A novel sol-gel-based amino-functionalized fiber for headspace solid-phase microextraction of phenol and chlorophenols from environmental samples

A novel amino-functionalized polymer was synthesized using 3-(trimethoxysilyl) propyl amine (TMSPA) as precursor and hydroxy-terminated polydimethylsiloxane (OH-PDMS) by sol–gel technology and coated on fused-silica fiber. The synthesis was designed in a way to impart polar moiety into the coating network. The scanning electron microscopy (SEM) images of this new coating showed the homogeneity and the porous surface structure of the film. The efficiency of new coating was investigated for headspace solid-phase microextraction (SPME) of some environmentally important chlorophenols from aqueous samples followed by gas chromatography–mass spectrometry (GC–MS) analysis. Effect of different parameters influencing the extraction efficiency such as extraction temperature, extraction time, ionic strength and pH was investigated and optimized. In order to improve the separation efficiency of phenolic compounds on chromatography column all the analytes were derivatized prior to extraction using acetic anhydride at alkaline condition. The detection limits of the method under optimized conditions were in the range of 0.02–0.05 ng mL⁻¹. The relative standard deviations (R.S.D.) (n = 6) at a concentration level of 0.5 ng mL⁻¹ were obtained between 6.8 and 10%. The calibration curves of chlorophenols showed linearity in the range of 0.5–200 ng mL⁻¹. The proposed method was successfully applied to the extraction from spiked tap water samples and relative recoveries were higher than 90% for all the analytes.     

Determination of organochlorine pesticides in ground water samples using solid-phase microextraction by gas chromatography-electron capture detection

A direct immersion solid-phase microextraction coupled with gas chromatography-electron capture detection (SPME-GC-ECD) method was optimized and validated for the quantitative determination of 18 organochlorine pesticides in ground water. Ionic strength, stirring speed, adsorption and desorption time and pH were some of the parameters investigated in order to select the optimum conditions for SPME with a 50/30 DVB/CAR/PDMS fiber coating. The SPME-GC/ECD method showed good linear response below 10 ng L⁻¹ with R² values in the range of 0.9950–0.9997. The repeatability of the measurements were lower than 10%. Values of relative recoveries located within the acceptable range (80–120%). Limits of quantification (LOQ) from 4.5 × 10⁻³ to 1.5 ng L⁻¹ were obtained. On average 8 organochlorines were found per sample, even so all the 18 organochlorines were quantified among them. Substances such as endrin ketone, γ-BHC and β-BHC were the pesticides determined in larger concentration (0.06–305 ng L⁻¹), while methoxychlor and aldrin in smaller amounts (0.151–1.55 ng L⁻¹). Measured levels of organochlorine pesticides were above the limits established by Brazilian regulations.     

Speciation analysis of arsenic and selenium compounds in environmental and biological samples by ion chromatography-inductively coupled plasma dynamic reaction cell mass spectrometer

An inductively coupled plasma mass spectrometer (ICP-MS) was used as an ion chromatographic (IC) detector for the speciation analysis of arsenic and selenium. The arsenic and selenium species studied included arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenobetaine (AsB), selenite [Se(IV)] and selenate [Se(VI)]. Gradient elution using (NH₄)₂CO₃ and methanol at pH 9 allowed the chromatographic separation of all species in less than 12 min. Effluents from the IC column were delivered to the nebulization system of ICP-DRC-MS for the determination of arsenic and selenium. The potentially interfering ³⁸Ar⁴⁰Ar⁺ and ⁴⁰Ar⁴⁰Ar⁺ at the selenium masses m/z 78 and 80 were reduced in intensity by approximately 3 orders of magnitude by using 0.6 mL min⁻¹ CH₄ as reactive cell gas in the DRC while an Rpq value of 0.3 was used. Meanwhile, arsenic was determined as the adduct ion ⁷⁵As¹²CHH⁺ at m/z 89, which is more sensitive than ⁷⁵As. The limits of detection for arsenic and selenium were in the range of 0.002–0.01 ng mL⁻¹ and 0.01–0.02 ng mL⁻¹, respectively, based on peak height. The relative standard deviation of the peak areas for five injections of 5 ng mL⁻¹ As and Se mixture was in the range of 2–4%. The concentrations of arsenic and selenium species have been determined in urine samples collected locally. The major As and Se species in urines were AsB, DMA and probably selenosugar at concentration of 20–40, 15–19 and 17–31 ng mL⁻¹, respectively. The recoveries were in the range of 94–105% for all the determinations. This method has also been applied to determine various arsenic compounds in two fish samples. In this study, a simple and rapid microwave-assisted extraction method was used for the extraction of arsenic compounds from fish. The arsenic species were quantitatively leached with an 80% v/v methanol solution in a focused microwave field during a period of 5 min.     

