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TALANTA · 2022 · IF 6.556

Development of a highly sensitive and specific monoclonal antibody-based ELISA coupled with immuno-affinity extraction for the detection of anticancer drug 5-fluorouracil in blood samples

Ting Zhou,Guangzhao He,Changjia Hu,Kang Wu,Ying Liu,Jianguo Li,Anping Deng

5-Fluorouracil (5-FU) is an effective anticancer drug widely used in cancer treatment. In this study, two 5-FU derivatives containing a spacer arm with the carboxylic group at the end were synthesized, which were linked to the carrier proteins to form 5-FU-protein conjugates used as the immunogens for the production of monoclonal antibody (mAb). Based on the produced mAb, the highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) for 5-FU detection was established. The IC 50 and LOD values of the assay were found to be 19.5 ng mL −1 and 0.5 ng mL −1 , respectively. There was no cross-reactivity (CR) of the ELISA with cytosine , thymine and uracil , which avoided the interference from inherent pyrimidines. The CR values of the assay with three substitutes of 5-FU (tegafur, 5-fluoro-2′-deoxyuridine, carmofur) were within 9.7%–17.6%. The produced mAb was also applied in sample extraction. The immuno-affinity column capable specific capturing 5-FU was prepared by immobilizing the mAb on Sepharose-4B gel and filling into a SPE column. The recoveries of 5-FU in spiked samples measured by ELISA were 72.4%–90.7% with RSD of 3.6%–8.3%. Five blood samples collected from patients were extracted by immuno-affinity column, then measured by ELISA and confirmed by HPLC-MS/MS. There was a good correlation between HPLC-MS/MS and ELISA. It is demonstrated that the developed ELISA combined with immuno-affinity extraction can be a powerful alternative method for the detection of 5-FU in blood samples.

✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF CHROMATOGRAPHY A · 2022 · IF 4.601

Hydrophobic and hydrophilic selectivity of a multifunctional carbonyldiimidazolium/dodecyl modified silica stationary phase

Kaixing Luo,Yu Luo,Yanjuan Liu,Yuefei Zhang,Wei Chen,Zhengwu Bai,Sheng Tang

s In this work, a new strategy of simultaneously grafting two different functional ligands onto the silica surface was adopted to prepare a carbonyldiimidazolium/dodecyl dual-functionalized silica stationary phase (Sil-CDI-DD) by recycling the discarded commercial C18 columns as the raw materials. A full characterization of the waste silica-C18, the calcined silica, the acidified silica and the new packing materials was done to explore the feasibility of recycling the waste chromatographic columns. Hydrophobic and hydrophilic selectivity of the obtained Sil-CDI-DD was specifically investigated, and retention mechanisms in reversed-phase and hydrophilic interaction chromatography modes were discussed. The experimental results showed that the perfect combination of carbonyldiimidazolium and long-chain alkyl groups commendably enhanced the separation selectivity and efficiency of the versatile Sil-CDI-DD, which could provide multiple interaction sites for diverse target analytes, thereby contributing to the favorable resolution of hydrophilic nucleosides/nucleobases and weak polar environmental endocrine disruptors , including alkylphenols, bisphenols , steroid hormones and phthalates .

✓ 已验证 产品关联 DOI PubMed 高影响因子
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CHEMICO-BIOLOGICAL INTERACTIONS · 2022 · IF 5.168

Hepatic RNA adduction derived from metabolic activation of retrorsine in vitro and in vivo

Weiwei Li,Ting Cheng,Tingting Jiang,Mengyue Zhou,Bowen Gong,Guode Zhao,Jing Li,Rong Tan,Xiaojing Yang,Kandarp Joshi,Ying Peng,Maosheng Cheng,Ting Liu,Dan Ohtan Wang,Jiang Zheng

Pyrrolizidine alkaloids (PAs) are among the most significant hepatotoxins widely distributed in plant species. Incidence of liver injuries caused by PAs has been reported worldwide, and the reactive metabolites of PAs are known to play a critical role in causing the hepatotoxicity. To better understand the toxicity-induction mechanisms, we explored the interactions of PA metabolites with cellular RNA molecules, and examined their effects on the biochemical and metabolic properties of hepatic RNAs. After exposure to retrorsine, adduction on adenosine and guanosine were detected in mouse liver microsomal incubations, cultured mouse primary hepatocytes, and mouse liver tissues. NMR analysis showed that the exocyclic amino group participated in the adduction. We found drastically altered properties and metabolism of the adducted RNA such as reverse-transcriptability, translatability, and RNase-susceptibility. In addition, endogenous modification of N 6 -methyladenosine (m6A) was remarkably reduced.

