Non-destructive Extraction Research Articles

Petrous bones versus tooth cementum for genetic analysis of aged skeletal remains.

A proper sampling strategy is important to obtain sufficient DNA for successful identification of aged skeletal remains. The petrous bone is the highest DNA-yielding bone in the human body. Because DNA extraction from the petrous bone is very destructive, the demand for other DNA sources is significant. When investigating aged skeletal remains, teeth are usually preserved, and recent studies have shown that DNA in teeth can be best preserved in the dental cementum that surrounds the surface of the tooth root. To extract DNA from the surface of the tooth root, a nondestructive method without grinding was used. Petrous bones and teeth from 60 archaeological adult skeletons were analyzed. The DNA yield, degree of DNA degradation, and STR typing success were compared, and the results showed higher DNA yield and higher amplification success in petrous bones, despite higher degradation of petrous bones' DNA. The greater success of petrous bones is associated with poorly preserved DNA in a quarter of the teeth analyzed. When teeth with badly preserved DNA were excluded from the statistical analysis, no differences in the success of STR loci amplification were observed even if DNA yield was higher in petrous bones, which can be explained by greater degradation of petrous bones' DNA. When teeth are well preserved, they can be used for genetically analyzing aged skeletal remains instead of petrous bones, and a rapid nondestructive extraction method can be applied to shorten the identification process and to physically preserve the biological specimen.

Design and Test of Seedling-Picking Mechanism of Fully Automatic Transplanting Machine

The seedling retrieval mechanism is a crucial component of fully automatic transplanting machines, significantly influencing the quality, reliability, and efficiency of the transplanting process. Nonetheless, the existing seedling retrieval mechanisms in current transplanting machines exhibit several deficiencies, including substantial damage to seedlings and inadequate retrieval accuracy. To overcome these challenges, we propose an integrated approach combining pneumatic and mechanical techniques to further improve performance. By employing a lower thimble elevation and clamping mechanism, alongside a mathematical model based on the seedling removal process, this method ensures precise seedling extraction and minimizes damage to the root system and substrate. The novelty of this study lies in its ability to reduce the adhesion between seedlings and the holes of the plug plate, thereby minimizing non-destructive extraction of the seedlings and preserving the integrity of the matrix, which is essential for ensuring healthy seedling growth. Moreover, the optimization of the seedling retrieval trajectory enhances the accuracy of the seedling retrieval mechanism while also meeting the requisite speed requirements. Experimental results indicate that at a rate of 72 seedlings per minute, the extraction success rate reached 94.90%, and the casting success rate was 98.53%. The seedling injury rate was only 1.95%, resulting in an overall success rate of 91.69%. These findings confirm that the device meets operational efficiency requirements and delivers effective performance.

A novel method for non-destructive efficient green recovery of multiple high value products from microalgal cells with enzymatically enhanced permeability

A method is developed for simultaneous extraction of lipids, hydrocarbons and carotenoids from primary algal biomass without the need for cell sacrifice. Enzymatically predigested cells (from previous experiments) with enhanced permeability were used, for ‘milking’ with biocompatible solvent. The continuous non-destructive extraction of lipids and carotenoids from an improved permeabilized microalgal cell using crude enzyme concoctions and biocompatible solvents has not been reported so far. Oocystis sp. cells showed considerable viability following enzymatic pretreatment and extraction using dodecane which was verified through cell viability assays, and chlorophyll content. HPLC analysis confirmed the presence of lutein and zeaxanthin. GC-MS analysis showed the presence of omega-3 alpha linolenic acid, PUFAs, and phenolics, in enzymatically digested microalgal cells. The content of fatty acids and esters in enzyme treated samples was 57.18 %, and the enzyme with dodecane treated samples showed 31.4 % fatty acids and esters content. Omega-3 linolenic acid comprised 11.66 % in enzyme treated cells and showed a marginal increase of 12.5 % in enzyme with dodecane treated cells. The method enables recovery of multiple compounds from a single biomass. In the present study, a cultivation strategy of microalgal cells with enhanced permeability, grown in biocompatible solvent is proposed, whereby maximum yield of multiple valuable metabolites can be obtained from a single biomass. The method simplifies downstream processing operations by circumventing cell harvesting and drying. The use of green extraction solvents reduces the overall carbon footprint of the process.