Comparison of AFS and ICP-MS detection coupled with gas chromatography for the determination of methylmercury in marine samples

Inductively coupled plasma mass spectrometry (ICP-MS) and atomic fluorescence spectrometry (AFS) coupled with gas chromatography (GC) have been evaluated as element specific detectors for the determination of methylmercury in marine samples. Detection limits for methylmercury chloride, obtained using ICP-MS and AFS, were 0.9 and 0.25 pg as Hg, respectively. Methylmercury was determined in marine tissue reference materials IAEA 142 and NIST 8044 mussel homogenate, and DOLT-2 dogfish liver by GC–AFS, with found values of 45±7, 26±4, and 671±41 ng g⁻¹, compared with certified values of 47±4, 28±2, and 693±53 ng g⁻¹. The analyses of IAEA 142 and NIST 8044 were repeated using GC–ICP-MS, with found values of 48±9 and 30±3 ng g⁻¹, respectively. Methylmercury was determined in real samples of ringed seal and beluga whale, with found values of 801±62 and 2830±113 ng g⁻¹, respectively.     

Determination of alkylphenol and bisphenol A in beverages using liquid chromatography/electrospray ionization tandem mass spectrometry

A comprehensive analytical method based on liquid chromatography electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS) with negative ionization mode has been developed for measuring of alkylphenols and bisphenol A in beverage samples. Concentration and clean up of samples were performed on 200 mg OASIS HLB solid extraction cartridges. The effects of mobile phases and additives on ionization were assessed. The recoveries for each compound ranged from 76.7 to 96.9% and reproducibilities were represented as having relative standard deviation (R.S.D.) below 10%. The limits of quantification (LOQ) of the method under multiple-reaction monitoring (MRM) acquisition mode were 0.04, 0.03 and 0.2 ng L⁻¹ for 2 L of mineral drinking water and 2.0, 1.8 and 8.0 ng L⁻¹ for 50 mL of soda beverages.     

Analysis for chloroanisoles and chlorophenols in cork by stir bar sorptive extraction and gas chromatography–mass spectrometry

A complete methodology for the determination of chloroanisoles and chlorophenols in cork material is proposed. The determination is accomplished by means of a previous liquid–solid extraction followed by stir bar sorptive extraction (SBSE) coupled to gas chromatography–mass spectrometry (GC–MS). Two different liquid–solid extraction experiments were conducted and eight compounds considered (2,6-dichloroanisole, 2,4-dichloroanisole, 2,4,6-trichloroanisole, 2,4,6-trichlorophenol, 2,3,4,6-tetrachloroanisole, 2,3,4,6-tetrachlorophenol, pentachloroanisole and pentachlorophenol). From the results obtained we can conclude that high volume extraction extending extraction time up to 24 h is the best choice if we have to release compounds from the inner surfaces of cork stoppers. Recovery percentages ranged from 51% for pentachloroanisole to 81% for 2,4-dichloroanisole. This method allows the determination of an array of compounds involved in cork taint at very low levels from 1.2 ng g⁻¹ for 2,4,6-tricholoroanisole to 23.03 ng g⁻¹ for 2,3,4,6-tetrachlorophenol.     

Extraction and determination of papaverin in pericarpium papaveris using aqueous two-phase system of poly(ethylene glycol)-(NH4)2SO4 coupled with high-performance liquid chromatography

Based on aqueous two-phase system (ATPS) of poly(ethylene glycol) (PEG)–(NH₄)₂SO₄, a simple pretreatment approach was developed for the extraction and determination of papaverin in pericarpium papaveris. The influence factors on phase behavior of the ATPS and partition behavior of papaverin was investigated, and partition mechanism based on the hydrophobic interaction between PEG and analyte molecules was proposed. Under the optimal conditions, the extraction efficiencies for papaverin were 93–96%, and the recoveries of the added standard were 97–106% with relative standard deviations of 1.8–2.5%. Combined with a high-performance liquid chromatography (HPLC) method, this extraction technique has been successfully applied to the determination of papaverin in pericarpium papaveris with the detection limit of 2 ng mL⁻¹ and the linear range of 0.10–10 μg mL⁻¹. Compared with the conventional liquid–liquid extraction or solid-phase extraction, this method was more environmentally benign, more cost effective and much simpler due to the direct injection of the upper phase into HPLC system.     

