Novel immunosensor based on electrochemiluminescence inner filter effect and static quenching between fibrillary Ag-MOGs and SiO2@PANI@AuNPs for enabling the sensitive detection of neuron-specific enolase.

Metal-organic gels (MOGs) are unique supramolecular gels that are convenient to synthesize. In this work, a cathodic electrochemiluminescence (ECL) system based on Ag-MOGs as a luminophore and K2S2O8 as a co-reactor was developed. The ECL spectrum of the Ag-MOGs overlapped significantly with the strong UV-Vis spectrum of the SiO2@PANI@AuNPs, which effectively quenched the ECL luminescence of the Ag-MOGs. Relying on the inner filter effect between Ag-MOGs and SiO2@PANI@AuNPs, a novel ECL-IFE immunosensor was developed for the detection of neuron-specific enolase (NSE). Under optimal conditions, the ECL signal of the immunosensor displayed excellent linearity over the NSE concentration range of 10 fg/mL-100 ng/mL. The limit of detection (LOD) was 2.6 fg/mL (S/N = 3) with a correlation coefficient R2 of 0.9975. The ECL immunosensor also exhibited excellent stability and reproducibility for the detection of NSE. The results reported provide a feasible concept for the development analytical methods for the detection of other clinically relevant biomarkers.

Aggregation-Induced Electrochemiluminescence Based on a Zinc-Based Metal-Organic Framework and a Double Quencher Au@UiO-66-NH2 for the Sensitive Detection of Amyloid ß 42 via Resonance Energy Transfer.

A novel sandwich electrochemiluminescence (ECL) biosensor based on aggregation-induced electrochemiluminescence resonance energy transfer (AIECL-RET) was designed for the sensitive detection of amyloid ß42 (Aß42). The synthesized silver nanoparticle-functionalized zinc metal-organic framework (Ag@ZnPTC) and gold nanoparticle-functionalized zirconium organic framework (Au@UiO-66-NH2) were used as the ECL donor and acceptor, respectively. AgNPs were generated in situ on the surface of ZnPTC, which further improved the ECL intensity and the loading of antibody 1 (Ab1). Under the optimized experimental conditions, the linear detection range of Aß42 concentration was 10 fg/mL to 100 ng/mL, and the detection limit was 2.4 fg/mL (S/N = 3). The recoveries of Aß42 were 99.5-104%. The method has good stability, repeatability, and specificity. Ag@ZnPTC/Au@UiO-66-NH2 provides an assay for the sensitive detection of disease biomarkers.

Electrochemiluminescence resonance energy transfer between a Ru-ZnMOF self-enhanced luminophore and a double quencher ZnONF@PDA to detect NSE.

The construction of advanced systems capable of accurately detecting neuron-specific enolase (NSE) is essential for rapidly diagnosing small-cell lung cancer. In this study, an electrochemiluminescence (ECL) resonance energy transfer immunosensor was proposed for the ultra-sensitive detection of NSE. The co-reactants C2O42- and Ru(bpy)32+ were integrated to form a self-enhanced ECL luminophore (Ru-ZnMOF) as the ECL donor. The abundant carboxyl functional groups of Ru-ZnMOF supported antibody 1 via an amidation reaction. Polydopamine-modified zinc dioxide nanoflowers, as ECL acceptors, inhibited Ru-ZnMOF ECL signaling. The linear range of NSE was 10 fg mL-1 to 100 ng mL-1 with a detection limit of 3.3 fg mL-1 (S/N = 3), which is suitably low for determining NSE in real samples.

A dual-emitting immunosensor based on manganese dioxide nanoflowers and zinc sulfide quantum dots with enhanced electrochemiluminescence performance for the ultrasensitive detection of procalcitonin.

A dual-emitting electrochemiluminescence (ECL) immunosensor based on manganese dioxide nanoflowers (MnO2NFs) and zinc sulfide quantum dots (ZnSQDs) was constructed for the first time to sensitively detect procalcitonin (PCT) in human serum. rGO@Ag functioned not only to adsorb primary antibodies (Ab1) but also to improve the electrical conductivity of the immunosensor. The MnO2NFs and ZnSQDs in the nanocomposite, synergistically with silver nanoparticles, simultaneously functioned as cathodic ECL emitters to enhance the detection sensitivity of PCT by shortening the electron-transfer path, thereby reducing energy loss. Under the optimized experimental conditions, the ECL immunosensor was capable of quantitatively detecting PCT in the linear range of 0.1 pg mL-1 to 100 ng mL-1 and over the scanning potential range of -2.0-0 V, with a detection limit of 0.033 pg mL-1. Furthermore, the ECL immunosensor demonstrated high specificity for PCT in the presence of other competing antigens, excellent stability over 10 cycles, and excellent reproducibility, corroborating its potential for measuring PCT concentrations in clinical samples.