Non-destructive strategy to extract sustainable helix and high-strength Musa core fibers for rapid water conduction and evaporation

The reuse and development of natural waste resources is a hotspots and challenges in the research of new fiber materials and the resolution of environmental concern globally. Herein, this study aimed to develop a simple and direct manual extraction process to extract Musa core fibers (MCFs) for rapid water conduction and evaporation. Through simple processes such as ring cutting and stretching, this green and non-destructive inside-out extraction strategy enabled Musa fibers to be naturally and harmlessly degummed from natural Musa stems, with good maintenance of the fiber structure and highly helical morphology. The extracted fibers are composed of regularly and closely arranged cellulose nanofibrils in the shape of ribbon spirally arranged multi-filaments, and the single filament is about 2.65 μm. The high-purity fibers exhibit ultra-high tensile strength under a non-destructive extraction process, and the ultimate tensile strength in dry state is as high as 742.95 MPa. The tensile strength is affected by the number of fiber bundles, which shows that tensile strength and tensile modulus is higher than those of vascular bundle fibers in dry or wet condition. In addition, the MCFs membrane indicates good water conductivity, with a water absorption height of 50 mm for the sample in only 60 s. Moreover, the water evaporation rate of MCFs reaches 1.37 kg m−2 h−1 in 30 min, which shows that MCFs have excellent water conductivity and evaporation rate compared with ordinary cotton fibers. These results indicate that MCFs have great potential in replacing the use of chemical methods to extract fibers from vascular bundles, providing an effective way to achieve sustainability in quick-drying applications, as well as in the sustainable development of natural waste resources.

An innovative microplastic extraction technique: The switchable calcium chloride density separation column tested for biodegradable polymers, polyethylene, and polyamide

Extracting microplastics from complex matrices poses challenges due to the potential impact of harsh chemical treatments on microplastic properties. For fate and hazard assessment reliable techniques are needed to not only quantify the particle number but also to assess the physicochemical properties of environmental microplastics with minimum changes induced by extraction. Here we present the method development for an innovative and non-destructive extraction protocol based on a switchable calcium chloride density separation column. In contrast to commonly reported extraction protocols, the presented technique is suitable for targeted microplastic property analysis (e.g., surface chemistry and texture) by keeping chemical treatments (such as oxidation and enzymatic digestion) to a minimum. By adjusting the temperature we can control the aggregate state of the highly concentrated salt solution, allowing to separate the microplastics from matrix by cutting of purified, solidified samples. Harsh chemical treatments are avoided, as well as obstruction of microplastic extraction by adsorption to matrix components when passing the tap at the bottom of traditional density separation funnels.The use of microplastics that were prelabeled with a fluorescence dye helped to solve difficulties observed during method development by visual inspection before measurement of extraction efficiency: We spiked a blank compost with low-density polyethylene (LDPE) and polyamide (PA). Additionally, UV aged LDPE was used to demonstrate applicability to more hydrophilic, more environmentally relevant microplastics. The obtained initial results show high recovery of both unaged and aged LDPE over 97 wt.-% and an efficient compost removal but a lower and less robust recovery (between 68 and 18 wt.-%) for PA particles that are more challenging to extract due to an unfortunate synergistic combination of smaller particle size and higher density. Method adaptation to other microplastic types may still be necessary. In short:•A low-cost and simple approach without oxidation to extract (pre-aged) microplastics from compost•Method development by visual observation using fluorescent labelled microplastics and method validation by spike-recovery tests

Ultrasound-assisted extraction of vegetable dyes and mordants from wool dyed with Curcuma longa and Reseda luteola

With the increasing demand to reduce the use of synthetic dyes, driven by environmental and health concerns, natural dyes are emerging as a promising alternative. Their environmental friendliness and sustainability are consistent with the global move towards greener practices in various industries, including textiles. However, the widespread adoption of natural dyes has also exposed their vulnerability to adulteration and fraudulent practices. As demand for natural dyes continues to grow, addressing these challenges is essential to ensuring the integrity and authenticity of naturally dyed fibers. This paper aims to establish a mild extraction method for the dye, mordant and wool in naturally dyed fabrics. Wool dyed with a mixture of Reseda luteola and Curcuma longa, was subjected to extraction using various additives, including HCl, acetic acid, NaOH, citric acid, formic acid and EDTA. In this study, two extraction techniques - heating and ultrasound extraction - were compared. The effect of the geometry of the extraction reactor was first studied, then the optimized conditions were applied to different solutions and reagents. Characterization of the wool fabric was carried out using SEM and FTIR, while HPLC and LC/MS were applied to identify dye markers. In addition, ICP analysis was used to characterize the metallic mordant involved in the dyeing process. The results demonstrated that ultrasound allowed an efficient extraction of dyes from wool under mild conditions. This study highlights the potential of ultrasound as a valuable tool for non-destructive extraction in the analysis of dyed wool, providing a thorough characterization of dye, mordant and fiber components. The studied technique contributes significantly to the field of textile analysis, enabling accurate identification of naturally dyed fibers.