Selected ion storage versus tandem MS/MS for organochlorine pesticides determination in drinking waters with SPME and GC-MS

The use of two modes for mass spectrometry (MS) detection with an ion trap instrument, selected ion storage (SIS) and tandem mass spectrometry (MS/MS), are compared for the solid-phase microextraction (SPME)–gas chromatography (GC) coupled to mass spectrometry (GC-MS) determination of 16 priority organochlorine pesticides (OCPs) in drinking water samples at the ultratrace levels (ng?L^?1) required by official guidelines in the European legislation. Experimental parameters investigated for the SPME sample preparation were: the type of coating (100?µm polydimethylsiloxane, PDMS, and 65?µm poly(dimethylsiloxane)–divinylbenzene, PDMS/DVB), SPME modality, extraction and desorption times and desorption temperature and the methanol percentage in the SPME working solution. Under the calculated optimal conditions two methodologies were developed, one for SIS and the other for MS/MS modes. The detection limits, precision and accuracy were evaluated for both alternatives and were appropriate to the official guidelines requirements. The SPME–GC-MS(SIS) methodology offered LODs from 0.2–6.6?ng?L^?1, precision below 13% and recoveries between 83 and 110%. The SPME–GC–MS/MS methodology provided limits of detection (LODs) ranging from 0.3 to 7.6 ng?L^?1, % RSD were ≤14% and recoveries of 79–108% were achieved. After the results observed within an Interlaboratory Exercise, the latest MS methodology was selected for the pursued analysis in real drinking water samples. Also, the good results in this round-robin exercise validate the proposed SPME–GC–MS/MS methodology.     

Flow-through optosensor combined with photochemically induced fluorescence for simultaneous determination of binary mixtures of sulfonamides in pharmaceuticals, milk and urine

A sensitive and selective flow-through optosensor implemented with photochemically induced fluorescence (PIF) is proposed for the simultaneous determination of mixtures sulfamethoxazole/sulfanilamide and sulfathiazole/sulfanilamide. The resolution was accomplished by placing in the flow system a minicolumn filled with an appropriate solid support. Whereas one of the sulfonamides is not retained in the minicolumn and is determined by measuring its native fluorescence on the solid surface of the sensing microbeads in the detection area, the other one is retained and, after its elution, it is photochemically converted into a strongly fluorescent photoproduct which is transitorily retained on the sensing support in the flow cell and monitored. Linear calibration graphs were obtained over a concentration range of 2–3 orders of magnitude. The detection limits for the determination of sulfamethoxazole, sulfanilamide and sulfathiazole are 8.1, 2.9 and 5.7 ng mL⁻¹, respectively. The method was applied to pharmaceuticals, milk and human urine. The recovery of sulfamethoxazole from pharmaceuticals was 102.5% indicating no interference from trimethoprim which is not photochemically active. The recoveries for urine and milk samples fortified with sulfonamides at levels between 0.1 and 0.7 μg mL⁻¹ agreed within 95.0–107.5% of spiked levels.     

Separation and determination of amitriptyline and nortriptyline by dispersive liquid-liquid microextraction combined with gas chromatography flame ionization detection

Dispersive liquid–liquid microextraction (DLLME) coupled with gas chromatography–flame ionization detection (GC–FID) was applied for the determination of two tricyclic antidepressant drugs (TCAs), amitriptyline and nortriptyline, from water samples. This method is a very simple and rapid method for the extraction and preconcentration of these drugs from environmental sample solutions. In this method, the appropriate mixture of extraction solvent (18 μL Carbon tetrachloride) and disperser solvent (1 mL methanol) are injected rapidly into the aqueous sample (5.0 mL) by syringe. Therefore, cloudy solution is formed. In fact, it is consisted of fine particles of extraction solvent which is dispersed entirely into aqueous phase. The mixture was centrifuged and the extraction solvent is sedimented on the bottom of the conical test tube. 2.0 μL of the sedimented phase is injected into the GC for separation and determination of TCAs. Some important parameters, such as kind of extraction and disperser solvent and volume of them, extraction time, pH and ionic strength of the aqueous feed solution were optimized. Under the optimal conditions, the enrichment factors and extraction recoveries were between 740.04–1000.25 and 54.76–74.02%, respectively. The linear range was (0.005–16 μg mL⁻¹) and limits of detection were between 0.005 and 0.01 μg mL⁻¹ for each of the analytes. The relative standard deviations (R.S.D.) for 4 μg mL⁻¹ of TCAs in water were in the range of 5.6–6.4 (n = 6). The performance of the proposed technique was evaluated for determination of TCAs in blood plasma.     

