Ultrasensitive SERS quantitative detection of antioxidants via diazo derivatization reaction and deep learning for signal fluctuation mitigation.

Synthetic antioxidants serve as essential protectors against oxidation and deterioration of edible oils, however, prudent evaluation is necessary regarding potential health risks associated with excessive intake. The direct adsorption of antioxidants onto conventional surface-enhanced Raman scattering (SERS) substrates is challenging due to the presence of phenolic hydroxyl groups in their molecular structures, resulting in weak Raman scattering signals and rendering direct SERS detection difficult. In this study, a diazo derivatization reaction was employed to enhance SERS signals by converting antioxidant molecules into azo derivatives, enabling the amplification of the weak Raman scattering signals through the strong vibrational modes induced by the N = N double bond. The resulting diazo derivatives were characterized using UV-visible absorption and infrared spectroscopy, confirming the occurrence of diazo derivatization of the antioxidants. The proposed method successfully achieved the rapid detection of three commonly used synthetic antioxidants, namely butylated hydroxyanisole (BHA), tert-butylhydroquinone (TBHQ), and propyl gallate (PG) on interfacial self-assembled gold nanoparticles. Furthermore, rapid predictions of BHA, PG, and TBHQ within the concentration range of 1 × 10-6 to 2 × 10-3 mol/L were achieved by integrating a convolutional neural network model. The predictive range of this model surpassed the traditional quantitative method of manually selecting characteristic peaks, with linear coefficients (R2) of 0.9992, 0.9997, and 0.9997, respectively. The recovery of antioxidants in real soybean oil samples ranged from 73.0 % to 126.4 %. Based on diazo derivatization, the proposed SERS method eliminates the need for complex substrates and enables the analysis and determination of synthetic antioxidants in edible oils within 20 min, providing a convenient analytical approach for quality control in the food industry.

Spectrofluorimetric determination of butylated hydroxytoluene and butylated hydroxyanisole in their combined formulation: application to butylated hydroxyanisole residual analysis in milk and butter.

Butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) are two antioxidants that have been extensively used in many applications. Both are well known for their debatable health risks due to their multiple intake sources. Therefore, conservative limits are set for them in different regulations adapted to the matrices in which they exist. Here we present a simple spectrofluorimetric method for the determination of BHT and BHA based on their native fluorescence and synchronous scanning mode. The type of solvent and the interval between emission and excitation wavelengths were carefully optimized. Under the optimized conditions, good linearities were obtained between the emission intensity and the corresponding concentrations of BHT and BHA over the range of 3-18 µg/mL and 0.1-7 µg/mL, respectively with a good correlation coefficient (r > 0.99). The limits of detection were 0.9 and 0.02 µg/mL, and the quantification limits were 3 and 0.05 µg/mL for BHT and BHA, respectively. The suggested procedure was validated according to ICH guidelines Q2 (R1). Furthermore, the method's greenness was assessed by three different methods, and it proved to be eco-reasonable. The method was successfully applied to the determination of BHT and BHA in pharmaceutical formulations. We also applied the suggested method for monitoring the residual BHA in conventional, powdered milk and butter, with good recovery in spiked samples.

Preparation and structural characterization of epoxidized soybean oils-based pressure sensitive adhesive grafted with tea polyphenol palmitate.

Vegetable oils-based pressure sensitive adhesives (PSAs) are green and sustainable but face unsatisfactory adhesion strengths and are prone to aging during storage and application due to the existence of residual double bonds and massive ester bonds. Nine common antioxidants (tea polyphenol palmitate (TPP), caffeic acid, ferulic acid, gallic acid, butylated hydroxytoluene, tertiary butylhydroquinone, butylated hydroxyanisole, propyl gallate, and tea polyphenols) were grafted into epoxidized soybean oils-PSA (ESO-PSA) system to enhance antiaging properties and adhesion strengths. Results showed ESO-PSAs grafted with caffeic acid, tertiary butylhydroquinone, butylated hydroxyanisole, propyl gallate, tea polyphenols, or TPP didn't occur failure with TPP having best performance. The optimal conditions were ESO reacted with 0.9 % TPP, 70 % rosin ester, and 7.0 % phosphoric acid at 50 °C for 5 min, under which peel strength and loop tack increased to 2.460 N/cm and 1.66 N, respectively, but peel strength residue reduced to 138.09 %, compared with control (0.407 N/cm, 0.43 N, and 1669.99 %). Differential scanning calorimetry and thermogravimetric results showed TPP grafting increased the glass transition temperature of ESO-PSA slightly but improved its thermal stability significantly. Fourier transform infrared spectroscopy and 1H nuclear magnetic resonance results showed TPP, phosphoric acid, and rosin ester all partially participated in the covalently crosslinking polymerization of ESO-PSAs and the rest existed in the network structures in the free form.

