Arsenic and its speciation analysis using high-performance liquid chromatography and inductively coupled plasma mass spectrometry

It is known that arsenic has different toxicological properties dependent upon both its oxidation state for inorganic compounds, as well as the different toxicity levels exhibited for organic arsenic compounds. The field of arsenic speciation analysis has grown rapidly in recent years, especially with the utilization of high-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS), a highly sensitive and robust detector system. Complete characterization of arsenic compounds is necessary to understand intake, accumulation, transport, storage, detoxification and activation of this element in the natural environment and living systems. This review describes the essential background and toxicity of arsenic in the environment, and more importantly, some currently used chromatographic applications and sample handling procedures necessary to accurately detect and quantify arsenic in its various chemical forms. Applications and work using only HPLC-ICP-MS for arsenic speciation of environmental and biological samples are presented in this review.     

Determination of pharmaceuticals in biosolids using accelerated solvent extraction and liquid chromatography/tandem mass spectrometry

An analytical method was developed to quantitatively determine pharmaceuticals in biosolid (treated sewage sludge) from wastewater treatment plants (WWTPs). The collected biosolid samples were initially freeze dried, and grounded to obtain relatively homogenized powders. Pharmaceuticals were extracted using accelerated solvent extraction (ASE) under the optimized conditions. The optimal operation parameters, including extraction solvent, temperature, pressure, extraction time and cycles, were identified to be acetonitrile/water mixture (v/v 7:3) as extraction solvent with 3 extraction cycles (15 min for each cycle) at 100 °C and 100 bars. The extracts were cleaned up using solid-phase extraction followed by determination by liquid chromatography coupled with tandem mass spectrometry. For the 15 target pharmaceuticals commonly found in the environment, the overall method recoveries ranged from 49% to 68% for tetracyclines, 64% to 95% for sulfonamides, and 77% to 88% for other pharmaceuticals (i.e. acetaminophen, caffeine, carbamazepine, erythromycin, lincomycin and tylosin). The developed method was successfully validated and applied to the biosolid samples collected from WWTPs located in six cities in Michigan. Among the 15 target pharmaceuticals, 14 pharmaceuticals were detected in the collected biosolid samples. The average concentrations ranged from 2.6 μg/kg for lincomycin to 743.6 μg/kg for oxytetracycline. These results indicated that pharmaceuticals could survive wastewater treatment processes, and accumulate in sewage sludge and biosolids. Subsequent land application of the contaminated biosolids could lead to the dissemination of pharmaceuticals in soil and water environment, which poses potential threats to at-risk populations in the receiving ecosystems.     

Arsenic speciation by coupling high-performance liquid chromatography with inductively coupled plasma mass spectrometry

Summary An ICP-MS detector in combination with HPLC has been evaluated for the analysis of six arsenic compounds. The influence of the presence of an organic modifier in the mobile phase on arsenic response and the quality parameters of the analysis are discussed. Detection limits for arsenic species under study range from 10 to 30 pg. The determination of arsenic compounds in solutions simulating fish or sediment extracts has been used to demonstrate the applicability of this technique.     

加速溶剂萃取-超高效液相色谱-串联质谱法测定茶叶中拟除虫菊酯类农药残留

建立了加速溶剂萃取（ASE）-超高效液相色谱（UPLC）-串联质谱法（MS/MS）测定茶叶中拟除虫菊酯类农药残留的方法。ASE萃取温度为80 ℃，萃取压力为10.34 MPa，以正己烷-丙酮（2∶1，v/v）为溶剂静态萃取5 min，循环一次。萃取液浓缩后经GCB/NH₂-Florisil柱净化，UPLC分离，MS/MS正离子扫描（ESI⁺）、多反应离子监测（MRM）模式进行分析，外标法定量。线性回归分析表明：10种拟除虫菊酯的浓度与其峰面积的线性关系显著，相关系数（r）均不小于0.9995，检出限（LOD）在0.5~5.0 μg/kg之间，定量限（LOQ）在1.6~16.6 μg/kg之间；在定量限、0.4 mg/kg以及最高残留限量（MRL，无MRL的加入1 mg/kg）3个水平进行添加回收试验（n=7），回收率为68.7%~103.8%，RSD为0.8%~13.2%。该方法前处理简单，耗时短，灵敏度和准确度高，可满足茶叶中痕量拟除虫菊酯类农药残留测定的要求。     

