Construction of a Dual-Mode Sensing Platform for Ultra-fast and Real-time Detection of Bisulfite in Food and Environmental Systems

Construction of a Dual-Mode Sensing Platform for Ultra-fast and Real-time Detection of Bisulfite in Food and Environmental Systems | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Construction of a Dual-Mode Sensing Platform for Ultra-fast and Real-time Detection of Bisulfite in Food and Environmental Systems Xiaoyu Huang, Jiaxing Li, Qiutong Chen, Mingyu Tian, Tianyu Liang, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5116472/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 20 Nov, 2024 Read the published version in Journal of Fluorescence → Version 1 posted 13 You are reading this latest preprint version Abstract Sulfur dioxide (SO 2 ) is widely used in food processing to extend the shelf life of food. However, excessive intake of SO 2 and its derivatives (HSO 3 - and SO 3 2- ) can cause oxidative damage to the body, resulting in several diseases. How to construct probes for rapid real-time detection of HSO 3 - in the field is beneficial to the developmental needs of practical applications, but it is also very challenging. Here we report a dual-mode fluorescent probe Rh-QL for ultrafast detection of HSO 3 - , which undergoes a specific 1,4-Michael addition reaction with sulfite to achieve Near-infrared fluorescence turn-on. Probe Rh-QL was able to detect HSO 3 - within 5 s with a significant color change from violet to green and a strong fluorescence signal at 700 nm. The probe Rh-QL has been used for the detection of HSO 3 - in real sugar samples and can be prepared as a portable sensing kit for the detection of HSO 3 - in the environment due to its high efficiency, rapidity and accuracy. In addition, the probe Rh-QL is able to target label Gram-negative bacteria after reacting with HSO 3 - , which has the potential to identify the type of pathogenic bacteria. SO2 derivatives Near-infrared fluorescence Environmental analysis bacteria. Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Sulfur dioxide (SO 2 ) as a gas signaling biomarker molecule that is strongly oxidizing and highly reactive, which are widely used in the processing of food to extend the shelf life of food.[ 1 – 5 ] Low concentrations (< 450 µM) of SO 2 and its derivatives (HSO 3 − and SO 3 2− ) are formed to maintain redox and biothiol homeostasis in organisms.[ 2 , 6 , 7 ] However, excessive intake of exogenous SO 2 can be metabolically converted to different types of sulfur/oxygen radicals that cause oxidative damage to the human body, leading to respiratory, cardiovascular, asthma, and lung cancer diseases.[ 8 , 9 ] The World Health Organization (WHO) recommends that the daily intake of SO 2 and its derivatives should be less than 0.7 mg/kg of body weight.[ 10 , 11 ] Therefore, the analysis and detection of harmful factors in food have become a key area of current research. In addition, bacteria are ubiquitous in human life, and serious bacterial infections will threaten human life and health.[ 12 , 13 ] It is worth noting that the massive abuse of antibiotics has led to increasing bacterial resistance.[ 14 , 15 ] Distinguishing between Gram-negative and Gram-positive bacteria to enable selective elimination of drug-resistant bacteria and prevent uncertainty in the inhibition process needs to be resolved. Therefore, it is urgent to find and develop tools that can rapidly discriminate between Gram-negative and Gram-positive bacteria. Currently, the traditional analytical methods for the detection of sulfur dioxide include fluorescent probe method, colorimetric method, chromatographic method and titrimetric method.[ 16 – 19 ] Among them, fluorescent probe-based detection methods have been widely used in the fields of bioimaging, food safety and bacterial detection due to their advantages of simple operation, selective specificity, high sensitivity, low background interference and real-time quantitative detection.[ 20 – 22 ] Up to now, a variety of fluorescent probes have been developed for the detection of SO 2 and its derivatives. However, most of the probes exhibit short fluorescence emission wavelengths or fluorescence quenching after interaction with HSO 3 − , which is often interfered by background fluorescence in food.[ 23 – 25 ] At the same time, some probes only exhibited a change in fluorescence characteristics without significant change in the UV spectrum, indicating that it is difficult to distinguished the levels of HSO 3 − in food and the environment with the naked eye. Therefore, these probes cannot be used as "indicator labels" for direct on-site detection, limiting the practical application value of the probes. In addition, utilizing the structural properties of the post-reaction probes to develop applications for the rapid identification of Gram-negative and positive bacteria would be beneficial for the timely diagnosis of bacterial infections and the development of subsequent therapeutic regimens. To address the above issues, a dual cationic fluorescent probe Rh-QL was designed and used as a selective multifunctional reagent to achieve a specific response that was successfully employed for indicator labeling, food detection, and bacterial target identification (Scheme 1 ). Experimental results showed that nucleophilic sulfites attacked the quinolinium backbone to generate an electron donor, leading to a structural change from the original acceptor-π-acceptor (A-π-A) to donor-π-acceptor (D-π-A), facilitating the ICT mechanism, accompanied by a substantial fluorescence enhancement and the change of solution color (purple to green). The ability of Rh-QL to achieve a dual-mode UV-fluorescence response to HSO 3 − within seconds (<5 s) prompted us to fabricate indicator labels for real-time monitoring of HSO 3 − levels in the field, while its near-infrared emission overcame background interference from other substances in the food samples to ensure detection accuracy. In addition, Rh-QL can be an excellent targeting agent for rapid identification of Gram-negative bacteria due to the intrinsic property of its cationic structure that enhances its interaction with anion-membrane bacteria. Therefore, the probe Rh-QL acts as an intelligent assay for rapid, sensitive, real-time monitoring of HSO 3 − and provides a valuable reference in the identification of Gram-negative bacteria. Result and discussion HSO 3 − response comparison of probes Rh-QL The synthetic route of the probe Rh-QL was summarized (Scheme 1 ) and structurally characterized by 1 H NMR, 13 C NMR and HRMS, the relevant spectra are shown in the Supporting Information. The optical response of the probe Rh-QL to HSO 3 − was first systematically evaluated. As shown in Fig. 1 a, different concentrations of HSO 3 − were added to the PBS solution containing the probe Rh-QL . The absorbance of the probe Rh-QL at 560 nm decreased rapidly with the increase of the HSO 3 − content, which was accompanied by the gradually increase of the absorbance at 690 nm, and the color of the solution of Rh-QL changed from purple to green. As shown in Fig. 1 b, the probe Rh-QL was in the dark state in PBS solution due to the typical A-π-A structure. Upon stepwise addition of HSO 3 − , a new fluorescence emission band centered at 725 nm appeared. When adding 200 µM of HSO 3 − , the fluorescence intensity of the probe Rh-QL increased about 55-fold and the curve stabilized, indicating that the saturation state was reached at this point. It is noteworthy that a satisfactory linear relationship was shown between the fluorescence intensity and HSO 3 − concentration (R 2 = 0.9997) (Fig. 1 c). The detection limit of the probe for HSO 3 − was calculated to be 2.61 µM according to the detection line equation σ/k.