Analytica Chimica Acta (v.882, #C)
Editorial board (iii).
Characterisation and determination of fullerenes: A critical review by Alina Astefanei; Oscar Núñez; Maria Teresa Galceran (1-21).
Display OmittedA prominent sector of nanotechnology is occupied by a class of carbon-based nanoparticles known as fullerenes. Fullerene particle size and shape impact in how easily these particles are transported into and throughout the environment and living tissues. Currently, there is a lack of adequate methodology for their size and shape characterisation, identification and quantitative detection in environmental and biological samples. The most commonly used methods for their size measurements (aggregation, size distribution, shape, etc.), the effect of sampling and sample treatment on these characteristics and the analytical methods proposed for their determination in complex matrices are discussed in this review. For the characterisation and analysis of fullerenes in real samples, different analytical techniques including microscopy, spectroscopy, flow field-flow fractionation, electrophoresis, light scattering, liquid chromatography and mass spectrometry have been reported. The existing limitations and knowledge gaps in the use of these techniques are discussed and the necessity to hyphenate complementary ones for the accurate characterisation, identification and quantitation of these nanoparticles is highlighted.
Keywords: Fullerenes; Aggregates; Particle size and shape analysis; Identification; Detection and quantitation; Complex matrices;
A label-free and self-assembled electrochemical biosensor for highly sensitive detection of cyclic diguanylate monophosphate (c-di-GMP) based on RNA riboswitch by Qingyun Xie; Fulin Zhao; Hongrui Liu; Yanke Shan; Fei Liu (22-26).
Display OmittedCyclic diguanylate monophosphate (c-di-GMP) is an important second messenger that regulates a variety of complex physiological processes involved in motility, virulence, biofilm formation and cell cycle progression in several bacteria. Herein we report a simple label-free and self-assembled RNA riboswitch-based biosensor for sensitive and selective detection of c-di-GMP. The detectable concentration range of c-di-GMP is from 50 nM to 1 μM with a detection limit of 50 nM.
Keywords: Cyclic diguanylate monophosphate (c-di-GMP); Riboswitch; Biosensor; Label-free; Self-assembled;
Sensitive detection of a serum biomarker based on peptide nucleic acid-coupled dual cycling reactions by Yuanyuan Zhang; Hao Li; Yue Huang; Tingting Yin; Lizhou Sun; Genxi Li (27-31).
Display OmittedSerum level of disease markers may provide important guidance for diagnosis and prognosis. In this work, a sensitive and specific method suitable for direct serum detection of biomarkers is developed based on peptide nucleic acid (PNA)-coupled DNA cycling reactions with dual amplification. In this method, PNA released from a target-triggered homogeneous DNA cycling is employed to initiate an interface DNA cycling, and both of the cycling reactions are based on polymerase-assisted strand displacement reaction. Consequently, two PNA-coupled DNA cycling steps can take place simultaneously in one-pot, leading to greatly enhanced limit of detection and simplified operation. This method has also been successfully applied for evaluating serum insulin in pregnant women as an indicator of gestational diabetes mellitus. So the application of this method in real bio-samples may allow it to hold considerable potential in clinical practice. In addition, since there is no requirement for specific sequence of aptamer, the strategy proposed can be extended for the detection of many other protein markers and peptide-hormones in the future.
Keywords: Peptide nucleic acid; Strand displacement; Dual amplification; Insulin; Gestational diabetes mellitus;
Label-free electrochemical aptasensor constructed by layer-by-layer technology for sensitive and selective detection of cancer cells by Tianshu Wang; Jiyang Liu; Xiaoxiao Gu; Dan Li; Jin Wang; Erkang Wang (32-37).
Display OmittedHere, a cytosensor was constructed with ferrocene-appended poly(allylamine hydrochloride) (Fc-PAH) functionalized graphene (Fc-PAH-G), poly(sodium-p-styrenesulfonate) (PSS) and aptamer (AS1411) by layer-by-layer assembly technology. The hybrid nanocomposite Fc-PAH-G not only brings probes on the electrode and also promotes electron transfer between the probes and the substrate electrode. Meanwhile, LBL technology provides more effective probes to enhance amplified signal for improving the sensitivity of the detection. While AS1411 forming G-quardruplex structure and binding cancer cells, the current response of the sensing electrode decreased due to the insulating properties of cellular membrane. Differential pulse voltammetry (DPV) was performed to investigate the electrochemical detection of HeLa cells attributing to its sensitivity of the current signal change. The as-prepared aptasensor showed a high sensitivity and good stability, a widely detection range from 10 to 106 cells/mL with a detection limit as low as 10 cells/mL for the detection of cancer cells.
