Dispersion of multi-wall carbon nanotubes in polyhistidine: Characterization and analytical applications

We report for the first time the use of polyhistidine (Polyhis) to efficiently disperse multiwall carbon nanotubes (MWCNTs). The optimum dispersion MWCNT–Polyhis was obtained by sonicating for 30 min 1.0 mg mL⁻¹ MWCNTs in 0.25 mg mL⁻¹ Polyhis solution prepared in 75:25 (v/v) ethanol/0.200 M acetate buffer solution pH 5.00. The dispersion was characterized by scanning electron microscopy, and by cyclic voltammetry and amperometry using ascorbic acid as redox marker. The modification of glassy carbon electrodes with MWCNT–Polyhis produces a drastic decrease in the overvoltage for the oxidation of ascorbic acid (580 mV) at variance with the response observed at glassy carbon electrodes modified just with Polyhis, where the charge transfer is more difficult due to the blocking effect of the polymer. The reproducibility for the sensitivities obtained after 10 successive calibration plots using the same surface was 6.3%. The MWCNT-modified glassy carbon electrode demonstrated to be highly stable since after 45 days storage at room temperature the response was 94.0% of the original. The glassy carbon electrode modified with MWCNT–Polyhis dispersion was successfully used to quantify dopamine or uric acid at nanomolar levels, even in the presence of large excess of ascorbic acid. Determinations of uric acid in human blood serum samples demonstrated a very good correlation with the value reported by Wienner laboratory.     

Determination of selenium in water and soil by hydride generation atomic absorption spectrometry using solid reagents

A hydride generation method for the determination of traces of selenium at ng mL⁻¹ concentration ranges has been introduced using a solid mixture of tartaric acid and sodium tetrahydroborate. Atomic absorption spectrometry (AAS) has been used as the detection system. Several parameters such as the ratio of tartaric acid to sodium tetrahydroborate, type and amount of acid, and the reaction temperature were optimized by using 640 ng mL⁻¹ (16 ng per 25 μL) of Se(IV) standard solution. The calibration curve was linear from 20 to 1200 ng mL⁻¹ (0.5-30 ng Se(IV) per 25 μL). The relative standard deviation (%R.S.D.) of the determination was 1.93% and the detection limit was 10.6 ng mL⁻¹ (265 pg per 25 μL) of Se(IV). The reliability of the method was checked using different types of environmental samples, such as several types of water, a sample of soil and also in a kind of calcium phosphate sample by standard addition method. For conversion of Se(VI) present in real samples to Se(IV), l-cysteine was added to NaBH₄ and tartaric acid mixture. The results showed good agreement between this method and other hydride generation techniques.     

Development of a chemiluminescent immunosensor for chloramphenicol

A direct competitive chemiluminescent immunosensor system that exploits the competition between chloramphenicol (CAP) as an analyte and CAP-horseradish peroxidase conjugate as a tracer for binding to an anti-CAP antibody on a solid support was devised by installing a flow-through cell which was connected to an injector and a peristaltic pump inside a dark box, followed by positioning a photomultiplier tube as light detector in front of it. The anti-CAP antibody was immobilized onto positively charged Biodyne B membrane pieces by a dipping procedure. The operating conditions for the immunosensor were selected with respect to substrate composition (0.25, 13.3 and 0.66 mM for luminol, H₂O₂ and p-iodophenol, respectively), injection volume of the substrate solution (200 μL) and the concentrations of antibody for immobilization (0.10 mg mL⁻¹) and tracer (0.030 mg mL⁻¹). At these conditions, sensor response according to analyte concentration was well fitted to a linear equation when plotted in semi-logarithmic scale, with the limit of detection for CAP of 10⁻⁸ M. By using the immunosensor, CAP measurement in the model samples prepared from five food materials was conducted.     

