This article presents a novel method
for selective acquisition
of Fourier transform infrared (FT-IR) spectra of microorganisms in-line
during fermentation, using Saccharomyces cerevisiae as an example. The position of the cells relative to the sensitive
region of the attenuated total reflection (ATR) FT-IR probe was controlled
by combing a commercially available ATR in-line probe with contact-free,
gentle particle manipulation by ultrasonic standing waves. A prototype
probe was successfully constructed, assembled, and tested in-line
during fed-batch fermentations of S. cerevisiae. Control over the position of the cells was achieved by tuning the
ultrasound frequency: 2.41 MHz was used for acquisition of spectra
of the cells (pushing frequency fp) and
1.87 MHz, for retracting the cells from the ATR element, therefore
allowing spectra of the medium to be acquired. Accumulation of storage
carbohydrates (trehalose and glycogen) inside the cells was induced
by a lack of a nitrogen source in the feed medium. These changes in
biochemical composition were visible in the spectra of the cells recorded
in-line during the application of fp and
could be verified by reference spectra of dried cell samples recorded
off-line with a FT-IR microscope. Comparison of the cell spectra with
spectra of trehalose, glycogen, glucose, and mannan, i.e., the major
carbohydrates present in S. cerevisiae, and principal components analysis revealed that the changes observed
in the cell spectra correlated well with the bands specific for trehalose
and glycogen. This proves the applicability and capability of ultrasound-enhanced
in-line ATR mid-IR spectroscopy as a real-time PAT method for the
in situ monitoring of cellular biochemistry during fermentation.
A fast and simple method to control variations in carbohydrate composition of Saccharomyces cerevisiae, baker's yeast, during fermentation was developed using mid-infrared (mid-IR) spectroscopy. The method allows for precise and accurate determinations with minimal or no sample preparation and reagent consumption based on mid-IR spectra and partial least squares (PLS) regression. The PLS models were developed employing the results from reference analysis of the yeast cells. The reference analyses quantify the amount of trehalose, glucose, glycogen, and mannan in S. cerevisiae. The selection and optimization of pretreatment steps of samples such as the disruption of the yeast cells and the hydrolysis of mannan and glycogen to obtain monosaccharides were carried out. Trehalose, glucose, and mannose were determined using high-performance liquid chromatography coupled with a refractive index detector and total carbohydrates were measured using the phenol–sulfuric method. Linear concentration range, accuracy, precision, LOD and LOQ were examined to check the reliability of the chromatographic method for each analyte.FigureComparison of workflows for carbohydrate determination in S.cerevisiae by FT-IR spectroscopy and HPLC-RIElectronic supplementary materialThe online version of this article (doi:10.1007/s00216-013-7239-9) contains supplementary material, which is available to authorized users.
A sensitive and reliable method based on MEKC has been developed and validated for trace determination of neonicotinoid insecticides (thiamethoxam, acetamiprid, and imidacloprid) and the metabolite 6-chloronicotinic acid in water and soil matrices. Optimum separation of the neonicotinoid insecticides was obtained on a 58 cm long capillary (75 μm id) using as the running electrolyte 40 mM SDS, 5 mM borate (pH 10.4), and 5% (v/v) methanol at a temperature of 25°C, a voltage of 25 kV and with hydrodynamic injection (10 s). The analysis time was less than 7 min. Prior to MEKC determination, the samples were purified and enriched by carrying out extraction-preconcentration steps. For aqueous samples, off-line SPE with a sorptive material such as Strata-X (polymeric hydrophobic sorbent) and octadecylsilane (C₁₈) was carried out to clean up and preconcentrate the insecticides. However, for soil samples, matrix solid-phase dispersion (MSPD) was applied with C₁₈ used as the dispersant. Good linearity, accuracy, and precision were obtained and the detection limits were in the range between 0.01 and 0.07 μg mL⁻¹ for river water and 0.17 and 0.37 μg g⁻¹ for soil samples. Recovery levels reached greater than 92% for all of the assayed neonicotinoids in river water samples with Strata-X. In soil matrices, the best recoveries (63-99%) were obtained with MSPD.
(Bio)chemical sensors are one of the most exciting fields in analytical chemistry today. The development of these analytical devices simplifies and miniaturizes the whole analytical process. Although the initial expectation of the massive incorporation of sensors in routine analytical work has been truncated to some extent, in many other cases analytical methods based on sensor technology have solved important analytical problems. Many research groups are working in this field world-wide, reporting interesting results so far. Modestly, Spanish researchers have contributed to these recent developments. In this review, we summarize the more representative achievements carried out for these groups. They cover a wide variety of sensors, including optical, electrochemical, piezoelectric or electro-mechanical devices, used for laboratory or field analyses. The capabilities to be used in different applied areas are also critically discussed.
Characterization of PEGs with average molecular masses of up to 2000 has been achieved using MEKC with UV detection. A rapid derivatization procedure with phenyl isocyanate using microwave radiation, in order to introduce chromophore groups in PEGs, has been developed involving a reaction time of 60 s. Different optimized conditions in accordance with the molecular weight have been studied to obtain the oligomer separation. The weight-average molecular mass the number-average molecular mass and the degree of polydispersity (molecular mass distribution) were calculated for the different PEGs obtaining similar results with those certified for standards. A good precision was obtained for characterizing the different oligomers. Ethylene glycol was used as the internal standard for the analysis of low-molecular-weight PEGs. The developed method was satisfactorily applied to the characterization of these polymers in several real samples, such as lubricant eye drops, toothpaste, tap water and eye make-up remover.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.