Determination of polybrominated diphenyl ethers in human milk samples in the Czech Republic: Comparative study of negative chemical ionisation mass spectrometry and time-of-flight high-resolution mass spectrometry

In the Czech Republic no study on the levels of brominated flame retardants in human milk has been conducted, yet. In the first step analytical method for determination of PBDEs in this bioindicator matrix was implemented. Liquid–liquid extraction (LLE) (hexane, diethyl ether), followed by gel permeation chromatography was employed for isolation of PBDEs. Identification and quantification of PBDEs was carried out by GC–MS operated in negative chemical ionisation (NCI). Two mass spectrometric technologies, one employing quadrupole and the other one high resolution (HR) time-of-flight (TOF) analyzer, etc. were used in our study. Detection limits (LODs) obtained by quadrupole analyzer ranged from 0.02 to 0.05 ng g⁻¹ lipid weight, using high resolution time-of-flight analyzer LODs were significantly lower, ranging from 0.002–0.005 ng g⁻¹ lipid weight, what enabled detection of minor PBDE congeners.

Within this pilot study 103 breast milk samples, obtained from mothers living in Olomouc region, were examined. Ten PBDE congeners were determined. All samples examined till now contained PBDEs residues, the dominating contaminant representing this group was congener BDE 47. In most of analysed samples levels of this compound ranged from 0.2 to 2 ng g⁻¹ of lipid weight. Three exceptionally contaminated samples, containing levels of PBDEs 5–10 times higher than other samples, were found.     

Analysis of acrylamide in different foodstuffs using liquid chromatography–tandem mass spectrometry and gas chromatography–tandem mass spectrometry

Acrylamide levels over a wide range of different food products were analysed using both liquid chromatography–tandem mass spectrometry (HPLC–MS–MS) and gas chromatography–tandem mass spectrometry (GC–MS–MS). Two different sample preparation methods for HPLC–MS–MS analysis were developed and optimised with respect to a high sample throughput on the one hand, and a robust and reliable analysis of difficult matrices on the other hand. The first method is applicable to various foods like potato chips, French fries, cereals, bread, and roasted coffee, allowing the analysis of up to 60 samples per technician and day. The second preparation method is not as simple and fast but enables analysis of difficult matrices like cacao, soluble coffee, molasses, or malt. In addition, this method produces extracts which are also well suited for GC–MS–MS analysis. GC–MS–MS has proven to be a sensitive and selective method offering two transitions for acrylamide even at low levels up to 1 μg kg⁻¹. For the respective methods the repeatability (n=10), given as coefficient of variation, ranged from 3% (acrylamide content of 550 μg kg⁻¹) to 12% (acrylamide content of 8 μg kg⁻¹) depending on the food matrix. The repeatability (n=3) for different food samples spiked with acrylamide (5–1500 μg kg⁻¹) ranged from 1 to 20% depending on the spiking level and the food matrix. The limit of quantification (referred to a signal-to-noise ratio of 9:1) was 30 μg kg⁻¹ for HPLC–MS–MS and 5 μg kg⁻¹ for GC–MS–MS. It could be demonstrated that measurement uncertainties were not only a result of analytical variability but also of inhomogeneity and stability of the acrylamide in food.     

Gas chromatographic determination of aromatic amines in water samples after solid-phase extraction and derivatization with iodine II. Enrichment

A procedure for the enrichment of aromatic amines via solid-phase extraction was developed. A HR-P phase based on styrene–divinylbenzene was used for the investigations, generally followed by derivatization with iodine and determination via GC–ECD. The recoveries of 53 aromatic amines in a drinking water matrix at pH 9 were determined. Most anilines showed relative recoveries between 80–120% with relative standard deviations of ≤5% at concentration levels between 10 and 20 μg l⁻¹. The comparison with a wastewater matrix led to similar results. The enrichment procedure was applied to real samples, e.g., samples of ammunition wastewater.     

Phthalates determination in physiological saline solutions by HPLC-ES-MS

Phthalates are a group of chemical compounds with increasing interest from the analytical point of view. The risks for human health associated with some of these compounds have unleashed the necessity to develop analytical methods with great sensitivity that allow us to detect their presence at trace levels in order to assure protection for the population.