A polypyrrole-cotton pad sorbent as micro-solid phase extractor enclosed in tea bag envelope for determination of synthetic antioxidants in non-alcoholic beverage products.

A polypyrrole (PPy)-cotton pad sorbent enclosed in tea bag envelope was developed and used in micro-solid phase extraction (µ-SPE) for the determination of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). After extraction, the extract was qualified and quantified by a gas chromatograph equipped with a flame ionization detector (GC-FID). Parameters influencing this developed method and the efficiency of µ-SPE were studied and optimized. Under the optimal conditions, the developed method provided good linearity in a concentration range of 0.100-100 µg L-1 for BHA and 0.050-50 µg L-1 for BHT, respectively. The limits of detection were 39.27 ± 0.52 ng L-1 for BHA and 16.96 ± 0.17 ng L-1 for BHT. Satisfactory relative recoveries of BHA and BHT were achieved in the range from 86.8 ± 1.9 to 117.1 ± 2.3% with acceptable relative standard deviation (RSD) below 8.1%. Good reproducibility was obtained with RSDs < 3.1%, for n = 6. The developed adsorbent is easy to operate, low cost, eco-friendly, reusable, with high extraction efficiency, and was successfully applied in the simultaneous synthetic antioxidant determination of non-alcoholic beverage samples.

Development of a voltammetric electronic tongue for the simultaneous determination of synthetic antioxidants in edible olive oils.

A voltammetric electronic tongue (E-tongue) is "a multisensor system, which consists of a number of low-selective sensors and uses advanced mathematical procedures for signal processing based on pattern recognition and/or data multivariate analysis such as artificial neural networks (ANNs), principal component analysis (PCA), among others". Thus, E-tongues in combination with chemometrics tools result in more accurate and selective analytical methods. In this work, we report results of a simple and reliable electroanalytical method to determine butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT) and propyl gallate (PG) in edible olive oils (EOO). Therefore, the square wave voltammetry (SWV) was used on platinum and carbon fiber disk ultramicroelectrodes (E-tongue configuration) combined with chemometrics tools to perform these studies. On the other hand, two data fusion strategies were used in order to combine electrochemical data obtained for each working electrode in the E-tongue: low-level data fusion (LLDF) and mid-level data fusion (MLDF). In addition, to reduce the dimensionality of the dataset in MLDF, the discrete wavelet transform (DWT) was used. Finally, to assert the predictive capability of the method for BHA, BHT, and PG determination in real samples, a recovery study for the antioxidants in EOO samples was performed, demonstrating the analytical accuracy of the proposed method. Moreover, from the comparison between the proposed electrochemical method with the AOAC reference method and others found in the literature in terms of the quality of the model (REP %) and the percent recovery assays (%) in different samples, our results were better than other reported previously for the simultaneous determination of BHA, BHT, and PG in real samples. Moreover, the percent recovery assays obtained with the proposed electrochemical method were in good agreement with those obtained by the chromatographic method.

A MWCNTs-COOH/PSS nanocomposite-modified screen-printed electrode for the determination of synthetic phenolic antioxidants by HPLC with amperometric detection.

New sensing platforms based on screen-printed carbon electrodes modified with composites based on polystyrene sulfonate and oxidized multi-walled carbon nanotubes (PSS/MWCNTs-COOH/SPCE) have been used to develop a novel HPLC method with electrochemical detection (ECD) for the determination of the most used synthetic phenolic antioxidants in cosmetics: butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), tert-butylhydroquinone (TBHQ) and propyl gallate (PG). Optimal separation conditions were achieved using methanol: 0.10 mol L-1 acetate solution at pH 6 as mobile phase with a gradient elution program from 60 to 90% of methanol percentage in 15 min. The electrochemical detection was carried out in amperometric mode using the PSS/MWCNTs-COOH/SPCE at + 0.80 V vs. Ag. Under these optimal separation and detection conditions, the limits of detection (LOD) were between 0.11 and 0.25 mg L-1. These LOD values were better, especially for BHT, than those previously published in other HPLC methods. Linear ranges from 0.37 mg L-1, 0.83 mg L-1, 0.69 mg L-1 and 0.56 mg L-1 to 10 mg L-1 were obtained for PG, TBHQ, BHA and BHT, respectively. RSD values equal or lower than 5% and 8% were achieved for repeatability and reproducibility, respectively. The HPLC-ECD method was successfully applied to analyze different cosmetic samples. Recovery values within 83-109% were obtained in the validation studies.

Colorimetric sensor array based on CoOOH nanoflakes for rapid discrimination of antioxidants in food.

