Determination of Flavonoid Markers in Honey with SPE and LC using Experimental Design

Determination of flavonoid markers quercetin, hesperetin, and chrysin, found in north Iranian citrus honey samples, was carried out by solid phase extraction (SPE) and isocratic liquid chromatographic separation using central composite design. Optimum conditions for SPE were achieved using 10 mL methanol/water (13:87, v/v, pH = 7) as the washing solvent and 4 mL methanol for elution. Good clean-up and high recovery >90% were observed for all analytes. The use of water/ACN/THF/AcOH (54:36:5:5, v/v) was found to serve as the optimum mobile phase composition and allowed for the separation of analytes from endogenous compounds present in honey. SPE parameters, such as maximum loading capacity and breakthrough volume, were also determined for each analyte. Limit of detection, linear range, recovery, repeatability of retention times, and peak heights were 3.11 × 10^?8–4.44 × 10^?8 g g^?1, 0.50–50.0 μg mL^?1 (R ² > 0.99), 90.7–96.9%, 3.0–3.6%, and 1.0–2.6%, respectively. Precision of the overall analytical procedure, estimated by five replicate measurements for quercetin, hesperetin and chrysin in citrus honey, as well as the relative standard deviations were 4.3%, 3.8%, and 5.5%, respectively.     

Multivariate optimization of the hollow fiber-based liquid phase microextraction of lead in human blood and urine samples using graphite furnace atomic absorption spectrometry

The present study developed a liquid-phase microextraction based on hollow fiber coupled with graphite furnace atomic absorption spectrometry for the effective extraction and quantitation of lead from urine and blood samples. A multivariate design was used for the optimization of the experimental conditions to ensure high extraction efficiency. Six factors (solvent type, chelating agent, time extraction, temperature, donor phase pH, and acceptor phase pH) were obtained by screening eleven factors of the Plackett–Burman design; these were optimized using the central composite design of response surface methodology. The optimum conditions of donor phase pH, acceptor phase pH, temperature, and extraction time were 5, 9.5, 40 °C, and 120 min, respectively. In addition, oleic acid containing dicyclohexyl-18-krone-6 was used for the membrane phase. Under optimal conditions, the enrichment factor, limit of detection, and limit of quantification were obtained in the ranges of 21.3–18.7, 0.001–0.002 ng mL^?1, and 0.008–0.01 ng mL^?1, respectively, in urine and blood samples. The linearity of the calibration curve was established for the concentration of Pb in the range of 1–50 ng mL^?1 (r²?=?0.9983). Finally, the performance of the developed method was evaluated for the determination of lead in urine and blood samples, and satisfactory results were obtained (RSDs <?10% with recovery >?95).     

Determination of Flavonoid Markers in Honey with SPE and LC using Experimental Design

Determination of flavonoid markers quercetin, hesperetin, and chrysin, found in north Iranian citrus honey samples, was carried out by solid phase extraction (SPE) and isocratic liquid chromatographic separation using central composite design. Optimum conditions for SPE were achieved using 10 mL methanol/water (13:87, v/v, pH = 7) as the washing solvent and 4 mL methanol for elution. Good clean-up and high recovery >90% were observed for all analytes. The use of water/ACN/THF/AcOH (54:36:5:5, v/v) was found to serve as the optimum mobile phase composition and allowed for the separation of analytes from endogenous compounds present in honey. SPE parameters, such as maximum loading capacity and breakthrough volume, were also determined for each analyte. Limit of detection, linear range, recovery, repeatability of retention times, and peak heights were 3.11 × 10⁻⁸–4.44 × 10⁻⁸ g g⁻¹, 0.50–50.0 μg mL⁻¹ (R ² > 0.99), 90.7–96.9%, 3.0–3.6%, and 1.0–2.6%, respectively. Precision of the overall analytical procedure, estimated by five replicate measurements for quercetin, hesperetin and chrysin in citrus honey, as well as the relative standard deviations were 4.3%, 3.8%, and 5.5%, respectively.