Development and validation of an HPLC-PDA method for the determination of myrsinoic acid B in the extracts of Rapanea ferruginea Mez

New, simple, rapid and precise HPLC-PDA method has been developed and validated for quantification of biomarker myrsinoic acid B in stem bark extracts of Rapanea ferruginea Mez. The method employs a Phenomenex C18 column (250 mm × 4.6 mm I.D., 5 μm) with acetonitrile:methanol:water (pH 2.6 with phosphoric acid) at 48:30:22 as mobile phase, at a flow rate of 0.7 mL min⁻¹ and photo diode array (PDA) detection at 270 nm. The validation data show that the method is specific, accurate, precise and robust. The method was linear, over a range of 5-100.0 μg mL⁻¹, with a limit of detection of 0.369 μg mL⁻¹ and limit of quantification of 1.233 μg mL⁻¹. The method has also shown consistent recoveries (average of 101.3% and 0.12% RSD) of the biomarker, with low intra and inter-day relative standard deviation (1.26% and 1.62%, respectively). The evaluated hydroethanolic extract and dry extract presented MAB values of 63.53 and 36.07 mg g⁻¹, respectively.     

Electrospun nanofiber based colorimetric probe for rapid detection of Fe in water

An imidazole derivative, 2-(2′-pyridyl)imidazole (PIMH), was developed as a colorimetric probe for the qualitative analysis of Fe²⁺ in aqueous solution. PIMH was then used to post-functionalize poly(vinylbenzyl chloride) (PVBC) nanofibers after electrospinning so as to afford a solid state colorimetric probe. Upon treatment with Fe²⁺ the probe displayed a distinctive color change both in liquid and solid platforms. The linear dynamic range for the colorimetric determination of Fe²⁺ was 0.0988–3.5 μg mL⁻¹. The ligand showed a high chromogenic selectivity for Fe²⁺ over other cations with a detection limit of 0.102 μg mL⁻¹ in solution (lower than the WHO drinking water guideline limit of 2 mg L⁻¹), and 2 μg mL⁻¹ in the solid state. The concentration of Fe²⁺ in a certified reference material (Iron, Ferrous, 1072) was found to be 2.39 ± 0.01 mg L⁻¹, which was comparable with the certified value of 2.44 ± 0.12 mg L⁻¹. Application of the probe to real samples spiked with Fe²⁺ achieved recoveries of over 97% confirming accuracy of the method and its potential for on-site monitoring.     

Disposable biosensor based on cathodic electrochemiluminescence of tris(2,2-bipyridine)ruthenium(II) for uric acid determination

A new method for uric acid (UA) determination based on the quenching of the cathodic ECL of the tris(2,2-bipyridine)ruthenium(II)–uricase system is described. The biosensor is based on a double-layer design containing first tris(2,2-bipyridine)ruthenium(II) (Ru(bpy)₃²⁺) electrochemically immobilized on graphite screen-printed cells and uricase in chitosan as a second layer. The uric acid biosensing is based on the ECL quenching produced by uric acid over the cathodic ECL caused by immobilized Ru(bpy)₃²⁺ in the presence of uricase. The use of a −1.1 V pulse for 1 s with a dwelling time of 10 s makes it possible to estimate the initial enzymatic rate, which is used as the analytical signal. The Stern–Volmer type calibration function shows a dynamic range from 1.0 × 10⁻⁵ to 1.0 × 10⁻³ M with a limit of detection of 3.1 × 10⁻⁶ M and an accuracy of 13.6% (1.0 × 10⁻⁴ M, n = 5) as relative standard deviation. Satisfactory results were obtained for urine samples, creating an affordable alternative for uric acid determination.     

ZnS quantum dots derived a reagentless uric acid biosensor

A reagentless amperometric uric acid biosensor based on zinc sulfide (ZnS) quantum dots (QDs) was firstly developed. It could detect uric acid without the presence of an electron mediator. The carboxyl group functionalized ZnS QDs were synthesized, and they were soluble biocompatible and conductive. ZnS QDs conjugates could provide increased enzyme binding sites, which may result in higher enzyme loading. Thus, the proposed uricase/ZnS QDs/l-cys biosensor exhibited higher amperometric response compared to the one without QDs (uricase/l-cys biosensor). In addition, there was little AA interference. It showed a linear dependence on the uric acid concentration ranging from 5.0 × 10⁻⁶ to 2.0 × 10⁻³ mol L⁻¹ with a detection limit of 2.0 × 10⁻⁶ mol L⁻¹ at 3σ.     

