A Raman waveguide detector for liquid chromatography

A novel real-time liquid core Raman waveguide detector designed for liquid chromatographic applications is described. The Raman waveguide detector provides enhanced selectivity over typical high-performance liquid chromatography (HPLC) detectors. The waveguide detector also greatly improves the sensitivity of a typical Raman measurement without resorting to surface enhancement or resonance approaches and is compatible with the typical peak width volumes eluted by microbore and minibore HPLC (packed 1-2-mm-i.d. columns). Detection limit enhancements of over 1000-fold have been achieved for the measurement of alcohols in the aqueous phase with the Raman cell utilizing liquid core waveguide technology. The liquid core waveguides demonstrated in this study were constructed using Teflon AF 2400 tubing with a refractive index of 1.29. The low refractive index of the polymer material allowed HPLC separations with Raman detection to be performed with an aqueous mobile phase. A calibration curve for aqueous solutions of 2-propanol was generated and a limit of detection (LOD) of 2 ppm was determined. The Raman waveguide detector is demonstrated for the HPLC analysis of alcohol test mixtures, with LODs in the low-ppm range at the detector. By coupling the temporal separation achieved by HPLC with the vibrational information gleaned from Raman detection, an information-rich multivariate data matrix is obtained that can be deconvoluted to provide chemical speciation even when the HPLC resolution is poor. In this paper, we will discuss the physical and optical design of the Raman waveguide detector and the demonstration of the detector for HPLC detection.     

A blind-deconvolution approach for chromatographic andspectroscopic peak restoration

In this paper a blind deconvolution method is proposed which allows the deconvolution of broad-band signals that pass through a low-pass device by combining homomorphic deconvolution and Van Cittert iterative deconvolution techniques. The method has been successfully applied on both synthetic and experimental data obtained from chromatography and deep level transient spectroscopy (DLTS)     

Application of rhodamine 800 for reversed phase liquid chromatographic detection using visible diode laser-induced fluorescence

This study reports the application of rhodamine 800, a far-red dye, suitable for excitation using visible diode laser-induced fluorescence (VDLIF) detection. A reagent synthesized from rhodamine 800 was evaluated as a precolumn reagent for derivatization with amino-containing analytes. The derivative of this reagent with primary amine analytes showed a loss of fluorescence. Rhodamine 800 was then applied as a mobile phase additive in the indirect mode for quantitation of valproic acid in plasma using reversed phase HPLC in combination with VDLIF detection. A visible diode laser (output power 8.50 mW) temperature-tuned to oscillate at 674.70 nm was used as a light source for a laboratory constructed HPLC fluorescence detector. A liquid/liquid extraction procedure was applied to human blank plasma. The selectivity of this method was validated by demonstration of a lack of interfering peaks in extracts of plasma (n = 3 sources). A calibration curve for valproic acid between 40 and 200 μg/mL was shown to be linear (r = 0.9932). The recoveries of valproic acid at concentrations of 50 and 100 μg/mL were evaluated and determined to be 73 and 72%, respectively. The precision and accuracy (n = 5) of the assay was within 7.0% RSD and 8.0% difference from the spiked concentration, respectively. The limits of detection (S/N = 3) for extracted and unextracted valproic acid were 15.0 and 11.54 μg/mL, respectively. The theoretical (C(lim)) and practical (C(det)) limits of detection in the detector flow cell for unextracted valproic acid at a S/N = 1 were found to be within 15%.     

High-performance liquid chromatography interfaced with fluorescence line-narrowing spectroscopy for on-line analysis

We have demonstrated, for the first time, that high-performance liquid chromatography (HPLC) can be interfaced with fluorescence line-narrowing spectroscopy (FLNS) for on-line identification and characterization of analytes. Interfacing centered primarily on the design and construction of a novel liquid helium cryostat that accommodates variable-sized quartz tubes/capillaries suitable for HPLC as well as capillary electrophoresis/electrochromatography. In addition to the high spectral resolution afforded by FLNS, analyzing the separated components at 4.2 K minimizes photodegradation from the excitation source and provides indefinite detection times for signal averaging. The proof-of-principle for the HPLC-FLNS system is first demonstrated with a mixture of four structurally similar polycyclic aromatic hydrocarbons and then applied to the analysis of DNA adducts from mouse skin exposed to the carcinogen dibenzo[a,l]pyrene. With femtomole detection limits, HPLC-FLNS can be used for real-world analyses of complex mixtures.     

A computer system for analysis of chromatographic data

Marshall, R.J., Bleasby, A.J., Turner, R. and Cooper, E.H., 1987. A computer system for analysis of chromatographic data. Chemometrics and Intelligent Laboratory Systems, 1: 285–295.An interactive computer program (CHAS) for chromatogram processing is described. CHAS is a FORTRAN program which has three basic functions: (a) for data management, (b) for graphical display, and (c) for chromatogram analysis. The program is designed to run off-line by accessing data from a library of chromatograms. Various types of graphical displays are available and its analytical procedures incorporate new algorithms to detect chromatogram peaks, to remove baseline drift and to compute similarities between chromatograms. We present some illustrative uses of the program for data generated by high-pressure liquid chromatography.     