Simultaneous co-extraction of organometallic species of different elements by accelerated solvent extraction and analysis by inductively coupled plasma mass spectrometry coupled to liquid and gas chromatography

This paper describes the development of an accelerated solvent extraction methodology that is capable of simultaneously extracting organometallic species of As, Sn and Hg in a semi-automated manner. Accelerated solvent extraction (ASE) methods based on previous research on the separate extraction of organotin and -arsenic species in our laboratory were adapted for the co-extraction of six different species from an oyster tissue certified reference material (BCR 710). For the first time, the extraction of MeHg by this technique is also investigated. The proposed ASE conditions employed 50% acetic acid in methanol at a temperature of 100 degrees C with up to five consecutive extraction cycles of 3 minutes. Extraction efficiencies for organoarsenic species ranged from 80% (dimethylarsinic acid, DMA) to 99% for arsenobetaine (AsB). Species of toxicological interest, such as dibutyltin (DBT), tributyltin (TBT) and methylmercury (MeHg), were extracted with mean recoveries of 81, 84 and 76%, respectively. The extracted species were analysed by gas chromatography/inductively coupled plasma mass spectrometry (GC/ICPMS; DBT, TBT and MeHg) and liquid chromatography/inductively coupled plasma mass spectrometry (LC/ICPMS; MMA, DMA, AsB) after ethylation with sodium tetraethylborate or dilution with water, respectively. In addition to those species for which the extraction efficiency was assessed during this study, a further five arsenic species (arsenite, arsenate and three unidentified species), as well as monobutyltin (MBT) and mono-, di- and triphenyltin, could also be extracted from other matrices. The developed ASE method provides a significant improvement over many currently available routine monitoring methods for trace element speciation due to the fact that it is capable of extracting several species of toxicological interest simultaneously and quantitatively.     

Development of a method based on accelerated solvent extraction and liquid chromatography/mass spectrometry for determination of arylphenoxypropionic herbicides in soil

A sensitive and specific analytical procedure for determining arylphenoxypropionic herbicides in soil samples, using Ionspray ionization (ISI) liquid chromatography/mass spectrometry (LC/MS), is presented. Arylphenoxypropionic acids are a new class of herbicides used for selective removal of most grass species from any non-grass crop, commercialized as herbicide esters. Previous studies have shown that the esters undergo fast hydrolysis in the presence of vegetable tissues and soil bacteria, yelding the corresponding free acid. The feasibility of rapidly extracting arylphenoxypropionic herbicides from soil by accelerated solvent extraction (ASE) techniques was evaluated. Four different soil samples were fortified with target compounds at levels of 5 and 20 ng/g by following a procedure able to mimic weathered soils. Herbicides were extracted by a methanol/water (80:20 v/v) solution (0.12 M) of NaCl at 90 degrees C. After clean-up using graphitized carbon black (GCB) as absorbent, the extract was analyzed by HPLC/ISI-MS. The effect of concentration of acid in the mobile phase on the response of ISI-MS was investigated. The effects of varying the orifice plate voltage on the production of diagnostic fragment ions, and on the response of the MS detector, were also investigated. The ISI-MS response was linearly related to the amounts of analytes injected between 1 and 200 ng. The limit of detection (signal-to-noise ratio = 3) of the method for the pesticides in soil samples was estimated to be less than 1 ng/g.     