[ 21 , 26 ] The above experimental results indicate that the probe exhibits UV-fluorescence dual-mode response to HSO 3 − with high sensitivity, and is capable of visually identifying HSO 3 − in real samples. Portable fluorescence sensors can realize real-time detection of real samples in the field, and the response efficiency of the probe is an important index for evaluating portable fluorescence sensors. The response time of the probe Rh-QL with HSO 3 − was explored as shown in Fig. 1 c. After adding 200 µM of HSO 3 − to the probe Rh-QL solution, the color instantly changed from purple to green, which is conducive to real-time detection of HSO 3 − in the field. In addition, the excellent sensing properties of the probe Rh-QL were further demonstrated using selective experiments, as shown in Fig. 1 d, various anions were added to the PBS solution containing the probe Rh-QL , and under the excitation of 605 nm, only the probe solution with the addition of HSO 3 − showed an obvious fluorescence enhancement phenomenon at 725 nm, while the addition of other relevant anions could not cause the fluorescence intensity to change. It indicates that the probe Rh-QL can specifically recognize HSO 3 − . Meanwhile, the UV absorption spectra showed the same experimental results. Only the addition of HSO 3 − and SO 3 2− (12 µM) significantly increased the absorbance at 690 nm compared to other potential interferents. The complexity of the ecological environment prompted us to further validate the anti-interference ability of the probe Rh-QL . As shown in Fig. 1 d, even in the presence of other analytes, the probe Rh-QL still responded specifically only to HSO 3 − and the fluorescence intensity was significantly enhanced, which further demonstrated the excellent anti-interference ability of the probe Rh-QL . The application potential of the probe Rh-QL was further evaluated by determining its pH range. As shown in Fig. 1 F, Rh-QL showed almost no fluorescence emission in PBS solution at pH 1–13, indicating the good stability of Rh-QL . In contrast, the addition of HSO 3 − resulted in a significant increase of fluorescence in the pH range of 5–12, indicating that the probe is capable of detecting HSO 3 − over a wide pH range. The probe was further validated to have application potential for rapid, real-time detection of sulfite in complex ecological environments. Sensing mechanism of probe Rh-QL for HSO To further validate the aforementioned hypothesis, we employ density functional theory (DFT) and time-dependent density functional theory (TD-DFT) methods for quantum chemical calculations. As shown in Fig. 2 b, the probe Rh-QL undergoes a significant charge transfer from the lowest unoccupied molecular orbital (LUMO) to the highest occupied molecular orbital (HOMO), with the charge primarily moving from the left group to the upper group, resulting in fluorescence quenching. Assuming two possible products after the interaction of probe Rh-QL with HSO 3 − , as depicted in Fig. 2 a and 2 b, Product 1 exhibits a complete charge separation from the LUMO to the HOMO, with the charge shifting from the left group to the upper group. The photoinduced electron transfer (PET) effect leads to fluorescence quenching, which does not align with our experimental observations. In contrast, Product 2 shows a localized transition from the LUMO to the HOMO, emitting fluorescence, which fully conforms to our experimental findings. This further confirms that Product 2 is the adduct obtained after the probe Rh-QL recognizes HSO 3 − . Determination of HSO 3 − Rh-QL-based test strip and identification of Rh-QL anti-counterfeiting seals for HSO 3 − . Inspired by the excellent performance of Rh-QL , we explored a portable sensing platform for on-site detection of HSO 3 − . As shown in Fig. 3 a, the Portable sensing kits are prepared by immersing test paper into Rh-QL (500 µM) solution, which appears purple under natural light. After adding different concentrations of HSO 3 − , the portable kit displayed obvious color change from purple to green (Fig. 3 b). The above experimental results demonstrated that the Rh-QL could detect HSO 3 − in real samples by naked eye on-site. This apparent color change prompted us to further evaluate application of the probe Rh-QL in anti-counterfeiting technology. As shown in Fig. 3 c, the probe Rh-QL was dissolved in anhydrous ethanol and glycerol to produce the specific solution. Then the anti-counterfeit inks with “❀” pattern was printed on the thin-layer chromatographic plate and sprayed with an aqueous solution containing HSO 3 − , which was observed to change the inks pattern from purple to green under natural light. This confirms that the probe Rh-QL can be applied to anti-counterfeit inks. Detection of HSO3- in real food Detection of HSO 3 − levels in food samples is crucial in the field of food safety. The excellent recognition of HSO 3 − by Rh-QL in the spectral data encouraged us to further explore its ability to detect HSO 3 − in real samples. As shown in Table 1 , we spiked different concentrations of HSO 3 − into HEPES buffer solutions of white sugar, granulated sugar, brown sugar and crystal sugar, and then measured the HSO 3 − content using the previously obtained working curve plots. The results showed good recovery and relative standard deviation data for this assay. In order to verify the accuracy of Rh-QL , spectrophotometric method (GB 5009.34–2022, National Standard for Food Safety) was used as a standard method to detect the content of SO 2 in the same food samples without extra addition, the results were very similar to those of the fluorescence method of probe Rh-QL , indicating that probe Rh-QL could be applied to quantitatively detect HSO 3 − in real food samples without the help of any special instrument. Table 1 Detection results of HSO 3 − in white sugar, granulated sugar, brown sugar and crystal sugar determined by Rh-QL and spectrophotometry method Sample HSO 3 − Spiked (µM) HSO 3 − recovered (µM) Recovery (%) RSD (%) SO 2 recovered (mg/kg) results of SO 2 detection(mg/kg) [1] SO 2 residue requirements(g/kg) [2] White sugar 0 0.53 ± 0.22 - 41.02 0.044900 0.343994 0.1 5 5.49 ± 1.04 99.13 18.97 10 10.59 ± 0.15 100.57 1.39 Granulated sugar 0 8.06 ± 0.26 - 3.21 0.026299 0.343994 0.1 5 13.08 ± 0.12 100.37 0.92 10 17.49 ± 0.47 94.23 2.70 Brown sugar 0 5.30 ± 0.51 - 2.03 0.323925 0.343994 0.1 5 10.18 ± 0.16 97.58 0.75 10 15.36 ± 0.84 100.55 1.48 Crystal sugar 0 8.83 ± 0.09 - 0.99 0.566389 0.343994 0.1 5 13.91 ± 0.56 101.68 4.03 10 19.18 ± 0.69 103.46 3.58 Bacterial imaging with Rh-QL Specific identification