Keywords: Graphene-based nanocomposite; Electrochemical aptasensor; Layer-by-layer technology; Differential pulse voltammetry; Cancer cells;
Quantitative analysis of low-abundance serological proteins with peptide affinity-based enrichment and pseudo-multiple reaction monitoring by hybrid quadrupole time-of-flight mass spectrometry by Kwang Hoe Kim; Yeong Hee Ahn; Eun Sun Ji; Ju Yeon Lee; Jin Young Kim; Hyun Joo An; Jong Shin Yoo (38-48).
Display OmittedMultiple reaction monitoring (MRM) is commonly used for the quantitative analysis of proteins during mass pectrometry (MS), and has excellent specificity and sensitivity for an analyte in a complex sample. In this study, a pseudo-MRM method for the quantitative analysis of low-abundance serological proteins was developed using hybrid quadrupole time-of-flight (hybrid Q-TOF) MS and peptide affinity-based enrichment. First, a pseudo-MRM-based analysis using hybrid Q-TOF MS was performed for synthetic peptides selected as targets and spiked into tryptic digests of human serum. By integrating multiple transition signals corresponding to fragment ions in the full scan MS/MS spectrum of a precursor ion of the target peptide, a pseudo-MRM MS analysis of the target peptide showed an increased signal-to-noise (S/N) ratio and sensitivity, as well as an improved reproducibility. The pseudo-MRM method was then used for the quantitative analysis of the tryptic peptides of two low-abundance serological proteins, tissue inhibitor of metalloproteinase 1 (TIMP1) and tissue-type protein tyrosine phosphatase kappa (PTPκ), which were prepared with peptide affinity-based enrichment from human serum. Finally, this method was used to detect femtomolar amounts of target peptides derived from TIMP1 and PTPκ, with good coefficients of variation (CV 2.7% and 9.8%, respectively), using a few microliters of human serum from colorectal cancer patients. The results suggest that pseudo-MRM using hybrid Q-TOF MS, combined with peptide affinity-based enrichment, could become a promising alternative for the quantitative analysis of low-abundance target proteins of interest in complex serum samples that avoids protein depletion.
Keywords: Low-abundance serological protein; Peptide affinity-based enrichment; Pseudo-multiple reaction monitoring; Hybrid quadrupole time-of-flight mass spectrometer;
Coumarins as new matrices for matrix-assisted laser-desorption/ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of hydrophobic compounds by Hang Wang; Bona Dai; Bin Liu; Han Lu (49-57).
Display OmittedHydrophobic compounds with hydroxyl, aldehyde or ketone groups are generally difficult to detect using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), because these compounds have low proton affinity and are poorly ionized by MALDI. Herein, coumarins have been used as new matrices for MALDI-MS analysis of a variety of hydrophobic compounds with low ionization efficiency, including steroids, coenzyme Q10, a cyclic lipopeptide and cholesterol oleate. Five coumarins, including coumarin, umbelliferone, esculetin, 7-hydroxycoumarin-3-carboxylic acid (HCA) and 6,7-dihydroxycoumarin-3-carboxylic acid (DCA), were compared with the conventional matrices of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA). Coumarins with hydroxyl or carboxylic acid groups enabled detection. Taking DCA as an example, this matrix proved to be superior to DHB or CHCA in detection sensitivity, stability, spot-to-spot and sample-to-sample reproducibility, and accuracy. DCA increased the stability of the target compounds and decreased the loss of water. The [M + Na]+ peaks were observed for all target compounds by adding NaCl as an additive, and the [M − H2O + H]+ and [M + H]+ peaks decreased. DCA was selected for the identification of sterols in yeast cells, and thirteen sterols were detected by Fourier transform ion cyclotron resonance (FT ICR) mass spectrometry. This work demonstrates the potential of DCA as a new matrix for detection of hydrophobic molecules by MALDI-MS and provides an alternative tool for screening sterols in antifungal research.
