Separation Resolution Research Articles

Adsorption differences and mechanism of chitooligosaccharides with specific degree of polymerization on macroporous resins with different functional groups

Ion exchange resins are promising materials for separating chitooligosaccharides (COSs) with different degrees of polymerization (DP). The adsorbent functional groups significantly determine the interactions with COSs and affect their adsorption and separation efficiency. This study investigated the adsorption behaviors of COSs on macroporous cation exchange resins with different functional groups to clarify the functional group effect on the adsorption of COSs with different DP, which can also help to provide a theoretical basis for the selection of separation medium and optimization of the separation technics of COSs. The results showed that the adsorption of COSs on sulfonic-acid type resin DX-10 and carboxylic-acid type resin DC-10 were favorable, and the adsorption isotherms were in accord with the Sips model. Adsorption thermodynamics showed that temperature had opposite effects on the two resins' equilibrium adsorption capacity of COSs. The diffusion coefficient of COSs on DX-10 resin was greater, and the affinity gap between COSs on DX-10 resin was larger than that of DC-10 resin, which is beneficial for improving the separation resolution of COSs. In contrast, the interaction energy between COSs with high DP and DC-10 resin was weaker compared with DX-10, which made the elution of COSs with high DP easier.

Simultaneous Differentiation of C═C Position Isomerism in Fatty Acids through Ion Mobility and Theoretical Calculations.

Fatty acids play a pivotal role in biological processes and have many isomers, particularly at the C═C position, that influence their biological function. Distinguishing between isomers is crucial to investigating their role in health and disease. However, separating the isomers poses a significant analytical challenge. In this study, we developed a simple and rapid strategy combining ion mobility spectrometry and theoretical chemical calculations to differentiate and quantify the C═C positional isomers in 2-/3-butenoic acid (BA), 2-/3-/4-pentenoic acid (PA), and 2-/3-/5-hexenoic acid (HA). C═C positional isomerism was mobility-differentiated by simple complexation with crown ethers (12C4, 15C5, and 18C6) and divalent metal ions (Mg2+, Ca2+, Mn2+, Fe2+, Co2+, Ni2+, Zn2+, Sr2+, and Ba2+), that is, converting C═C positional isomers with small structural differences into complexes with large structural differences through the interaction with metal ions and crown ethers. Metallized isomers were formed but could not be differentiated due to their complex and overlapping extracted ion mobiliograms (EIMs). Binary crown ether-isomer complexes were not observed, indicating that C═C positional isomers could not be separated by simple mixing with crown ethers. However, significant EIM differences were obtained for the formed ternary complexes, allowing baseline separation for the isomers. Notably, all crown ethers and metal ions have a separation effect with the isomers, with a calculated separation resolution (Rp-p) of 0.07-2.44. Theoretical chemical calculations were performed to provide in-depth structural information for the complexes and explain the separation principle. Theoretical conformational space showed that the divalent metal ions act as a bridge connecting the crown ether and the isomer. Additionally, the ternary complex becomes more compact as the distance between C═C and -COOH increases. Theoretical results can reflect the features of mobility experiments, with relative errors between the experiment collision cross-section (CCS) and theoretical CCS of no more than ±8.06%. This method was also evaluated in terms of quantification, accuracy, and precision repeatability. Overall, this study establishes that the crown ether-metal ion pair can function as a robust unit for differentiating C═C positional isomerism.

Synchro-Squeezed Adaptive Wavelet Transform-Based Optimized Multiple Analytical Mode Decomposition: Parameter Identification of Cable-Stayed Bridge under Earthquake Input

Deriving critical parametric information from recorded signals for system identification is critical in structural health monitoring and damage detection, while the time-varying nature of most signals often requires significant processing efforts due to structural nonlinearity. In this study, synchro-squeezed adaptive wavelet transform-based optimized multiple analytical mode decomposition (SSAWT-oMAMD) is proposed. The SSAWT algorithm acts as the preprocessing algorithm for clear signal component separation, high temporal and frequency resolution, and accurate time–frequency representation. Optimized MAMD is then utilized for signal denoising, decomposition, and identification, with the help of AWT for bisecting frequency determination. The SSAWT-oMAMD is first verified by the analytical model of two Duffing systems, where clear separation of the two signals is presented and identification of complex time-varying stiffness is achieved with errors less than 2.9%. The algorithm is then applied to system identification of a cable-stayed bridge model subjected to earthquake loading. Based on both numerical and experimental results, the proposed method is effective in identifying the structural state and viscous damping coefficient.