of Gram-negative and Gram-positive bacteria enables selective elimination of drug-resistant bacteria and facilitates timely diagnosis of the type of bacterial infection and development of subsequent treatment regimens. As shown in Fig. 4 , the probe Rh-QL was co-incubated with S.putrefaciens and S.aureus , no obvious red fluorescence was observed in the red channel. After the probe Rh-QL interacted with HSO 3 − and co-cultured with S.putrefaciens and S.aureus respectively, there was still no obvious red fluorescence in the S.aureus , while a bright red fluorescence signal was produced in the S.putrefaciens , indicating that Rh-QL - HSO 3 − was able to successfully illuminate the S.putrefaciens . The main reason for this phenomenon is that the intrinsic properties of the Rh-QL -HSO 3 − cationic structure interacts with the bacterial anionic membrane, which enables it to act as a targeting agent to rapidly recognize Gram-negative bacteria emitting bright red fluorescence. Thus, the ability of Rh-QL -HSO 3 − to specifically recognize pathogenic bacteria contributes to the rational selection of subsequent treatment regimens. Conclusions In summary, we report a colorimetric and fluorescent dual-mode sensing platform Rh-QL for ultrafast detection of HSO 3 − . The probe Rh-QL exhibited exclusive selectivity, excellent immunity to interference, ultrafast response time, and high sensitivity to HSO 3 − . In addition, Rh-QL achieves quantitative detection of HSO 3 − in real sugar samples and specifically recognizes pathogenic bacteria types, contributing to the selective elimination of bacteria. The probe Rh-QL can be used as a portable sensing platform to accurately, rapidly, and cost-effectively achieve the detection of HSO 3 − , which is potentially valuable in food safety and other fields. Declarations Author Contribution Xiaoyu Huang, performed the experiment and wrote the manuscript;Jiaxing Li, Investigation and performed the data analyses;Qiutong Chen, performed the data analyses;Mingyu Tian, contributed to the conception of the study and imaging work;Tianyu, Liang, Investigation and validation;Lijun Tang, contributed to the conception of the study and funding acquisition. Acknowledgement This work was supported by the National Natural Science Foundation of China (No. 22278038), the Program for Distinguished Professor of Liaoning Province, and the Open Project of the Institute of Ocean of Bohai University (No. BDHYYJY2024010). References Shang ZY, Wu MM, Meng QT, Jiao Y, Zhang ZQ, Zhang R (2024) A near-infrared fluorescent probe for rapid and on-site detection of sulfur dioxide derivative in biological, food and environmental systems. 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Anal Chem 87609-616 Li Y, Sun XF, Zhou LL, Tian L, Zhong KL, Zhang JL, et al (2022) Novel Colorimetric and NIR Fluorescent Probe for Bisulfite/Sulfite Detection in Food and Water Samples and Living Cells Based on the PET Mechanism. J Agr Food Chem 70:10899-906 Scheme Scheme 1 is available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Supportinginformation.docx GraphicalAbstract.png Graphical Abstract We report a new quinoline-based dual-mode sensor (Rh-QL) for selective recognition of Bisulfite with near-infrared emission Scheme1.png Cite Share Download PDF Status: Published Journal Publication published 20 Nov, 2024 Read the published version in Journal of Fluorescence → Version 1 posted Editorial decision: Revision requested 08 Oct, 2024 Reviews received at journal 08 Oct, 2024 Reviews received at journal 07 Oct, 2024 Reviewers agreed at journal 06 Oct, 2024 Reviewers agreed at journal 06 Oct, 2024 Reviews received at journal 04 Oct, 2024 Reviewers agreed at journal 03 Oct, 2024 Reviewers agreed at journal 02 Oct, 2024 Reviewers agreed at journal 01 Oct, 2024 Reviewers invited by journal 01 Oct, 2024 Editor assigned by journal 20 Sep, 2024 Submission checks completed at journal 20 Sep, 2024 First submitted to journal 19 Sep, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5116472","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":363633530,"identity":"2bcc1500-b46c-406d-8241-198a483ca547","order_by":0,"name":"Xiaoyu Huang","email":"","orcid":"","institution":"Bohai University","correspondingAuthor":false,"prefix":"","firstName":"Xiaoyu","middleName":"","lastName":"Huang","suffix":""},{"id":363633531,"identity":"458c3c36-9d74-477b-b49d-7241bcd1df0a","order_by":1,"name":"Jiaxing Li","email":"","orcid":"","institution":"Bohai University","correspondingAuthor":false,"prefix":"","firstName":"Jiaxing","middleName":"","lastName":"Li","suffix":""},{"id":363633532,"identity":"b20bb792-ca44-49e0-91a4-7e66345c5ad4","order_by":2,"name":"Qiutong Chen","email":"","orcid":"","institution":"Bohai University","correspondingAuthor":false,"prefix":"","firstName":"Qiutong","middleName":"","lastName":"Chen","suffix":""},{"id":363633533,"identity":"199d7bd6-2a60-4684-a107-75be55551f7e","order_by":3,"name":"Mingyu Tian","email":"","orcid":"","institution":"Bohai University","correspondingAuthor":false,"prefix":"","firstName":"Mingyu","middleName":"","lastName":"Tian","suffix":""},{"id":363633534,"identity":"a80ea107-5f63-4394-ba1c-c7c53bbe8e12","order_by":4,"name":"Tianyu Liang","email":"","orcid":"","institution":"Bohai University","correspondingAuthor":false,"prefix":"","firstName":"Tianyu","middleName":"","lastName":"Liang","suffix":""},{"id":363633535,"identity":"36cab4e6-8370-463d-bf48-7a5bab4f1c15","order_by":5,"name":"Lijun Tang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAw0lEQVRIiWNgGAWjYBACPoYDIMomwQBE8RCjhY3hMIhKI0kLM4g6TIoWxvMHPxf8Op9nLpHA+OBtG4O8OREOY5ae2Xe72HJGArPh3DYGw50NhLUwSPP23E7ccCOBTZq3jSHB4AARtvzm7TkH0sL+m1gtbNI8Pw6AbWEmVouZNW9DcrHBmYfNknPOSRhuIKSFX+Lg49s8f+zyDI4nH/zwpsxGnqAtDBJAFYxtIBZjA4hLSD3IGpDCP0QoHAWjYBSMgpELAF3oQGSOsgzMAAAAAElFTkSuQmCC","orcid":"","institution":"Bohai University","correspondingAuthor":true,"prefix":"","firstName":"Lijun","middleName":"","lastName":"Tang","suffix":""}],"badges":[],"createdAt":"2024-09-19 11:08:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5116472/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5116472/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10895-024-04031-x","type":"published","date":"2024-11-20T15:57:43+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":68358062,"identity":"7d1e4b15-2f2f-44a7-8f68-0d740d066ac2","added_by":"auto","created_at":"2024-11-06 11:44:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":637099,"visible":true,"origin":"","legend":"\u003cp\u003e(a) UV–vis absorption spectrum changes of \u003cstrong\u003eRh-QL\u003c/strong\u003e (50 μM) with different concentrations of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e from 0 μM to 200 μM; (b) Fluorescence-emission spectra of \u003cstrong\u003eRh-QL\u003c/strong\u003e (50 μM) reduced by different concentrations of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e from 0 μM to 200 μM; (c) Time-trace heliographs of \u003cstrong\u003eRh-QL\u003c/strong\u003e (50 μM) in aqueous solution after addition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e (50 μM); (d) The fluorescence intensity (at 725 nm) of the probe \u003cstrong\u003eRh-QL\u003c/strong\u003e (10 μM) in PBS solution was determined by addition of various analytes (200 μM) and HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e- \u003c/sup\u003e(200 μM); (e) The absorbance (at 690 nm) of the probe \u003cstrong\u003eRh-QL\u003c/strong\u003e (10 μM) in PBS solution was determined by addition of various analytes (200 μM) and HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e(200 μM); (Analytes include HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, SO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e, HS\u003csup\u003e-\u003c/sup\u003e, CN\u003csup\u003e-\u003c/sup\u003e, CH\u003csub\u003e3\u003c/sub\u003eCOO\u003csup\u003e-\u003c/sup\u003e, PPI, GSH, Cys, Hcy, Cl\u003csup\u003e-\u003c/sup\u003e, Br\u003csup\u003e-\u003c/sup\u003e, I\u003csup\u003e-\u003c/sup\u003e, F\u003csup\u003e-\u003c/sup\u003e, SCN\u003csup\u003e-\u003c/sup\u003e, HPO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e, S\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e, SO\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e, S\u003csup\u003e2-\u003c/sup\u003e, N\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, HCO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, CO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e); (f) Fluorescence response (630 nm) of \u003cstrong\u003eRh-QL\u003c/strong\u003e (10 μM) to HSO\u003csup\u003e3-\u003c/sup\u003e at different pH conditions.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/50034af6c8a8bf410a6d7718.png"},{"id":68357086,"identity":"a2d95f07-06f3-4fbb-a07f-826e168a78af","added_by":"auto","created_at":"2024-11-06 11:36:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":738546,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Proposed mechanism of probe \u003cstrong\u003eRh-QL \u003c/strong\u003eresponse to HSO\u003csub\u003e3\u003c/sub\u003e; (b) The optimal structure, HOMO and LUMO distributions of \u003cstrong\u003eRh-QL\u003c/strong\u003e, \u003cstrong\u003eRh-QL\u003c/strong\u003e - HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/1cea2700efde212df57c6aea.png"},{"id":68357088,"identity":"487ace68-f9b5-4214-9326-e58c3cb46ecd","added_by":"auto","created_at":"2024-11-06 11:36:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":318593,"visible":true,"origin":"","legend":"\u003cp\u003e(a) Schematic representation of the preparation and testing procedures of \u003cstrong\u003eRh-QL\u003c/strong\u003e-based paper strips. (b) Colorimetric recognition of \u003cstrong\u003eRh-QL\u003c/strong\u003e-based test strips toward HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e (0 μM、50 μM、100 μM、150 μM、200 μM); (c) The Color change of the pattern on the TLC plate after spraying HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e solution\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/c38bd5dd8325b05b01e2e31d.png"},{"id":68357090,"identity":"19bd7936-e9f6-4886-bd65-532405b896e6","added_by":"auto","created_at":"2024-11-06 11:36:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":560803,"visible":true,"origin":"","legend":"\u003cp\u003eThe confocal images of S. putrefaciens and S. aureus bacteria under \u003cstrong\u003eRh-QL\u003c/strong\u003e (10 μM) and \u003cstrong\u003eRh-QL\u003c/strong\u003e-HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e (10 μM) incubation. Fluorescence (\u003cem\u003eλ\u003c/em\u003e\u003csub\u003e\u003cem\u003eex\u003c/em\u003e\u003c/sub\u003e = 600 nm, \u003cem\u003eλ\u003c/em\u003e\u003csub\u003e\u003cem\u003eem\u003c/em\u003e\u003c/sub\u003e = 660 - 740 nm), Scale bars: 10 μm.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/16959a97620adf78bcd896b1.png"},{"id":69835865,"identity":"f344041d-53ad-4fad-88d2-e3117412a9b7","added_by":"auto","created_at":"2024-11-25 16:14:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3222288,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/fe2ae00d-5189-4654-aa30-36a1432d3e60.pdf"},{"id":68357091,"identity":"d1ae5397-2c29-4495-aa7b-198bb18fb067","added_by":"auto","created_at":"2024-11-06 11:36:39","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":636428,"visible":true,"origin":"","legend":"","description":"","filename":"Supportinginformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/34441cf8a6727de4fb1bb9f0.docx"},{"id":68358063,"identity":"7ed5362d-a6af-4d2d-b8f9-98364931f168","added_by":"auto","created_at":"2024-11-06 11:44:39","extension":"png","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":59561,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical Abstract\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe report a new quinoline-based dual-mode sensor (\u003cstrong\u003eRh-QL\u003c/strong\u003e) for selective recognition of Bisulfite with near-infrared emission\u003c/p\u003e","description":"","filename":"GraphicalAbstract.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/d5a8ff7051423a71f10d1559.png"},{"id":68357092,"identity":"392cce24-a4f2-40e9-9116-426dc66ffba8","added_by":"auto","created_at":"2024-11-06 11:36:39","extension":"png","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":197265,"visible":true,"origin":"","legend":"","description":"","filename":"Scheme1.png","url":"https://assets-eu.researchsquare.com/files/rs-5116472/v1/3f00d0a378c9325e1590635e.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Construction of a Dual-Mode Sensing Platform for Ultra-fast and Real-time Detection of Bisulfite in Food and Environmental Systems","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSulfur dioxide (SO\u003csub\u003e2\u003c/sub\u003e) as a gas signaling biomarker molecule that is strongly oxidizing and highly reactive, which are widely used in the processing of food to extend the shelf life of food.[\u003cspan additionalcitationids=\"CR2 CR3 CR4\" citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] Low concentrations (\u0026lt;\u0026thinsp;450 \u0026micro;M) of SO\u003csub\u003e2\u003c/sub\u003e and its derivatives (HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and SO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2\u0026minus;\u003c/sup\u003e) are formed to maintain redox and biothiol homeostasis in organisms.[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] However, excessive intake of exogenous SO\u003csub\u003e2\u003c/sub\u003e can be metabolically converted to different types of sulfur/oxygen radicals that cause oxidative damage to the human body, leading to respiratory, cardiovascular, asthma, and lung cancer diseases.[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e] The World Health Organization (WHO) recommends that the daily intake of SO\u003csub\u003e2\u003c/sub\u003e and its derivatives should be less than 0.7 mg/kg of body weight.[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] Therefore, the analysis and detection of harmful factors in food have become a key area of current research. In addition, bacteria are ubiquitous in human life, and serious bacterial infections will threaten human life and health.[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] It is worth noting that the massive abuse of antibiotics has led to increasing bacterial resistance.