Keywords: Coumarins; Hydrophobic compounds; Matrix-assisted laser desorption/ionization; Mass spectrometry; Yeast sterols;
A colorimetric sensor array for detection and discrimination of biothiols based on aggregation of gold nanoparticles by Forough Ghasemi; M. Reza Hormozi-Nezhad; Morteza Mahmoudi (58-67).
Display OmittedDevelopments of sensitive, rapid, and cheap systems for identification of a wide range of biomolecules have been recognized as a critical need in the biology field. Here, we introduce a simple colorimetric sensor array for detection of biological thiols, based on aggregation of three types of surface engineered gold nanoparticles (AuNPs). The low-molecular-weight biological thiols show high affinity to the surface of AuNPs; this causes replacement of AuNPs’ shells with thiol containing target molecules leading to the aggregation of the AuNPs through intermolecular electrostatic interaction or hydrogen-bonding. As a result of the predetermined aggregation, color and UV–vis spectra of AuNPs are changed. We employed the digital mapping approach to analyze the spectral variations with statistical and chemometric methods, including hierarchical cluster analysis (HCA) and principal component analysis (PCA). The proposed array could successfully differentiate biological molecules (e.g., cysteine, glutathione and glutathione disulfide) from other potential interferences such as amino acids in the concentration range of 10–800 μmol L−1.
Keywords: Gold nanoparticle; Biothiol; Aggregation; Colorimetric sensor array; Chemometric method;
A highly selective turn-on fluorescent probe for hypochlorous acid based on hypochlorous acid-induced oxidative intramolecular cyclization of boron dipyrromethene-hydrazone by Wei-Chieh Chen; Parthiban Venkatesan; Shu-Pao Wu (68-75).
Display OmittedA BODIPY-based fluorescent probe, HBP, was developed for the detection of hypochlorous acid based on the specific hypochlorous acid-promoted oxidative intramolecular cyclization of heterocyclic hydrazone in response to the amount of HOCl. The reaction is accompanied by a 41-fold increase in the fluorescent quantum yield (from 0.004 to 0.164). The fluorescence intensity of the reaction between HOCl and HBP is linear in the HOCl concentration range of 1–8 μM with a detection limit of 2.4 nM (S/N = 3). Confocal fluorescence microscopy imaging using RAW264.7 cells showed that the new probe HBP could be used as an effective fluorescent probe for detecting HOCl in living cells.
Keywords: Hypochlorous acid; Fluorescent probe; Boron dipyrromethene; Bioimaging;
A near-infrared emissive Al3+ sensing platform for specific detection in solution, cells and probing DNase activity by Barun K. Datta; Durairaj Thiyagarajan; Chirantan Kar; Aiyagari Ramesh; Gopal Das (76-82).
Display OmittedA new tricarbocyanine-based chemosensor exhibited a dramatic Al3+-specific fluorescence turn-on response in the near-infrared (NIR) region. The receptor was found to be highly selective towards Al3+ over other metal ions in physiological condition. The sensor was non-toxic and could thus be employed as an imaging probe for detecting intracellular Al3+ in live cells. Interestingly, upon interaction with DNA in solution, the L–Al3+ ensemble rendered tracking of DNase activity in solution through a systematic reduction in the fluorescence emission intensity.
Keywords: Near-infrared fluorescence; Al3+ sensor; Cell imaging; DNase activity sensing;
Single-step, paper-based concentration and detection of a malaria biomarker by David Y. Pereira; Ricky Y.T. Chiu; Samantha C.L. Zhang; Benjamin M. Wu; Daniel T. Kamei (83-89).