Analysis of human tear fluid by Raman spectroscopy

Tear fluid is a complex aqueous solution containing proteins, metabolites, electrolytes and lipids. This study uses Raman spectroscopy to analyse the composition of human tear fluid from three healthy volunteers. Two different methods are used to obtain Raman spectra from the 3 μL tear samples: (i) solution-phase Raman spectroscopy, and (ii) drop coating deposition Raman spectroscopy (DCDRS). Tear samples were either basal fluid, or yawn reflex secreted fluid. Calibration of the solution technique with standard protein solutions (5-15 mg mL⁻¹) showed that this method could predict the protein concentration (cross-validation) with an error of less than 1 mg mL⁻¹. The Raman signals from the tear fluid were very weak but signals due to protein and urea were clearly observable in all samples. The drop coating deposition technique was shown to produce very high signal-to-noise spectra for relatively short acquisition times, and small sample volumes. Raman point mapping combined with principal components analysis showed that the protein, urea, bicarbonate and lipid could all be detected in the tear samples and that the distribution of these components was inhomogeneous. Their position within the drying pattern was shown to depend on their relative solubilities. The results of this study suggest that solution Raman measurements may be calibrated to give the total tear protein concentration and DCDRS could be used to give a fingerprint of the tear protein (and lipid) composition.     

Sensitive determination of fluoride in biological samples by gas chromatography–mass spectrometry after derivatization with 2-(bromomethyl)naphthalene

A gas chromatography–mass spectrometric method was developed in this study in order to determine fluoride in plasma and urine after derivatization with 2-(bromomethyl)naphthalene. 2-Fluoronaphthalene was chosen as the internal standard. The derivatization of fluoride was performed in the biological sample and the best reaction conditions (10.0 mg mL⁻¹ of 2-(bromomethyl)naphthalene, 1.0 mg mL⁻¹ of 15-crown-5-ether as a phase transfer catalyst, pH of 7.0, reaction temperature of 70 °C, and heating time of 70 min) were established. The organic derivative was extracted with dichloromethane and then measured by a gas chromatography–mass spectrometry. Under the established condition, the detection limits were 11 μg L⁻¹ and 7 μg L⁻¹ by using 0.2 mL of plasma or urine, respectively. The accuracy was in a range of 100.8–107.6%, and the precision of the assay was less than 4.3% in plasma or urine. Fluoride was detected in a concentration range of 0.12–0.53 mg L⁻¹ in six urine samples after intake of natural mineral water containing 0.7 mg L⁻¹ of fluoride.     

Biosensors for determination of glucose with glucose oxidase immobilized on an eggshell membrane

A glucose biosensor using an enzyme-immobilized eggshell membrane and oxygen electrode for glucose determination has been fabricated. Glucose oxidase was covalently immobilized on an eggshell membrane with glutaraldehyde as a cross-linking agent. The glucose biosensor was fabricated by positioning the enzyme-immobilized eggshell membrane on the surface of a dissolved oxygen sensor. The detection scheme was based on the depletion of dissolved oxygen content upon exposure to glucose solution and the decrease in the oxygen level was monitored and related to the glucose concentration. The effect of glutaraldehyde concentration, pH, phosphate buffer concentration and temperature on the response of the glucose biosensor has been studied in detail. Common matrix interferents such as ethanol, d-fructose, citric acid, sodium benzoate, sucrose and l-ascorbic acid did not give significant interference. The resulting sensor exhibited a fast response (100 s), high sensitivity (8.3409 mg L⁻¹ oxygen depletion/mmol L⁻¹ glucose) and good storage stability (85.2% of its initial sensitivity after 4 months). The linear response is 1.0×10⁻⁵ to 1.3×10⁻³ mol L⁻¹ glucose. The glucose content in real samples such as commercial glucose injection preparations and wines was determined, and the results were comparable to the values obtained from a commercial glucose assay kit based on a spectrophotometric method.     