Seed to seed variation of proteins of the yellow pea (Pisum sativum L.).

The existing variation among pea protein isolates’ functionality limits their application in food formulations. The source and extent of variations among yellow pea protein profiles was assessed in 10 single seeds of two varieties with different size and weight. A new approach was developed to analyze proteins of yellow pea combining three analytical methods of size exclusion chromatography (SEC), reverse phase high performance liquid chromatography (RP-HPLC), and microfluidic SDS-PAGE, to achieve the highest separation resolution. A high variation of protein concentration was observed not only between varieties, but also among seeds of the same variety. Vicilin to legumin ratio was between 2.72–4.19, and 1.70–2.22 among the individual seeds of AC Agassiz and CDC Saffron varieties, respectively. V/L ratio was significantly different among the individual seeds for both varieties. The amount of some protein fractions/subunits were correlated with seeds’ size and weight for AC Agassiz, while such correlations were not observed for CDC Saffron.

Are preserved coastal water bodies in Spanish Mediterranean basin impacted by human activity? Water quality evaluation using chemical and biological analyses.

The Spanish Mediterranean basin is particularly susceptible to climate change and human activities, making it vulnerable to the influence of anthropogenic contaminants. Therefore, conducting comprehensive and exhaustive water quality assessment in relevant water bodies of this basin is pivotal. In this work, surface water samples from coastal lagoons or estuaries were collected across the Spanish Mediterranean coastline and subjected to target and suspect screening of 1,585 organic micropollutants by liquid chromatography coupled to ion mobility separation and high resolution mass spectrometry. In total, 91 organic micropollutants could be confirmed and 5 were tentatively identified, with pharmaceuticals and pesticides being the most prevalent groups of chemicals. Chemical analysis data was compared with data on bioanalysis of those samples (recurrent aryl hydrocarbon receptor (AhR) activation, and estrogenic receptor (ER) inhibition in wetland samples affected by wastewater streams). The number of identified organic contaminants containing aromatic rings could explain the AhR activation observed. For the ER antagonistic effects, predictions on estrogenic inhibition potency for the detected compounds were used to explain the activities observed. The integration of chemical analysis with bioanalytical observations allowed a comprehensive overview of the quality of the water bodies under study.

Automated Capillary Isoelectric Focusing-Mass Spectrometry with Ultrahigh Resolution for Characterizing Microheterogeneity and Isoelectric Points of Intact Protein Complexes

Protein complexes are the functional machines in the cell and are heterogeneous due to protein sequence variations and post-translational modifications (PTMs). Here, we present an automated nondenaturing capillary isoelectric focusing-mass spectrometry (ncIEF-MS) methodology for uncovering the microheterogeneity of intact protein complexes. The method exhibited superior separation resolution for protein complexes than conventional native capillary zone electrophoresis (nCZE-MS). In our study, ncIEF-MS achieved liquid-phase separations and MS characterization of seven different forms of a streptavidin homotetramer with variations of N-terminal methionine removal, acetylation, and formylation and four forms of the carbonic anhydrase-zinc complex arising from variations of PTMs (succinimide, deamidation, etc.). In addition, ncIEF-MS resolved different states of an interchain cysteine-linked antibody-drug conjugate (ADC1) as a new class of anticancer therapeutic agents that bears a distribution of varied drug-to-antibody ratio (DAR) species. More importantly, ncIEF-MS enabled precise measurements of isoelectric points (pIs) of protein complexes, which reflect the surface electrostatic properties of protein complexes. We studied how protein sequence variations/PTMs modulate the pIs of protein complexes and how drug loading affects the pIs of antibodies. We discovered that keeping the N-terminal methionine residue of one subunit of the streptavidin homotetramer decreased its pI by 0.1, adding one acetyl group onto the streptavidin homotetramer reduced its pI by nearly 0.4, incorporating one formyl group onto the streptavidin homotetramer reduced its pI by around 0.3, and loading two more drug molecules on one ADC1 molecule increased its pI by 0.1. The data render the ncIEF-MS method a valuable tool for delineating protein complexes.