Progress of non-destructive hydrocarbon extraction technology of Botryococcus braunii

Coal, petroleum and natural gas have remained the premium energy sources driving the economy and sustaining population growth for years. Global human population has shown a 7-fold increase in the last two centuries and, consequently, an astronomical increase in the global economy and energy demand. Interestingly, rising global warming, climate change and destruction of the natural environment are trends parallel with the world population profile and continual dependence on fossil fuels. Actions to mitigate these challenges have ignited interest in autotrophic production of hydrocarbons from microalgae as an alternative to fossil-based fuels. The microalgal strain, Botryococcus braunii, offers an excellent platform for renewable hydrocarbons production. B. braunii has unusual and fascinating attributes, and by producing liquid hydrocarbons extracellularly, a classical non-invasive, non-destructive extraction technique known as milking can be applied. In the milking process, hydrocarbons in the extracellular matrix of the algae are repetitively extracted without the need for cell harvesting, dewatering, disruption, and purification of the metabolites, thereby reducing downstream processing costs. In this way, the algae do not need to grow for cellular biomass production but can act only as light-driven catalysts to convert CO2 into hydrocarbons that accumulate externally in the medium. In this work, the different lipid extraction techniques from B. braunii are discussed in depth, taking critical analysis of the effect of each process on hydrocarbon recovery and purity as well as their economic viability. Moreover, hydrocarbon composition of B. braunii and the factors influencing the hydrocarbon production is comprehensively reviewed. The milking process is critically discussed, and the cultivation protocols suitable for milking as well as strategies to improve this process are highlighted. We indicate that switchable solvent-based milking has exceptional potential and biofilm-based cultivation holds great promise to reduce downstream costs. However, more research is needed to improve their large-scale implementation and feasibility.

A Method to Replace NaCl as a Flotation Solution for Extracting MPs in Soil: A Case Study of the Jiaxing Agricultural Soil from China.

Microplastics (MPs) have become an important global issue in recent years. However, MPs in the soil have received far less attention than water. Effective and nondestructive extraction of MPs is important for studying MPs in agricultural soils. This study uses different floatation solutions as experiments and uses MgCl2 as the floatation solution of the density extraction method. Five types of standard MPs (PE, PP, PS, PVC, and PET) are used as the objects of this experiment. The recovery of the two particle sizes was between 90.82% and 109.69%. The extracted standard MPs were then subjected to IR and Raman spectroscopic analysis, and the results showed that Raman spectroscopy was more suitable for the identification of the extracted MPs. Finally, this method collected and verified a vast number of soil samples and further analyzed the abundance and characteristics of the collected MPs.

DNA barcoding of exuviae for species identification of Central European damselflies and dragonflies (Insecta: Odonata)

Monitoring of odonates has become an important instrument for ecological status assessment of (semi-)aquatic habitats. Besides information on presence and abundance, knowledge about a species´ autochthony at the surveyed waterbody is a significant information within the assessment process. Here, the finding of exuviae represents the ultimate proof of successful reproduction. Although feasible for most odonate species, morphological identification of exuviae is often time consuming, as it relies on small, fragile structures. To facilitate species identification of exuviae, a DNA barcoding approach was developed, including (1) non-destructive extraction of DNA using whole exuviae or their tracheal tubes, and (2) primer systems for long (< 600 bp) and short (< 200 bp) CO1 fragments. A total of 85 exuviae from 33 species were analysed and compared to results of morphological identification. Additionally, factors potentially influencing DNA quality and quantity, as well as PCR and sequencing success were investigated. Eighty-two exuviae matched the morphologically identified genus, and 60 matched at species level. Of the 33 species present in the data set, 82% could be identified to species level via DNA barcoding. The results show how DNA-based approaches can support fast and accurate species identification and therefore enhance monitoring of an ecologically important taxonomic group, with high relevance for conservation and habitat restoration. Moreover, the use of exuviae as DNA resource once more shows that non-invasive sampling offers great potential for molecular species identification, which is essential when studying rare and endangered species.Implications for insect conservationOur results show how molecular tools, here DNA barcoding of odonate exuviae, can support species monitoring without the need of catching individuals, harming, or even killing them. Obtaining DNA from non-invasive sources can thus be a direct advantage to the conservation of insects, especially when dealing with rare and endangered species and/or populations. Using the example of odonates as bioindicator organisms for aquatic and semi-aquatic habitats, we highlight the importance of non-invasive genetic approaches for population studies and monitoring of insect species and/or species communities for ecosystem assessments and conservation management.