Electro membrane extraction of sodium diclofenac as an acidic compound from wastewater,urine, bovine milk,and plasma samples and quantification by high-performance liquid chromatography

Electro membrane extraction (EME) as a new microextraction method was applied for extraction of sodium diclofenac (SDF) as an acidic compound from wastewater, urine, bovine milk and plasma samples. Under applied potential of 20 V during the extraction, SDF migrated from a 2.1 mL of sample solution (1 mM NaOH), through a supported liquid membrane (SLM), into a 30 μL acceptor solution (10 mM NaOH), exist inside the lumen of the hollow fiber. The negative electrode was placed in the donor solution, and the positive electrode was placed in the acceptor solution. 1-octanol was immobilized in the pores of a porous hollow fiber of polypropylene as SLM. Then the extract was analyzed by means of high-performance liquid chromatography (HPLC) with UV-detection for quantification of SDF. Best results were obtained using a phosphate running electrolyte (10 mM, pH 2.5). The ranges of quantitation for different samples were 8–500 ng mL⁻¹. Intra- and inter-day RSDs were less than 14.5%. Under the optimized conditions, the preconcentration factors were between 31 and 66 and also the limit of detections (LODs) ranged from 2.7 ng mL⁻¹ to 5 ng mL⁻¹ in different samples. This procedure was applied to determine SDF in wastewater, bovine milk, urine and plasma samples (spiked and real samples). Extraction recoveries for different samples were between 44–95% after 5 min of extraction.     

Development of enzyme-linked immunosorbent assays for the detection of deacetoxycephalosporin C and isopenicillin N synthase activity

Although there are a number of existing assays for monitoring the activity of both isopenicillin N synthase (IPNS) and deacetoxycephalosporin C synthase (DAOCS), none have demonstrated the qualities required for screening a mutant library. Hence, enzyme-linked immunosorbent assays (ELISAs) for IPNS and DAOCS were developed based on the detection of the catalytic turnover products isopenicillin N and cephalexin/phenylacetyl-7-aminodeacetoxycephalosporanic acid (G-7-ADCA), respectively. These assays are relatively fast compared to existing assays, such as the hole-plate bioassay, and are amenable with high-throughput screening. Both the IPNS and DAOCS-ELISAs were optimised for use with crude protein extracts rather than purified protein, thereby eliminating any additional time required for purification. The ELISA developed for the detection of cephalexin had an IC₅₀ value of 154 ± 9 ng mL⁻¹ and LOD of 7.2 ± 2.2 ng mL⁻¹ under conditions required for the assay. Good recoveries and correlation was observed for spiked samples when the concentration of crude protein was kept below 1 mg mL⁻¹. The DAOCS-ELISA was found to have increased sensitivity compared to the hole-plate bioassay (10.3 μg mL⁻¹). The IPNS-ELISA did not significantly increase the sensitivity (approximately 5 μg mL⁻¹) compared to that of the hole-plate bioassay (16 μg mL⁻¹) for isopenicillin N. The minimum amount of crude protein extract required for producing detectable amounts of product for both assays was below 0.5% of the maximum amount of protein that the assay could contain without any effect on the ELISA. This suggests that when screening a mutant library, mutants producing low amounts of the product could still be detected using these assays.     

Flame atomic absorption spectrometry for the determination of trace amount of rhodium after separation and preconcentration onto modified multiwalled carbon nanotubes as a new solid sorbent

