High-Performance Liquid Chromatography with Fluorescence Detection for Simultaneous Analysis of Retinoids (Retinyl Palmitate,Retinyl Acetate,and Free Retinol) and α-, β-, γ-, and δ-Tocopherols in Foods

A method based on high-performance liquid chromatography with fluorescence detection for the simultaneous analysis of retinoids (vitamin A) and tocopherols (vitamin E) was developed. This method consists of an isocratic solution using hexane/ethyl acetate (85:15, v/v) as the mobile phase and fluorescence detection using a time program that sets the excitation (Ex) and emission (Em) wavelengths at adequate elution times for retinoids (Ex 342 nm, Em 476 nm) and tocopherols (Ex 298 nm, Em 325 nm), respectively. The separation of three retinoids (retinyl palmitate, retinyl acetate, and free retinol) and four tocopherol homologs was achieved with sufficient reproducibility and quantitative ability. Additionally, the necessity of saponification was considered. As a result, saponification was not used in this method because of the complexity of the procedure and the loss of free retinol. The retinoid and tocopherol contents of various foods were evaluated using the developed method. Our method could evaluate the retinoid and tocopherol contents of fish (eel, Anguilla japonica, and amberjack, Seriola dumerili) muscle and liver, roasted soybean (Glycine max) flour, and Japanese torreya seed (Torreya nucifera). Additionally, our method could be applied to the determination of retinoids and tocopherols not only in foods but also in supplements and cosmetics.     

Simultaneous Determination of Squalene, α-Tocopherol and β-Carotene in Table Olives by Solid Phase Extraction and High-Performance Liquid Chromatography with Diode Array Detection

Olives, the fruit of the Olea europaea tree, are highly appreciated in olive oil and table olives (20 % of crops) not only for their flavor but also for their nutritional properties, especially for antioxidant compounds such as squalling (SQ), α-tocopherol (TH) and β-carotene (BC). This paper presents a new analytical method for simultaneously determining SQ, TH and BC in table olives by using solid phase extraction (SPE) and high performance-liquid chromatography with diode array detection (HPLC-DAD), avoiding the classic saponification process. The correlation coefficients of calibration curves of the analyzed compounds ranged from 0.998 to 0.999, and the recoveries were in the range of 89.4–99.6 %. The validated method was used to analyze 30 table olive samples from Italy for their content of SQ (537–1,583 mg kg^?1), TH (21–90 mg kg^?1) and BC (0.4–2.6 mg kg^?1). Finally, experiments with HPLC-MS were conducted to compare this novel method with the classic saponification procedure.     

Validation of an Electrochemical Detection–High-Performance Liquid Chromatography Method for Simultaneous Determination of Lignans in Flaxseed (Linum usitatissimum L.)

Flaxseed is a major source of lignans, which are important bioactive compounds. The aims of this work were to validate a liquid chromatographic method for the rapid and simultaneous determination of the main lignans in flaxseed using high-performance liquid chromatography coupled with electrochemical detection and to analyze the composition of commercial samples of flaxseed. The performance criteria of the proposed method demonstrate that the method can be used for the analysis of secoisolariciresinol diglucoside (SDG), secoisolariciresinol (SECO), matairesinol (MATA), pinoresinol (PINO), lariciresinol (LARI), hydroxymatairesinol (HYDROXY), and isolariciresinol (ISOLARI) in flaxseeds at suitable levels. Calibration curves were determined for six different concentrations of standard solutions injected in triplicate. The sensitivity of the calibration curve was evaluated considering the confidence intervals of the intercept and slope. The limit of detection and limit of quantification were 7.4 and 10.9 μg/l, respectively, for LARI and 17.7 and 37.5 μg/l, respectively, for MATA. The relative standard deviation of repeatability values were lower than 2.59 %, which are acceptable because the Horwitz ratio values were 0.1 for all of the lignans. The recoveries of lignans were in the range of 74–100 % of SECO, which are consistent with the literature. The precision of the proposed method was determined by analyzing four flaxseed samples of different years and varieties. SDG was the main lignan present in all the samples, followed by ISOLARI and HYDROXY.     