A nanoparticle-coated microfluidic chip for automated, non-destructive extraction of encapsulated DNA in data storage.

DNA data storage based on tubes as physical storage carriers has been developed to solve the problem of the exponential growth of information. Compact disk (CD)-microfluidics that employs centrifugal forces for fluidic manipulation offers an attractive alternative that integrates complex assays onto a miniaturized platform to result in automation for DNA data storage. In this work, we develop a CD microfluidic chip modified with nanoparticles for accurate fluid flow control. The nanoparticle coating turns microchannels into valves or pumps, which reduces the error in fluidic control from 62% to 6%. Based on the nanoparticle coating, the chip integrates demineralization, nucleic acid amplification, and re-mineralization functions for automated, non-destructive information extraction. We prove the functionality of the chip with mineralized DNA data. Compared to purely manual operation and traditional amplification techniques, the microfluidic chip saves human power and time consumption, demonstrating an essential contribution to the development of DNA data storage.

Non-Destructive Extraction and Separation of Nano- and Microplastics from Environmental Samples by Density Gradient Ultracentrifugation.

Nano-/microplastics (NMPs, particle diameter < 5 mm) are widespread emerging pollutants causing diverse impacts on organisms due to their sizes, shapes, and chemical properties. Despite the fast increase in NMP research, an effective method to separate and identify NMP types from environmental samples is still lacking. Here, we developed a simple and effective approach for the non-destructive extraction and separation of various types of NMPs from environmental samples by density gradient ultracentrifugation (DGU). For the first time, DGU was capable to separate various NMPs from the complex matrix with high selectivity (100%), purity (93%), and applicability. Through a gradually changing density of the density gradient medium by changing the concentrations or volumes of CsCl/water solution (from 0.00065 to 0.01989 g cm-3 mm-1), various NMPs (with particle sizes as little as 50 nm) could be extracted and separated from soil samples with high recovery (78.5-96.0%). We confirmed the effectiveness and compatibility of DGU through a correct identification of all types of NMPs separated from artificial soil samples with Raman spectroscopy, simultaneous thermal analysis (STA), and pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). DGU is compatible with all analytical processes compared to other existing methods with much less sample pretreatment time (0.5 h). Overall, DGU is an effective and cheap method (2.2 USD/sample) to separate NMPs from environmental samples such as soil and water and, hence, can facilitate research on NMPs related to terrestrial and marine environments as well as human health.

Three-dimensional reconstruction and phenotype measurement of maize seedlings based on multi-view image sequences.

As an important method for crop phenotype quantification, three-dimensional (3D) reconstruction is of critical importance for exploring the phenotypic characteristics of crops. In this study, maize seedlings were subjected to 3D reconstruction based on the imaging technology, and their phenotypic characters were analyzed. In the first stage, a multi-view image sequence was acquired via an RGB camera and video frame extraction method, followed by 3D reconstruction of maize based on structure from motion algorithm. Next, the original point cloud data of maize were preprocessed through Euclidean clustering algorithm, color filtering algorithm and point cloud voxel filtering algorithm to obtain a point cloud model of maize. In the second stage, the phenotypic parameters in the development process of maize seedlings were analyzed, and the maize plant height, leaf length, relative leaf area and leaf width measured through point cloud were compared with the corresponding manually measured values, and the two were highly correlated, with the coefficient of determination (R2) of 0.991, 0.989, 0.926 and 0.963, respectively. In addition, the errors generated between the two were also analyzed, and results reflected that the proposed method was capable of rapid, accurate and nondestructive extraction. In the third stage, maize stem leaves were segmented and identified through the region growing segmentation algorithm, and the expected segmentation effect was achieved. In general, the proposed method could accurately construct the 3D morphology of maize plants, segment maize leaves, and nondestructively and accurately extract the phenotypic parameters of maize plants, thus providing a data support for the research on maize phenotypes.