[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] Distinguishing between Gram-negative and Gram-positive bacteria to enable selective elimination of drug-resistant bacteria and prevent uncertainty in the inhibition process needs to be resolved. Therefore, it is urgent to find and develop tools that can rapidly discriminate between Gram-negative and Gram-positive bacteria.\u003c/p\u003e \u003cp\u003eCurrently, the traditional analytical methods for the detection of sulfur dioxide include fluorescent probe method, colorimetric method, chromatographic method and titrimetric method.[\u003cspan additionalcitationids=\"CR17 CR18\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] Among them, fluorescent probe-based detection methods have been widely used in the fields of bioimaging, food safety and bacterial detection due to their advantages of simple operation, selective specificity, high sensitivity, low background interference and real-time quantitative detection.[\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e] Up to now, a variety of fluorescent probes have been developed for the detection of SO\u003csub\u003e2\u003c/sub\u003e and its derivatives. However, most of the probes exhibit short fluorescence emission wavelengths or fluorescence quenching after interaction with HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, which is often interfered by background fluorescence in food.[\u003cspan additionalcitationids=\"CR24\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e] At the same time, some probes only exhibited a change in fluorescence characteristics without significant change in the UV spectrum, indicating that it is difficult to distinguished the levels of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in food and the environment with the naked eye. Therefore, these probes cannot be used as \"indicator labels\" for direct on-site detection, limiting the practical application value of the probes. In addition, utilizing the structural properties of the post-reaction probes to develop applications for the rapid identification of Gram-negative and positive bacteria would be beneficial for the timely diagnosis of bacterial infections and the development of subsequent therapeutic regimens.\u003c/p\u003e \u003cp\u003eTo address the above issues, a dual cationic fluorescent probe \u003cb\u003eRh-QL\u003c/b\u003e was designed and used as a selective multifunctional reagent to achieve a specific response that was successfully employed for indicator labeling, food detection, and bacterial target identification (Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Experimental results showed that nucleophilic sulfites attacked the quinolinium backbone to generate an electron donor, leading to a structural change from the original acceptor-π-acceptor (A-π-A) to donor-π-acceptor (D-π-A), facilitating the ICT mechanism, accompanied by a substantial fluorescence enhancement and the change of solution color (purple to green). The ability of \u003cb\u003eRh-QL\u003c/b\u003e to achieve a dual-mode UV-fluorescence response to HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e within seconds (\u0026lt;5 s) prompted us to fabricate indicator labels for real-time monitoring of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e levels in the field, while its near-infrared emission overcame background interference from other substances in the food samples to ensure detection accuracy. In addition, \u003cb\u003eRh-QL\u003c/b\u003e can be an excellent targeting agent for rapid identification of Gram-negative bacteria due to the intrinsic property of its cationic structure that enhances its interaction with anion-membrane bacteria. Therefore, the probe \u003cb\u003eRh-QL\u003c/b\u003e acts as an intelligent assay for rapid, sensitive, real-time monitoring of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003eand provides a valuable reference in the identification of Gram-negative bacteria.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Result and discussion","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eHSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e response comparison of probes Rh-QL\u003c/h2\u003e \u003cp\u003eThe synthetic route of the probe \u003cb\u003eRh-QL\u003c/b\u003e was summarized (Scheme \u003cspan refid=\"Sch1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) and structurally characterized by \u003csup\u003e1\u003c/sup\u003eH NMR, \u003csup\u003e13\u003c/sup\u003eC NMR and HRMS, the relevant spectra are shown in the Supporting Information.\u003c/p\u003e \u003cp\u003eThe optical response of the probe \u003cb\u003eRh-QL\u003c/b\u003e to HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e was first systematically evaluated. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, different concentrations of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e were added to the PBS solution containing the probe \u003cb\u003eRh-QL\u003c/b\u003e. The absorbance of the probe \u003cb\u003eRh-QL\u003c/b\u003e at 560 nm decreased rapidly with the increase of the HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e content, which was accompanied by the gradually increase of the absorbance at 690 nm, and the color of the solution of \u003cb\u003eRh-QL\u003c/b\u003e changed from purple to green. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb, the probe \u003cb\u003eRh-QL\u003c/b\u003e was in the dark state in PBS solution due to the typical A-π-A structure. Upon stepwise addition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, a new fluorescence emission band centered at 725 nm appeared. When adding 200 \u0026micro;M of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, the fluorescence intensity of the probe \u003cb\u003eRh-QL\u003c/b\u003e increased about 55-fold and the curve stabilized, indicating that the saturation state was reached at this point. It is noteworthy that a satisfactory linear relationship was shown between the fluorescence intensity and HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e concentration (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9997) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). The detection limit of the probe for HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e was calculated to be 2.61 \u0026micro;M according to the detection line equation σ/k.[\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] The above experimental results indicate that the probe exhibits UV-fluorescence dual-mode response to HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e with high sensitivity, and is capable of visually identifying HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in real samples.