The identification of synthetic antioxidants has considerable significance in food safety. Here, we described the development of a colorimetric sensor array for rapid detection of eight antioxidants in food through the redox reaction between CoOOH and antioxidants in the presence of colorimetric signal indicators. The CoOOH nanoflakes exhibited high catalytic oxidation activity and can independently catalyze oxidation signal indicators showing different colors. The color reaction was inhibited to different degrees in the presence of antioxidants, which resulted in distinct signal response patterns for their discrimination. The method showed good linearity in the range from 50 to 1000 nM for butylated hydroxytoluene (BHT), butylhydroxyanisole (BHA), propyl gallate (PG) and tert-butyl hydroquinone (TBHQ). Moreover, different proportions of antioxidants were located in the middle pattern of each single antioxidant, and showed certain linear relationships among different concentration ratios. Finally, the proposed colorimetric sensor array was used for practical applications where TBHQ and BHT were detected in biscuits and sausages, and BHA and PG were detected in fried pork kebabs, respectively. The results were further confirmed by high-performance liquid chromatography, which demonstrated the great potential of the colorimetry sensor array for practical applications.

Using Physical Organic Chemistry Knowledge to Predict Unusual Metabolites of Synthetic Phenolic Antioxidants by Cytochrome P450.

Biotransformation, especially by human CYP450 enzymes, plays a crucial role in regulating the toxicity of organic compounds in organisms, but is poorly understood for most emerging pollutants, as their numerous "unusual" biotransformation reactions cannot retrieve examples from the textbooks. Therefore, in order to predict the unknown metabolites with altering toxicological profiles, there is a realistic need to develop efficient methods to reveal the "unusual" metabolic mechanism of emerging pollutants. Combining experimental work with computational predictions has been widely accepted as an effective approach in studying complex metabolic reactions; however, the full quantum chemical computations may not be easily accessible for most environmentalists. Alternatively, this work practiced using the concepts from physical organic chemistry for studying the interrelationships between structure and reactivity of organic molecules, to reveal the "unusual" metabolic mechanism of synthetic phenolic antioxidants catalyzed by CYP450, for which the simple pencil-and-paper and property-computation methods based on physical organic chemistry were performed. The phenol-coupling product of butylated hydroxyanisole (BHA) (based on spin aromatic delocalization) and ipso-addition quinol metabolite of butylated hydroxytoluene (BHT) (based on hyperconjugative effect) were predicted as two "unusual" metabolites, which were further confirmed by our in vitro analysis. We hope this easily handled approach will promote environmentalists to attach importance to physical organic chemistry, with an eye to being able to use the knowledge gained to efficiently predict the fates of substantial unknown synthesized organic compounds in the future.

2,6-Dichloro-1,4-benzoquinone formation from chlorination of substituted aromatic antioxidants and its control by pre-ozonation in drinking water treatment plant.

Halobenzoquinones are frequently detected as disinfection by-products in drinking water. Among identified halobenzoquinones, 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ) is particularly toxic and is frequently detected in drinking water. Synthetic aromatic antioxidants discharged to source water may increase the risk of 2,6-DCBQ formation, as many studies suggest that aromatic compounds are the most likely precursors to 2,6-DCBQ. Herein, we investigated the formation of 2,6-DCBQ from chlorination of three model aromatic antioxidants, including 3-tert-butyl-4-hydroxyanisole (BHA), 2,6-di-tert-butyl-4-methylphenol (BHT) and bis(4-tert-butylphenyl)amine (BBPA). Only BBPA produced 2,6-DCBQ under chlorination, while chlorination of BHA and BHT formed α, ß-unsaturated C4-dicarbonyl ring-opening products and phenolic compounds. Based on mass balance and intermediate transformation analysis, mechanisms for the formation of 2,6-DCBQ from BBPA chlorination involved hydrolysis, tert-butyl group cleavage, chlorine substitution, desamination and oxidation. Mitigating aromatic compounds will be an efficient method for 2,6-DCBQ control, such as pre-ozonation, because the intermediates involved in 2,6-DCBQ formation were aromatic compounds. Real water samples from two drinking water treatment plants (DWTPs), one with pre-ozonation (DWTP 2) and the other without pre-ozonation (DWTP1), were analyzed. The two DWTPs were built along the Yangtse river in Nanjing city. Raw water parameters from the two DWTPs, including dissolved organic carbon (DOC), UV absorbance at 254 nm (UV254) and NH3-N, indicated the water quality between these sources was similar. Pre-ozonation in DWTP 2 vanished 2,6-DCBQ in raw water. Concentrations of 2,6-DCBQ in finished water from DWTP 1 (5.69 ng/L) was higher than concentrations generated from DWTP 2 (1.31 ng/L). These results demonstrate that pre-ozonation, granular activated carbon (GAC) and quartz sand treatments at DWTP 2 remove more 2,6-DCBQ precursors than the conventional quartz sand and GAC treatments in DWTP 1. These results suggest the pre-ozonation, GAC and quartz sand treatments can help minimize concentrations of 2,6-DCBQ generated in DWTPs.