A simple and rapid method for determining a group of phthalate esters in aqueous samples was developed. The method was based on high-performance liquid chromatography–(electrospray)-mass spectrometry (HPLC–ES-MS), working in positive ionisation (PI) mode. A gradient elution was performed with acetonitrile–ultrapure water starting from 5 to 75% acetonitrile in 5 min followed by isocratic elution during 5 min. Standard calibration curves were linear for all the analytes over the concentration range 10–500 ng mL⁻¹ .The LOD values found for DMP, DEP, BBP and DBP were 0.8, 3.4, 0.6 and 1.2 ng mL⁻¹ respectively. The relative standard deviation ranged from 0.8 to 1.7%, which indicated good method precision.

The proposed analytical method has been applied to the analysis of commercial physiological saline solutions in order to check the presence of phthalates and to determine their concentration.     

Determination of cadmium in paint samples by graphite furnace atomic absorption spectrometry with optical temperature control

A graphite furnace atomic absorption spectrometry (GFAAS) method with optical temperature control for the determination of trace cadmium in paint samples is described. Optical temperature control was superior in many respects to current temperature control. The sensibility increased by 60%, the linear range widened by 60%, and the life of graphite tube showed a 200–300% increase because atomization temperature was lowered distinctly and atomization time was shortened. Use of lanthanum chloride as a matrix modifier was investigated. The linear range of calibration curve was 0–24 ng mL⁻¹. The detection limit was 9.6 ng L⁻¹. The characteristic mass was 3.0 pg. The method also resulted in excellent reproducibility (≤2.5% R.S.D.) at such low levels, and the recovery of added cadmium in paint samples was from 94.6% to 102%. This method is readily applicable to the determination of cadmium in paint samples.     

Efficiency and mechanism of new poly(acryl-phenylamidrazone phenylhydrazide) chelating fiber for adsorbing trace Ga, In, Bi, V and Ti from solution

A new po1y(acrylphenylamidrazone phenylhydrazide) chelating fiber is synthesized from polyacrylonitrile fiber and used for preconcentration and separation of trace Ga(III), In(III), Bi(III), V(V) and Ti(IV) from solution (5–50 ng ml⁻¹ Ti(IV) or V(V) and 50–500 ng ml⁻¹ Ga(III), In (III) or Bi(III) in 1000–100 ml of solution can be enriched quantitatively by 0.15 g of fiber at a 4 ml min⁻¹ flow rate in the pH range 5–7 with recoveries >95%). These ions can be desorbed quantitatively with 20 ml of 4 M hydrochloric acid at 2 ml min⁻¹ from the fiber column. When the fiber which had been treated with concentrated hydrochloric acid and washed with distilled water until neutral was reused eight times, the recoveries of the above ions by enrichment were still >95%. Two-hundred-fold to 10 000-fold excesses of Cu(II), Zn(II), Ca(II), Mn(II), Cr(III), Fe(III), Ba(II) and Al(III) caused little interference in the determination of these ions by inductively coupled plasma-atomic emission spectrometers (ICP-AES). The relative standard deviations for enrichment and determination of 50 ng ml⁻¹ Ga, In or Bi and 10 ng ml⁻¹ V or Ti are in the range 1.2–2.7%. The contents of these ions in real solution samples determined by this method were in agreement with the certified values of the samples with average errors <3.7%.     

Applications of solid-phase microextraction in food analysis

Food analysis is important for the evaluation of the nutritional value and quality of fresh and processed products, and for monitoring food additives and other toxic contaminants. Sample preparation, such as extraction, concentration and isolation of analytes, greatly influences the reliable and accurate analysis of food. Solid-phase microextraction (SPME) is a new sample preparation technique using a fused-silica fiber that is coated on the outside with an appropriate stationary phase. Analyte in the sample is directly extracted to the fiber coating. The SPME technique can be used routinely in combination with gas chromatography (GC), GC–mass spectrometry (GC–MS), high-performance liquid chromatography (HPLC) or LC–MS. Furthermore, another SPME technique known as in-tube SPME has also been developed for combination with LC or LC–MS using an open tubular fused-silica capillary column as an SPME device instead of SPME fiber. These methods using SPME techniques save preparation time, solvent purchase and disposal costs, and can improve the detection limits. This review summarizes the SPME techniques for coupling with various analytical instruments and the applications of these techniques to food analysis.     