Electrochemiluminescence immunosensor based on Cu3(PO4)2 hybrid nanoflowers as a novel luminophore for the sensitive detection of prostate-specific antigen.

Copper phosphate hybrid nanoflowers (Cu3(PO4)2HNFs) were demonstrated to produce cathodic ECL emission in the presence of potassium persulfate (K2S2O8) and then used as a carrier due to their large specific surface area. AgNPs modified on Cu3(PO4)2HNFs provided more binding sites for immobilizing secondary antibodies and accelerating the electron transfer rate to enhance the ECL signal. In addition, FONDs-Au was used to capture primary antibodies due to its good biocompatibility and large specific surface area. A sandwich electrochemiluminescence (ECL) immunosensor based on copper phosphate hybrid nanoflower/Ag nanoparticle (Cu3(PO4)2HNFs@Ag) composite and Au NPs-functionalized Fe2O3 nanodendrites (FONDs-Au) was constructed to detect prostate-specific antigen (PSA) in real samples. Under optimal conditions, the constructed sandwich ECL immunosensor was sensitive to PSA with a detection limit of 0.037 pg/mL (S/N = 3), a linear detection concentration range of 0.0001-50 ng/mL, and a recovery range of 97.33-102.5%. This immunosensor is expected to provide a method to detect PSA or other biomarkers.

An electrochemiluminescence immunosensor with double co-reaction accelerators based on Ag3PO4@EuPO4-AgNP for detecting squamous cell carcinoma antigen.

This study aimed to design a sandwich electrochemiluminescence (ECL) immunosensor with double co-reaction accelerators for sensitively detecting squamous cell carcinoma antigen (SCCA). First, silver orthophosphate (Ag3PO4) nanoparticles were modified on the surface of EuPO4 nanowires to improve their poor dispersibility/solubility. At the same time, EuPO4 was used as a co-reaction accelerator to catalyze S2O82- to produce more intermediates (SO4•-), significantly enhancing the ECL signal of Ag3PO4. Ag nanoparticles (AgNP) modified on Ag3PO4@EuPO4 composite nanomaterials were used not only as linkers of luminescence groups and biomarkers but also as a co-reaction accelerator to effectively enhance ECL signal. The designed ECL immunosensor displayed several advantages, including good stability and reproducibility. Under the optimal conditions, its linear range in detecting SCCA was 0.0001-50 ng·mL-1, the detection limit was 25 fg·mL-1 (S/N = 3), the recovery was 96.6-100.4%, and the relative standard deviation was less than 4.8%. It was successfully applied to detect SCCA in human serum.

Dual-potential electrochemiluminescence cytosensor based on a metal-organic framework and ABEI-PEI-Au@AgNPs for the simultaneous determination of phosphatidylserine and epidermal growth factor receptors on an apoptotic cell surface.

A new electrochemiluminescence (ECL) cytosensor is proposed for the simultaneous determination of phosphatidylserine (PS) and epidermal growth factor receptor (EGFR) based on the ECL signals of metal-organic framework-5 (MOF-5) loaded CdS quantum dots and N-(aminobutyl)-N-(ethylisoluminol)-polyethylenimine capped Au and Ag nanoparticles. Apoptosis promotes the exposure of PS and reduces the expression of EGFR in cell membranes. Two spatially resolved areas on dual-disk glassy carbon electrodes were designed to eliminate the interference from different ECL probes. Using HepG2 cells treated with resveratrol to induce apoptosis, the cytosensor exhibited high sensitivity, simplicity, and high reproducibility, demonstrating its potential in drug screening and rapid apoptotic cell detection. The strategy reported provides a promising platform for the highly sensitive cytosensing and convenient screening of clinically relevant anticancer drugs.

Carbon quantum dots with blue/near infrared emissions for ratiometric fluorescent lornoxicam sensing and bio-imaging.