CAS / MDL: 58-61-7 58-96-8
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MOLECULES · 2022 ·Vol. 27 ·期 13 ·4130 · IF 4.927

Study on Quality Control of Compound Anoectochilus roxburghii (Wall.) Lindl. by Liquid Chromatography–Tandem Mass Spectrometry

Qiuhua Zhang,Lingyi Huang,Youjia Wu,Liying Huang,Xiaowen Xu,Renyi Lin

CompoundAnoectochilus roxburghii(Wall.) Lindl. (A. roxburghii) oral liquid (CAROL) is a hospital preparation ofA. roxburghiiandGanoderma lucidum(G. lucidum),which have hepatoprotective effects. Eight active components (five nucleosides/nucleobases and three triterpenoid acids) in CAROL,A. roxburghii, andG. lucidumwere simultaneously detected by high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS). The multiple reaction monitoring (MRM) mode was applied for the detection of analytes. These eight compounds were separated well within 12 min and quantified using the internal standard working curve method. The method showed good linearity (R2> 0.9935) and high sensitivity (limit of detection = 0.29 ng/mL). The analyte recovery ranged from 85.07% to 97.50% (relative standard deviation < 3.31%). The content of the target analytes in four batches of CAROL, and the raw materials ofG. lucidumandA. roxburghiifrom the five regions was determined using this method. The contents of guanosine and ganoderic acid A in four batches of oral liquid were high and stabilized and could be recommended as quality markers (Q-marker) for CAROL. Simultaneous qualitative and quantitative analysis of nucleosides and triterpenoid acids in CAROL,A. roxburghii, andG. lucidumby LC–MS/MS based on the MRM model was reported for the first time. The proposed method provides a sensitive, rapid, and reliable approach for the quality control of Chinese medicinal products.Keywords:LC–MS/MS;Anoectochilus roxburghii(Wall.) Lindl. and its compound oral liquid;Ganoderma lucidum;analysis;quality control

✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF CHROMATOGRAPHY A · 2022 · IF 4.601

Boronic acid grafted metal-organic framework for selective enrichment of cis-diol-containing compounds

Shitao Feng,An Zhang,Fengshou Wu,Xiaogang Luo,Juan Zhang

This study constructed boronic acid grafted Zr-MOF (BA-Zr-MOF) by a simple pre-installation strategy through mixed organic ligands. Typically, BA-Zr-MOF was prepared by one-step hydrothermal method used for enrichment of cis -diol-containing nucleosides through pipette tip solid-phase extraction (PT-SPE) followed by detection of high-performance liquid chromatography. It is worth mentioning that BA is well assembled into MOF and cis -diol-containing compounds can be efficiently and selectively enriched by abundant boronic acid groups. Three groups of different types of compounds were used to evaluate their selectivity and the results showed the excellent selectivity to cis -diol-containing compounds of as-prepared adsorbent. The BA-Zr-MOF adsorbent possesses a high adsorption capacity, which can reach 86.40 mg g −1 for adenosine. Under the optimal extraction condition, a PT-SPE-HPLC method based on BA-Zr-MOF for analysis of nucleosides was established. The linear range of the four nucleosides is 0.01 to 50 µg mL −1 with R 2 ≥ 0.99 and the detection limits (LODs) are estimated at between 0.005-0.012 µg mL −1 . The recoveries in urine were used to test the reliability of the analytical methods, which ranged from 82.8% to 117.1%, with intra-day relative standard deviations (RSDs) ranged from 0.1% to 4.2% and the inter-day RSDs ranged from 0.2% to 6.2%. All the results show that the pre-installation strategy based on dual ligands is an alternative to fabricate MOF composite material and BA-Zr-MOF is a promising material for the analysis of cis -diol-containing biomolecules.