加速溶剂萃取-磁固相萃取-高效液相色谱-串联质谱法测定水产品中10种氟喹诺酮类药物残留

养殖过程中的不合理使用和滥用使得水产品中氟喹诺酮抗生素残留累积量呈递增趋势,对人类的健康造成潜在的风险。建立高效、灵敏和可靠的水产品中氟喹诺酮类药物的同时分析方法十分重要。该研究建立了加速溶剂萃取-磁固相萃取-高效液相色谱-串联质谱法(ASE-MSPE-HPLC-MS/MS)测定水产品中沙拉沙星、氧氟沙星、恩诺沙星、丹氟沙星、洛美沙星、培氟沙星、环丙沙星、依诺沙星、诺氟沙星和双氟沙星残留的检测方法。采用室温自组装法,即在室温(25 ℃)下,将氧化石墨烯和零价纳米铁储备液快速涡旋混合,磁性分离收集沉淀物,得到GO@nZVI磁性复合材料。以扫描电镜、傅里叶变换红外光谱和X-射线衍射对磁性复合材料进行表征,结果显示GO@nZVI成功制备。该材料应用于MSPE净化中,ASE萃取温度为70 ℃,萃取溶剂为甲醇,萃取压力为10.34 MPa,静态萃取时间为5 min,循环萃取3次。萃取液浓缩后经MSPE有效净化后,采用Agilent ZORBAX Eclipse Plus C18色谱柱(100 mm×3.0 mm, 1.8 μm)梯度洗脱分离,MS/MS电喷雾正离子(ESI⁺)扫描、多反应监测(MRM)模式进行定量分析。氟喹诺酮的线性范围为1~100 μg/kg,线性相关系数(r²)均大于0.9995;目标物的检出限(S/N=3)为0.02~0.29 μg/kg,定量限(S/N=10)为0.07~0.98 μg/kg。所建方法成功用于黄鱼、草鱼、黑鱼、明虾和沼虾中氟喹诺酮的测定,在1倍、2倍、10倍定量限加标水平下得到的加标回收率为81.6%~105.8%,相对标准偏差(RSD)为4.2%~13.6%。研究表明,将ASE与MSPE有机结合,利用外部磁场实现萃取液的净化,无需离心和过滤操作,溶剂用量少,并且该方法所使用磁性萃取材料制备方法简单,具有自动化程度高、简便快速、方法灵敏度高、准确度高、重复性好等特点,可满足对水产品中FQs残留限量检测要求,具有实际应用前景。     

Antimony speciation by inductively coupled plasma mass spectrometry using solid phase extraction cartridges

A novel and simple method for inorganic antimony speciation is described based on selective solid phase extraction (SPE) separation of antimony(III) and highly sensitive inductively coupled plasma mass spectrometric (ICP-MS) detection of total antimony and antimony(V) in the aqueous phase of the sample. Non-polar SPE cartridges, such as the Isolute silica-based octyl (C8) sorbent-containing cartridge, selectively retained the Sb(III) complex with ammonium pyrrolidine dithiocarbamate (APDC), while the uncomplexed Sb(V) remained as a free species in the solution and passed through the cartridge. The Sb(III) concentration was calculated as the difference between total antimony and Sb(V) concentrations. The detection limit was 1 ng L(-1) antimony. Factors affecting the separation and detection of antimony species were investigated. Acidification of samples led to partial or complete retention of Sb(V) on C8 cartridge. Foreign ions tending to complex with Sb(III) or APDC did not interfere with the retention behavior of the Sb(III)-APDC complex. This method has been successfully applied to antimony speciation of various types of water samples.     

Arsenic speciation in waste waters by extraction chromatography followed by atomic absorption spectrometry

Summary A method is described for the determination of arsenic(III) and arsenic(V) in water samples. The sample (adjusted to pH 2.5 to 3.5 with HCl or NaOH) is passed through a chromatographic column filled with inert support modified with the organotin reagent (C₈H₁₇)₂SnCl₂. Under these conditions arsenate is quantitatively retained, while arsenite is not. Arsenate is eluted from the column with 2 mol/l HCl (1–2 ml). Both effluate [As(III)] and eluate [As(V)] are analyzed by flame or graphite furnace AAS. The method was applied to the analysis of waste waters from a metallurgical plant and potable water from the same region. Recoveries are in the range of 85–115%.     