\u003c/p\u003e \u003cp\u003ePortable fluorescence sensors can realize real-time detection of real samples in the field, and the response efficiency of the probe is an important index for evaluating portable fluorescence sensors. The response time of the probe \u003cb\u003eRh-QL\u003c/b\u003e with HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e was explored as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec. After adding 200 \u0026micro;M of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e to the probe \u003cb\u003eRh-QL\u003c/b\u003e solution, the color instantly changed from purple to green, which is conducive to real-time detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in the field. In addition, the excellent sensing properties of the probe \u003cb\u003eRh-QL\u003c/b\u003e were further demonstrated using selective experiments, as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed, various anions were added to the PBS solution containing the probe \u003cb\u003eRh-QL\u003c/b\u003e, and under the excitation of 605 nm, only the probe solution with the addition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e showed an obvious fluorescence enhancement phenomenon at 725 nm, while the addition of other relevant anions could not cause the fluorescence intensity to change. It indicates that the probe \u003cb\u003eRh-QL\u003c/b\u003e can specifically recognize HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. Meanwhile, the UV absorption spectra showed the same experimental results. Only the addition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and SO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2\u0026minus;\u003c/sup\u003e (12 \u0026micro;M) significantly increased the absorbance at 690 nm compared to other potential interferents. The complexity of the ecological environment prompted us to further validate the anti-interference ability of the probe \u003cb\u003eRh-QL\u003c/b\u003e. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed, even in the presence of other analytes, the probe \u003cb\u003eRh-QL\u003c/b\u003e still responded specifically only to HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and the fluorescence intensity was significantly enhanced, which further demonstrated the excellent anti-interference ability of the probe \u003cb\u003eRh-QL\u003c/b\u003e. The application potential of the probe \u003cb\u003eRh-QL\u003c/b\u003e was further evaluated by determining its pH range. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eF, \u003cb\u003eRh-QL\u003c/b\u003e showed almost no fluorescence emission in PBS solution at pH 1\u0026ndash;13, indicating the good stability of \u003cb\u003eRh-QL\u003c/b\u003e. In contrast, the addition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e resulted in a significant increase of fluorescence in the pH range of 5\u0026ndash;12, indicating that the probe is capable of detecting HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e over a wide pH range. The probe was further validated to have application potential for rapid, real-time detection of sulfite in complex ecological environments.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSensing mechanism of probe Rh-QL for HSO\u003c/h3\u003e\n\u003cp\u003eTo further validate the aforementioned hypothesis, we employ density functional theory (DFT) and time-dependent density functional theory (TD-DFT) methods for quantum chemical calculations. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb, the probe \u003cb\u003eRh-QL\u003c/b\u003e undergoes a significant charge transfer from the lowest unoccupied molecular orbital (LUMO) to the highest occupied molecular orbital (HOMO), with the charge primarily moving from the left group to the upper group, resulting in fluorescence quenching. Assuming two possible products after the interaction of probe \u003cb\u003eRh-QL\u003c/b\u003e with HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, as depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb, Product 1 exhibits a complete charge separation from the LUMO to the HOMO, with the charge shifting from the left group to the upper group. The photoinduced electron transfer (PET) effect leads to fluorescence quenching, which does not align with our experimental observations. In contrast, Product 2 shows a localized transition from the LUMO to the HOMO, emitting fluorescence, which fully conforms to our experimental findings. This further confirms that Product 2 is the adduct obtained after the probe \u003cb\u003eRh-QL\u003c/b\u003e recognizes HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eDetermination of HSO\u003c/b\u003e \u003csub\u003e \u003cb\u003e3\u003c/b\u003e \u003c/sub\u003e \u003csup\u003e \u003cb\u003e\u0026minus;\u003c/b\u003e \u003c/sup\u003e \u003cb\u003eRh-QL-based test strip and identification of Rh-QL anti-counterfeiting seals for HSO\u003c/b\u003e\u003csub\u003e\u003cb\u003e3\u003c/b\u003e\u003c/sub\u003e\u003csup\u003e\u003cb\u003e\u0026minus;\u003c/b\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eInspired by the excellent performance of \u003cb\u003eRh-QL\u003c/b\u003e, we explored a portable sensing platform for on-site detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea, the Portable sensing kits are prepared by immersing test paper into \u003cb\u003eRh-QL\u003c/b\u003e (500 \u0026micro;M) solution, which appears purple under natural light. After adding different concentrations of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, the portable kit displayed obvious color change from purple to green (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). The above experimental results demonstrated that the \u003cb\u003eRh-QL\u003c/b\u003e could detect HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in real samples by naked eye on-site. This apparent color change prompted us to further evaluate application of the probe \u003cb\u003eRh-QL\u003c/b\u003e in anti-counterfeiting technology. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec, the probe \u003cb\u003eRh-QL\u003c/b\u003e was dissolved in anhydrous ethanol and glycerol to produce the specific solution. Then the anti-counterfeit inks with \u0026ldquo;❀\u0026rdquo; pattern was printed on the thin-layer chromatographic plate and sprayed with an aqueous solution containing HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, which was observed to change the inks pattern from purple to green under natural light. This confirms that the probe \u003cb\u003eRh-QL\u003c/b\u003e can be applied to anti-counterfeit inks.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003eDetection of HSO3- in real food\u003c/h3\u003e\n\u003cp\u003eDetection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e levels in food samples is crucial in the field of food safety. The excellent recognition of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e by \u003cb\u003eRh-QL\u003c/b\u003e in the spectral data encouraged us to further explore its ability to detect HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in real samples. As shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, we spiked different concentrations of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e into HEPES buffer solutions of white sugar, granulated sugar, brown sugar and crystal sugar, and then measured the HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e content using the previously obtained working curve plots. The results showed good recovery and relative standard deviation data for this assay. In order to verify the accuracy of \u003cb\u003eRh-QL\u003c/b\u003e, spectrophotometric method (GB 5009.34\u0026ndash;2022, National Standard for Food Safety) was used as a standard method to detect the content of SO\u003csub\u003e2\u003c/sub\u003e in the same food samples without extra addition, the results were very similar to those of the fluorescence method of probe \u003cb\u003eRh-QL\u003c/b\u003e, indicating that probe \u003cb\u003eRh-QL\u003c/b\u003e could be applied to quantitatively detect HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in real food samples without the help of any special instrument.