The present article reports on the application of modified multiwalled carbon nanotubes (MMWCNTs) as a new, easily prepared and stable solid sorbent for the preconcentration of trace rhodium ion in aqueous solution. Rhodium ions were complexed with 1-(2-pyridylazo)-2-naphthol (PAN) in the pH range of 3.2-4.7 and then the formed Rh-PAN complex was adsorbed on the oxidized MWCNTs. The adsorbed complex was eluted from MWCNTs sorbent with 5.0 mL of N,N-dimethylformamide (DMF). The rhodium in eluted solution was determined by flame atomic absorption spectrometry (FAAS). Linear range for the determination of rhodium was maintained between 0.16 ng mL⁻¹ and 25.0 μg mL⁻¹ in initial solution. Relative standard deviation for the 10 replicated determination of 4.0 μg mL⁻¹ of rhodium was ±0.97%. Detection limit was 0.010 ng mL⁻¹ in initial solution (3S_bl, n = 10) and preconcentration factor was 120. Sensitivity for 1% absorbance of rhodium (III) was 0.112 μg mL⁻¹. The sorption capacity of oxidized MWCNTs for Rh (III) was 6.6 mg g⁻¹. The effects of the experimental parameters, including the sample pH, flow rates of sample and eluent solution, eluent type, breakthrough volume and interference ions were studied for the preconcentration of Rh³⁺. The proposed method was successfully applied to the extraction and determination of rhodium in different samples.     

Determination of atrazine and simazine in water samples by high-performance liquid chromatography after preconcentration with heat-treated diatomaceous earth

A sensitive and selective column adsorption method is proposed for the preconcentration and determination of atrazine and simazine. Atrazine and simazine were preconcentrated on heat-treated diatomaceous earth as an adsorbent and then determined by high-performance liquid chromatography (HPLC). Several parameters on the recoveries of the analytes were investigated. The experimental results showed that it was possible to obtain quantitative analysis when the solution pH was 2 using 100 mL of validation solution containing 1.5 μg of triazines and 5 mL of ethanol as an eluent. Recoveries of atrazine and simazine were 95.7 ± 4.2% and 75.0 ± 1.9% with a relative standard deviation for seven determinations of 4.7% and 2.7% under optimum conditions. The maximum preconcentration factor was 100 for triazines when 500 mL of sample solution volume was used. The linear ranges of calibration curves for atrazine and simazine were 1-150 ng mL⁻¹ and 1-300 ng mL⁻¹, respectively, with correlation coefficients of 0.999 and the detection limits (3Signal-to-Noise) were 0.24 ng mL⁻¹ and 0.21 ng mL⁻¹ for atrazine and simazine. The capacity of the adsorbent was also examined and found to be 0.8 mg g⁻¹ and 1.3 mg g⁻¹ for atrazine and simazine, respectively. The proposed method was successfully applied to the determination of triazines in river water and tap water samples with high precision and accuracy.     

Development of a simple method for the determination of nitrite and nitrate in groundwater by high-resolution continuum source electrothermal molecular absorption spectrometry

In this work, it was developed a method for the determination of nitrite and nitrate in groundwater by high-resolution continuum source electrothermal molecular absorption spectrometry of NO produced by thermal decomposition of nitrate in a graphite furnace. The NO line at 215.360 nm was used for all analytical measurements and the signal obtained by integrated absorbance of three pixels. A volume of 20 μL of standard solution or groundwater sample was injected into graphite furnace and 5 μL of a 1% (m/v) Ca solution was co-injected as chemical modifier. The pyrolisis and vaporization temperatures established were of 150 and 1300 °C, respectively. Under these conditions, it was observed a difference of thermal stability among the two nitrogen species in the presence of hydrochloric acid co-injected. While that the nitrite signal was totally suppressed, nitrate signal remained nearly stable. This way, nitrogen can be quantified only as nitrate. The addition of hydrogen peroxide provided the oxidation of nitrite to nitrate, which allowed the total quantification of the species and nitrite obtained by difference. A volume of 5 μL of 0.3% (v/v) hydrochloric acid was co-injected for the elimination of nitrite, whereas that hydrogen peroxide in the concentration of 0.75% (v/v) was added to samples or standards for the oxidation of nitrite to nitrate. Analytical curve was established using standard solution of nitrate. The method described has limits of detection and quantification of 0.10 and 0.33 μg mL⁻¹ of nitrogen, respectively. The precision, estimated as relative standard deviation (RSD), was of 7.5 and 3.8% (n = 10) for groundwater samples containing nitrate–N concentrations of 1.9 and 15.2 μg mL⁻¹, respectively. The proposed method was applied to the analysis of 10 groundwater samples and the results were compared with those obtained by ion chromatography method. In all samples analyzed, the concentration of nitrite–N was always below of the limit of quantification of both the methods. The concentrations of nitrate–N varied from 0.58 to 15.5 μg mL⁻¹. No significant difference it was observed between the results obtained by both methods for nitrate–N, at the 95% confidence level.     