Display OmittedThe lateral-flow immunoassay (LFA) is an inexpensive and rapid paper-based assay that can potentially detect infectious disease biomarkers in resource-poor settings. Despite its many advantages that make it suitable for point-of-care diagnosis, LFA is limited by its inferior sensitivity relative to sophisticated laboratory-based assays. Our group previously introduced the use of a micellar aqueous two-phase system (ATPS), comprised of the nonionic Triton X-114 surfactant, to concentrate biomarkers in a sample and enhance their detection with LFA. However, achieving complete phase separation and target concentration using the Triton X-114 system required many hours, and the concentrated sample needed to be manually extracted and applied to LFA. Here, we successfully integrated the concentration and detection steps into a single step that occurs entirely within a portable paper-based diagnostic strip. In a novel approach, we applied the micellar ATPS to a 3-D paper design and effectively reduced the macroscopic phase separation time from 8 h to approximately 3 min. The 3-D design was integrated with LFA to simultaneously concentrate and detect Plasmodium lactate dehydrogenase (pLDH), a malaria biomarker, in both phosphate-buffered saline and fetal bovine serum within 20 min at room temperature. Compared to a conventional LFA setup with a pLDH detection limit of 10 ng μL−1, our single-step diagnostic successfully detected pLDH at 1.0 ng μL−1, demonstrating a 10-fold detection limit improvement and resulting in a sensitive and user-friendly assay that can be used at the point-of-care. The integration of a micellar ATPS and LFA represents a new platform that can improve and promote the use of paper-based diagnostic assays for malaria and other diseases within resource-poor settings.
Keywords: Lateral-flow immunoassay; Malaria; Micellar aqueous two-phase system; Triton X-114; Diagnostic;
Determination of carbohydrates in tobacco by pressurized liquid extraction combined with a novel ultrasound-assisted dispersive liquid–liquid microextraction method by Kai Cai; Deyu Hu; Bo Lei; Huina Zhao; Wenjie Pan; Baoan Song (90-100).
Display OmittedA novel derivatization-ultrasonic assisted-dispersive liquid–liquid microextraction (UA-DLLME) method for the simultaneous determination of 11 main carbohydrates in tobacco has been developed. The combined method involves pressurized liquid extraction (PLE), derivatization, and UA-DLLME, followed by the analysis of the main carbohydrates with a gas chromatography-flame ionization detector (GC-FID). First, the PLE conditions were optimized using a univariate approach. Then, the derivatization methods were properly compared and optimized. The aldononitrile acetate method combined with the O-methoxyoxime-trimethylsilyl method was used for derivatization. Finally, the critical variables affecting the UA-DLLME extraction efficiency were searched using fractional factorial design (FFD) and further optimized using Doehlert design (DD) of the response surface methodology. The optimum conditions were found to be 44 μL for CHCl3, 2.3 mL for H2O, 11% w/v for NaCl, 5 min for the extraction time and 5 min for the centrifugation time. Under the optimized experimental conditions, the detection limit of the method (LODs) and linear correlation coefficient were found to be in the range of 0.06–0.90 μg mL−1 and 0.9987–0.9999. The proposed method was successfully employed to analyze three flue-cured tobacco cultivars, among which the main carbohydrate concentrations were found to be very different.
Keywords: Tobacco; Pressurized liquid extraction; Derivatization; Fractional factorial design; Doehlert design; Ultrasonic assisted-dispersive liquid–liquid microextraction;
Advanced dress-up chiral columns: New removable chiral stationary phases for enantioseparation of chiral carboxylic acids by Kenichiro Todoroki; Yasuhiro Ishii; Takafumi Ide; Jun Zhe Min; Koichi Inoue; Xin Huang; Wei Zhang; Yoshitaka Hamashima; Toshimasa Toyo’oka (101-111).
Display OmittedThis paper describes the preparation of new dress-up columns featuring reproducibly removable and replaceable chiral stationary phases. After synthesizing perfluroalkylated quinine and quinidine derivatives as chiral stationary phase compounds (F-CSPs), we adsorbed them reversibly onto a fluorous LC column through pumping of their solutions. Using this dress-up chiral column and fluorophobic elution of aqueous ammonium formate/MeOH mixtures, we could enantioseparate four racemic N-acetyl amino acids, dichlorprop, and sixteen fluorescent 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC)-derivatized amino acids. Dressing and undressing of the coated F-CSPs could be controlled by varying the fluorophilicity and fluorophobicity of the eluent. The relative standard deviations of the retention times, the retention factors, the number of theoretical plates, the enantioseparation factors, and the resolutions of each of four preparations of such dress-up columns were all less than or equal to 5.26% (from 20 repeated analyses); the reproducibilities from four different preparations were all less than or equal to 10.6%. These columns also facilitated highly sensitive and selective analyses of AQC-amino acids when detected using LC–MS/MS.
