>\\n </section>\\n </div>\",\"PeriodicalId\":8861,\"journal\":{\"name\":\"Biomedical Chromatography\",\"volume\":\"39 7\",\"pages\":\"\"},\"PeriodicalIF\":1.8000,\"publicationDate\":\"2025-06-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bmc.70133\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biomedical Chromatography\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/bmc.70133\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedical Chromatography","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/bmc.70133","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Optimization and Validation of a High-Performance Liquid Chromatography (HPLC-UV) Method for Quantification of α-Lipoic Acid in Cyclodextrins Complex for Ocular Delivery
Alpha-lipoic acid (ALA) is a natural antioxidant with therapeutic potential, but no method currently quantifies ALA in ocular structures. This study utilizes beta-cyclodextrins (β-CD) and (2-hydroxypropyl)-β-cyclodextrins (HP-β-CD) to enhance ALA solubility and stability in 1:1 M ratio. This study aims to optimize and validate a method for quantifying ALA in CD complexes using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) and to perform a diffusion study to evaluate ALA's permeability in ocular tissue. The HPLC method employed a C18 column (150 × 4.6 mm, 5 μm particle size) with an isocratic mobile phase of acetonitrile and water (50:50, v/v) at pH 4.0, a flow rate of 1 mL/min, detected ALA after 4.8 min at a wavelength of 334 nm. Validation of the method was conducted according to the ICH guideline M10 on bioanalytical method validation. The stability study revealed that better stability was obtained for ALA in β-CD inclusion in comparison with HP-β-CD. The ex vivo study using Franz diffusion cells was performed to confirm that ALA was able to permeate through the cornea and sclera via ocular administration. This accurate and precise HPLC-UV method paves the way for future research into ALA's potential to alleviate ocular conditions.
Summary
This study optimized and validated a precise and accurate HPLC-UV method for quantifying ALA in CD complexes, following ICH guideline M10 on bioanalytical method validation.
Stability studies revealed that β-CD complexes provided better thermal and solubility stability for ALA compared to HP-β-CD complexes.
The ex vivo diffusion study confirmed that ALA successfully permeated the cornea and sclera via ocular administration.
The validated method lays the groundwork for future research on ALA's potential to alleviate complications associated with ocular diseases.
期刊介绍：
Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.
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