An economical and eco-friendly hydrothermal method for the preparation of nitrogen-doped carbon quantum dots (N-CQDs) was studied with rambutan peel and lysine. The morphology, structure, and optical properties of N-CQDs were characterized by transmission electron microscopy, Fourier transform infrared spectrometry, X-ray powder diffractometer, X-ray photoelectron spectrometry, and UV spectrophotometry. The synthesized N-CQDs have excellent characteristics such as strong fluorescence, good dispersion, high stability, and excellent water solubility. The absolute fluorescence quantum yield is 1.02%, the average particle size is 1.63 nm, and the maximum excitation wavelength is 340 nm. The maximum emission wavelengths are 430 nm and 800 nm. As a quencher, lornoxicam (LNX) was used to quench the fluorescence of N-CQDs with the mechanism of inner filter effect. The fluorescence ratio of N-CQDs (F430/F800) shows a good linear relationship to the concentration of LNX. The linear range and the detection limit of LNX are 0.01‒100 and 0.003 µmol/L, respectively. An effective ratiometric fluorescence probe for the detection of LNX was constructed. The method has the advantages of low detection limit, high sensitivity, wide linear range, and can be applied to the determination of LNX in real samples. Moreover, according to the excitation-dependent fluorescence behavior, dual-wavelength emission, and biocompatibility of N-CQDs, it has been applied to cell imaging.

Construction of efficient electrochemiluminescence resonance energy transfer sensor based on SnO2/SnS2QDs-Ru@IRMOF-3 composite for sensitive detection of procalcitonin.

An efficient electrochemiluminescence resonance energy transfer (ECL-RET) method is proposed which combines the luminescent materials of tris(4,4'-dicarboxylicacid-2,2'-bipyridyl) ruthenium(II) (energy donor) and tin dioxide and tin disulfide quantum dots (SnO2/SnS2QDs) (energy acceptor) into the isoreticular metal - organic framework-3 (IRMOF-3) material to form a composite. In this mode, the distance between the energy donor and the acceptor was greatly shortened, reducing the energy loss, and thereby effectively improving RET efficiency and further significantly improving the ECL signal. The obtained composite (SnO2/SnS2QDs-Ru@IRMOF-3) was combined with sandwich immunoreaction to construct an ECL immunosensor for the sensitive detection of procalcitonin (PCT). Under the optimized experimental conditions with a working potential of - 1.48 V (vs Ag/AgCl), the proposed PCT biosensor exhibited a linear concentration range of 1 × 10-4-200 ng mL-1, with a detection limit of 0.029 pg mL-1 (S/N = 3). The biosensor was used to detect PCT in actual samples. The biosensor has broad application prospects in biological analysis and clinical diagnosis due to its high sensitivity, good selectivity, and good stability.

A potential-resolved electrochemiluminescence resonance energy transfer strategy for the simultaneous detection of neuron-specific enolase and the cytokeratin 19 fragment.

An electrochemiluminescence resonance energy transfer (ECL-RET) immunosensor was developed based on the potential-resolved technology for the simultaneous detection of neuron-specific enolase (NSE) and the cytokeratin 19 fragment (CYFRA21-1). The absorption spectrum of gold nanorods (AuNRs) perfectly overlapped with the ECL spectra of SnS2@Pt and Ru(bpy)32+/Zn-MOF, so they exhibited an excellent ECL-RET effect with high efficiency. Zn-MOF possesses a large surface area, which allows for the loading of Ru(bpy)32+. This results in a signal probe of Ru(bpy)32+/Zn-MOF/Ab1 showing a strong ECL emission. Simultaneously, owing to the excellent electronic conductivity of PtNPs, they can increase the electron transfer rate between S2O82- and tin disulfide nanoflowers (SnS2NFs). Hence, the ECL signal of SnS2NFs can be enhanced. Under the optimal conditions, the linear range for NSE is 0.2 pg mL-1-20 ng mL-1 with a detection limit of 79 fg mL-1. The linear range for CYFRA21-1 is 1.25 pg mL-1-12.5 ng mL-1 with a detection limit of 0.43 pg mL-1. The proposed immunosensor can be used for the sensitive simultaneous detection of NSE and CYFRA21-1 in human serum and has promise for clinical diagnostics.

Electrochemiluminescence immunoassay of human chorionic gonadotropin using silver carbon quantum dots and functionalized polymer nanospheres.