✓ 已验证 产品关联 DOI PubMed 高影响因子
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MOLECULES · 2022 ·Vol. 27 ·期 13 ·4019 · IF 4.927

Ultrasensitive and Specific Detection of Anticancer Drug 5-Fluorouracil in Blood Samples by a Surface-Enhanced Raman Scattering (SERS)-Based Lateral Flow Immunochromatographic Assay

Hanwen Liu,Ying Liu,Ting Zhou,Penghui Zhou,Jianguo Li,Anping Deng

5-Fluorouracil (5-FU) is an effective anticancer drug widely used in the world. To improve therapy efficiency and reduce side effects, it is very important to frequently detect the concentration of 5-FU in blood samples of patients. In this work, a new type of lateral flow immunochromatographic assay (LFIA) based on surface-enhanced Raman scattering (SERS) for ultrasensitive and specific detection of 5-FU in blood samples was developed. Au@Ag/Au nanoparticles (NPs) employing Au particles as the core and Ag/Au alloy as the shell were synthesized, characterized and used as the substrate in SERS-LFIA due to their high SERS enhancement and biocompatibility. The immunoprobe was made in the form of AuMBA@Ag/Au-Ab in which mercaptobenzoic acid (MBA, a common Raman active reporter) was embedded in the core–shell layer and the monoclonal antibody (mAb) against 5-FU was immobilized on the surface. The performance of SERS-LFIA was similar to that in colloidal gold based-LFIA, and the entire assay time was within 20 min. According to the color intensity on the testing (T) lines of LFIA strips visualized by eyes, the contents of 5-FU in the samples could be qualitatively or semi-quantitatively identified. Furthermore, by measuring the characteristic Raman intensities of MBA on T lines, quantitative detection of 5-FU in the samples were achieved. The IC50and limit of detection (LOD) of the LFIA for 5-FU were found to be 20.9 pg mL−1and 4.4 pg mL−1, respectively. There was no cross-reactivity (CR) of the LFIA with nine relative compounds, and the CR with cytosine, tegafur and carmofur were less than 4.5%. The recoveries of 5-FU from spiked blood samples were in the range of 78.6~86.4% with the relative standard deviation (RSD) of 2.69~4.42%. Five blood samples containing 5-FU collected from the Cancer Hospital were measured by SERS-LFIA, and the results were confirmed by LC-MS/MS. It was proven that the proposed method was able to simply and rapidly detect 5-FU in blood samples with high sensitivity, specificity, accuracy and precision.Keywords:surface-enhanced Raman scattering (SERS);lateral flow immunochromatographic assay (LFIA);5-fluorouracil (5-FU);anticancer drug;blood samples

✓ 已验证 产品关联 DOI PubMed 高影响因子
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Genes · 2022 ·Vol. 13 ·期 5 ·878 · IF 4.141

Potential Misidentification of Natural Isomers and Mass-Analogs of Modified Nucleosides by Liquid Chromatography–Triple Quadrupole Mass Spectrometry

Xiuying Lin,Qianhui Zhang,Yichao Qin,Qisheng Zhong,Daizhu Lv,Xiaopeng Wu,Pengcheng Fu,Huan Lin

Triple quadrupole mass spectrometry coupled to liquid chromatography (LC-TQ-MS) can detect and quantify modified nucleosides present in various types of RNA, and is being used increasingly in epitranscriptomics. However, due to the low resolution of TQ-MS and the structural complexity of the many naturally modified nucleosides identified to date (>160), the discrimination of isomers and mass-analogs can be problematic and is often overlooked. This study analyzes 17 nucleoside standards by LC-TQ-MS with separation on three different analytical columns and discusses, with examples, three major causes of analyte misidentification: structural isomers, mass-analogs, and isotopic crosstalk. It is hoped that this overview and practical examples will help to strengthen the accuracy of the identification of modified nucleosides by LC-TQ-MS.Keywords:nucleoside misidentification;RNA modification;epitranscriptomics;LC-MS/MS;tRNA;HILIC

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JOURNAL OF APPLIED PHYCOLOGY · 2022 ·Vol. 34 ·期 3 ·1497-1511 · IF 3.404

In silico and in vitro assessment of bioactive peptides from Arthrospira platensis phycobiliproteins for DPP-IV inhibitory activity, ACE inhibitory activity, and antioxidant activity