Simultaneous determination of some food additives in soft drinks and other liquid foods by flow injection on-line dialysis coupled to high performance liquid chromatography

Flow injection on-line dialysis was developed for sample pretreatment prior to the simultaneous determination of some food additives by high performance liquid chromatography (FID-HPLC). A liquid sample or mixed standard solution (900 μL) was injected into a donor stream (5%, w/v, sucrose) of FID system and was pushed further through a dialysis cell, while an acceptor solution (0.025 mol L⁻¹ phosphate buffer, pH 3.75) was held in the opposite side of the dialysis membrane. The dialysate was then flowed to an injection loop of the HPLC valve, where it was further injected into the HPLC system and analyzed under isocratic reverse-phase HPLC conditions and UV detection (230 nm). The order of elution of five food additives was acesulfame-K, saccharin, caffeine, benzoic acid and sorbic acid, respectively, with the analysis time of 14 min. On-line dialysis and HPLC analysis could be performed in parallel, providing sample throughput of 4.3 h⁻¹. Dialysis efficiencies of five food additives were in ranges of 5-11%. Linear calibration graphs were in ranges of 10-100 mg L⁻¹ for acesulfame-K and saccharin, 10-250 mg L⁻¹ for benzoic acid and 10-500 mg L⁻¹ for caffeine and sorbic acid. Good precisions (RSD < 5%) for all the additives were obtained. The proposed system was applied to soft drink and other liquid food samples. Acceptable percentage recoveries could be obtained by appropriate dilution of the sample before injecting into the system. The developed system has advantages of high degrees of automation for sample pretreatment, i.e., on-line sample separation and dilution and low consumption of chemicals and materials.     

Simultaneous detection of uric acid and glucose on a dual-enzyme chip using scanning electrochemical microscopy/scanning chemiluminescence microscopy

Scanning electrochemical microscopy (SECM) and scanning chemiluminescence microscopy (SCLM) were used for imaging an enzyme chip with spatially-addressed spots for glucose oxidase (GOD) and uricase microspots. For the SECM imaging, hydrogen peroxide generated from the GOD and/or uricase spots was directly oxidized at the tip microelectrode in a solution containing glucose and/or uric acid (electrochemical (EC) detection). For the SCLM imaging, a tapered glass capillary (i.d. of 1∼2 μm) filled with luminol and horseradish peroxidase (HRP) was used as the scanning probe for generating the chemiluminescence (CL). The inner solution was injected from the capillary tip at 78 pl s⁻¹ while scanning above the enzyme-immobilized chip. The CL generated when the capillary tip was scanned above the enzyme spots was detected using a photon-counter (CL detection). Two-dimensional mapping of the oxidation current and photon-counting intensity against the tip position affords images of which their contrast reflects the activity of the immobilized GOD and uricase. For both the EC and CL detections, the signal responses were plotted as a function of the glucose and uric acid concentrations in solution. The sensitivities for the EC and CL detection were found to be comparable.     