A composite, reduced graphene oxide (rGO) doped with silver nanoparticles (Ag NPs), was prepared by using binary reductants of sodium citrate and hydrazine hydrate. Carbon quantum dots (CQDs) synthesized by papaya peel combined with silver ions to form a CQDs-loaded silver nanoparticle (AgCQDs) nanocomposite. Polymer nanospheres (PNS) were generated via the infinite coordination polymer of ferrocene dicarboxylic acid and employed as carriers to load AgCQDs. The prepared AgCQDs@PNS-PEI has good biocompatibility and electrical conductivity and can be used as a matrix for the immobilization of a secondary antibody (Ab2). A sandwich-type electrochemiluminescence (ECL) immunosensor using AgCQDs@PNS-PEI nanocomposite as probe has been developed for the detection of human chorionic gonadotropin (HCG). The proposed immunosensor exhibits a linear range from 0.00100 to 500 mIU mL-1 and the detection limit is 0.33 µIU mL-1 (S/N = 3) under optimal conditions. The sensor exhibits excellent selectivity, good reproducibility, and high stability. These features demonstrate that the proposed method has promising potential for clinical protein detection and displays a new strategy to fabricate an immunosensor. Graphical abstract.

Facile Preparation of Boron and Nitrogen Codoped Green Emission Carbon Quantum Dots for Detection of Permanganate and Captopril.

A hydrothermal strategy for preparing boron and nitrogen codoped carbon quantum dots was studied using the precursors of p-amino salicylic acid, boric acid and ethylene glycol dimethacrylate. The boron and nitrogen codoped carbon quantum dots have high fluorescence intensity, good monodispersity, high stability, superior water solubility, and a fluorescence quantum yield of 19.6%. Their average size is 5 nm. Their maximum excitation and emission wavelengths are 380 and 520 nm, respectively. Permanganate (MnO4-) quenched boron and nitrogen codoped carbon quantum dots fluorescence through inner filter effect and static quenching effects. The linear relation between quenching efficiency and MnO4- concentration ranged from 0.05 to 60 µmol/L with a detection limit of 13 nmol/L. In the presence of captopril, MnO4- was reduced to Mn2+ and the fluorescence of boron and nitrogen codoped carbon quantum dots was recovered. The linear range between recovery and captopril concentration was from 0.1 to 60 µmol/L. The limit of detection was 0.03 µmol/L. The developed method can be employed as a sensitive fluorescence sensing platform for MnO4-. It has been successfully used for captopril detection in mouse plasma.

Electrochemiluminescent immunoassay for neuron specific enolase by using amino-modified reduced graphene oxide loaded with N-doped carbon quantum dots.

An ultrasensitive electrochemiluminescence based sandwich immunoassay is presented for determination of neuron specific enolase. The method uses silver-cysteine nanowires as the capture probe and a composite made of amino-modified reduced graphene oxide and nitrogen-doped carbon quantum dots as the signal probe. It was synthesized by covalent coupling of amino-modified reduced graphene oxide to the carboxy groups of nitrogen-doped carbon quantum dots. The nanowires possess a large specific surface and abundant functional groups which facilitate immobilizing the primary antibody (Ab1). The amino-modified reduced graphene oxide is employed as a carrier for loading a large number of the quantum dots and secondary antibody (Ab2). This increases the electrochemiluminescence intensity of quantum dots. Response to neuron specific enolase is linear in the 0.55 fg·mL-1 to 5.5 ng·mL-1 concentration range. It has a detection limit of 0.18 fg·mL-1 (at S/N = 3). The relative standard deviation (for n = 6) is less than 2.9%. The assay is highly sensitive, reproducible, selective and stable. Graphical abstractA novel electrochemiluminescence immunosensor is described that uses amino-modified reduced graphene oxide (amino-rGO), nitrogen-doped carbon quantum dots (N-CQDs) and silver-cysteine nanowires (SCNWs). It was applied to the determination of neuron specific enolase (NSE). Bovine serum albumin: BSA;1-ethyl-3-(3-dimethylaminopropyl)carbodiimide: (EDC;, N-hydroxysuccinimide: NHS.

Electrochemiluminescence resonance energy transfer of MnCO3 for ultrasensitive amyloid-ß protein detection.