Decorated magnetic nanoparticles with polyvinyl alcohol brushes modified with metal chelate affinity groups for purification of proteins.

Refining is a serious challenge in the production of recombinant proteins, and this study proposes a new and easy strategy for the synthesis of magnetic affinity nanoparticles. First, the nanoparticles attached to polyvinyl alcohol brushes were synthesized, and then the hydroxyl groups of this polymer were converted to chelate groups. The nanoparticles were examined by (SEM) scanning electron microscopy, (TEM) transmission electron microscopy, (DLS) dynamic light scattering, (FT-IR) Fourier transform infrared, (XRD) X-ray powder diffraction, (VSM) vibrating sample magnetometry, and (TGA) thermal gravimetric analysis. The results confirm that uniform and spherical magnetic polymer nanoparticles with high magnetization and superparamagnetic properties were successfully synthesized. The S100A9 protein, a His-tagged recombinant protein, was expressed and purified using the synthesized nanoparticles. According to the (SDS-PAGE) sulfate-polyacrylamide gel electrophoresis results, there is a high degree of resolution in protein separation. The synthesized nanoparticles have a high protein binding capacity of about 208 mg of protein per gram of nanoparticles.

Synthesis of C8F13-SiO2 stationary phase for chromatographic separation of highly polar compounds

Up to now, the chromatographic separation and analysis of highly polar compounds are still challenging because of low retention ability and selectivity of conventional stationary phases (SPs). Developing new SPs to achieve high-resolution and high-efficiency separation of polar compounds is of great importance for health and medicine, food safety, environmental protection, etc.. Herein, an influential multi-step modification strategy (modification with 1H,1H,2H,2H-perfluorooctyltriethoxysilane on the silica twice and end-capped with trimethylchlorosilane once) is introduced to prepare C8F13-SiO2 SP. The physical properties of C8F13-SiO2 SP, including morphologies, pore structures and carbon content were fully characterized by various instrumental techniques. The packed HPLC column with C8F13-SiO2 SP exhibited high column efficiency, good peak shape (tailing factors for most analytes are between 0.9 and 1.3), and resolution for simultaneous separation of ginsenosides, sulfonamides, nucleosides, and benzoylurea insecticides (BUs); meanwhile, the retention mechanism of typical reversed-phase liquid chromatography (RPLC) mode was observed. A column efficiency of 30,220 plates/m for separation of ginsenoside Re was obtained. Furthermore, the resulting columns benefit from the advantages of supreme reproducibility, repeatability and stability. Importantly, the chromatographic properties (e.g. column efficiencies, chromatographic peak shapes, retention ability and resolution) of the C8F13-SiO2 SP for the polar analytes were superior to those of the commercial SPs (including commercial amino-modified silica SP, diol-modified silica SP and commercial PFP-SiO2 SP). Thus, the C8F13-SiO2 SP is able to satisfy the strict requirements of highly polar compounds separation.

Multilayered graphene oxide membrane with precisely controlled interlayer spacing for separation of molecules with very close molecular weights

Precise interlayer spacing control and stability improvement had been two main challenges for preparation of high performance multilayered graphene based membrane (MGM). In this study, UV irradiation was adopted to precisely regulate and stabilize the interlayer spacing of a nanofiltration MGM. The UV irradiation on the GO suspension would homogenously and precisely tune the content of the oxygen containing groups on the rGO nanosheets, which should be responsible for uniformly formed interlayer spacing of MGM after assembly. The actual pore size of MGM was estimated through the probability density function model, which was highly uniform and logarithmically decreasing with the increasing UV reduction time. Owing to the uniformly and precisely (in sub-nanometer scale) controlled pore size of MGM, two molecules with a very low molecular weight difference at about 130 Da were completely separated by the MGM prepared from rGO reduced by UV for 8 h. The in-situ AFM nanoindentation as well as the long-term filtration test proved the stability of MGM was dramatically enhanced through the UV reduction and could ensure the practical applications of MGM. This study proposed a facial and scalable protocol to prepare high separation resolution multilayered graphene based NF membrane, which indicated great potential for separation of molecules with very close molecular weights in practical.