     

Supramolecular solvent microextraction of Sudan blue II in environmental samples prior to its spectrophotometric determination

A liquid phase microextraction method-based conformation of supramolecular assembly was developed for the separation and preconcentration of trace levels of Sudan blue II. Various analytical parameters such as pH, supramolecular solvent type and volume, sample volume and matrix effect etc. were optimised. Sudan blue II concentration in the extraction phase was determined by UV-visible spectrophotometer. Under the optimised conditions, detection limit and preconcentration factor was found as 2.16 µg L^?1 and 80, respectively. Relative standard deviation value was found 5%. The developed procedure was successfully applied for the determination of trace levels of Sudan blue II in environmental samples.     

Homogeneous Liquid–Liquid Microextraction for Determination of Organochlorine Pesticides in Water and Fruit Samples

A fast and effective preconcentration method for extraction of organochlorine pesticides (OCPs) was developed using a homogeneous liquid–liquid extraction based on phase separation phenomenon in a ternary solvent (water/methanol/chloroform) system. The phase separation phenomenon occurred by salt addition. After centrifugation, the extraction solvent was sedimented in the bottom of the conical test tube. The OCPs were transferred into the sedimented phase during the phase separation step. The extracted OCPs were determined using gas chromatography–electron capture detector. Several factors influencing the extraction efficiency were investigated and optimized. Optimal results were obtained at the following conditions: volume of the consolute solvent (methanol), 1.0 mL; volume of the extraction solvent (chloroform), 55 μL; volume of the sample, 5 mL; and concentration of NaCl, 5 % (w/v). Under optimal conditions, the preconcentration factors in the range of 486–1,090, the dynamic linear range of 0.01–100 μg L^?1, and the limits of detection of 0.001–0.03 μg L^?1 were obtained for the OCPs. Using internal standard, the relative standard deviations for 1 μg L^?1 of the OCPs in the water samples were obtained in the range of 4.9–8.6 % (n = 5). Finally, the proposed method was successfully applied for extraction and determination of the OCPs in water and fruit samples.     

Syringe-to-syringe-dispersive liquid–phase microextraction combined with flame atomic absorption spectrometry for pre-concentration and determination of cobalt with the aid of experimental design

A rapid and selective technique has been proposed for the extraction, pre-concentration and determination of trace amounts of cobalt in water and pharmaceutical samples by syringe-to-syringe-dispersive liquid–phase microextraction (SS-DLPME) combined with flame atomic absorption spectrometry (FAAS). In the developed method, 1-nitroso-2-naphthol was used as a selective complexing agent and 1-octanol was selected as the extraction solvent. Factors such as pH of the sample solution, concentration of the complexing agent, volume of the extraction solvent, number of injections and centrifugation time affecting the extraction efficiency were screened using a Plackett–Burman design (PBD) and optimised using a Box–Behnken design (BBD). Under optimum conditions, a dynamic linear range of 2.5–650 μg L^?1 with the coefficient of determination r² = 0.997 was obtained. The resultant limit of detection (LOD) was 0.68 μg L^?1, whereas the enrichment factor (EF), intraday precision and inter-day precision were 281, 1.43% and 1.93%, respectively. This method was used successfully for pre-concentration and determination of the analyte in environmental water and drug samples.     

Optimization of parameters for the alcoholic-assisted dispersive liquid–liquid microextraction of estrogens in water

Extraction and determination of estrogens in water samples were performed using alcoholic-assisted dispersive liquid–liquid microextraction (AA-DLLME) and high-performance liquid chromatography (UV/Vis detection). A Plackett–Burman design and a central composite design were applied to evaluate the AA-DLLME procedure. The effect of six parameters on extraction efficiency was investigated. The factors studied were volume of extraction and dispersive solvents, extraction time, pH, amount of salt and agitation rate. According to Plackett–Burman design results, the effective parameters were volume of extraction solvent and pH. Next, a central composite design was applied to obtain optimal condition. The optimized conditions were obtained at 220 μL 1-octanol as extraction solvent, 700 μL ethanol as dispersive solvent, pH 6 and 200 μL sample volume. Linearity was observed in the range of 1–500 μg L^?1 for E2 and 0.1–100 μg L^?1 for E1. Limits of detection were 0.1 μg L^?1 for E2 and 0.01 μg L^?1 for E1. The enrichment factors and extraction recoveries were 42.2, 46.4 and 80.4, 86.7, respectively. The relative standard deviations for determination of estrogens in water were in the range of 3.9–7.2 % (n = 3). The developed method was successfully applied for the determination of estrogens in environmental water samples.     