Keywords: Dress-up chiral column; Fluorous separation; Chiral carboxylic acids; Cinchona alkaloids;
Determination of chiral pharmaceuticals and illicit drugs in wastewater and sludge using microwave assisted extraction, solid-phase extraction and chiral liquid chromatography coupled with tandem mass spectrometry by Sian E. Evans; Paul Davies; Anneke Lubben; Barbara Kasprzyk-Hordern (112-126).
Display OmittedThis is the first study presenting a multi-residue method allowing for comprehensive analysis of several chiral pharmacologically active compounds (cPACs) including beta-blockers, antidepressants and amphetamines in wastewater and digested sludge at the enantiomeric level. Analysis of both the liquid and solid matrices within wastewater treatment is crucial to being able to carry out mass balance within these systems. The method developed comprises filtration, microwave assisted extraction and solid phase extraction followed by chiral liquid chromatography coupled with tandem mass spectrometry to analyse the enantiomers of 18 compounds within all three matrices. The method was successfully validated for 10 compounds within all three matrices (amphetamine, methamphetamine, MDMA, MDA, venlafaxine, desmethylvenlafaxine, citalopram, metoprolol, propranolol and sotalol), 7 compounds validated for the liquid matrices only (mirtazapine, salbutamol, fluoxetine, desmethylcitalopram, atenolol, ephedrine and pseudoephedrine) and 1 compound (alprenolol) passing the criteria for solid samples only. The method was then applied to wastewater samples; cPACs were found at concentration ranges in liquid matrices of: 1.7 ng L−1 (metoprolol) – 1321 ng L−1 (tramadol) in influent, <LOD (desmethylcitalopram and metoprolol) – 506 ng L−1 in effluent, and in solid matrix digested sludge: 0.4 ng g−1 (metoprolol) – 275 ng g−1 (citalopram). Enantiomeric profiling revealed that studied compounds were present in analysed samples in non-racemic composition. Furthermore, enantiomeric composition of studied analytes differed in liquid and solid matrices. This demonstrates that not analysing the solid fraction of wastewater may lead to over-estimation of the removal rates of cPACs as well as possible misrepresentation of the enantiomeric fraction of the compounds as they leave the wastewater treatment plant. Consequently risks from cPACs entering the environment might be higher than anticipated.
Keywords: Chiral drugs; Pharmaceuticals; Illicit; Wastewater; Sludge; Chiral chromatography coupled with mass spectrometry;
Aminoglycoside analysis in food of animal origin with a zwitterionic stationary phase and liquid chromatography–tandem mass spectrometry by Cristina Díez; Davy Guillarme; Aline Staub Spörri; Emmanuelle Cognard; Didier Ortelli; Patrick Edder; Serge Rudaz (127-139).
Display OmittedIn this study, fourteen highly polar aminoglycoside (AGs) antibiotics were selected. Various stationary phases were tested, including Obelisc R, ZIC-HILIC, BEH amide and aminopropyl. The nature of the stationary phase, mobile phase (water or buffer solutions and acetonitrile), pH (percentage of formic acid), gradient conditions and injection solvents were systematically studied as relevant parameters for tuning retention selectivity and detectability of AGs in liquid chromatography electrospray tandem mass spectrometry (LC–(ESI)–MS/MS). Only the two zwitterionic phases (Obelisc R and ZIC-HILIC) achieved a proper chromatographic separation considering interferences due to the crosstalk effect in low resolution mass spectrometers. The water/acetonitrile mobile phase containing 1% formic acid used with Obelisc R provided more sensitivity than the highly concentrated buffered mobile phases required for ZIC-HILIC. A solid phase extraction (SPE) clean-up procedure with polymeric weak cation exchange (WCX) cartridges was optimized for honey, milk and liver samples. Different brands of cartridges and elution solvents were tested, and the Taurus WCX offered the best recovery rate with a buffer elution at pH 3. The final optimized method was validated in these matrices according to Decision 2002/657/EC. A monitoring campaign for sixty honey, milk and liver samples was carried out at the Food Authority Control in Geneva. The concentration of dihydrostreptomycin (DSTP) found in one ovine liver exceeded the established maximum residue levels (MRLs) within the European and Swiss legislations but it was compliant taking into account the validation data.
Keywords: Aminoglycosides; Zwitterionic; Honey; Milk; Liver;