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDetection results of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in white sugar, granulated sugar, brown sugar and crystal sugar determined by \u003cb\u003eRh-QL\u003c/b\u003e and spectrophotometry method\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eSpiked\u003c/p\u003e \u003cp\u003e(\u0026micro;M)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eHSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/p\u003e \u003cp\u003erecovered\u003c/p\u003e \u003cp\u003e(\u0026micro;M)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRecovery (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRSD (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSO\u003csub\u003e2\u003c/sub\u003e recovered\u003c/p\u003e \u003cp\u003e(mg/kg)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eresults of SO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003cp\u003edetection(mg/kg)\u003csup\u003e[1]\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eSO\u003csub\u003e2\u003c/sub\u003e residue requirements(g/kg)\u003csup\u003e[2]\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eWhite sugar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e0.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e41.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.044900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.343994\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e5.49\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e99.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e10.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eGranulated\u003c/p\u003e \u003cp\u003esugar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e8.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.026299\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.343994\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e13.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e17.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e94.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eBrown sugar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e5.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.323925\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.343994\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e10.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e97.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e15.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eCrystal sugar\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e8.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.566389\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.343994\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e13.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e101.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e19.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e103.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e\n\u003ch3\u003eBacterial imaging with Rh-QL\u003c/h3\u003e\n\u003cp\u003eSpecific identification of Gram-negative and Gram-positive bacteria enables selective elimination of drug-resistant bacteria and facilitates timely diagnosis of the type of bacterial infection and development of subsequent treatment regimens. As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, the probe \u003cb\u003eRh-QL\u003c/b\u003e was co-incubated with \u003cem\u003eS.putrefaciens\u003c/em\u003e and \u003cem\u003eS.aureus\u003c/em\u003e, no obvious red fluorescence was observed in the red channel. After the probe \u003cb\u003eRh-QL\u003c/b\u003e interacted with HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and co-cultured with \u003cem\u003eS.putrefaciens\u003c/em\u003e and \u003cem\u003eS.aureus\u003c/em\u003e respectively, there was still no obvious red fluorescence in the \u003cem\u003eS.aureus\u003c/em\u003e, while a bright red fluorescence signal was produced in the \u003cem\u003eS.putrefaciens\u003c/em\u003e, indicating that \u003cb\u003eRh-QL\u003c/b\u003e- HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e was able to successfully illuminate the \u003cem\u003eS.putrefaciens\u003c/em\u003e. The main reason for this phenomenon is that the intrinsic properties of the \u003cb\u003eRh-QL\u003c/b\u003e-HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e cationic structure interacts with the bacterial anionic membrane, which enables it to act as a targeting agent to rapidly recognize Gram-negative bacteria emitting bright red fluorescence. Thus, the ability of \u003cb\u003eRh-QL\u003c/b\u003e-HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e to specifically recognize pathogenic bacteria contributes to the rational selection of subsequent treatment regimens.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eIn summary, we report a colorimetric and fluorescent dual-mode sensing platform \u003cb\u003eRh-QL\u003c/b\u003e for ultrafast detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. The probe \u003cb\u003eRh-QL\u003c/b\u003e exhibited exclusive selectivity, excellent immunity to interference, ultrafast response time, and high sensitivity to HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. In addition, \u003cb\u003eRh-QL\u003c/b\u003e achieves quantitative detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in real sugar samples and specifically recognizes pathogenic bacteria types, contributing to the selective elimination of bacteria. The probe \u003cb\u003eRh-QL\u003c/b\u003e can be used as a portable sensing platform to accurately, rapidly, and cost-effectively achieve the detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, which is potentially valuable in food safety and other fields.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eXiaoyu Huang, performed the experiment and wrote the manuscript;Jiaxing Li, Investigation and performed the data analyses;Qiutong Chen, performed the data analyses;Mingyu Tian, contributed to the conception of the study and imaging work;Tianyu, Liang, Investigation and validation;Lijun Tang, contributed to the conception of the study and funding acquisition.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis work was supported by the National Natural Science Foundation of China (No. 22278038), the Program for Distinguished Professor of Liaoning Province, and the Open Project of the Institute of Ocean of Bohai University (No. BDHYYJY2024010).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eShang ZY, Wu MM, Meng QT, Jiao Y, Zhang ZQ, Zhang R (2024) A near-infrared fluorescent probe for rapid and on-site detection of sulfur dioxide derivative in biological, food and environmental systems. 