LC Determination of Chloramphenicol in Honey Using Dispersive Liquid–Liquid Microextraction

A novel method, dispersive liquid–liquid microextraction coupled with liquid chromatography-variable wavelength detector (LC-VWD), has been developed for the determination of chloramphenicol (CAP) in honey. A mixture of extraction solvent (30 μL 1,1,2,2-tetrachloroethane) and dispersive solvent (1.00 mL acetonitrile) were rapidly injected by syringe into a 5.0 mL real sample for the formation of cloudy solution, the analyte in the sample was extracted into the fine droplets of C₂H₂Cl₄. After extraction, phase separation was performed by centrifugation and the enriched analyte in the sedimented phase was determined by LC-VWD. Some important parameters, such as the kind and volume of extraction solvent and dispersive solvent, extraction time, sample solution pH, sample volume and salt effect were investigated and optimized. Under the optimum extraction condition, the method yields a linear calibration curve in the concentration range from 3 to 2,000 μg kg^?1 for target analyte. The enrichment factor for CAP was 68.2, and the limit of detection (S/N = 3) were 0.6 μg kg^?1. The relative standard deviation (RSD) for the extraction of 10 μg kg^?1 of CAP was 4.3% (n = 6). The main advantages of method are high speed, high enrichment factor, high recovery, good repeatability and extraction solvent volume at μL level. Honey samples were successfully analyzed using the proposed method.     

Determination of ultra traces of lead in water samples after combined solid-phase extraction–dispersive liquid–liquid microextraction by graphite furnace atomic absorption spectrometry

A solid-phase extraction coupled with dispersive liquid–liquid microextraction (DLLME) method followed by graphite furnace atomic absorption spectrometry (GFAAS) was developed for the extraction, preconcentration, and determination of ultra trace amounts of lead in water samples. Variables affecting the performance of both steps were thoroughly investigated. Under optimized conditions, 100 mL of lead solution were first concentrated using a solid phase sorbent. The extracts were collected in 1.50 mL of THF and 18 μL of carbon tetrachloride was dissolved in the collecting solvent. Then 5.0 mL pure water was injected rapidly into the mixture of THF and carbon tetrachloride for DLLME, followed by GFAAS determination of lead. The analytical figures of merit of method developed were determined. With an enrichment factor of 1,800, a linear calibration of 3–60 ng L^?1 and a limit of detection of 1.0 ng L^?1 were obtained. The relative standard deviation for seven replicate measurements of 30 ng L^?1 of lead was 5.2 %. The relative recoveries of lead in mineral, tap, well, and river water samples at spiking level of 10 and 20 ng L^?1 are in the range 94–106 %.     

Determination of carbamazepine in formulation samples using dispersive liquid–liquid microextraction method followed by ion mobility spectrometry

A simple, sensitive, fast and efficient method based on dispersive liquid–liquid microextraction (DLLME) followed by ion mobility spectrometry (IMS) has been proposed for preconcentration and trace detection of carbamazepine (CBZ) in formulation samples. In this method, 1 mL of methanol (disperser solvent) containing 80 μL of chloroform (extraction solvent) was rapidly injected by a syringe into a sample. After 5 min centrifugation, the preconcentrated carbamazepine in the organic phase was determined by IMS. Development of DLLME procedure includes optimization of parameters influencing the extraction efficiencies such as kind and volume of extraction solvent, disperser solvent and salt addition, centrifugation time and pH of the sample solution. The proposed method presented good linearity in the range of 0.05–10 μg mL^?1 and the detection limit was 0.025 μg mL^?1. The repeatability of the method expressed as relative standard deviation was 6 % (n = 5). This method has been applied to the analysis of carbamazepine formulation samples with satisfactory relative recoveries ≤75 %.     