Let’s talk about the (lady)birds and the bees: how insects can whisper a multitude of stories

If you have watched A Bug’s Life, you would have seen that insects come in an assortment of colours, shapes, and sizes. They are the perfect organism that could be used to describe the myriad diversity of all life on earth. These six-legged creatures are one of the most diverse groups of species, accounting for more than 80% of all documented living animals Ødegaard 2008. This huge diversity also makes it extremely time- and labour-intensive to carry out large-scale monitoring of insects. High-throughput sequencing technologies and the use of DNA barcodes for species identification have paved the way for the rapid biomonitoring of insects Hebert et al. 2003. However, millions of insect species are not well-represented in DNA reference databases, making species-level identification challenging for molecular research. Projects such as the Darwin Tree of Life (DToL) aim to cover this gap and generate DNA barcodes for all eukaryotic species found in the UK Blaxter et al. 2022. At the Wellcome Sanger Institute, the BIOSCAN UK for Flying Insects project has two main aims; Documenting the diversity of UK flying insects Discovering insect-cobiont interactions Documenting the diversity of UK flying insects Discovering insect-cobiont interactions To meet these aims, we will be documenting the diversity of one million malaise-caught insects from 100 sites across the UK in the next five years. We will be using a non-destructive DNA extraction technique to preserve insect specimen integrity for museum collections or educational purposes Korlević et al. 2021. COI barcoding will be carried out using ONT and/or PacBio long-read technology to identify each insect specimen. To tease apart insect conbiont interactions, we will carry out mini barcoding using primers targeting microbes, parasites, vertebrates, invertebrates, and plants Fig. 1. Together, this molecular dataset consisting of one million specimens collected over space and time in the next five years will allow us to discover how insects interact with the ecosystem and advance insect biomonitoring research in the UK.

Disentangling bias for non-destructive insect metabarcoding.

A fast and reliable method for obtaining a species-level identification is a fundamental requirement for a wide range of activities, from plant protection and invasive species management to biodiversity assessments and ecological studies. For insects, novel molecular techniques such as DNA metabarcoding have emerged as a rapid alternative to traditional morphological identification, reducing the dependence on limited taxonomic experts. Until recently, molecular techniques have required a destructive DNA extraction, precluding the possibility of preserving voucher specimens for future studies, or species descriptions. Here we paired insect metabarcoding with two recent non-destructive DNA extraction protocols, to obtain a rapid and high-throughput taxonomic identification of diverse insect taxa while retaining a physical voucher specimen. The aim of this work was to explore how non-destructive extraction protocols impact the semi-quantitative nature of metabarcoding, which alongside species presence/absence also provides a quantitative, but biased, representation of their relative abundances. By using a series of mock communities representing each stage of a typical metabarcoding workflow we were able to determine how different morphological (i.e., insect biomass and exoskeleton hardness) and molecular traits (i.e., primer mismatch and amplicon GC%), interact with different protocol steps to introduce quantitative bias into non-destructive metabarcoding results. We discuss the relevance of taxonomic bias to metabarcoding identification of insects and potential approaches to account for it.

Nondistractive extraction of intracellular molecules from yeast using PEF‐assisted autolysis

AbstractThis paper discusses the possibility of nondestructive extraction of intracellular proteins of yeast (Saccharomyces cerevisiae) by means of pulsed electric fields (PEFs) combined with autolysis. First, the effects of PEFs (20 kV/cm, 600 ns) on yeast and the subsequent cell wall collapse phenomenon were studied by morphological observation of fluorescence staining. Morphological observation shows that the PEFs increase the permeability of the cell membrane and also disrupt the intracellular vacuoles, whereas the cell wall morphology remains unchanged. However, the cell wall was collapse during the postPEF incubation. Next, the amount of extracted protein was evaluated by using the BCA assay. The amount of extracted protein in the PEF treated suspension was only a little, whereas the additional incubation promotes the extraction. Lastly, the size of the extracted proteins was investigated by using both Native‐PAGE and SDS‐PAGE. The PEF‐treated and incubated suspension contains the large protein over 250 kDa.

Production of Lectins from Marine Algae: Current Status, Challenges, and Opportunities for Non-Destructive Extraction.

Marine algae are an excellent source of novel lectins. The isolation of lectins from marine algae expands the diversity in structure and carbohydrate specificities of lectins isolated from other sources. Marine algal lectins have been reported to have antiviral, antitumor, and antibacterial activity. Lectins are typically isolated from marine algae by grinding the algal tissue with liquid nitrogen and extracting with buffer and alcohol. While this method produces higher yields, it may not be sustainable for large-scale production, because a large amount of biomass is required to produce a minute amount of compound, and a significant amount of waste is generated during the extraction process. Therefore, non-destructive extraction using algal culture water could be used to ensure a continuous supply of lectins without exclusively disrupting the marine algae. This review discusses the traditional and recent advancements in algal lectin extraction methods over the last decade, as well as the steps required for large-scale production. The challenges and prospects of various extraction methods (destructive and non-destructive) are also discussed.