A reversed-phase high performance liquid chromatography coupled with resonance Rayleigh scattering detection for the determination of four tetracycline antibiotics

A new reversed-phase high performance liquid chromatography with resonance Rayleigh scattering detection (HPLC-RRS) was developed for simultaneous separation and determination of four tetracycline antibiotics (TCs). A good chromatographic separation among the compounds was achieved using a Synergi Fusion-RP column (150 mm × 4.6 mm; 4 μm) and a mobile phase consisting of methanol-acetonitrile-oxalic acid (5 mM) at the flow rate of 0.8 mL min⁻¹. Column temperature was 30 °C. The RRS signal was detected at λ_ex = λ_em = 370 nm. The recoveries of sample added standard ranged from 95.3% to 103.5%, and the relative standard deviation was below 2.79%. A detection limit of 2.12-5.12 μg mL⁻¹ was reached and a linear range was found between peak height and concentration in the range of 10.36-518.0 μg mL⁻¹ for oxytetracycline (OTC), 12.11-605.5 μg mL⁻¹ for tetracycline (TC), 11.79-589.5 μg mL⁻¹ for chlortetracycline (CTC) and 10.32-516.0 μg mL⁻¹ for doxycycline (DC). The linear regression coefficients were all above 0.999. The method has been applied successfully to the determination of OTC, TC, CTC, DC in pharmaceutical formulations and in honey. The method was simple, rapid and showed a better linear relation and high repeatability.     

A comparison of solid-phase microextraction and stir bar sorptive extraction coupled to liquid chromatography for the rapid analysis of resveratrol isomers in wines, musts and fruit juices

A comparison of direct immersion solid-phase microextraction (DI-SPME) and stir bar sorptive extraction (SBSE) coupled to liquid chromatography (HPLC) with fluorimetric detection for the rapid analysis of resveratrol isomers is described. For DI-SPME, a polar Carbowax-template resin (CW/TPR) 50 μm fiber was the most efficient and optimum extraction conditions were 40 °C and an extraction time of 30 min, stirring in the presence of 5% (m/v) sodium chloride and 0.07 M acetate/acetic acid buffer (pH 6). Desorption was carried out using the static mode for 10 min. Linearity was obtained in the 5-150 and 2-150 ng mL⁻¹ ranges for trans- and cis-resveratrol, with detection limits of 2 and 0.5 ng mL⁻¹, respectively. When using SBSE, a polydimethylsiloxane (PDMS) twister provided best extraction by means of a derivatization reaction in the presence of acetic anhydride and potassium carbonate. The same time and temperature were used for the extraction step in the presence of 2.5% (m/v) sodium chloride, and liquid desorption was performed with 150 μL of a 50/50 (v/v) acetonitrile/1% (v/v) acetic acid solution in a desorption time of 15 min. Linearity was now between 0.5 and 50 ng mL⁻¹ for trans-resveratrol with a detection limit of 0.1 ng mL⁻¹, while cis-resveratrol could not be extracted. The proposed methods were successfully applied to determining the resveratrol isomer content of wine, must and fruit juices.     

Development of a validated liquid chromatographic method for the quality control of Prunellae Spica: Determination of triterpenic acids

A simple and rapid reversed-phase HPLC-UV method was developed for the determination of triterpenic acids in the crude extract of Prunellae Spica. Five triterpenic acids were extracted and isolated from P. Spica as marker compounds for use in the quality control of herbal medicines. Various solvent extraction techniques were evaluated, and the greatest efficiency was observed with sonication in 100% ethanol. Elemental compositions of the five marker compounds were determined by high-resolution mass spectroscopy. The dynamic range of the HPLC-UV method depended on the specific analyte, and acceptable quantitation was obtained between 10 and 250 μg mL⁻¹ for oleanolic acid, between 10 and 300 μg mL⁻¹ for ursolic acid, between 3 and 75 μg mL⁻¹ for 2α,3α,24-trihydroxyolean-12en-28oic acid, between 5 and 100 μg mL⁻¹ for euscaphic acid, and between 5 and 100 μg mL⁻¹ for 2α,3α-dihydroxyurs-12en-28oic acid. The method was deemed satisfactory by inter- and intra-day validation and exhibited both high accuracy and precision (relative standard deviation <9.4%). Overall limits of quantitation and detection were approximately 0.5-2.5 μg mL⁻¹ at a signal-to-noise ratio (S/N) of 3 and were about 3.0-10.0 μg mL⁻¹ at a S/N of 10. In addition, principal component analysis (PCA) was performed on the analytical data of 15 different P. Spica samples in order to classify samples collected from different regions.