Simultaneous determination of three bufadienolides in rat plasma after intravenous administration of bufadienolides extract by ultra performance liquid chromatography electrospray ionization tandem mass spectrometry

A novel, rapid and specific ultra performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) method has been developed for simultaneous determination and pharmacokinetic studies of three bufadienolides (bufalin, cinobufagin and resibufogenin) in rat plasma. The analytes, bufalin, cinobufagin, resibufogenin and the internal standard, diphenhydramine were extracted from rat plasma samples by a one-step liquid–liquid extraction and separated on an ACQUITY UPLC™ BEH C₁₈ column with gradient elution using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.20 mL min⁻¹. Detection was carried out on a triple-quadrupole tandem mass spectrometer in the multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) interface. The three bufadienolides could be simultaneously determined within 3.0 min. Linear calibration curves were obtained over the concentration ranges of 1.0–200 ng mL⁻¹ for all the analytes. The intra- and inter-day precisions (relative standard deviation (R.S.D.)) were less than 11.35 and 10.87%, respectively. The developed method was applied for the first time to the pharmacokinetic studies of bufadienolides in rats following a single intravenous administration of 2.10 mg kg⁻¹ bufadienolides.     

A new immune resonance scattering spectral assay for trace fibrinogen with gold nanoparticle label

An on-line stacking method based on moving reaction boundary (MRB) was developed for the sensitive determination of barbital and phenobarbital in human urine via capillary electrophoresis (CE). The optimized conditions for the method are: 60 mmol L⁻¹ pH 11.0 Gly–NaOH as the background electrolyte, 10 mmol L⁻¹ pH 5.5 Gly–HCl as sample buffer, secobarbital as the internal standard (IS), 12.5 kV, 1.4 psi 10 s sample injection, 75 μm ID 60.2 cm total length (50 cm effective length) capillary and 214 nm detect wavelength. Under the optimized conditions, the method can well stack and separate barbital and phenobarbital in urine samples and result in 20.5-fold and 22.6-fold improvement in concentration sensitivity for barbital and phenobarbital, respectively. Furthermore, the method holds: (1) good linear calibration functions for the two target compounds (correlation coefficients r > 0.999), (2) low limits of detection (0.27 μg mL⁻¹ for barbital and 0.26 μg mL⁻¹ for phenobarbital), (3) low limits of quantification (0.92 μg mL⁻¹ for barbital and 0.87 μg mL⁻¹ for phenobarbital), (4) good precision (R.S.D. of intra-day and inter-day less than 5.38% for barbital and 1.67% for phenobarbital, respectively) and (5) high recoveries at three concentration levels (90.27–106.36% for barbital and 93.05–113.60% for phenobarbital in urine). The method is simple, sensitive and efficient, and can fit to the need of clinical and forensic toxicology.     

Comparative study of the instrumental couplings of high performance liquid chromatography with microwave-assisted digestion hydride generation atomic fluorescence spectrometry and inductively coupled plasma mass spectrometry for chiral speciation of selenomethionine in breast and formula milk

A comparison of chiral separation and analysis of selenomethionine in breast and formula milk, using high performance liquid chromatography (HPLC) on a glycopeptide teicoplanin-based chiral stationary phase (Chirobiotic T), coupled to atomic fluorescence spectrometry (AFS) and inductively coupled plasma (ICP) MS detectors has been performed. The coupling HPLC-microwave-assisted digestion hydride generation requires on-line post-column analytes treatment, and a severe sample clean-up for fat and proteins elimination using centrifugation and ultrafiltration. Underivatized -selenomethionine enantiomers were completely resolved in 10 min using unbuffered water mobile phase at 1 ml min⁻¹ flow. Good selectivity and sensitivities (detection limits 3.1 and 3.5 ng ml⁻¹ as Se for - and -selenomethionine, respectively) were obtained, and method robustness and simplicity, together to the low cost of AFS detector, makes it suitable for infant milk routine analysis. HPLC–ICP-MS coupling exhibits very low detection limits (0.9 ng ml⁻¹, as Se) for each -selenomethionine enantiomers, but the method suffers from matrix influence, that produces a poor S/N ratio and low reliability.

The methods were applied to breast and formula milk samples with recoveries of 80% of the total selenium presence, which is attributable to the existence of other unknown species. -Selenomethionine was the only isomer present in breast milk, but a 30% of -selenomethionine was also detected in formula milk.