A composite material MnCO3/poly(diallyl dimethyl ammonium chloride) (PDDA)/Ag with excellent electrochemiluminescence (ECL) performance and high biocompatibility was prepared by adding MnCO3 and PDDA to silver nanoparticles (AgNPs). MnCO3/PDDA/Ag and Au@SiO2NPs were used as ECL donors and acceptors, respectively. Thus, an effective ECL-resonance energy transfer (RET) sensing platform was established. In a potassium persulfate (K2S2O8) medium, MnCO3 exhibited ECL emission with an ECL band appearing at 500-600 nm. In addition, Au@SiO2 nanoparticles showed a UV-visible absorption at 450-650 nm. The ECL emission spectra of MnCO3 overlapped with the absorption spectra of Au@SiO2NPs. The effective ECL quenching resulted in a good response to the concentration of Aß42 in serum samples. The linear range was 5 fg ⋅ mL-1 to 100 ng ⋅ mL-1, and the detection limit was 2 fg ⋅ mL-1. The recovery ranged from 97.7% to 104%. The high-efficiency ECL-RET immunosensor has potential application in detecting human serum Aß42 and other biomarkers, and can be used for the early screening of diseases.

Au@NiFeMOFs as the signal quencher of Au@g-C3N4NSs composite for sensitive "on-off" electrochemiluminescence immunosensing of beta-2-microglobulin.

In this study, an electrochemiluminescence resonance energy transfer (ECL-RET) immunosensor was constructed to detect beta-2-microglobulin (B2M). As a donor-acceptor pair, a carbon nitride nanosheet modified with gold nanoparticles (Au@g-C3N4NSs) and a nickel- and iron-based organic framework modified with gold nanoparticles (Au@NiFeMOFs) were prepared. The sandwich immunosensor was successfully constructed so that ECL-RET occurred between Au@NiFeMOFs and Au@g-C3N4NSs. The ECL intensity of the immunosensor decreased with the increase the B2M concentration due to the low conductivity of B2M. The linear range of the ECL-RET immunosensor was from 10 fg/mL to 10 ng/mL, and the limit of detection was 2.3 fg/mL (S/N = 3). The developed immunosensor had high sensitivity, high specificity, and excellent stability. It could realize the sensitivity test of B2M and provide a novel idea for the detection of biomarkers.

Aggregation-induced electrochemiluminescence enhancement of Ag-MOG for amyloid ß 42 sensing.

This study aimed to introduce an immunosensor for measuring amyloid ß 42 (Aß42) levels by aggregation-induced enhanced electrochemiluminescence (ECL). Metal-organic gels (MOGs) are novel soft materials with advantages such as high gel stability, good light-emitting properties, and easy preparation. This study used silver nanoparticle metal-organic gel (Ag-MOG) as a substrate to connect Aß42-Ab2 and the cathodoluminescent probe. Potassium persulfate was used as a co-reactant that could emit a high ECL signal. CuS@Au had the benefits of a relatively large surface area with excellent carrier function; therefore, it was used as a substrate to load a large amount of Aß42-Ab1, significantly improving the immunosensor sensitivity. The ECL intensity of Aß42 was linear in the range of 0.01 pg/mL to 250 ng/mL with a detection limit of 2.2 fg/mL (S/N = 3) under optimized detection conditions. This ECL immunosensor has been successfully applied to detect Aß42 in human serum with the advantages of excellent stability and high selectivity. This method not only expands the potential applications of ECL immunosensors based on biological testing and clinical diagnosis but also provides a viable approach to basic clinical testing.

An electrochemiluminescence immunosensor based on ABEI-GO-AgNPs as a double-amplified luminophore for the ultra-sensitive detection of prostate-specific antigen.

A sandwich electrochemiluminescence (ECL) immunosensor based on an N-(4-aminobutyl)-N-ethylisoluminol-graphene oxide-Ag nanoparticle (ABEI-GO-AgNPs) complex and cysteine silver nanowires (AgCysNWs) was prepared to detect prostate-specific antigen (PSA). Our results showed that an ECL signal probe, ABEI-GO-AgNPs, with an ultrahigh specific surface area, favorable catalytic properties, and electrical conductivity was prepared by a one-step synthesis method. ABEI-GO-AgNPs with good biocompatibility immobilized secondary antibody (Ab2) via AgN bonds. Furthermore, AgCysNWs containing many -COOH groups were prepared and used to enrich primary antibody (Ab1), which could be used as an affinity probe for the selective capture of PSA. Lastly, through layer-by-layer assembly, we established an ECL immunosensing platform for the sensitive detection of PSA. Under the optimized conditions, the designed ECL immunosensor showed promising sensitivity and selectivity for the detection of PSA in the linear range of 5.5 × 10-7-5.5 ng/mL, with a detection limit of 1.2 × 10-7 ng/mL. The constructed ECL sensing platform possessed good specificity, reproducibility, and stability and could detect PSA in actual human serum samples.