Liquid–Liquid–Liquid Micro Extraction Combined with CE for the Determination of Rare Earth Elements in Water Samples

A simple method for determination of rare earth elements (REEs) by liquid–liquid–liquid microextraction (LLLME) coupled with capillary electrophoresis and ultraviolet technique was developed. In the LLLME system, 40 mmol L^?1 4-benzoyl-3-methy-1-phenyl-5-pyrazolinone (PMBP) acted as extractant and 4% (v/v) formic acid was used as back-extraction solution. The parameters influencing the LLLME, including the type of the organic solvent, sample pH, formic acid concentration, PMBP concentration, extraction time, volume of organic solvent, stirring rate and phase volume ratio, were investigated. Under the optimized conditions, the detection limits (S/N = 3) of REEs were in the range of 0.19–0.70 ng mL^?1. The developed method was successfully applied to the determination of trace amounts of REEs in water samples.     

Dispersive liquid–liquid microextraction using the freezed floating organic drop for rapid,fast, and sensitive determination of lead

A rapid, simple, and sensitive method was developed for lead preconcentration and separation in various real samples by dispersive liquid–liquid microextraction based on the freezing of floating organic drop. In this method, a suitable extraction solvent dissolved in a dispersive solvent was quickly syringed into the water sample so that the solution became turbid. Then, two phases were separated by centrifugation. The floating extractant droplet can be easily solidified on an ice bath and taken out of the water sample. Then, it can be liquefied instantly at room temperature, and analyte can be determined in it. In the creation of a hydrophobic complex with lead, 1-(2-pyridylazo)-2-naphthole (PAN) was used as the chelating agent. 1-Undecanol and acetone were used as extraction and disperser solvent. To achieve the highest recovery, some factors (type and volume of dispersive and extraction solvent, pH, PAN concentration, and salt concentration) were optimised. Under optimised conditions (pH = 9, 1.0 × 10^–3 mol L^?1 PAN, 15% w/v NaCl, 100 µL 1-undecanol, and 0.3 mL acetone), the lead calibration graph was linear from 1.5 to 80 μg L^?1. The detection limit and preconcentration factor were 0.5 μg L^?1 and 50, respectively. Lead was successfully determined in water and food (spinach, rice, potato, carrot, and black tea bag) samples by this method.     

QbD-driven development and validation of an efficient bioanalytical UPLC method for estimation of olmesartan medoxomil

The present studies describe quality by design-based development of bioanalytical ultra performance liquid chromatography method of olmesartan medoxomil. Initially, method objectives were defined and critical analytical attributes (CAAs) earmarked. Method optimization was conducted using a central composite design for optimizing mobile phase ratio and injection volume as the critical method parameters (CMPs) identified from risk assessment and factor screening studies, and evaluated for their influence on peak area, theoretical plates, and asymmetry factor as CAAs. Chromatographic separation was achieved using acetonitrile:water solvent system containing 0.1% orthophosphoric acid (54:46, v/v) as the mobile phase with UV detection at 243 nm. Further optimization of bioanalytical extraction process was accomplished using a Box–Behnken design selecting extraction time, centrifugation speed, and centrifugation time as the CMPs identified from failure mode and effect analysis, and evaluated for percent recovery, peak asymmetry, and theoretical plate count as the CAAs. Establishment of calibration curve indicated linearity between concentration range of 100 and 800 ng mL^?1, excellent accuracy and precision with limit of detection and limit of quantification as 6.2 and 19.0 ng mL^?1, respectively. Drug stability studies indicated mean percent recovery ranging between 92.4 and 97.3% under various stress conditions.     

