Determination of Dextromethorphan and its Metabolite Dextrorphan in Human Hair by Gas Chromatography–Mass Spectrometry

An analytical method has been developed for determination of dextromethorphan (DMP) and dextrorphan (DRP) in human hair by gas chromatography–mass spectrometry (GC–MS). Hair samples (30 mg) were washed with distilled water and acetone and cut into small fragments (<1 mm) before extraction with methanol. The extracts were evaporated to dryness and then derivatized by use of BSTFA containing 1% TMCS, for preparation of the trimethylsilyl (TMS) derivative of DRP. The extract (1 μL) was then injected into the GC–MS. Recoveries of DMP and DRP at 7.0 and 22.5 ng mg^?1 were in the range 90.6–97.2% with intra-assay and inter-assay precision of less than 5.7% and 4.7%, respectively. LOD and LOQ were, respectively, 0.67 and 2.13 ng mg^?1 for DMP and 0.14 and 0.47 ng mg^?1 for DRP. The responses were linear with correlation coefficients (r > 0.9995) for the drugs studied. The applicability of the method was proven by analysis of authentic hair samples.     

Determination of Fluoride in Water by Reversed-Phase High-Performance Liquid Chromatography using F−-La3+-Alizarin Complexone Ternary Complex

A method for the determination of fluoride by reversed-phase, high-performance liquid chromatography (RP-HPLC) is described. Fluoride, La³⁺ and alizarin complexone form F-La³⁺-alizarin complexone ternary complex, which is separated from the matrix on a RP, Ultrasphere C₁₈ column (250 × 4.6 mm, 5 μm) using methanol-water (19:81, v/v) mobile phase at 1.00 mL min^?1; detection at 568 nm. The calibration graph was linear from 1.0–150 ng mL^?1 for fluoride with a correlation coefficient: 0.9993 (n=6). The detection limit was 0.2 ng mL^?1. The method was successfully applied to the determination of fluoride in river and tap water. Recovery was: 94–102%, RSD in the range: 1.9 –3.6%.     

Performance of hollow-fiber flow field-flow fractionation in protein separation

Since hollow-fiber flow field-flow fractionation (HF FIFFF) utilizes a cylindrical channel made of a hollow-fiber membrane, which is inexpensive and simple in channel assembly and thus disposable, interests are increasing as a potential separation device in cells, proteins, and macromolecules. In this study, performance of HF FIFFF of proteins is described by examining the influence of flow rate conditions and length of fiber (polyacrylonitrile or PAN in this work) on sample recovery as well as experimental plate heights. The interfiber reproducibility in terms of separation time and recovery was also studied. Experiments showed that sample recovery was consistent regardless of the length of fiber when the effective field strength (equivalent to the mean flow velocity at the fiber wall) and the channel void time were adjusted to be equivalent for channels of various fiber lengths. This supported that the majority of sample loss in HF FIFFF separation of apoferritin and their aggregates may occur before the migration process. It is finally demonstrated that HF FIFFF can be applied for characterizing the reduction in Stokes' size of low density lipoproteins from blood plasma samples obtained from patients having coronary artery disease and from healthy donors.     

Calix[2]furano[2]pyrrole and related compounds as the neutral carrier in silver ion-selective electrode

The performance of calix[2]furano[2]pyrrole and related compounds used as neutral carriers for silver selective polymeric membrane electrode was investigated. The silver ion-selective electrode based on calix[2]furano[2]pyrroles gave a good Nernstian response of 57.1 mV per decade for silver ion in the activity range 1×10⁻⁶ to 1×10⁻² M. The present silver ion-selective electrode displayed very good selectivity for Ag⁺ ion against alkali and alkaline earth metal ions, NH₄⁺, and H⁺. In particular, the present Ag⁺-selective electrode exhibited very low responses towards Hg²⁺ and Pb²⁺ ions. The potentiometric selectivity coefficients of the silver ion-selective electrode exhibited a strong dependence on the solution pH. In particular, the response of the electrode to the Hg²⁺ activity was greatly diminished at pH 2.5 compared to that at pH 5.0. Overall, the performance of the present silver ion-selective electrode based on the ionophore, calix[2]furano[2]pyrrole, is very comparable to that of the electrode prepared with the commercially available neutral carrier in terms of slope, linear range, and detection limits.     

Influence of cell packing by centrifugation on 40-MHz ultrasound backscatter

High-frequency ultrasound (HFUS) signals backscattered from RBL-2H3 cell pellets prepared under different centrifugal forces were analyzed to investigate the packing effect of cell aggregates. The measurements were performed in a pulse-echo setup with a 40-MHz transducer. The changes of ultrasound signals from cell pellet in backscattered power, statistical parameter, and pellet thickness were monitored after centrifugation at between 100g and 1600g. Experimental results showed that the HFUS backscattered power from cell pellets was inversely proportional to centrifugal force and increased to a plateau within 1-2 h after centrifugation. The initial thickness of cell pellets decreased with higher centrifugal force, but the changes in thickness and time that took to reach a plateau increased at higher centrifugal force. The envelope statistics of backscattered signals with Nakagami distribution indicates that the centrifugal force and elapsed time after centrifugation affected the backscattering characteristics. The present study suggests that centrifugal force and data acquisition time after cell pellet formation should be considered in in vitro cell packing method with centrifugation to emulate the tissue in vivo.     

