Analytical Methods最新文献

{"title":"Colorimetric loop-mediated isothermal amplification (cLAMP) assay for the genotyping of a thrombophilia genetic risk factor, MTHFR (C677T)†","authors":"Reham Altwayan, Huseyin Tombuloglu, Moneerah Alsaeed and Abdulrahman Alhusil","doi":"10.1039/D5AY00215J","DOIUrl":"10.1039/D5AY00215J","url":null,"abstract":"<p >Methylenetetrahydrofolate reductase (MTHFR) is an enzyme involved in regulating serum homocysteine and folate levels. A single nucleotide polymorphism (SNP) of <em>MTHFR</em> (rs1801133; C677T) is strongly associated with an increased risk of thrombophilia, heart attack, stroke, and dementia. Genotyping this SNP is vital; however, techniques need to be simplified. This study developed a colorimetric loop-mediated isothermal amplification (cLAMP) technique to genotype the C677T in thrombophilia-susceptible individuals. The results were compared with those of kompetitive allele-specific PCR (KASP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays. The cLAMP assay successfully detected the C677T SNPs in the <em>MTHFR</em> gene. The results agreed with the KASP and PCR-RFLP results. The MTHFR-cLAMP assay is a simple, rapid, and affordable technique for colorimetric genotyping. This is the first study describing visual detection of a SNP associated with thrombophilia. The assay can also be used in screening other SNPs at low-resource centers and hospitals for point-of-care testing.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 21","pages":" 4432-4442"},"PeriodicalIF":2.7,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144109017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A water-soluble fluorescent probe based on porphyrin derivatives for Cu2+ detection in aqueous solution and living cells†","authors":"Jiajun Chen, Chenyang Zhang, Weiye Nie, Lidan Liu, Qi Zhou and Wenwu Cao","doi":"10.1039/D5AY00524H","DOIUrl":"10.1039/D5AY00524H","url":null,"abstract":"<p >Accurately tracing copper ions (Cu<small>²⁺</small>) is critically important in the fields of environmental protection and human body monitoring. Excessive intake of Cu<small>²⁺</small> not only easily leads to diseases, but also affects human health. Therefore, it is particularly important to find an easy-to-use, effective, and highly selective probe for <em>in situ</em> Cu<small>²⁺</small> detection in living cells. Herein, a water-soluble porphyrin derivative (WSPD) was successfully synthesized and used as a highly sensitive and selective fluorescent probe for detecting Cu<small>²⁺</small> in aqueous solutions and living cells. The WSPD possesses good water solubility and photostability, large Stokes shift (&gt;200 nm), appreciable red fluorescence and low cytotoxicity. The fluorescence signal could be rapidly quenched by Cu<small>²⁺</small> with high selectivity due to the strong affinity of Cu<small>²⁺</small> to the porphyrin core of the WSPD. The proposed fluorescent probe exhibited a low detection limit of 6.43 nM and a fast response time (&lt;5 min) for Cu<small>²⁺</small> detection in aqueous solution. Furthermore, it is successfully employed for the fluorescence imaging of intracellular Cu<small>²⁺</small> in living cells, demonstrating its great potential in the fields of biomedical imaging and biosensing.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 21","pages":" 4351-4358"},"PeriodicalIF":2.7,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144074953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Classification of Lu'an Gua Pian tea before and after Qingming Festival using HPLC-DAD analysis: a comparison of different data analysis strategies†","authors":"Dehuan Yang, Lilin Lin, Zhangfeng Tang, Zengping Chen, Yao Chen, Tong Wang and Ruqin Yu","doi":"10.1039/D5AY00382B","DOIUrl":"10.1039/D5AY00382B","url":null,"abstract":"<p >Lu'an Gua Pian tea (LAGP) is a traditional Chinese historical tea and one of the top ten famous teas in China. The price of LAGP from the same place of origin varies greatly in the market depending on the harvest time, with the LAGP harvested before the Qingming Festival (between April 4th and 6th every year) being regarded as a precious product. To accurately determine the harvest time of LAGP, especially around Qingming, HPLC-DAD combined with three different data analysis strategies (including targeted component analysis, non-targeted component analysis, and non-targeted fingerprint analysis) was evaluated and compared. Four machine learning algorithms were used to build the corresponding classification models, among which principal component analysis-linear discriminant analysis (PCA-LDA), partial least squares discriminant analysis (PLS-DA) and <em>K</em>-nearest neighbors (<em>K</em>-NN) achieved more than 95% classification accuracy. In addition, the non-targeted component analysis can classify LAGP picking time to a much smaller extent with an accuracy of 100% for PLS-DA. The advantages and disadvantages of the three data analysis strategies were compared. Regardless of the data analysis strategy used, the final classification accuracy was satisfactory. The appropriate data analysis strategy can be selected according to the specific experimental purpose. This work provides a variety of alternative solutions based on HPLC-DAD for identifying the picking time of Lu'an Gua Pian.