Arsenic speciation in edible alga samples by microwave-assisted extraction and high performance liquid chromatography coupled to atomic fluorescence spectrometry

Twelve commercially available edible marine algae from France, Japan and Spain and the certified reference material (CRM) NIES No. 9 Sargassum fulvellum were analyzed for total arsenic and arsenic species. Total arsenic concentrations were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) after microwave digestion and ranged from 23 to 126 μg g⁻¹. Arsenic species in alga samples were extracted with deionized water by microwave-assisted extraction and showed extraction efficiencies from 49 to 98%, in terms of total arsenic. The presence of eleven arsenic species was studied by high performance liquid chromatography–ultraviolet photo-oxidation–hydride generation atomic–fluorescence spectrometry (HPLC–(UV)–HG–AFS) developed methods, using both anion and cation exchange chromatography. Glycerol and phosphate sugars were found in all alga samples analyzed, at concentrations between 0.11 and 22 μg g⁻¹, whereas sulfonate and sulfate sugars were only detected in three of them (0.6-7.2 μg g⁻¹). Regarding arsenic toxic species, low concentration levels of dimethylarsinic acid (DMA) (<0.9 μg g⁻¹) and generally high arsenate (As(V)) concentrations (up to 77 μg g⁻¹) were found in most of the algae studied. The results obtained are of interest to highlight the need to perform speciation analysis and to introduce appropriate legislation to limit toxic arsenic species content in these food products.     

Determination of four major saponins in the seeds of Aesculus chinensis Bunge using accelerated solvent extraction followed by high-performance liquid chromatography and electrospray-time of flight mass spectrometry

A new method based on accelerated solvent extraction (ASE) followed by a reliable high-performance liquid chromatography-diode array detector (HPLC-DAD) and positive ion electrospray-time of flight mass spectrometry (ESI-TOF/MS) analysis has been developed for the characterization and quantification of four major saponins in extracts of the seeds of Aesculus chinensis Bunge (Semen Aesculi). The saponins escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted from seeds of A. chinesis Bunge via ASE, and the operational parameters of ASE were optimized, such as extraction solvent, extraction temperature, static extraction time and extraction cycles. The optimized procedure employed 70% MeOH as extraction solvent, 120 °C of extraction temperature, 7 min of static extraction time, 60% flush volume and the extraction recoveries of the four compounds were nearly to 100% for two cycles. The HPLC conditions are as follows: SinoChrom ODS BP C₁₈ (4.6 mm × 200 mm, 5 μm) column, acetonitrile and 0.10% phosphoric acid solution as mobile phase, flow rate is 1.0 mL min⁻¹, detection length of UV is 203 nm, injection volume is 10 μL. The results indicated that the developed HPLC method is simple, sensitive and reliable for the determination of four major saponins in seeds of A. chinesis Bunge with a good linearity (r² > 0.9994), precision (relative standard deviation (R.S.D.) <1.5%) and the recovery ranges of 95.2-97.3%. The limits of detection (LOD) of the four compounds were in the range of 0.40-0.75 μg mL⁻¹. This assay can be readily utilized as a quality control method for Semen Aesculi and other related medicinal plants.     