Compressed High Resolution Satellite Image Processing to Detect Water Bodies with Combined Bilateral Filtering and Threshold Techniques

Water body extraction has become an increasingly important source of information for the prediction of heavy flood situations over the last few decades. Several approaches for identifying water bodies from separate High Resolution satellite data with varying spatial, spectral, and temporal characteristics have been developed. The vast amount of data available, supported by open access regulations adopted by many agencies, is transforming the remote sensing environment, underlining the necessity of data processing speed and efficiency. The current study presents an autonomous method for extracting water bodies from satellite pictures that use a single-band threshold approach with bilateral filtering. One important goal is to use low-cost software to understand raw picture data to evaluate multi-temporal and multi-sensor images. Using the proposed framework, the workflow is constructed on a free and open-source software processing framework. Water was detected properly and quickly from other land cover features.

3D printed microfluidic device for automated, pressure-driven, valve-injected microchip electrophoresis of preterm birth biomarkers.

A 3D printed, automated, pressure-driven injection microfluidic system for microchip electrophoresis (µCE) of preterm birth (PTB)-related peptides and proteinshas been developed. Functional microvalves were formed, either with a membrane thickness of 5µm and a layer exposure time of 450ms or with a membrane thickness of 10µm and layer exposure times of 300-350ms. These valves allowed for control of fluid flow in device microchannels during sample injection for µCE separation. Device design and µCE conditions using fluorescently labeled amino acidswere optimized. A sample injection time of 0.5s and a separation voltage of 450V (460V/cm) yielded the best separation efficiency and resolution. We demonstrated the first µCE separation with pressure-driven injection in a 3D printed microfluidic device using fluorescently labeled PTB biomarkers and 532nm laser excitation. Detection limits for two PTB biomarkers, peptide 1 and peptide 2, for an injection time of 1.5s were 400pM and 15nM, respectively, and the linear detection range for peptide 2 was 50-400nM. This 3D printed microfluidic system holds promise for future integration of on-chip sample preparation processes with µCE, offering promising possibilities for PTB risk assessment.

Enhanced capillary zone electrophoresis in cyclic olefin copolymer microchannels using the combination of dynamic and static coatings for rapid analysis of carnosine and niacinamide in cosmetics

Cosmetics that have medicinal effects, including anti-inflammatory and antioxidant, have become a daily care routine consumption. The peptide additives, such as carnosine and nicotinamide, were frequently used to realize these medicinal effects. To accomplish rapid and effective quantitation of carnosine and niacinamide in cosmetics, capillary zone electrophoresis was executed in cyclic olefin copolymer microchips having both dynamic and static coatings. The static coating of cyclic olefin copolymer microchannel was constructed from bovine serum albumin adsorption, immobilization, and active site closure, while the dynamic coating was formed by adding surfactant into running buffer of capillary zone electrophoresis. The static coating can improve the hydrophilicity of cyclic olefin copolymer surface and avoid nonspecific peptide adsorption. The dynamic coating of sodium dodecyl sulfate in running buffer proved to be useful in flow velocity adjustment and the column efficiency enhancement in the capillary zone electrophoresis separation channel of the cyclic olefin copolymer microchip device. A separation resolution up to 4.24 on the mixture of carnosine and nicotinamide was obtained. Moreover, an analysis method was established and applied to simultaneous carnosine and nicotinamide determination in a liquid whitening essence and a solid antiglycation pill, and the results were verified by comparison with high-performance liquid chromatography methods, indicating its potential in complex sample analysis.

Comparison between one- and two-way coupling approaches for estimating effective transport properties of suspended particles undergoing Brownian sieving hydrodynamic chromatography

Simplified one-way coupling approaches are often used to model transport properties of diluted particle suspensions for predicting the performance of microcapillary hydrodynamic chromatography (MHDC). Recently, a one-way coupling approach was exploited to optimize the geometry and operating conditions of an unconventional double-channel geometry with a square cross section, where a Brownian sieving mechanism acting alongside the MHDC separation drive (BS-MHDC) is enforced to boost separation resolution. In this article, a cylindrical geometry enforcing the same BS-MHDC separation drive is thoroughly investigated by following a two-way coupling, fully three-dimensional approach, and results are compared with those obtained enforcing the one-way coupling analysis. Device geometry and operating conditions are optimized by maximizing the separation resolution. The effective velocity and dispersion coefficient of spherical, finite-sized particles of different diameters are computed, and two-phase effects are discussed in detail. Similar to the square channel device, the cylindrical double-channel geometry allows for a sizable reduction in the column length and in the analysis time (a factor above 12 for the length and a factor larger than 3 for the processing time) when compared to the standard MHDC configuration ensuring the same separation resolution. As expected, the one-way coupling approach overestimates the separation performance of both the BS-MHDC and the standard MHDC devices with respect to the two-way coupling analysis. But, surprisingly, the enhancement factor of the BS-MHDC over the standard MHDC is underestimated by the single-phase approximation as it doubles when wall/particle interactions are properly accounted for with a two-phase description.