Liu Jing,Bai Xinpeng,Fu Pengcheng

Arthrospira platensis proteins are considered as viable ingredients for functional foods with nutraceutical properties and health-promoting effects. In this study, we used five different proteases including pepsin, trypsin, alcalase, papain, and bromelain to hydrolyze A. platensis phycobiliprotein. LC–MS/MS (liquid chromatography-tandem mass spectrometry) was used to determine the composition of the hydrolysates with thousands of bioactive peptides. Based on peptide sequencing, a candidate list of 1,333 bioactive peptides was constructed for the first time for which bioinformatics tools were used to assess the properties of the bioactive peptides. The inhibitory activity of dipeptidyl peptidase IV (DPP-IV) and angiotensin-converting enzyme (ACE), as well as the antioxidant activity of five phycobiliprotein hydrolysates (PBPHs), were verified in vitro. The IC 50 values of PBPH-pepsin, PBPH-trypsin, PBPH-alcalase, PBPH-papain, and PBPH-bromelain for DPP-IV inhibited activity were determined to be 4.059, 5.603, 5.257, 3.819, and 4.195 mg mL −1 , respectively. All the above five PBPHs were also seen to significantly inhibit ACE activity ( P  < 0.0001) in the range of 0.1–1.0 mg mL −1 . The activity of five PBPHs fraction was found for the 2-azino-bis (3-ethylbenzothiazoline6-sulfonic acid) diammonium salt (ABTS) assay (the highest was 1.37 mM Trolox equivalent of 30 mg mL −1 of PBPH-trypsin), 1,1diphenyl-2-picrylhydrazyl (DPPH) assay (the highest was 204.67 µM Trolox equivalent of 10 mg mL −1 of PBPH-pepsin), and Fe 2+ chelating ability (the highest was 639.73 µM FeSO 4 equivalent of 10 mg mL −1 of PBPH-trypsin). This study indicated that it was feasible to utilize A. platensis phycobiliprotein as a source of bioactive peptides which may be used as functional food and/or nutritional supplements for human health.

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MICROCHEMICAL JOURNAL · 2023 · IF 4.8

Preparation and characterization of mixed-mode hydrophilic stationary phase based on triple hydrophilic functionalized moieties and its application to traditional Chinese herbal compound

Ye Shu,Tong Chen,Tao Liu,Hechen Xu,Jinjin He,Zhihui Cai,Wanning Zhu,Zihan Zhao,Jun Jiang

In this paper, the surface modification of spherical silica was sequentially grafted with functional groups including imidazole, amide and sulfonic acid. The novel surface polymerization type stationary phases (SPs) for hydrophilic interaction liquid chromatography (HILIC) were obtained in three procedures. Elemental analysis, Fourier transform infrared spectroscopy, transmission scanning electron microscopy, Brunauer Emmett Teller and Barrett Joyner Halenda were carried out for the characterization of the SPs. The bonding rate of the functionalized group (1-acrylamide propyl sulfonic) was calculated to be 2.02 μmol/m 2 . Positive outcomes were attained for the surface grafting of the SPs. The effect of various chromatographic conditions on the retention time of various analytes was investigated on high performance liquid chromatography. The dual retention mechanism of hydrophilic interaction and ion exchange mechanism of the prepared SPs was confirmed. The polymeric HILIC SPs had more abundant functionalized groups on the surface, which provided superior hydrophilic properties, ion exchange and thus higher selectivity. Compared with the C 18 column, the resolution of the HILIC column was better. The HILIC SPs were used for the separation of sulfonamides and flavonoid natural components, and had achieved better baseline separations. In addition, the HILIC mode was used for the separation of the extract of the Fructus Ligustri Lucidi , in which the quality markers had satisfactory peak shape and baseline separation. In conclusion, the prepared surface polymeric SPs were competent for the analysis of polar components and applicable to complex samples.