Determination of MK-0431 in human plasma using high turbulence liquid chromatography online extraction and tandem mass spectrometry

A robust and sensitive method using high turbulence liquid chromatography (HTLC) online extraction with tandem mass spectrometry (MS/MS) for the determination of MK-0431 in human plasma was developed and validated to support the clinical studies. This HTLC online extraction method eliminated the time-consuming offline sample extraction procedures and significantly increased productivity. A narrow bore large particle size reversed-phase column (Cyclone, 50 x 1.0 mm, 60 microm) and a BDS Hypersil C18 column (30 x 2.1 mm, 3 microm) were used as extraction and analytical columns, respectively. The linear dynamic range of the calibration curve was 0.5 to 1000 ng/mL. Intraday validation was conducted using five calibration curves prepared in five lots of human control plasma, and the intraday precision (RSD%) was from 2.4 to 9.0% and the accuracy was from 98.0 to 103% of the nominal value. The intraday precision (RSD%, n = 5) for plasma quality control (QC) samples varied from 2.0 to 5.3% and accuracy from 103 to 105% of the nominal value. The interday precision (RSD%) for 100 sets of plasma QC samples in 29 analytical runs varied from 6.3 to 9.0% and the accuracy from 98.8 to 104% of the nominal value. No significant difference was observed between the interday and intraday precision and accuracy of the QC samples.     

Determination of organochlorine pesticides and chlorobenzenes in strawberries by using accelerated solvent extraction combined with sorptive enrichment and gas chromatography/mass spectrometry

An analytical scheme for the determination of several organochlorine pesticides like hexachlorocyclohexanes (HCHs) and DDX compounds (p,p'-DDE, p,p'-DDD, and p,p'-DDT) as well as chlorobenzenes in strawberries has been developed. The procedure is based on aqueous accelerated solvent extraction (ASE) followed by solid-phase microextraction (SPME) or stir bar sorptive extraction (SBSE) and subsequent thermodesorption-gas chromatography/mass spectrometry analysis. A 65 microm polydimethylsiloxane/ divinylbenzene fiber was chosen for the SPME experiments. Significant SPME and ASE parameters were optimized using spiked water and strawberry samples. For the ASE of the organochlorine compounds, a water-acetone mixture (90 + 10, v/v) as the extraction solvent, an extraction temperature of 120 degrees C, and 2 cycles of 10 min extraction proved optimal. The developed method was evaluated with respect to precision and limits of detection (LOD). The relative standard deviations of replicate ASE-SPME determinations (n = 5) were in the range of 4-24%. LOD values between 1 and 10 microg/kg were achieved with the exception of DDT and DDE (40 microg/kg). Using SBSE, the LOD of these compounds could be improved (2 and 5 microg/kg). The main advantages of this method are the avoidance of cleanup and concentration procedures as well as the significant reduction of the required volume of organic solvents. The described method was applied to the determination of the pollutants in strawberry samples collected from different allotment gardens in a potentially polluted area, the Bitterfeld-Wolfen region (Germany).     

Azithromycin extraction from municipal wastewater and quantitation using liquid chromatography/mass spectrometry

Azithromycin is a broad spectrum antimicrobial agent that is approved in the United States for use in humans. Due to azithromycin's low rate of metabolism it is likely to be found in wastewater treatment plants, where its broad spectrum of antimicrobial activity could lead to development of resistance in bacteria. A liquid-liquid extraction using K2CO3 and methyl-t-butyl ether (MTBE) was used to extract azithromycin from 10 ml aliquots of wastewater. Liquid chromatography was performed using a Luna C18(2) (30 mm x 2.0 mm) column with a mass spectrometer friendly mobile phase containing 50:24:2:24 acetonitrile, methanol, tetrahydrofuran, and 0.04 M ammonium hydroxide. The mass spectrometer used an electrospray source with positive ionization and an ion trap detector. A linear standard curve from 5 to 200pg/ml was validated and used to quantitate azithromycin in wastewater.     