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Coordin Chem Rev 388:310-33\u003c/li\u003e\n \u003cli\u003eLi YF, Wang Y, Lei XM, Guo KT, Ai Q, Zhang FF, et al. Development of a responsive probe for colorimetric and fluorescent detection of bisulfite in food and animal serum samples in 100% aqueous solution. Food Chem, 407 (2023) 135146\u003c/li\u003e\n \u003cli\u003eXu JC, Pan J, Jiang XM, Qin CQ, Zeng LT, Zhang H, et al (2016) A mitochondria-targeted ratiometric fluorescent probe for rapid, sensitive and specific detection of biological SO derivatives in living cells. Biosens Bioelectron 77:725-732\u003c/li\u003e\n \u003cli\u003eFang Y, Wang J, Yu H, Zhang Q, Chen SJ, Wang KP, et al (2022) A sequential dual-key-dual-lock fluorescent probe for detection of SO\u003csub\u003e2\u003c/sub\u003e and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in cells and mice. 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J Mol Struct 1299:137168\u003c/li\u003e\n \u003cli\u003eSun W, Guo SG, Hu C, Fan JL, Peng XJ (2016) Recent Development of Chemosensors Based on Cyanine Platforms. Chem Rev 116:7768-7817\u003c/li\u003e\n \u003cli\u003eZhang SY, Yang XP, Xu Y, Wang HY, Luo F, Fu GM, et al (2024) Rational design of a rapidly responsive and highly selective fluorescent probe for SO\u003csub\u003e2\u003c/sub\u003e derivatives detection and imaging. Food Chem 439:138151\u003c/li\u003e\n \u003cli\u003eFang YY, Zheng DB, Zhang TR, Cao ZX, Zhou HC, Deng Y, et al (2024) A rationally designed fluorescent probe for sulfur dioxide and its derivatives: applications in food analysis and bioimaging. Anal Bioanal Chem 416:533-543\u003c/li\u003e\n \u003cli\u003eChen WQ, Fang Q, Yang DL, Zhang HY, Song XZ, Foley J (2015) Selective, Highly Sensitive Fluorescent Probe for the Detection of Sulfur Dioxide Derivatives in Aqueous and Biological Environments. Anal Chem 87609-616\u003c/li\u003e\n \u003cli\u003eLi Y, Sun XF, Zhou LL, Tian L, Zhong KL, Zhang JL, et al (2022) Novel Colorimetric and NIR Fluorescent Probe for Bisulfite/Sulfite Detection in Food and Water Samples and Living Cells Based on the PET Mechanism. J Agr Food Chem 70:10899-906\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Scheme","content":"\u003cp\u003eScheme 1 is available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-fluorescence","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jofl","sideBox":"Learn more about [Journal of Fluorescence](https://www.springer.com/journal/10895)","snPcode":"10895","submissionUrl":"https://submission.nature.com/new-submission/10895/3","title":"Journal of Fluorescence","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"SO2 derivatives, Near-infrared fluorescence, Environmental analysis, bacteria.","lastPublishedDoi":"10.21203/rs.3.rs-5116472/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5116472/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSulfur dioxide (SO\u003csub\u003e2\u003c/sub\u003e) is widely used in food processing to extend the shelf life of food. However, excessive intake of SO\u003csub\u003e2\u003c/sub\u003e and its derivatives (HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e and SO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e2-\u003c/sup\u003e) can cause oxidative damage to the body, resulting in several diseases. How to construct probes for rapid real-time detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e in the field is beneficial to the developmental needs of practical applications, but it is also very challenging. Here we report a dual-mode fluorescent probe \u003cstrong\u003eRh-QL\u003c/strong\u003e for ultrafast detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, which undergoes a specific 1,4-Michael addition reaction with sulfite to achieve Near-infrared fluorescence turn-on. Probe \u003cstrong\u003eRh-QL\u003c/strong\u003e was able to detect HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e within 5 s with a significant color change from violet to green and a strong fluorescence signal at 700 nm. The probe \u003cstrong\u003eRh-QL\u003c/strong\u003e has been used for the detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e- \u003c/sup\u003ein real sugar samples and can be prepared as a portable sensing kit for the detection of HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e in the environment due to its high efficiency, rapidity and accuracy. In addition, the probe \u003cstrong\u003eRh-QL\u003c/strong\u003e is able to target label Gram-negative bacteria after reacting with HSO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e, which has the potential to identify the type of pathogenic bacteria.\u003c/p\u003e","manuscriptTitle":"Construction of a Dual-Mode Sensing Platform for Ultra-fast and Real-time Detection of Bisulfite in Food and Environmental Systems","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-06 11:36:34","doi":"10.21203/rs.3.rs-5116472/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-10-08T11:45:25+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-10-08T08:48:20+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-10-07T04:03:01+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"122279669948488522407040372308408491085","date":"2024-10-07T02:33:03+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"268442533233382557743943114102069246864","date":"2024-10-07T00:47:00+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-10-05T03:17:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"298829488492144007351016141502082897398","date":"2024-10-03T14:40:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"202222024123757507494204468842980765741","date":"2024-10-02T12:28:46+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"6464804163476453262907303008185499951","date":"2024-10-01T22:42:14+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-10-01T14:31:58+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-09-20T20:47:22+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-09-20T20:46:58+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Fluorescence","date":"2024-09-19T11:05:49+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-fluorescence","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jofl","sideBox":"Learn more about [Journal of Fluorescence](https://www.springer.com/journal/10895)","snPcode":"10895","submissionUrl":"https://submission.nature.com/new-submission/10895/3","title":"Journal of Fluorescence","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"057813e8-aa28-4c72-9b2e-8310c6a52f9e","owner":[],"postedDate":"November 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-11-25T16:13:02+00:00","versionOfRecord":{"articleIdentity":"rs-5116472","link":"https://doi.org/10.1007/s10895-024-04031-x","journal":{"identity":"journal-of-fluorescence","isVorOnly":false,"title":"Journal of Fluorescence"},"publishedOn":"2024-11-20 15:57:43","publishedOnDateReadable":"November 20th, 2024"},"versionCreatedAt":"2024-11-06 11:36:34","video":"","vorDoi":"10.1007/s10895-024-04031-x","vorDoiUrl":"https://doi.org/10.1007/s10895-024-04031-x","workflowStages":[]},"version":"v1","identity":"rs-5116472","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5116472","identity":"rs-5116472","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}