Experimental methods in chemical engineering: Gas chromatography—GC

AbstractGas chromatography (GC) separates volatile and semi‐volatile liquids and gases based on their affinity for a stationary phase, either a thin liquid or a polymer layer on a solid. A gaseous mobile phase carries the volatile compounds from an injection port to a column with the stationary phase and then to an amdetector. The most common detectors are based on mass spectrometry (MS), thermal conductivity, and flame ionization. A furnace houses the columns and maintains or ramps temperatures to increase the separation efficiency—reduce run time or increase peak separation (resolution). The analyst chooses the chemistry of the stationary phase and the physical structure of the columns based on the polarity and functional groups of the solute. Packed columns tolerate impurities better than capillary columns that have a greater separating power and bleed the functional groups less. Efficiency is best when the sample polarity matches the column polarity. High temperatures and contamination cause columns to bleed the active components, which reduces the resolution but might also introduce ghost peaks to the chromatogram. The Web of Science assigned 15 000 articles to chemical engineering that mention GC and gas chromatography as keywords. Since 2017, it has indexed close to 1000 new articles every year. A bibliometric analysis classifies these contributions into four clusters: (1) pyrolysis and biomass, (2) GC–MS and extraction, (3) lignin and aromatics, and (4) decomposition and kinetics.

Improvement of chiral separation efficiency through temperature control during one time high performance liquid chromatography analysis.

This study examined how manual temperature control affects the separation of two enantiomers of five racemic mixtures using high-performance liquid chromatography (HPLC) in terms of separation factor (α), resolution (Rs), and the number of theoretical plates (N). The results showed that heating/cooling during one time HPLC analysis improved the separation factor and resolution.

Multiplexed miRNA Quantitation Using Injectionless Microfluidic Thermal Gel Electrophoresis.

MicroRNAs (miRNAs) are a class of biomolecules that have high clinical and pharmaceutical significance because of their ability to regulate protein expression. Better methods are needed to quantify target miRNAs, but their similar sequence lengths and low concentrations in biomedical samples impede analysis. This report aimed to develop a simple, rapid method to directly quantify multiple miRNAs using microfluidic thermal gel electrophoresis (TGE). Fluorescent probes were designed complementarily in sequence to four target miRNAs that also contained variable DNA overhangs to alter their electrophoretic mobilities. Samples and probes were directly added into thermal gel and loaded throughout a microchannel. Applying voltage resulted in an inline preconcentration and separation of the miRNAs that did not require a sample injection nor user intervention to switch between modes. Baseline resolution was achieved between four double-stranded miRNA-probe hybrids and four excess single-stranded probes. Analytical performance was then improved by designing an innovative microfluidic device with a tapered channel geometry. This device exhibited superior detection limits and separation resolution compared to standard channel devices without increasing the complexity of microfabrication or device operation. A proof-of-concept demonstration was then performed, showing that target miRNAs could be detected from cell extracts. These results demonstrate that TGE provides a simple, inexpensive means of conducting multiplexed miRNA measurements, with the potential for automation to facilitate future clinical and pharmaceutical analyses.

Three-dimensional feature matching improves coverage for single-cell proteomics based on ion mobility filtering.

SUMMARYSingle-cell proteomics (scProteomics) promises to advance our understanding of cell functions within complex biological systems. However, a major challenge of current methods is their inability to identify and provide accurate quantitative information for low-abundance proteins. Herein, we describe an ion-mobility-enhanced mass spectrometry acquisition and peptide identification method, transferring identification based on FAIMS filtering (TIFF), to improve the sensitivity and accuracy of label-free scProteomics. TIFF extends the ion accumulation times for peptide ions by filtering out singly charged ions. The peptide identities are assigned by a three-dimensional MS1 feature matching approach (retention time, accurate mass, and FAIMS compensation voltage). The TIFF method enabled unbiased proteome analysis to a depth of >1,700 proteins in single HeLa cells, with >1,100 proteins consistently identified. As a demonstration, we applied the TIFF method to obtain temporal proteome profiles of >150 single murine macrophage cells during lipopolysaccharide stimulation and identified time-dependent proteome changes. A record of this paper’s transparent peer review process is included in the supplemental information.