Biotin-avidin mediated competitive enzyme-linked immunosorbent assay to detect residues of tetracyclines in milk

Tetracycline (TC) antibiotics are widely used for prevention and control of disease because they inhibit the growth of bacteria. However, the presence of TC antibiotics residues in food causes harmful effects on consumer's health such as allergic reactions, liver damage and gastrointestinal disturbance, so that many countries have set MRLs (maximum residue levels). Therefore, it is necessary to detect tetracycline residues, to develop suitable analytical techniques to be used as routine screens and field detection.A new approach to the biotin-avidin mediated competitive ELISA is developed to determine tetracycline residues in milk. After optimization, the LOD and LOQ were 1.0 × 10^− 10 M (0.048 μg/L) and 1.0 × 10^− 9 M, respectively, and the working range from 3.16 × 10^− 10 M to 3.16 × 10^− 7 M toward TC in milk. No cross-reactivity was observed with the structurally similar compounds; chlortetracycline (13.7%), oxytetracycline (10%) and doxytetracycline (< 1%). Additionally percent recoveries of TC spiked in milk were quite satisfactory (∼ 90%). Comparing our results obtained in this work with others, it shows with the capability to detect TC ranging in MRLs (100 μg/L in milk) sufficiently with highly sensitivity in milk, and with simple pre-treatment. In addition, this method can apply to developing useful ELISA test kit for determination of TCs in milk.     

Improvement of ultrasound speckle image velocimetry using image enhancement techniques

Ultrasound-based techniques have been developed and widely used in noninvasive measurement of blood velocity. Speckle image velocimetry (SIV), which applies a cross-correlation algorithm to consecutive B-mode images of blood flow has often been employed owing to its better spatial resolution compared with conventional Doppler-based measurement techniques. The SIV technique utilizes speckles backscattered from red blood cell (RBC) aggregates as flow tracers. Hence, the intensity and size of such speckles are highly dependent on hemodynamic conditions. The grayscale intensity of speckle images varies along the radial direction of blood vessels because of the shear rate dependence of RBC aggregation. This inhomogeneous distribution of echo speckles decreases the signal-to-noise ratio (SNR) of a cross-correlation analysis and produces spurious results. In the present study, image-enhancement techniques such as contrast-limited adaptive histogram equalization (CLAHE), min/max technique, and subtraction of background image (SB) method were applied to speckle images to achieve a more accurate SIV measurement. A mechanical sector ultrasound scanner was used to obtain ultrasound speckle images from rat blood under steady and pulsatile flows. The effects of the image-enhancement techniques on SIV analysis were evaluated by comparing image intensities, velocities, and cross-correlation maps. The velocity profiles and wall shear rate (WSR) obtained from RBC suspension images were compared with the analytical solution for validation. In addition, the image-enhancement techniques were applied to in vivo measurement of blood flow in human vein. The experimental results of both in vitro and in vivo SIV measurements show that the intensity gradient in heterogeneous speckles has substantial influence on the cross-correlation analysis. The image-enhancement techniques used in this study can minimize errors encountered in ultrasound SIV measurement in which RBCs are used as flow tracers instead of exogenous contrast agents.     

Asymmetric radial expansion and contraction of rat carotid artery observed using a high-resolution ultrasound imaging system

The geometry of carotid artery bifurcation is of high clinical interest because it determines the characteristics of blood flow that is closely related to the formation and development of atherosclerotic plaque. However, information on the dynamic changes in the vessel wall of carotid artery bifurcation during a pulsatile cycle is limited. This pilot study investigated the cyclic changes in carotid artery geometry caused by blood flow pulsation in rats. A high-resolution ultrasound imaging system with a broadband scanhead centered at 40 MHz was used to obtain longitudinal images of the rat carotid artery. A high frame rate retrospective B-scan imaging technique based on the use of electrocardiogram to trigger signal acquisition was used to examine precisely the fast arterial wall motion. Two-dimensional geometry data obtained from nine rats showed that the rat carotid artery asymmetrically contracts and dilates during each cardiac cycle. Systolic/diastolic vessel diameters near the upstream and downstream regions from the bifurcation were 0.976 ± 0.011/0.825 ± 0.015 mm and 0.766 ± 0.015/0.650 ± 0.016 mm, respectively. Their posterior/anterior wall displacement ratios in the radial direction were 41.0 ± 14.9% and 2.9 ± 1.6%, respectively. These results indicate that in the vicinity of bifurcation, the carotid artery favorably expands to the anterior side during the systolic phase. This phenomenon was observed to be more prominent in the downstream region near the bifurcation. The cyclic variation pattern in wall movement varies depending on the measurement site, which shows different patterns at far upstream and downstream of the bifurcation. The asymmetric radial expansion and contraction of the rat carotid artery observed in this study may be useful in studying the hemodynamic etiology of cardiovascular diseases because the pulsatile changes in vessel geometry may affect the local hemodynamics that determines the spatial distribution of wall shear stress, one of important cardiovascular risk factors. Further systematic study is needed to clarify the effects of wall elasticity, branch angle and vessel diameter ratio on the asymmetric wall motion of carotid artery bifurcation.