Reversed-Phase High-Performance Liquid Chromatography–Fluorescence Detection for the Analysis of Glutathione and Its Precursor γ-Glutamyl Cysteine in Wines and Model Wines Supplemented with Oenological Inactive Dry Yeast Preparations

A reversed-phase high-performance liquid chromatography–fluorescence detection methodology involving a pre-column derivatization procedure using 2,3-naphtalenedialdehyde in the presence of 5 and 0.5 mM of dithiothreitol to determine total and reduced glutathione (GSH) and γ-glutamyl-cysteine (γ-glu-cys) in musts and wines has been set up and validated. The proposed method showed good linearity (R ² >99% for reduced and total GSH, and R ² >98% for γ-glu-cys) in synthetic wines, over a wide range of concentration (0–10 mg L⁻¹). The limits of detection for reduced GSH in synthetic and real wines were almost the same (0.13 and 0.15 mg L⁻¹, respectively) and slightly higher for γ-glu-cys (0.24 mg L⁻¹). The application of the method allowed knowing, for the first time, the amount of total and reduced GSH and γ-glu-cys released into synthetic wines by oenological preparations of commercial inactive dry yeast (IDY). In addition, the evolution of these three compounds during the winemaking and shelf life (0–9 months) of an industrially manufactured rosé wine supplemented with a GSH-enriched IDY showed that although GSH is effectively released from IDY, it is rapidly oxidized during alcoholic fermentation, contributing to the higher total GSH content determined in wines supplemented with GSH-enriched IDYs compared to control wines.     

Analysis of Pesticides in Tomato Combining QuEChERS and Dispersive Liquid–Liquid Microextraction Followed by High-Performance Liquid Chromatography

A new sample preparation procedure combining QuEChERS and dispersive liquid–liquid microextraction (DLLME) was optimized for the determination at trace levels of 13 pesticides from different chemical families (i.e. 2,4-D, acetamiprid, bentazone, cymoxanil, deltamethrin, dicamba, diuron, foramsulfuron, mesotrione, metalaxyl-M, methomyl, pyraclostrobin and tembotrione) in tomato by high-performance liquid chromatography with diode array detection. Target pesticides from tomato samples were isolated by liquid partitioning with acetonitrile and salts and cleaned up by dispersive solid-phase extraction (d-SPE); the analytes were concentrated in trichloromethane by the DLLME procedure. The disperser solvent from DLLME was used at the same time as carrier of analytes form extraction in QuEChERS method. The main factors affecting sample cleanup by d-SPE in QuEChERS and DLLME yield were optimized by means of an experimental design. Under the optimum conditions, good linearity was obtained, the recoveries of pesticides in tomato samples at spiking levels between 0.01 and 1.00 mg/kg ranged from 86 to 116 % (for foramsulfuron and cymoxanil, respectively). Precision was within 15.0 % (RSD) except at the LQ for tembotrione, which was 17.4 %. Limits of quantification achieved (ranging from 0.0058 to 0.15 mg/kg) were below the maximum residue limits established by the European Union.     

Simultaneous Determination of 18 Bioactive Compounds in Italian Bitter Liqueurs by Reversed-Phase High-Performance Liquid Chromatography–Diode Array Detection

A simple, fast and accurate method has been developed to simultaneously determine 18 bioactive compounds in Italian bitter liqueurs containing gentian, cinchona, cinnamon, rhubarb, clove, star anise or orange, by reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with diode array detection (DAD). HPLC analysis was performed with a C18 column using methanol and aqueous phosphoric acid (pH 2.5) as mobile phase. Selected wavelengths, i.e. 210, 232, 275, 285, 291, 310 and 368 nm, were used for quantification of compounds. The column temperature was controlled at 30 °C. The correlation coefficients (R ²) of the calibration curves of the analysed compounds were ≥0.9999 in a relatively wide concentration range (0.5–50 μg/ml). The proposed method proved successful in simultaneously analysing 18 bitter liqueurs produced in Italy. The concentration of the most important bitter principles, gentiopicroside, amarogentin, quinine and naringin, ranged as follows: 1.17–299.20, 0.25–32.24, 1.44–6.93 and 0.28–39.99 μg/ml, respectively.     