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 19","pages":" 4061-4069"},"PeriodicalIF":2.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143953209","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancements in diagnostic approaches for Wilson's disease","authors":"Jitender Kumar and Indrajit Roy","doi":"10.1039/D5AY00118H","DOIUrl":"10.1039/D5AY00118H","url":null,"abstract":"<p >Wilson's disease (WD) is a genetic disorder that results in excessive copper build-up in tissues, causing significant liver and neurological damage. Early and accurate diagnosis is crucial for effective management and treatment. Traditional diagnostic methods, including serum ceruloplasmin and urinary copper level monitoring, liver biopsy and genetic testing, are limited by sensitivity, specificity, invasiveness, and accessibility. Recent advances in diagnostic technologies offer new hope for more accurate, rapid, and non-invasive detection of WD. In this regard, nanotechnology-driven formulations hold significant promise for both the early diagnosis and treatment of WD. Through the innovative use of advanced nanomaterials, researchers are developing more effective therapeutic options and highly sensitive diagnostic tools, potentially transforming the management and prognosis of this genetic disorder. This review provides a comprehensive analysis of recent advancements in WD diagnostics, focusing on research published since 2016. It explores the development and application of novel biomarkers, advanced imaging modalities, innovative biosensors, and emerging nanotechnology-based approaches. By integrating these cutting-edge methodologies, the review highlights their potential to enhance early and accurate detection of WD, addressing current diagnostic challenges and improving clinical outcomes.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 21","pages":" 4228-4250"},"PeriodicalIF":2.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144074967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of H2O2 and redox potential of an acute wound of rabbits with different sugar intakes based on a SERS-active microneedle","authors":"Xiulei Xiong, Haihua Shen, Songnan Liu, Fei Rong and Jian Dong","doi":"10.1039/D5AY00203F","DOIUrl":"10.1039/D5AY00203F","url":null,"abstract":"<p >To detect H<small>₂</small>O<small>₂</small> and redox potential of an acute wound simultaneously, a SERS-active microneedle was fabricated by integrating H<small>₂</small>O<small>₂</small> and redox potential SERS probes into two grooves of an acupuncture needle, respectively. When the SERS-active microneedle was inserted into tissues, an acute wound was formed and the two SERS probes were introduced into the wound to sense H<small>₂</small>O<small>₂</small> and redox potential of the wound. The feasibility of the SERS-active microneedle was evaluated <em>ex vivo</em> and <em>in vivo</em>. Both sugar deficiency and sugar excess induce increases in H<small>₂</small>O<small>₂</small> and redox potential in most tissues. Short-term physiological disturbances eventually returned to normal levels in healthy organisms, and abnormal sugar intakes disrupt the normal physiological environment and induce stress responses in all tissues. The SERS-active microneedle for detection of H<small>₂</small>O<small>₂</small> and redox potential would become a powerful tool, used for collecting physiological signals in different tissues to elucidate the mechanisms of wound healing and redox-related diseases.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 19","pages":" 4080-4086"},"PeriodicalIF":2.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143953107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cu-MOFs/GO composite-based electrochemical sensor for the simultaneous measurement of xanthine, hypoxanthine and caffeine at trace levels†","authors":"K. M. Supritha and M. Pandurangappa","doi":"10.1039/D5AY00308C","DOIUrl":"https://doi.org/10.1039/D5AY00308C","url":null,"abstract":"<p >Copper-MOFs were prepared <em>via</em> a simple solvothermal route using copper metal ion as the source and trimesic acid as the linker molecule at room temperature. The composite was then prepared by mixing the copper-MOFs with graphene oxide and used as a modifier for the simultaneous measurement of xanthine derivatives, such as xanthine, hypoxanthine and caffeine. The