Non-Destructive Extraction of DNA from Preserved Tissues in Medical Collections

Museum specimens and histologically fixed material are valuable samples for the study of historical soft tissues and represent a possible pathogen-specific source for retrospective molecular investigations. However, current methods for molecular analysis are inherently destructive, posing a dilemma between performing a study with the available technology, thus damaging the sample, and conserving the material for future investigations. Here the authors present the first tests of a non-destructive alternative that facilitates genetic analysis of fixed wet tissues while avoiding tissue damage. The authors extracted DNA from the fixed tissues as well as their embedding fixative solution, to quantify the DNA that was transferred to the liquid component. The results show that human historical DNA can be retrieved from the fixative material of medical specimens and provide new options for sampling valuable collections.

Measurement techniques for sulfur trioxide concentration in coal-fired flue gas: a review

Under the extensive implementation of ultra-low emission facilities, sulfur trioxide (SO3) has received increasing attention. This article reviews the measurement techniques for SO3 in flue gas, which include controlled condensation method (CCM), isopropanol absorption method (IPA), salt method, tunable diode laser absorption spectroscopy (TDLAS), ultraviolet absorption spectroscopy (UVs), and Fourier transform infrared spectroscopy (FTIR). The first three methods are chemical methods, which focus on the extraction of SO3 from flue gas. With highly reactive nature and relatively low concentrations, which are about 5 mg/m3 even lower, achieving high-fidelity flue gas sampling and non-destructive extraction of SO3 is the key to SO3 measurement. The latter three methods belong to spectroscopic methods, which focus on the principle, system composition, and influencing factor analysis. With real-time response and 1-ppm detection limit, attention is attracted to spectroscopic methods on online measurement. This article comprehensively introduces the measurement techniques for SO3 concentration in flue gas and presents conclusions so as to enable researchers to decide the direction of further investigation.

Turberas en Valle del Alto Mayo, Perú: importancia, amenazas y perspectivas de conservación

Los palmares de Mauritia flexuosa (aguajales) son considerados las principales turberas tropicales amazónicas que contribuyen al mantenimiento de la biodiversidad y del ciclo hidrológico, además de su aporte en el secuestro y almacenamiento de carbono. Mediante una revisión bibliográfica y observaciones de campo, se identifica la importancia, factores de amenaza y oportunidad para estos ecosistemas. Las turberas almacenan más del doble de carbono que todos los bosques del mundo en conjunto y conforman un grupo objetivo para el estudio del cambio climático. El acceso a sistemas de extracción no destructiva para la cosecha de los frutos debe estar articulado con planes socioeconómicos de desarrollo. Es imprescindible la aplicación de políticas participativas de educación ambiental e incentivos económicos a las comunidades que protegen estos territorios.

Porous carbon nanosphere-based imprinted composite membrane for selective and effective separation of dibenzothiophene

In spite of the extensive efforts in selective separation membranes, the preparation of molecularly imprinted membranes with both enhanced adsorption capacities and high selectivity still remains strong challenges. Herein, a stepwise imprinting strategy was adopted to construct functionally modified ordered mesoporous carbon nanospheres (f-OMCNs) based molecularly imprinted composite membrane (f-OMCNs@MIM) for the specific and non-destructive extraction of dibenzothiophene (DBT). The key design of f-OMCNs@MIM was integrating f-OMCNs into porous polyvinylidene fluoride (PVDF) membrane to increase the porosity and imprinted efficiency of imprinted composite membrane. Attributed to the design of the uniform f-OMCNs-based composite structure, as-prepared f-OMCNs@MIM exhibited an outstanding adsorption capacity (101.65 mg g−1), as well as an excellent relative selectivity factor (2.01 and 1.92 with respect to biphenyl and fluorine, respectively). Moreover, 1H NMR spectra and Materials Studio simulation verified that the enhanced selective separation efficiency of f-OMCNs@MIM towards DBT could be mainly originated from the interaction between DBT and functional groups from f-OMCNs-based composite structure. Our synthesis strategy of f-OMCNs@MIM in this work shows strong potential for the efficient selective adsorption and separation of more target molecules.