Surfactant–Solvent-Based Quaternary Component Emulsification Microextraction Followed by High-Performance Liquid Chromatography for the Simultaneous Analysis of Benzimidazole Anthelmintics in Milk Samples

A simple surfactant-solvent-based quaternary component emulsification microextraction (SSEME) method combined with high-performance liquid chromatography–photodiode array detection has been developed for the extraction, preconcentration, and determination of four benzimidazole anthelmintic (i.e., oxfendazole, mebendazole, albendazole, and fenbendazole) residues in milk samples. The quaternary component solvent of SSEME carried out in 10 mL aqueous solution were Triton X-114 (emulsifier or carrier), acetonitrile (disperser solvent), and 1-octanol (extraction solvent). The surfactant has an important role in the enhancement of the extraction efficiency of the high polar analytes. For milk sample analyses, linearity was obtained in the range of 10–200 μg/L with the determination coefficients (R ²) higher than 0.996. Preconcentration factor was obtained in the range of 21–38, corresponding to limits of detection in the range of 2.6–9.9 μg/L. Intra-day (n?=?6) and inter-day (n?=?6?×?3) precisions in the sample studied were obtained with relative standard deviation below 8.8 %. The recoveries for the spiked target anthelmintics at different concentrations (25, 50, 100, and 150 μg/L) were obtained in the range 80.1–114.1 %. The proposed SSEME method has been demonstrated that is simple, effective, and reliable for the analysis of analytes in the samples studied and can be used as an alternative green analytical technique for benzimidazole analysis.     

Detection of Adulterants and Contaminants in Liquid Foods—A Review

Milk and fruit juices have paramount importance in human diet. Increasing demand of these liquid foods has made them vulnerable to economic adulteration during processing and in supply chain. Adulterants are difficult to detect by consumers and thus necessitating the requirement of rapid, accurate and sensitive detection. The potential adulterants in milk and fruit juices and their limits set by different regulatory bodies have been briefly described in this review. Potential advantages and limitations of various techniques such as physicochemical methods, chromatography, immunoassays, molecular, electrical, spectroscopy with chemometrics, electronic nose, and biosensors have been described. Spectroscopy in combination with chemometrics has shown potential for rapid, precise, and sensitive detection of potential adulterants in these liquid foods.     

Gas Purge–Microsyringe Extraction Coupled with Liquid Chromatography and Fluorescence Detection for the Determination of Aldehydes from Fried Meat

In this study, a green, simple, and sensitive method was developed for the analysis of aliphatic aldehydes from fried meat by using a modified gas purge–microsyringe extraction (GP–MSE) system in combination with high-performance liquid chromatography (HPLC) with fluorescence detection. The modified GP–MSE system possessed two gas channels and showed better recoveries for compounds with diverse density in comparison with one gas channel GP–MSE system. Target compounds in fried meat were effectively extracted without the traditional solvent extraction and lipid removing process, while the HPLC sensitivity of aldehydes was enhanced by introducing 2-(12-benzo[b]acridin-5(12H)-yl)-acetohydrazide (BAAH) with excellent fluorescence property into the molecules. Parameters influencing the extraction efficiency and HPLC sensitivity were optimized. The limits of detection (LODs) ranged from 0.30 to 0.45 μg/kg, and the limits of quantification (LOQs) ranged from 1.0 to 1.5 μg/kg. The recoveries of the target compounds were in the range of 86.9 to 95.6%. The proposed method was successfully applied to the analysis of aldehydes in fried meat samples. Formaldehyde, acetaldehyde, pentanal, hexanal, heptanal, octanal, nonaldehyde, and decanal were all found in fried meat samples with concentrations ranging from 0.05 to 17.8 mg/kg.     

Novel Binary Solvents-Dispersive Liquid—Liquid Microextraction (BS-DLLME) Method for Determination of Patulin in Apple Juice Using High-Performance Liquid Chromatography

A simple and rapid binary solvents-based dispersive liquid–liquid microextraction (BS-DLLME) method has been developed for determination of patulin (PAT) in apple juice followed by high-performance liquid chromatography. This method involves the use of an appropriate mixture of miscible binary extraction solvents and disperser solvent to form fine droplets of extractant in a sample solution. Parameters affecting extraction efficiency such as the type and volume of high-density extraction solvent, the volume of ethyl acetate, the kind and volume of disperser solvent, and salt addition were investigated and optimized. The detection and quantification limits were 2.0 and 10.0 μg L^?1, respectively. The relative standard deviation for five measurements of 25 μg L^?1 of PAT was 3.8 %. The relative recoveries of PAT from apple juice samples at spiking levels of 25, 50, and 75 ng mL^?1 were in the range of 91.3–95.2 %.     