A DEA-based approach for allocation of emission reduction tasks

Rapid economic growth has led to increasing pollution emission, leading governments to require emission reductions by specific amounts. The allocation of specific emission reduction tasks has become a significant issue and has drawn the attention of academia. Data envelopment analysis (DEA) has been extended to construct the allocation of emission reduction tasks model. These previous DEA-based approaches have strong assumptions about individual enterprise production. In this paper, we propose a new method to accurately assess the production, using each enterprise’s previously observed production to construct its own production technology plan. With emission permits decreased, the enterprise can have new production strategy based on its own technology. Assuming emission permits can be freely bought and sold, we show how each enterprise can determine the optimal amount of emission allowance that should be used for production, which may leave some allowance to be sold for extra profit or may require the purchase of permits from other firms. Considering the limitation on the total allowance from emission permits, we introduce the concept of satisfaction degree and use it in maximising the minimum enterprise satisfaction degree. Last, a numerical example is presented and an empirical application is given to verify the proposed approach.     

Temperature pulsing for controlling chromatographic resolution in capillary liquid chromatography

In this study we introduce the implementation of rapid temperature pulses for selectivity tuning in capillary liquid chromatography. Short temperature pulses improved resolution in discrete sections of chromatograms, demonstrated for ion-exchange chromatography (IC) and hydrophilic interaction chromatography (HILIC) modes. Using a resistively heated column module capable of accurate and rapid temperature changes, this concept is first illustrated with separations of small anions by IC using a packed capillary column as well as a series of nucleobases and nucleosides by HILIC using a silica monolithic column with zwitterionic functionality (ZIC-HILIC). Both positive (increasing temperature) and negative temperature pulses are demonstrated to produce significant changes in selectivity and are useful approaches for improving resolution between coeluted compounds. The approach was shown to be reproducible over a large number of replicates. Finally, the use of temperature gradients as well as other complex temperature profiles was also examined for both IC and HILIC separations.     

Confocal Raman microscopy of protein adsorbed in chromatographic particles

Confocal Raman microscopy is a nondestructive analytical technique that combines the chemical information from vibrational spectroscopy with the spatial resolution of confocal microscopy. It was applied, for the first time, to measure conformation and distribution of protein adsorbed in wetted chromatographic particles. Monoclonal antibody was loaded into the Fractogel EMD SO(3) (M) cation exchanger at 2 mS/cm or 10 mS/cm. Amide I and III frequencies in the Raman spectrum of the adsorbed protein suggest that there are no detectable changes of the original β-sheet conformation in the chromatographic particles. Protein depth profile measurements indicate that, when the conductivity is increased from 2 mS/cm to 10 mS/cm, there is a change in mass transport mechanism for protein adsorption, from the shrinking-core model to the homogeneous-diffusion model. In this study, the use of confocal Raman microscopy to measure protein distribution in chromatographic particles fundamentally agrees with previous confocal laser scanning microscopic investigations, but confocal Raman spectroscopy enjoys additional advantages: use of unlabeled protein to eliminate fluorescent labeling, ability for characterization of protein secondary structure, and ability for spectral normalization to provide a nondestructive experimental approach to correct light attenuation effects caused by refractive index (RI) mismatching in semiopaque chromatographic particles.     

Exciplex fluorescence imaging for liquid mixing studies

A diagnostic technique based on excited-state complex (exciplex) fluorescence is reported for visualization of diffusion layers formed between mixing liquids. High-spatial-resolution instantaneous images and time-sequenced images are presented. Single drops and jets are visualized mixing in a liquid pool. Time-resolved images of exciplex fluorescence have also been obtained that provide information on the duration of the mixing event and the length and time scales of the diffusion process. The results demonstrate that the technique has the potential to provide new information concerning the physics of liquid mixing processes.     

A combined molecular dynamics and Raman spectroscopy approach for designing ice-metal interfaces

Summary Understanding the structure and interatomic interactions of an ice-metal interface plays a fundamental role in the design of deicing coatings. This is demonstrated by a novel approach, combining vibrational results from laser Raman spectroscopy with molecular dynamics simulations to obtain insights into icing on solids which, in turn, lead to design criteria for minimizing adhesion. An atomistic model of ice-copper interaction is constructed based on electronic structure calculations and used to show that reasonable molecular geometry and binding energy at the interface can be obtained. Through molecular dynamics simulations we find that the ice layer adjacent to the copper surface is structurally more disordered than the layers further away, a result which is verified by the Raman spectra of vibrational frequencies. The primary adhesive bond is made by the adsorption of oxygen atoms at the lattice sites of the metal substrate. The information obtained by Raman spectroscopy and molecular dynamics is then exploited to arrive at specific recommendations for designing polymeric deicing coatings and materials.     