N-doped carbon quantum dots from osmanthus fragrans as a novel off-on fluorescent nanosensor for highly sensitive detection of quercetin and aluminium ion, and cell imaging.

In this work, fluorescent N-doped carbon quantum dots (N-CQDs) have been synthesized by simple hydrothermal heating of natural osmanthus fragrans, without any toxic ingredients or surface chemical modifications. The N-CQDs possess a high quantum yield of 21.9 %, outstanding blue fluorescence, good water dispersity, and excellent optical stability. Because the favorable inner filter effect (IFE) between N-CQDs and quercetin (QT) occurs, the addition of QT to N-CQDs can cause their fluorescence quenching. When Al3+ was added to the N-CQDs/QT system solution, it was found that the inhibition of IFE leads to the fluorescence intensity of N-CQDs/QT system enhancement by virtue of a specific binding of QT to aluminum ion (Al3+). Therefore, we used the N-CQDs as a novel off-on fluorescent nanosensor to detect QT and Al3+. Under optimal conditions, the fluorescent nanosensor can detect QT within the wide linear response in the range of 0.003-80 µmol/L with as low as 1 nmol/L detection limit. For the detection of Al3+, the N-CQDs/QT system showed linearity response toward Al3+ in a range of 0.1∼100 µmol/L and the limit of detection was found at 26 nmol/L. In addition, N-CQDs have been successfully used to efficient quantification QT in human plasma and monitor Al3+ in serum samples. Noteworthy, the N-CQDs demonstrated low toxicity toward T24 cells, which realized sensing QT and Al3+ in the living cells.

Spatially-resolved dual-potential sandwich electrochemiluminescence immunosensor for the simultaneous determination of carbohydrate antigen 19-9 and carbohydrate antigen 24-2.

A new electrochemiluminescence (ECL) immunosensor based on spatially-resolved dual-potential technology was designed for the simultaneous determination of carbohydrate antigen 19-9 (CA 19-9) and carbohydrate antigen 24-2 (CA 242). Luminol-AgNPs@ZIF-67 was used as the anodic probe, and Pt nanoparticle-functionalized graphitic carbon nitride nanosheets (g-C3N4@PtNPs) were used as the cathodic probe. Two spatially-resolved areas on the dual-disk glassy carbon electrode (DDGCE) were modified with a AuNPs film by electrodeposition to improve the conductivity of the sensing interface. By recording the ECL responses at two different excitation potentials, the linear range for CA 19-9 was determined to be 0.0001-10 U/mL, with a limit of detection of 31 µU/mL. The linear range for CA 242 was 0.0005-10 U/mL, with a limit of detection of 0.16 mU/mL. Moreover, the ECL immunosensor possessed high selectivity and stability and successfully detected CA 19-9 and CA 242 in real samples. This immunosensor provides a new platform for clinical immunoassays.

Ginkgo leaf-based synthesis of nitrogen-doped carbon quantum dots for highly sensitive detection of salazosulfapyridine in mouse plasma.

A simple, economical hydrothermal strategy for synthesizing nitrogen-doped carbon quantum dots (N-CQDs) was developed using Ginko leaves as a carbon source. These N-CQDs have strong blue fluorescence, excitation-relevant emissions, high monodispersity, good stability, good water solubility, and a 22.8% fluorescence quantum yield. They average 3 nm in size, and have maximum excitation and emission wavelengths of 350 and 436 nm, respectively. They are used as an effective fluorescent sensing platform for the label-free sensitive detection of salazosulfapyridine (SASP) due to the strong quenching effect of SASP. When SASP concentration is 0.1-80 µmol/L, there is a good linear relationship with a detection limit of 40 nmol/L. This method was successfully applied to detect SASP in mouse plasma. The results show that the SASP recovery range was 96%-101%. RSDs ranged from 2.6% to 3.1%.