Determination of Pydiflumetofen Residues in Some Foods of Plant and Animal Origin by QuEChERS Extraction Combined with Ultra-Performance Liquid Chromatography–Tandem Mass

A rapid and effective analytical method for determination of pydiflumetofen residues in some foods of plant and animal origin (grapes, tomatoes, wheat, pork, milk, and eggs) was developed using a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation procedure followed by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS). Acetonitrile was served as the extraction solvent, and an octadecylsilane-dispersive solid-phase extraction (C18-dSPE) was used to cleanup the analyte, and then detected by UPLC–MS/MS. Pydiflumetofen was eluted within 3.0 min from the HSS T3 chromatography column connected to an electrospray ionization source in positive mode. The linearity of the method was excellent (R²?≥?0.992) in the pydiflumetofen concentration range of 10–1000 μg kg^?1. The recoveries of spiked pydiflumetofen (10, 100, and 1000 μg kg^?1) from the matrices were satisfactory, being between 72.0 and 110.3%, and all with relative standard deviation values of <?15.1%. The limit of quantification for pydiflumetofen was 10 μg kg^?1. This study provides a method for the routine monitoring of pydiflumetofen.     

Simultaneous Determination of 15 Plant Growth Regulators in Bean Sprout and Tomato with Liquid Chromatography–Triple Quadrupole Tandem Mass Spectrometry

The present paper describes the development and validation of a new reversed-phase liquid chromatography–electrospray ionization tandem mass spectrometric method for determination of 15 plant growth regulators in bean sprout and tomato. Plant growth regulators were extracted with mixed solvents methanol/acetonitrile/water/acetic acid (v/v/v/v, 40/40/20/1) from 10 g vegetable and analyzed directly without further cleanup. The identification of each analyte was established by chromatographic retention time, analyte-specific fragmentation patterns, and relative peak area ratios of precursor/product ion pairs. The recoveries of 15 plant growth regulators in bean sprout and tomato were 70.3–110.0 %. The repeatability of this method is excellent, and relative standard deviation value is less than 10 %.     

Analysis of α-Cryptoxanthin, β-Cryptoxanthin, α -Carotene,and β-Carotene of Pandanus Conoideus Oil by High-performance Liquid Chromatography (HPLC)

Pandanus conoideus is an endemic plant of Papua, Indonesia, reported to be very rich in carotenoids. The purpose of this study was to develop method for the determination of carotenoids (α-cryptoxanthin, β-cryptoxanthin, α-carotene and β-carotene) in P. conoideus oil (PO) by high-performance liquid chromatography (HPLC). sing the proposed method in this research, carotenoids content of nine clones of PO were analyzed which ranged from 5.4-138.5 ng/mg for α-cryptoxanthin, 3.9-29.4 ng/mg for β-cryptoxanthin, 3.5-80.0 ng/mg for α-carotene, and 10.8-118.0 ng/mg for β-carotene. Our results showed that four carotenoids content was very small as compared to total carotenoids content (3027-19959 ng/mg). This suggests that those four carotenoids were not a major component of the PO carotenoids. Using the principal component analysis, nine clones of P. conoideus can be grouped based on the proximity of its carotenoid content into group A (Monsor, Mbarugum, Himbiak, Monsrus and Memeri), group B (Menjib Rumbai), and group C (Edewewits, Hibcau and Hityom).     

Aluminium and Its Complexes in Teas and Fruity Brew Samples,Speciation and Ions Determination by Ion Chromatography and High-Performance Liquid Chromatography–Fluorescence Analytical Methods

The paper presents results of aluminium determination in samples of black and fruit teas. Total aluminium concentration was determined along with the concentration of aluminium in a cup of tea in tea samples in two price groups (>1€ and <1€). Based on the conducted study, no differences were found in aluminium concentration in black and fruit teas depending on the price group. Developed ion chromatography method was applied to determine inorganic and organic ions in tea samples, especially those which may form complexes with aluminium: fluoride, sulphate, oxalate and citrate ions. Analysis by this method using ion chromatography allowed for the determination of 12 anions: F^?, HCOO^?, CH₃COO^?, NO₂ ^?, Br^?, NO₃ ^?; Cl^?, CH₂(COO)₂ ^2?, SO₄ ^2?, C₂O₄ ^2?, PO₄ ^3? and C₃H₅O(COO)₃ ^3? in the time of 40 min. Speciation analysis of aluminium was performed in optimised HPLC-fluorescence analytical system (with Lumogallion as a post-column reagent). It was observed that organic aluminium complexes are quickly degraded to form Al³⁺ which is the reason why speciation analysis in tea samples does not provide the full image of speciation distribution. Nevertheless, this developed method was successfully used in the determination of aluminium complexes with fluorides in tea samples.     