Ultrasonic-Assisted Drop-to-Drop Solvent Microextraction in a Capillary Tube for Analyzing Trace Benzene, Toluene, Xylene in One Drop of a Water Sample

A novel analytical technique termed ultrasonic-assisted drop-to-drop solvent microextraction (USA-DDSME) in a capillary tube was developed to determine trace benzene, toluene, xylene in one drop of a water sample, which was combined with gas chromatography–flame ionization detection (GC–FID). The advantages of this method are rapidity, convenience, ease of operation, simplicity of the device, and extremely little solvent and sample consumption. Extraction conditions including the type of extraction solvent, the volume of extraction solvent, the volume of sample, extraction time and effect of salt concentration were optimized. The best optimum parameters for extraction were achieved with 3 μL of extraction solvent. Chloroform was divided into four equal divisions in 20 μL water sample (without salt addition) in a capillary tube and ultrasonicated for 10 min, centrifugated at 2,500 rpm for 5 min to let the extraction solvent settle at the bottom of the capillary tube, then 1 μL of the separated extraction solvent was injected into the GC–FID for analysis. Linearity of the method was determined by analyzing spiked water samples over a concentration range of 0.1–50 μg mL^?1. Correspondingly, the LOD values were 0.01 μg mL^?1. All calibration curves were found to have good linearity with correlation coefficients (r ²) > 0.995. The precision (RSD) of the system, measured by six repeated determinations of the analytes at 1 μg mL^?1 were in the range of 1.6–3.5%.     

Preconcentration procedure using vortex-assisted liquid–liquid microextraction for the fast determination of trace levels of thorium in water samples

A new simple and rapid vortex-assisted liquid–liquid microextraction method was applied for the determination of thorium in water samples. In this method, chloroform used as extraction solvent was directly injected into the water sample solution. The extraction solvent was dispersed into the aqueous phase under vigorously shaking with the vortex. After centrifuging, the fine droplets of extractant phase were settled to the bottom of the conical-bottom centrifuge tube. The effect of different experimental parameters on the performance of the method were studied and discussed. Under the optimum conditions, the detection limit for Th(IV) was 7.5 ng mL^?1. The precision of the method, evaluated as the relative standard deviation obtained by analyzing of 10 replicates, was 2.8 %. The practical applicability of the developed method was examined using natural water and monazite sand samples.     

Development of a dispersive liquid–liquid microextraction method for determination of palladium in water samples using dicyclohexano-18- crown-6 as extracting agent

A new separation procedure for determination of palladium using dispersive liquid–liquid microextraction with dicyclohexano-18-crown-6 as complexing reagent was developed. In this method, potassium–dicyclohexano-18-crown-6 was used as a hydrophobic complex for the microextraction of palladium as PdCl₄ ^2? complex ion. The main factors affecting DLLME efficiency, such as type and volume of extractant and disperser solvent, concentration of chelating reagent, concentration of KCl and pH were optimized. Under the optimal conditions, the limit of detection for palladium was 16.0 ng mL^?1 with enrichment factor of 138. The present method was applied to the determination of palladium in water samples with satisfactory analytical results. The method was simple, rapid, cost efficient and sensitive for the extraction and preconcentration of palladium.     

Simultaneous separation and preconcentration of ultra-trace of Cu(II), Pb(II) and Zn(II) in water samples by air-assisted liquid–liquid microextraction prior to determination by graphite furnace atomic absorption spectrometry

A simple, rapid and efficient method has been developed for the extraction, preconcentration and determination of copper, lead and zinc ions in water samples by air-assisted liquid–liquid microextraction coupled with graphite furnace atomic absorption spectrometry (GFAAS). In the proposed method, much less volume of an organic solvent (in the order of some µL) was used as the extraction solvent in the absence of disperser solvent. Fine organic droplets were formed by sucking and injecting of the mixture of aqueous sample solution and extraction solvent with a syringe for several times in a conical test tube. After extraction, phase separation was achieved by centrifugation and the enriched analytes in the sedimented phase were determined by GFAAS. Several variables potentially affecting the extraction efficiency were investigated and optimized. Calibration graphs were linear in the concentration range of 45.0–1100 ng L^?1. Detection limits were in the range of 18.0–26.0 ng L^?1. The accuracy of the developed procedure was checked by analyzing NRCC-SLRS4 Riverine water as a certified reference material. Finally, the proposed method was successfully applied to determine the selected heavy metals in tap, surface and river water samples.     