Quantitative determination of DNA adducts using liquid chromatography/electrospray ionization mass spectrometry and liquid chromatography/high-resolution inductively coupled plasma mass spectrometry

The quantitative determination of nucleotides from DNA modified by styrene oxide is described using a combination of inductively coupled plasma high-resolution mass spectrometry (ICP-HRMS) and electrospray ionization mass spectrometry (ESI-MS), both interfaced to reversed-phase high-performance liquid chromatography (HPLC). LC/ICP-MS (resolution > 1500 to discriminate against 15N16O+ and 14N16OH+) was employed to determine quantitatively the content of modified nucleotides in standard solutions based on the signal of phosphorus; phosphoric acid served as an internal standard. By means of the standard addition technique the sensitivity of the LC/ESI-MS approach was subsequently determined. Since a comparison of UV, ICP and ESI-MS data suggested that in ESI-MS the ionization efficiency of the adducts is identical within the error limits, quantitative determination of all adducts is possible. For LC/ESI-MS with single ion monitoring, the detection limit for styrene oxide adducts of nucleotides was determined to be 20 pg absolute or 14 modified in 10(8) unmodified nucleotides in a 5 micrograms DNA sample, which comes close to the best methods available for the detection of chemical modifications in DNA.     

Method optimization and quality assurance in speciation analysis using high performance liquid chromatography with detection by inductively coupled plasma mass spectrometry

Achievement of optimum selectivity, sensitivity and robustness in speciation analysis using high performance liquid chromatography (HPLC) with inductively coupled mass spectrometry (ICP-MS) detection requires that each instrumental component is selected and optimized with a view to the ideal operating characteristics of the entire hyphenated system. An isocratic HPLC system, which employs an aqueous mobile phase with organic buffer constituents, is well suited for introduction into the ICP-MS because of the stability of the detector response and high degree of analyte sensitivity attained. Anion and cation exchange HPLC systems, which meet these requirements, were used for the seperation of selenium and arsenic species in crude extracts of biological samples. Furthermore, the signal-to-noise ratios obtained for these incompletely ionized elements in the argon ICP were further enhanced by a factor of four by continously introducing carbon as methanol via the mobile phase into the ICP. Sources of error in the HPLC system (column overload), in the sample introduction system (memory by organic solvents) and in the ICP-MS (spectroscopic interferences) and their prevention are also discussed. The optimized anion and cation exchange HPLC-ICP-MS systems were used for arsenic speciation in contaminated ground water and in an in-house shrimp reference sample. For the purpose of verification, HPLC coupled with tandem mass spectrometry with electrospray ionization was additionally used for arsenic speciation in the shrimp sample. With this analytical technique the HPLC retention time in combination with mass analysis of the molecular ions and their collision-induced fragments provide almost conclusive evidence of the identity of the analyte species. The speciation methods are validated by establishing a mass balance of the analytes in each fraction of the extraction procedure, by recovery of spikes and by employing and comparing independent techniques. The urgent need for reference materials certified for elemental species is stressed.     

Determination of polar organophosphorus pesticides in vegetables and fruits using liquid chromatography with tandem mass spectrometry: selection of extraction solvent

A method based on liquid chromatography (LC)-mass spectrometry (MS)/MS was developed for sensitive determination of a number of less gas chromatography (GC)-amenable organophosphorus pesticides (OPs; acephate, methamidophos, monocrotophos, omethoate, oxydemeton-methyl and vamidothion) in cabbage and grapes. For extraction, several solvents were evaluated with respect to the possibility of direct injection, matrix-induced suppression or enhancement of response, and extraction efficiency. Overall, ethyl acetate was the most favourable solvent for extraction, although a solvent switch was required. For some pesticide/matrix combinations, reconstitution of the residue after evaporation required special attention. Extracts were analysed on a C18 column with polar endcapping. The pesticides were ionised using atmospheric pressure chemical ionisation on a tandem mass spectrometer in multiple reaction monitoring mode. The final method is straightforward and involves extraction with ethyl acetate and a solvent switch to 0.1% acetic acid/water without further cleanup. The method was validated at the 0.01 and 0.5 mg/kg level, for both cabbage and grapes. Recoveries were between 80 and 101% with R.S.D. < 11% (n = 5). The limit of quantification was 0.01 mg/kg and limits of detection were between 0.001 and 0.004 mg/kg.     