composite was characterised by X-ray diffraction, infrared spectroscopy, thermogravimetry and BET surface area studies. The electrochemical behaviour of the composite was studied through cyclic voltammetry, impedance and square wave voltammetry. The analytical signal responses for all three analytes were linear in the concentration range of 1–400 μM, with limits of detection of 0.045, 0.052 and 0.02 μM for xanthine, hypoxanthine and caffeine, respectively. The sensor exhibited wide linearity, good stability and reproducibility and was successfully applied to measure target analyte molecules from real sample matrices at trace levels, even in the presence of common interferants. The fabricated sensor can be utilised as an alternative method for the simultaneous measurement of xanthine, hypoxanthine and caffeine in complex sample matrices at low concentration levels.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 18","pages":" 3720-3728"},"PeriodicalIF":2.7,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925398","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A CRISPR/Cas12a-coupled multiplexed amplification system for ultrasensitive detection of miRNA-155†","authors":"Zhengdong Ai, Wenjun Wang, Xiang Li, Xianfeng Wang, Jia Chen, Jianming Wu and Shiying Zhou","doi":"10.1039/D5AY00415B","DOIUrl":"10.1039/D5AY00415B","url":null,"abstract":"<p >miRNA plays an important role in gene regulation and can be an effective biomarker for disease diagnosis. Herein, a new miRNA detection platform based on the CRISPR/Cas12a-coupled multiplexed amplification system is developed. In this strategy, miRNA-155 acts as an intermediary to trigger the recombinase polymerase amplification (RPA). Due to the introduction of endonuclide recognition sites in the amplification template, the resulting double-stranded DNA (dsDNA) can in turn initiate a strand replacement reaction (SDA), generating a great deal of single-stranded DNA (ssDNA). The ssDNA can directly unlock the trans-cleavage activity of CRSIPR/Cas12a, and the process is independent of PAM sites. Subsequently, the activated Cas12a trans-cleaves nearby signaling molecules, outputting a fluorescence/visualization signal. This method achieves miRNA detection as low as 68.69 fM, with a linear range of 200 fM to 1 nM, and shows good selectivity and repeatability. Meanwhile, the target of 10 pM can be distinguished by the naked eye. Moreover, the proposed method can achieve miRNA-155 detection in complicated cell extracts. The excellent detection sensitivity is mainly due to the integration of two amplification techniques, while the CRISPR/Cas12a system enables fast and accurate visual detection. More importantly, the actual detection results are consistent with standard methods (RT-qPCR), indicating that the CRISPR/Cas12a-coupled multiplexed amplification system is reliable and has potential clinical application value.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 19","pages":" 4044-4050"},"PeriodicalIF":2.7,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143950746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigations into bioavailability and micelle-assisted spectroscopic quantification of brodifacoum in natural samples†","authors":"Ganesh Suku, Anupama Vijayan and John Prakash","doi":"10.1039/D4AY01228C","DOIUrl":"10.1039/D4AY01228C","url":null,"abstract":"<p >The ability of a pseudo-surfactant, aqueous humic acid (HA), to significantly enhance the solubility of hydrophobic brodifacoum (BDF) – a second-generation anticoagulant – and to enable micelle assisted spectroscopic quantification of BDF is explored. The solubilization efficiency of aqueous HA for BDF was determined to be 2.3000 ± 0.0010 mg ppm<small>⁻¹</small>. The potential impact of the increased bioavailability of BDF on human health was monitored by studying the interaction of BDF with Bovine Serum Albumin (BSA) protein. There is a strong affinity between BDF and BSA, as evidenced by the binding constant (<em>K</em>) and the number of binding sites (<em>n</em>), which were found to be 391 000 ± 3.03 L mol<small>⁻¹</small> and 1, respectively. Additionally, fluorescence lifetime studies were conducted that showed that BDF interacted with the hydrophobic domain IIA of BSA. These findings underscore the need for a simple and sensitive analytical technique for effectively quantifying BDF. A micelle assisted fluorescence-based quantification tool for BDF is introduced, demonstrating good fidelity and excellent analytical figures of merits. The micellar system is characterized using Dynamic Light Scattering studies and confocal microscopy (size ∼270 nm). The number of molecules per micelle is calculated using Poisson's distribution, and it is found to be no more than one (&lt;0.2%). Encouragingly, in this study, micelle encapsulation was found to