Analysis of lysozyme in human saliva by CE-UV: A new simple, fast, and reliable method

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2025-07-21 DOI:10.1016/j.talanta.2025.128605

Radovan Tomašovský , Martina Opetová , Michaela Polačková , Katarína Maráková

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引用次数: 0

Abstract

Many studies have shown a correlation between salivary lysozyme levels and the presence of various diseases or stress. Here, we developed a new, simple method for the analysis of lysozyme in human saliva by capillary electrophoresis (CE) and UV detection that has potential application in clinical settings. The optimized method used 1 M formic acid +10 % (v/v) isopropyl alcohol as a background electrolyte, transient isotachophoresis as a preconcentration method, deactivation of the glass insert surface by carbonic anhydrase as a blocking agent to eliminate non-specific protein adsorption, and only simple dilution as a sample pretreatment. The validation showed high linearity (r² = 0.9997), inter- and intra-day accuracy (−2.03 to +5.36 %) and precision (<5 % RSD), and limit of detection (0.5 μg/mL). The stability of lysozyme in human saliva stored at −80 °C for 7 days using protease inhibitors and the variations of lysozyme excretion during the day and over 3 consecutive days were also investigated. The optimized and validated method was applied for the determination of salivary lysozyme levels in 25 healthy volunteers and the results showed the mean salivary lysozyme concentration at 14.3 ± 10.5 μg/mL. To the best of our knowledge, this is the first CE-UV method for human salivary lysozyme determination. This method meets the criteria of simplicity (minimum steps in sample pretreatment and analysis), speed (the entire process from saliva collection to result takes less than 45 min), reliability (highly satisfactory validation data), and suitability for application to real-world samples.

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CE-UV法分析人唾液中溶菌酶：一种简单、快速、可靠的新方法

许多研究表明唾液溶菌酶水平与各种疾病或压力之间存在相关性。在此，我们建立了一种新的，简单的毛细管电泳（CE）和紫外检测方法来分析人唾液中的溶菌酶，具有潜在的应用前景。优化后的方法以1 M甲酸+ 10% （v/v）异丙醇为背景电解液，瞬态等速电泳为预浓缩法，以碳酸酐酶为阻断剂使玻璃插入表面失活，消除非特异性蛋白质吸附，仅简单稀释作为样品前处理。结果表明，该方法具有良好的线性关系（r2 = 0.9997），日内、日间准确度（- 2.03 ~ + 5.36%）和精密度（< 5% RSD），检出限（0.5 μg/mL）。用蛋白酶抑制剂测定人唾液中溶菌酶在- 80°C保存7天的稳定性，以及溶菌酶在白天和连续3天的排泄变化。将优化后的方法应用于25名健康志愿者唾液溶菌酶水平的测定，结果表明，唾液溶菌酶平均浓度为14.3±10.5 μg/mL。据我们所知，这是首个CE-UV测定人唾液溶菌酶的方法。该方法符合简单性（样品预处理和分析步骤最少）、速度（从唾液采集到结果的整个过程少于45分钟）、可靠性（高度满意的验证数据）和适用于实际样品的标准。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.