RESUMO
Cellulose thread substrates offer a platform for microsampling and reactive ionization of free fatty acid (FFA) isomers for direct differentiation by mass spectrometry. Ambient corona discharge forms when direct current high voltage is applied to the tiny subfibers on the thread substrate in the presence of a polar spray solvent (MeOH/H2O, 2:1, v/v), facilitating chemical reactions across a CâC bond of unsaturated fatty acids. The process was applied for diagnosis of obesity, which we observed to show better discriminatory power when compared to determinations based on body mass index. Overall, the integrated reactive thread-based platform is capable of (i) microsampling and dry-state, room-temperature storage (>30 days) of the biofluids, (ii) in-capillary liquid/liquid extraction, and (iii) in situ epoxidation reactions to locate the CâC bond position in unsaturated fatty acids via reactions with reactive oxygen species present in ambient corona discharge. The study showcased the ability to correctly characterize FFAs, including degree of unsaturation, and the determination of their relative concentrations in clinical biofluid samples.
Assuntos
Ácidos Graxos não Esterificados , Ácidos Graxos Insaturados , Ácidos Graxos Insaturados/química , Humanos , Isomerismo , Espectrometria de Massas/métodos , Obesidade/diagnósticoRESUMO
Direct analysis of whole blood on bloodstained textiles is achieved with thread spray mass spectrometry (MS). This capability satisfies investigators' first priority in crime scene investigations, which is determining if a stain is blood. This thread spray method explores the use of evidentiary fabric threads for rapid determination of hemoglobin directly from whole blood within textiles without prior extraction steps. The multiplicity of information that can be derived from the thread spray MS method distinguishes it from the current presumptive Bluestar® method, by enabling the detection of hemoglobin (both α- and ß-chains), the heme co-factor and lipids all from a single blood sample. Lipid composition was found to differ for blood samples originating from human, canine, and horse species. The robustness of the thread spray MS method as a forensic analytical platform was evaluated in three ways: (1) its successful applicability to samples previously tested by the Bluestar® presumptive method, offering a confirmatory test without prior sample pre-treatment, (2) successful detection of heme from previously washed fabrics, which demonstrated the unprecedented sensitivity of the thread spray method, and (3) the ability to analyze samples stored under ambient conditions for up to 30 days. These results attest to the potential capabilities of the thread spray MS platform in forensic serology, and its application for direct analysis of evidentiary garments, which confer the advantages of rapid analysis and the reduction of the false positive and negative identification rates for blood on textiles.
Assuntos
Manchas de Sangue , Medicina Legal , Animais , Cães , Cavalos , Espectrometria de Massas , TêxteisRESUMO
Ricin is a naturally occurring, highly potent toxin native to castor bean plants that has recently been used as a biological weapon in cases of bioterrorism and suicide attempts. Difficulties with direct detection arise from large heterogeneities in ricin glycosylation, which leads to markedly different bioactivity, and the fact that carefully developed and laborious sample preparation steps are required to maintaining the activity of the protein during analysis. Herein, we present an alternative, two-tiered approach to identify the presence of ricin by detecting ricinoleic acid and ricinine, which are co-extracted with the protein. This direct mass spectrometric-based technique takes as little as 2 minutes, and we determined its sensitivity to be in the parts-per-trillion range. Our method is applicable to paper substrates from suspected contaminated envelopes and biofluids from at-risk patients. The fact that prior sample preparations are not needed in this procedure means that analysis can be performed in the field for emergency cases.
RESUMO
Storage and quantitative analysis of small volumes of biofluids are challenging, especially when low concentrations of analytes are to be detected in the presence of complex matrices. In this study, we describe an integrated thread-based approach for stabilizing small blood volumes in the dry-state at room temperature, while also offering direct analysis capabilities via thread spray mass spectrometry. The analytical merits of this novel microsampling platform was demonstrated via the direct analysis of diazepam and cocaine in dried blood samples stored for 42 days. In-situ in-capillary blood processing from hydrophobic threads enabled limits of detection as low as parts-per-quadrillion to be reached. We validated this ultra-sensitivity by analyzing small tissue-like residues collected after pushing a thread through the sample once. The implications of this sample collection, storage, and analysis platform can be extensive with direct applications in forensics and clinical studies.