Ionic Liquid-Based Dispersive Liquid–Liquid Microextraction Following High-Performance Liquid Chromatography for the Determination of Fungicides in Fruit Juices

This paper described an ionic liquid-based dispersive liquid–liquid microextraction (IL-DLLME) combined with high-performance liquid chromatography (HPLC) method to determine fungicides in fruit juices. In this method, 1-hexyl-3-methyli-midazolium hexafluorophosphate (HMIMPF₆) was used as extraction solvent, which dispersed into the fruit juices under vigorously shaking with the vortex. The effects of experimental parameters, such as extraction solvent volume, disperser solvent and its volume, vortex time, centrifugation time, sample pH, on the extraction efficiency were investigated. Under the optimum conditions, the linear correlation coefficients ranged from 0.9902 to 0.9979 for concentration levels of 0.02–2 mg l^?1, the extraction recoveries were ranged 66.2–92.9 % except pyrimethanil (39.5–44.6 %), The relative standard deviations (RSDs; n?=?6) ranged from 2.2 % to 11.6 %, and the limits of detection (LODs) for the fungicides were between 3.1 and 10.2 μg l^?1. Two real samples including apple and grape juices, spiked at two concentration levels were analyzed and yielded recoveries ranging from 71.3–93.1 % and 65.4–87.7 %, respectively.     

Application and Optimization of Microwave-Assisted Extraction and Dispersive Liquid–Liquid Microextraction Followed by High-Performance Liquid Chromatography for the Determination of Oleuropein and Hydroxytyrosol in Olive Pomace

In the present study, a new method based on microwave-assisted extraction and dispersive liquid–liquid microextraction (MAE–DLLME) followed by high-performance liquid chromatography (HPLC) was proposed for the separation and determination of oleuropein (Ole) and hydroxytyrosol (HyT) from olive pomace samples. The effective factors in the MAE–DLLME process such as microwave power, extraction time, the type and volume of extraction, and dispersive solvents were studied and optimized with the aid of response surface methodology (RSM) based on a central composite design (CCD) to obtain the best condition for Ole and HyT extraction. At the optimized conditions, parameter values were 220 W microwave power, 12 min extraction time, 60 μL extracting solvent, and 500 μL dispersive solvent. The calibration graphs of the proposed method were linear in the range of 10–500,000 μg L^?1, with the coefficient of determination (R²) higher than 0.99 for Ole and HyT. Repeatability of the method, described as the relative standard deviation (RSD), was 4.12–5.63% (n?=?6). The limits of detection were 35 and 20 μg L^?1 for Ole and HyT, respectively. The recoveries of these compounds in the spiked olive pomace sample were from 93 to 98%. The proposed method, MAE–DLLME–HPLC–UV, was an accurate, rapid, and reliable method when compared with previous methods.     

Rapid Quantification of Soyasaponins I and βg in Italian Lentils by High-Performance Liquid Chromatography (HPLC)–Tandem Mass Spectrometry (MS/MS)

In this work, an innovative and fast analytical method for the quantification of soyasaponins I and βg in lentils has been developed. Samples were extracted using 70 % aqueous ethanol at room temperature and then injected into a high-performance liquid chromatography–tandem mass spectrometry system. The correlation coefficients of calibration curves of the analyzed compounds were ≥0.9997. The recoveries obtained by spiking the lentil samples with a standard mixture of soyasaponins I and βg at 50 and 100 mg l^?1 were in the range of 96–101 and 98–103 %, respectively. The validated method was applied to the analysis of 30 lentil samples from central Italy. Soyasaponins I and βg were present in these lentils in concentrations that ranged from 54 to 226 mg kg^?1 and from 436 to 1,272 mg kg^?1, respectively. Our data indicated that lentils cultivated in fields at intermediate altitudes (1,142–1,387 m) showed the highest levels of soyasaponins, a finding confirmed by principal component analysis.