Automated method for subtraction of fluorescence from biological Raman spectra

One of the challenges of using Raman spectroscopy for biological applications is the inherent fluorescence generated by many biological molecules that underlies the measured spectra. This fluorescence can sometimes be several orders of magnitude more intense than the weak Raman scatter, and its presence must be minimized in order to resolve and analyze the Raman spectrum. Several techniques involving hardware and software have been devised for this purpose; these include the use of wavelength shifting, time gating, frequency-domain filtering, first- and second-order derivatives, and simple curve fitting of the broadband variation with a high-order polynomial. Of these, polynomial fitting has been found to be a simple but effective method. However, this technique typically requires user intervention and thus is time consuming and prone to variability. An automated method for fluorescence subtraction, based on a modification to least-squares polynomial curve fitting, is described. Results indicate that the presented automated method is proficient in fluorescence subtraction, repeatability, and in retention of Raman spectral lineshapes.     

Coherent-interface-induced strain in large lattice-mismatched materials:A new approach for modeling Raman shift

Strain engineering as one of the most powerful techniques for tuning optical and electronic properties of Ⅲ-nitrides requires reliable methods for strain invest...     

A motion-based approach to abdominal clutter reduction

In ultrasound images, clutter is a noise artifact most easily observed in anechoic or hypoechoic regions. It appears as diffuse echoes overlying anatomical structures of diagnostic importance, obscuring tissue borders and reducing image contrast. A novel clutter reduction method for abdominal images is proposed, wherein the abdominal wall is displaced during successive-frame image acquisitions. A region of clutter distal to the abdominal wall was observed to move with the abdominal wall, and finite impulse response (FIR) and blind source separation (BSS) motion filters were implemented to reduce this clutter. The proposed clutter reduction method was tested in simulated and phantom data and applied to fundamental and harmonic in vivo bladder and liver images from 2 volunteers. Results show clutter reductions ranging from 0 to 18 dB in FIR-filtered images and 9 to 27 dB in BSS-filtered images. The contrast-to-noise ratio was improved by 21 to 68% and 44 to 108% in FIR- and BSS-filtered images, respectively. Improvements in contrast ranged from 4 to 12 dB. The method shows promise for reducing clutter in other abdominal images.     

Multiresidue method for pesticides in drinking water using a graphitized carbon black cartridge extraction and liquid chromatographic analysis

A general liquid-solid extraction procedure for the isolation of pesticides from groundwater and drinking water for high-performance liquid chromatography (HPLC) is presented. This simple and rapid procedure involved passing a 2-L sample through a 250-mg graphitized carbon black (Carbopack B) cartridge at a flow rate of 150-160 mL/min. By taking advantage of the presence of positively charged active centers on the Carbopack B surface, a stepwise elution system allowed the complete separation of base-neutral pesticides from acidic ones. After partial solvent removal, the components in the two fractions were separated and quantified by gradient elution, reversed-phase HPLC with ultraviolet (UV) detection. The performance of the Carbopack cartridge was compared with that of a 500-mg C-18 bonded silica cartridge. With the Carbopack cartridge, the grand mean measurement accuracy of the 35 pesticides considered was 95%. With the C-18 cartridge, the grand mean measurement accuracy of the analytes was 76%. Compared to the C-18 cartridge, additional advantages of using a Carbopack cartridge are that the extraction procedure is about 7 times shorter, no pH adjustment of the environmental sample is necessary for trapping acidic compounds, and one cartridge instead of two suffices to extract base-neutral and acidic pesticides, making the Carbopack cartridge more adaptable than the C-18 one for field use. The detection limits by this method of all the pesticides considered were between 0.003 and 0.07 micrograms/L.     

Simultaneous high-performance liquid chromatographic analysis for famotidine, ranitidine HCl, cimetidine, and nizatidine in commercial products

A high-performance liquid chromatographic (HPLC) procedure for the simultaneous determination of famotidine (FMT), ranitidine HCl (RNT), cimetidine (CMT), and nizatidine (NZT) was developed using a two-level, full-factorial design with three variables (volume of methanol, percentage of triethylamine, and concentration of phosphate buffer) to select an acceptable mobile phase. A column (15cm × 4.6 mm ID) of Inertsil ODS-2 (5 μm) was used, and 0.04M aqueous sodium dihydrogen phosphate/acetonitrile/methanol/TEA at a proportion of 345/20/35/0.7 (v/v/v/v) was the selected mobile phase (1 ml/min). The detection wavelength was set at 230 nm, and procaine HCl was used as the internal standard. Precision and linearity of the method were assessed. None of the commercial samples was found to be outside the compendial limits of 90.0% to 110.0% of the claim amount.     

A reversed-phase high-performance liquid chromatographic method for the determination of soya bean proteins in bovine milks

A reversed-phase high-performance liquid chromatographic method was designed for the quantitation of soya bean proteins in bovine milks. The method consisted of a linear binary gradient, acetonitrile-water-0.1% trifluoroacetic acid, at a flow rate of 1 mL/min and a temperature of 50 degrees C which resulted in a separation time for soya bean proteins of 11 min. Calibration by the external standard method using a soya bean protein isolate as standard was employed, and the method was validated by evaluating precision, accuracy, and robustness. This method was shown to be useful for the analysis of soya bean proteins in bovine milks spiked with soya bean protein isolate; soya bean protein concentrations of approximately 13 microg/g of bovine milk could be detected by using the optimized method. The results obtained for some of the bovine milks were compared with those obtained by the method of standard additions.