Assuntos
Fibra de Algodão , Teste em Amostras de Sangue Seco/métodos , Extração em Fase Sólida/métodos , Anfetamina/sangue , Cocaína/análogos & derivados , Cocaína/sangue , Diazepam/sangue , Teste em Amostras de Sangue Seco/instrumentação , Gossypium , Humanos , Limite de Detecção , Espectrometria de Massas/métodos , Metanfetamina/sangue , Sefarose/química , Extração em Fase Sólida/instrumentação , Manejo de EspécimesRESUMO
Accurate and rapid analysis of complex microsamples are challenging tasks in translational research. Nanoelectrospray ionization (nESI) is the method of choice for analyzing small sample volumes by mass spectrometry (MS), but this technique works well only for polar analytes. Herein, we describe a versatile dual noncontact nESI/nAPCI (nanoatmospheric pressure chemical ionization) source that allows simultaneous detection of both polar and nonpolar analytes in microliter quantities of samples under ambient conditions and without pretreatment. The same device can be activated to enable electrophoretic separation. The noncontact nESI/nAPCI MS platform was applied to analyze different samples, including high sensitive direct analysis of biofluids and the efficient detection of proteins in buffers with high concentration of nonvolatile salts. Excellent linearity, accuracy and limits of detection were achieved for compounds with different chemical properties in different matrices. The high sensitivity, universality, simplicity, and ease of operation make this MS technique promising for use in clinical and forensic applications.
Assuntos
Misturas Complexas/análise , Animais , Análise Química do Sangue , Bovinos , Eletroforese , Humanos , Limite de Detecção , Compostos Orgânicos/sangue , Compostos Orgânicos/urina , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
Thread spray ambient ionization uses single threads as a medium for sampling and ionization. This approach was demonstrated through the detection of various capsaicinoids from the interior of pepper fruits without destruction of the sample. Pepper residues present on the thread were analyzed by the application of DC voltage and solvent to cause field-induced charged droplet generation. Capsaicinoids extracted from the sample are contained in the electrosprayed droplets and transported to the mass spectrometer for characterization. The thread spray mass spectrometry method was optimized using commercially available materials like 100% cotton, cotton:polyester (35/65), 100% polyester, and nylon fabrics and subsequently applied for in-situ analysis of six different pepper fruits and pepper spray residues on fabrics. The results indicated that the special physico-chemical characteristics of threads allowed a rapid and convenient sampling and ionization of pepper products for analysis by mass spectrometry. The total capsaicinoid ion yields for the various pepper products correlated very well with that reported in Scoville Heat Units, suggesting that quantitative assessment of pungency levels may be achieved via the direct sample analysis without prior separation.
Assuntos
Capsaicina/análise , Capsicum/química , Frutas/química , Espectrometria de Massas , Estrutura MolecularRESUMO
Monomer sequence is demonstrated to be a primary factor in determining the hydrolytic degradation profile of poly(lactic-co-glycolic acid)s (PLGAs). Although many approaches have been used to tune the degradation of PLGAs, little effort has been expended in exploring the sequence-control strategy exploited by nature in biopolymers. Cylindrical matrices and films prepared from a series of sequenced and random PLGAs were subjected to hydrolysis in a pH 7.4 buffer at 37 °C. Swelling ranged from 107% for the random racemic PLGA with a 50:50 ratio of lactic (L) to glycolic (G) units to 6% for the sequenced alternating copolymer poly LG. Erosion followed an inverse trend with the random 50:50 PLGA showing an erosion half-life of 3-4 weeks while poly LG required ca. >10 weeks. Stereosequence was found to play a large role in determining swelling and erosion; stereopure analogs swelled less and were slower to lose mass. Molecular weight loss followed similar trends and increases in dispersity correlated with the onset of significant swelling. The relative proportion of rapidly cleavable G-G linkages relative to G-L/L-G (moderate) and L-L (slow) correlates strongly with the degree of swelling observed and the rate of erosion. The dramatic sequence-dependent variation in swelling, in the absence of a parallel hydrophilicity trend, suggest that osmotic pressure, driven by the differential accumulation of degradation products, plays an important role.