Preparation of mixed-mode stationary phase for separation of peptides and proteins in high performance liquid chromatography

Porous silica particles were prepared by sol–gel method with some modification to get wide-pore particles. These particles were derivatized with N-phenylmaleimide-methylvinylisocyanate (PMI) and styrene by reversible addition fragmentation chain transfer (RAFT) polymerization to prepare N-phenylmaleimide embedded polystyrene (PMP) stationary phases. Narrow bore stainless steel column (100 × 1.8 mm i.d) was packed by slurry packing method. The chromatographic performance of PMP column was evaluated for the separation of synthetic peptides mixture composed of five peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine enkephalin) and tryptic digest of human serum albumin (HAS) respectively. Number of theoretical plates as high as 280,000 plates/m were obtained for peptides mixture at optimum elution condition. Separation performance of the developed column was compared with commercial Ascentis Express RP-Amide column and it was observed that separation performance of PMP column was better than commercial column in terms of separation efficiency and resolution.

Additive-assisted preferential crystallization of racemic component: A case of norvaline

Herein, we report a successful resolution of L-norvaline, a racemic compound system, performed near the eutectic composition by coupling preferential crystallization with tailor-made additives. The structures of enantiomer and racemate were characterized by PXRD, DSC, and the enantiomeric purity were measured by Chiral HPLC. The preferential crystallization process was designed and established from binary melting phase diagram and ternary solution phase diagram. The effects of supersaturation, solution enantiomer composition, and seed quantity on the preferential crystallization process were investigated. However, the significant improvements of enantiomeric purity and product yield were demonstrated by additive-assisted preferential crystallization approach using tailor-made additives. These additives bearing similar structure motifs to norvaline a highly selective binding to the racemate rather than enantiomer and remarkably suppress nucleation of DL-norvaline, the key factor determining the resultant enantiomer purity and separation efficiency. The underly inhibition principle was revealed due to the centrosymmetric packing of the racemate while the polarity of enantiomer leads to only partial or slight crystallization suppression. The interesting inhibition mode by tailor-made additives is also believed to be applicable for other racemic crystallization systems. Our established additive-assisted preferential crystallization has showed great potential in the development of separation technology and resolution of enantiomer from racemic compounds.

Identification of cadmium containing metabolites in HepG2 cells after treatment with cadmium-selenium quantum dots

The transformation of quantum dots (QDs) by organisms has attracted broad attention but remains unclear. Understanding of the metabolites helps to reveal the transformation pathway of QDs. Cd containing-metallothionein (MT) are the main species formed by Cd released from CdSe QDs in HepG2 cells, while speciation analysis of Cd containing MTs remains a challenge because MTs has several subisoforms and can bind with several metals. Herein, we built a hyphenated platform for speciation analysis of QDs in HepG2 cells after treatment with CdSe/ZnS QDs. The Cd-containing MTs were separated in reversed phase high performance liquid chromatography (RP-HPLC) and subsequently online detected by inductively coupled plasma mass spectrometry (ICP-MS) and electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) parallelly. Four groups of Cd-containing metabolites were found by detecting Cd in ICP-MS. Their structures were identified in ESI-Q-TOF-MS and further confirmed with standards of four subisoforms of MT, including N-terminal acetylation MT2a, N-terminal acetylation MT1e, N-terminal acetylation MT1g and MT1m. Each group of them contains various stoichiometry of Cd/Zn. The metabolites of QDs remain same while the concentrations of each metabolite and its stoichiometry of Cd/Zn vary for different incubation concentration/time. This work provides a new parallel hyphenation technique of HPLC-ICP-MS/ESI-MS with high separation resolution and powerful detection ability, and the obtained results provide detailed metabolism information of QDs in HepG2 cells after treatment of CdSe/ZnS QDs, contributing to deep exploration of the functional mechanisms of QDs in organisms.