Arsenic and its speciation in water samples by high performance liquid chromatography inductively coupled plasma mass spectrometry--last decade review

Arsenic composes a danger for human health all over the world as it is responsible for water resources contamination. The toxicity of arsenic depends on its chemical form. However, occurrence of particular arsenic species is dependent on processes occurring in water. Nowadays, more arsenic species is detected and analyzed in different kind of water (mineral, tap, waste), mainly owing to great possibilities resulting from coupling high performance liquid chromatography (HPLC) with inductively coupled plasma mass spectrometry (ICP-MS). This review mainly describes arsenic speciation analysis by HPLC-ICP-MS technique on the basis of articles that have been published since 2000. Arsenic chemistry, occurrence in different kind of water, total arsenic determination with interferences elimination and its validation and analytical performance are also reviewed.     

Simultaneous determination of 405 pesticide residues in grain by accelerated solvent extraction then gas chromatography-mass spectrometry or liquid chromatography-tandem mass spectrometry

A new method has been established for simultaneous determination of 405 pesticide residues in grain, using accelerated solvent extraction (ASE), solid-phase extraction (SPE), and GC-MS and LC-MS-MS. The method was based on appraisal of the GC-MS and LC-MS-MS characteristics of 660 pesticides, their efficiency of extraction from grain, and their purification. Samples of grain (10 g) were mixed with Celite 545 (10 g) and the mixture was placed in a 34-mL cell of an accelerated solvent extractor and extracted with acetonitrile in the static state for 3 min with two cycles at 1,500 psig and at 80°C. For the 362 pesticides determined by GC-MS, half of the extracts were cleaned with an Envi-18 cartridge and then further cleaned with Envi-Carb and Sep-Pak NH₂ cartridges in series. The pesticides were eluted with acetonitrile-toluene, 3:1, and the eluates were concentrated and used for analysis after being exchanged with hexane twice. For the 43 pesticides determined by LC-MS-MS the other half of the extracts were cleaned with Sep-Pak Alumina N cartridge and further cleaned with Envi-Carb and Sep-Pak NH₂ cartridges. Pesticides were eluted with acetonitrile-toluene, 3:1. After evaporation to dryness the eluates were diluted with acetonitrile-water, 3:2, and used for analysis. In the linear range of each pesticide the linear correlation coefficient r was equal to or greater than 0.956 and 94% of linear correlation coefficients were greater than 0.990. At low, medium, and high fortification levels, at the limit of detection (LOD), twice the LOD and ten times LOD, respectively, recoveries ranged from 42 to 132%; for 382 pesticides, or 94.32%, recovery was from 60 to 120%. The relative standard deviation (RSD) was always below 38% and was below 30% for 391 pesticides, or 96.54%. The LOD was 0.0005–0.3000 mg kg⁻¹. The proposed method is suitable for determination of 405 pesticide residues in grain such as maize, wheat, oat, rice, and barley, etc.      

Determination of 2-methoxyestradiol in human plasma, using liquid chromatography/tandem mass spectrometry

A liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay was developed for the quantitative determination of 2-methoxyestradiol (2ME2) in human plasma. Sample pretreatment involved liquid-liquid extraction with ethyl acetate of 0.3-mL aliquots of plasma spiked with the internal standard, deuterated 2ME2 (2ME2-d5). Separation was achieved on a Zorbax Eclipse C18 column (2.1 x 50 mm, i.d., 5 microm) at room temperature using a gradient elution with methanol and water at a flow rate of 0.25 mL/min. Detection was performed using atmospheric pressure chemical ionization MS/MS by monitoring the ion transitions from m/z 303.1 --> 136.8 (2ME2) and m/z 308.1 --> 138.8 (2ME2-d5). Calibration curves were linear in the concentration range of 1-100 ng/mL. The accuracy and precision values, obtained from three different sets of quality control samples analyzed in quintuplicate on four separate occasions, ranged from 105-108% and from 3.62-5.68%, respectively. This assay was subsequently used for the determination of 2ME2 concentration in plasma of a patient with cancer after a single oral administration of 2ME2 at a dose of 2200 mg.