Rapid Enrichment and Sensitive Determination of Tetrabromobisphenol A in Environmental Water Samples with Ionic Liquid Dispersive Liquid-Phase Microextraction Prior to HPLC–ESI-MS–MS

A new microextraction method termed ionic liquid dispersive liquid-phase microextraction has been developed for the rapid enrichment and sensitive determination of tetrabromobisphenol A in environmental water samples prior to high-performance liquid chromatography–electrospray tandem mass spectrometry. Instead of using toxic organic solvents, green solvent ionic liquid was used as extraction solvent. Factors that may influence the enrichment efficiency, such as type and volume of ionic liquid, type and volume of disperser solvent, sample pH, extraction time and NaCl content were investigated and optimized in detail. Under optimum conditions, linearity of the method was observed over the range 1–100 μg L^?1 with correlation coefficient 0.9986. The proposed method has been found to have excellent sensitivity with limit of detection 0.06 μg L^?1 and precision 6.95% (RSD, n = 5). This method has been successfully applied to analyze real environmental water samples and satisfactory results were achieved. All these results indicated that the present method was an environmentally friendly method for the rapid enrichment and sensitive analysis of tetrabromobisphenol A at trace level in environmental water samples.     

Application of DLLME to Isolation and Concentration of Non-Steroidal Anti-Inflammatory Drugs in Environmental Water Samples

Dispersive liquid–liquid microextraction (DLLME) coupled with liquid chromatography-tandem mass spectrometry detection was applied for determination of selected anti-inflammatory pharmaceuticals: ibuprofen, ketoprofen, naproxen and diclofenac. Development of DLLME procedure included optimisation of several important parameters such as kind and volume of extracting and dispersive solvents as well as sample pH. Under optimised conditions a two-step extraction with sonication was used. Chloroform was applied as the extracting and acetone as dispersing solvent. Calibration curves ranges were 1–500 μg L^?1 for naproxen and ibuprofen and 0.25–500 μg L^?1 for ketoprofen and diclofenac with correlation coefficients at least 0.997. Limits of quantitation were from 0.5 to 10 ng L^?1. The developed analytical method was employed for determination of ibubrofen, ketoprofen, naproxen and diclofenac in river and tap water samples. The results showed that DLLME is a simple, rapid and sensitive analytical technique for the pre-concentration of trace amounts of pharmaceuticals in environmental water samples.     

Application of dispersive liquid–liquid microextraction with alcoholic solvents followed by HPLC–UV as a sensitive and efficient method for the extraction and determination of citalopram in biological samples using an experimental design

A sensitive and rapid method based on alcoholic-assisted dispersive liquid–liquid microextraction followed by high-performance liquid chromatography for determination of citalopram in human plasma and urine samples was developed. The effects of six parameters (extraction time, stirring speed, pH, volume of extraction and disperser solvents, and ionic strength) on the extraction recovery were investigated and optimized utilizing Plackett–Burman design and Box–Behnken design, respectively. According to Plackett–Burman design results, the volume of disperser solvent, stirring speed, and extraction time had no effect on the recovery of citalopram. The optimized condition was a mixture of 172 µL of 1-octanol as extraction solvent and 400 µL of methanol as disperser solvent, pH of 10.3 and 1% w/v of salt in the sample solution. Replicating the experiment in optimized condition for five times, gave the average extraction recoveries equal to 89.42%. The detection limit of citalopram in human plasma was obtained 4 ng/mL, and the linearity was in the range of 10–1200 ng/mL. The corresponding values for human urine were 5.4 ng/mL with the linearity in the range of 10–2000 ng/mL. Relative standard deviations for inter- and intraday extraction of citalopram were less than 7% for five measurements. The proposed method was successfully implemented for the determination of citalopram in human plasma and urine samples.