significantly enhance the fluorescence of BDF, allowing for its quantification at the nanomolar level. The analytical figures of merit such as the limit of detection (LOD) and limit of quantification (LOQ), were found to be 62 nM and 208 nM, respectively, with very good linearity (<em>R</em><small>²</small> = 0.994). The fidelity of the micelle assisted fluorescence quantification method was verified with high performance liquid chromatography using spiked natural water samples of varying HA content and the percentage recovery and Root Mean Square Error of Prediction (RMSEP) value were found to be about 100 and 0.1001, respectively. The low RMSEP indicates the potential utility of micelle assisted fluorescence quantification in developing sensitive assays for detecting BDF.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 20","pages":" 4158-4166"},"PeriodicalIF":2.7,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent advances in screen-printed carbon electrodes for food additive analysis","authors":"Gabriel Chitolina-Rodrigues, Devu Chandran, Rejithamol R. and Habdias A. Silva-Neto","doi":"10.1039/D5AY00236B","DOIUrl":"https://doi.org/10.1039/D5AY00236B","url":null,"abstract":"<p >Screen-printed carbon electrodes (SPCEs) are regarded as the actual and future sensing option for additive analysis in food samples; nonetheless, the sample preparation, selectivity, and detectability are key challenges to overcome for its technological development and wide application. In the present review, we inform, discuss, and compare some pivotal aspects associated with the fabrication of SPCEs, the presence of additives in foods, sample preparation, and voltammetric measurements of additives in food samples. Also, the proposed study has indicated that it is possible to develop suitable options for electroanalytical methodologies by using bare or modified SPCEs, which present affordable results in terms of selectivity, linear concentration range, and limit of detection for different classes of additives. Lastly, the review introduces challenging points that can be carefully evaluated for the next generation of SPCEs dedicated to additive analysis.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 18","pages":" 3613-3628"},"PeriodicalIF":2.7,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A RP-HPLC-UV method for the dual detection of fluconazole and clobetasol propionate and application to a model dual drug delivery hydrogel†","authors":"Robyn A. Macartney, Annabelle T. R. Fricker, Andrew M. Smith, Stefano Fedele, Ipsita Roy and Jonathan C. Knowles","doi":"10.1039/D4AY02219J","DOIUrl":"https://doi.org/10.1039/D4AY02219J","url":null,"abstract":"<p >Advanced drug delivery systems have become widely investigated to improve the efficacy of treatments for several diseases. These devices offer improved efficient, sustained, and targeted delivery which improves patient compliance, quality of life and minimises potential systemic side effects. As these therapeutic devices have advanced there is a potential for the development of products which deliver multiple drugs for simultaneous treatment of diseases. Given the interest in these dual-delivery devices it follows that new analytical methods need to be developed to detect and quantify different analytes during device development and validation. Here, for the first time, a reverse-phase high performance liquid chromatography (RP-HPLC) method is validated, utilising UV detection, for the dual detection of fluconazole and clobetasol propionate. The method is tested on a dual loaded model implant material intended as mucosal patches for the direct treatment of lichen planus and associated fungal infections. The method described here exhibited specificity and robustness with accurate and precise results. Good linearity was obtained between 0.25 and 2.5 mg mL<small>⁻¹</small> for fluconazole and 5 and 50 μg mL<small>⁻¹</small> for clobetasol propionate, with an <em>R</em><small>²</small> value of 0.9999 for the dual detection of fluconazole and clobetasol propionate. The developed method demonstrated selectivity and the solution containing both fluconazole and clobetasol propionate remained stable over a range of storage temperatures for up to 28 days. Within this validation study, the protocol was applied to a relevant dual loaded film showing the suitability of the method in studying drug release characteristics. The method described here also has a broader applicability for analysis and quantification of <em>in vitro</em> and <em>in vivo</em> drug release studies.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 18","pages":" 3694-3704"},"PeriodicalIF":2.7,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/ay/d4ay02219j?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143925395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}