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REACTIVE & FUNCTIONAL POLYMERS · 2023 · IF 5.1

Boronate affinity-based surface molecularly imprinted polymer microspheres using polyethyleneimine/dopamine coating for efficient selective recognition and separation of Ginsenoside Rb1

Yu Liu,Xinqi Qiu,Minhui Zhang,Yali Lin,Hui Lan,Xican Li,Quanzhou Wu,Jianfeng He

A novel approach combining boronate affinity with surface molecular imprinting was developed for the specific recognition and separation of ginsenoside Rb1 . Firstly, the boronic acid-functionalized polymer microspheres were prepared as inner cores via suspension polymerization using methyl methacrylate , ethyleneglycol dimethacrylate, and 4-vinylphenyl boronic pinacol ester (4-VBPE) as polymerization monomers . Then, the Ginsenoside Rb1 (GRb1) template was covalently pre-anchored onto the surface of microspheres through boronate affinity binding after the hydrolysis of 4-VBPE. Finally, dopamine and polyethyleneimine were used as co -coating monomers to form a thickness-controllable imprinting layer onto the polymer microspheres via in-water self-copolymerization. The key factors affecting specific recognition, including the boronic acid content, adsorption media, and polymerization time were carefully optimized. The resulting molecularly imprinted microspheres (MIMs) exhibited strong surface hydrophilicity . In the aqueous media, the optimal adsorption capacity for the template reached 81.45 μmol g −1 , and the imprinting factor was 5.32. The specific evaluation showed that the MIMs had a much higher selectivity for GRb1 than its analogues, and the selectivity coefficients were all higher than 2.4. In addition, the MIMs can be applied as adsorbents for solid-phase extraction to selectively separate ginsenosides from American ginseng extract. The eluting recovery ratios of GRb1, GRg2, and GRd reach 92.39%, 88.90%, and 92.49%, respectively.

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ANALYTICAL SCIENCES · 2023 · IF 1.6

A comprehensive evaluation of a polymeric zwitterionic hydrophilic monolith for nucleotide separation

Lai Liang,Zhang Mengyun,Liu Chusheng,Qu Jiahuan,Xu Dongsheng,Jiang Zhengjin

Rapid and effective separation of nucleotides (NTs) and their derivatives is crucial for studying their physiological functions. In this work, we comprehensively evaluated the separation ability of a zwitterionic hydrophilic monolith, i.e., poly(N,N-dimethyl-N-(3-methacrylamidopropyl)-N-(3-sulfopropyl)ammonium betaine- co –N,N’-methylenebisacrylamide) (poly(SPP- co -MBA)) for NTs analysis, including its selectivity, chemical stability under extremely basic condition and compatibility with hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (HILIC-MS). The poly(SPP- co -MBA) monolith exhibited excellent chemical stability, as evidenced by the low relative standard deviation of retention time (0.16–1.05%) after 4000 consecutive injections over one month under strong alkaline elution condition (pH 10). After optimizing the separation conditions, including buffer pH and concentration, organic solvent content and column temperature, four nucleoside triphosphates, five nucleoside diphosphates and five nucleoside monophosphates were baseline separated within 7 min. Additionally, the mixtures containing one nucleoside and its corresponding mono-, di-, and triphosphates were baseline separated within only 3 min, respectively. It is good HILIC-MS compatibility was also confirmed by the satisfactory peak shape and high response of nine NTs. Overall, the proposed poly(SPP- co -MBA) monolith exhibited good mechanical stability and compatibility of HILIC-MS, making it a promising technique for NTs analysis. Graphical Abstract

✓ 已验证 产品关联 DOI PubMed 高影响因子
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FOOD CHEMISTRY · 2023 · IF 8.8

Insights into the effects of steaming on organoleptic quality of salmon (Salmo salar) integrating multi-omics analysis and electronic sensory system

Meng Dong,Feng Liang,Shuang Cui,Bing-Bing Mao,Xu-Hui Huang,Lei Qin

The effect of steaming treatment on salmon quality was explored by different multi-omics and electronic sensory system in this study. A comparison between conventional steaming (CS) and anaerobic steaming (AS) was conducted in organoleptic quality of salmon. Twelve key volatile compounds were identified, which contributed to the flavor difference. The concentrations of hexanal, (E)-2-octen-1-al, and decanal in AS salmon were significantly lower than in CS salmon, which account for 68.9–80.5 % of the latter. During steaming, the fatty acids and diacylglycerols decreased significantly by 37.4 % and 57.9 %, respectively. Anaerobic steaming limited the degradation of some oxidized lipids, further reduced some volatile secondary oxidation products. Nucleotides and derivatives, succinic acid, glutamic acid, hydroxyproline and betaine contributed to the saltness, umami, richness of steamed salmon. Metabolomics data revealed that the higher creatinine, Ala-Ala and Ala-Leu provided more umami and less bitterness to AS salmon.

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