[Study of Rapid HPLC Method for the Determination of Antioxidants in Foods, and Standard Solution Stability during Storage].

In this study, we simultaneously determined three antioxidants, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and tert-butylhydroquinone (TBHQ), using HPLC equipped both with a photodiode array detector and a fluorescence detector in 25 minutes per sample. Due to the combined use of the two detectors, we could achieve improved target selectivity. Further, quantification at the specific wavelengths for each target substance particularly increased BHT detection sensitivity. This approach enabled us to avoid repeated measurements during daily inspections. Furthermore, detections were performed using LC-MS/MS instead of GC-MS to overcome the problem of helium gas shortage.In addition, we investigated antioxidant stability in standard solutions during storage. Although TBHQ was stable in methanol with ascorbic acid at -20℃, ascorbic acid storage has possibility to lead to decrease in BHT and BHA concentrations. We recognized that the mixture of BHT and BHA dissolved in methanol at 4℃ and that of BHT, BHA and TBHQ dissolved in methanol with ascorbic acid at -20℃ were suitable for about one year.

Electrochemical sensor for determination of butylated hydroxyanisole (BHA) in food products using poly O-cresolphthalein complexone coated multiwalledcarbon nanotubes electrode.

In this study, we have reported an electrochemical sensor for the determination of butylated hydroxyanisole (BHA) by electropolymerization of O-cresolphthalein complexone (OC) over the multiwalled carbon nanotubes (MWCNTs). In order to confirm the surface morphology, oxidation states, functional groups and charge transfer property of POC/MWCNTs electrode, the resulting POC film with MWCNTs electrode was characterized by spectroscopy, microscopy, and electrochemical techniques. The fabricated electrode was evaluated for its electrochemical performance in oxidation of BHA and the study showed that at POC/MWCNTs electrodes BHA oxidation occurred at 0.27 V. POC/MWCNTs electrode has shown a linear range for the detection of BHA from 0.33 µM to 110 µM with the detection limit of 0.11 µM (S/N = 3). Amperometric determination of BHA was also done using chronoamperometric techniques and the result was found to be linear. The real time analysis of sensors is also validated by analysing the packed potato chips samples.

Migration analysis, antioxidant, and mechanical characterization of polypropylene-based active food packaging films loaded with BHA, BHT, and TBHQ.

Polypropylene (PP) based active composite films were prepared by adding butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), and tertiary butylated hydroquinone (TBHQ) antioxidants using the extrusion molding process. All concentrations of BHT, 2% to 3% BHA, and 3% TBHQ significantly increased the tensile strength (TS) of the composite films compared with control films. Increasing antioxidant concentration decreased TS values for BHT films, whereas an opposite trend was observed for BHA and TBHQ films. BHA at < 2%, BHT at > 2%, and TBHQ at all added concentrations significantly reduced elongation at break (Eb ) of the composite films compared to control films. Water vapor permeability (WVP) of 1% BHT film was not significantly different from control. However, other antioxidants especially at increased concentrations significantly increased WVP values. TBHQ films with 300% to 662% increase had the highest WVP and BHT films with 5% to 81% increase had the lowest WVP among composite films. All three antioxidants had a negative effect on the transparency of the films; however the effect of BHA at higher concentrations was greater. The antioxidants did not change the color attributes of the films. Films containing all antioxidants showed 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity, which increased with increase in their concentration, especially for those containing 3 wt.% BHT and TBHQ. Overall, incorporating BHA and BHT into a PP matrix improved mechanical, barrier, antioxidant properties, and film appearance and consequently were proposed for the development of antioxidant active PP films. TBHQ film is not recommended for food packaging because of its weak mechanical properties (lower Eb and TS values, higher WVP, and greater migration).

Comparative study of rosemary extracts and several synthetic and natural food antioxidants. Relevance of carnosic acid/carnosol ratio.

The antiradical power, at equal concentrations of active principles, of the following antioxidants were studied using the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) assay: butylated-hydroxyanisole, butylated-hydroxytoluene, tert-butylhydroquinone, ascorbyl palmitate, tocopherol, grape seed extract, olive extract and five rosemary extracts with different concentrations of carnosic acid (CA) and carnosol (COL). The reaction kinetics of DPPH scavenging activity in each studied substance identified significant variations in the time needed to reach the steady state. Rosemary extracts were seen to be more effective than the other compounds. CA had higher antioxidant activity than COL, although COL seemed to react faster with DPPH. The relevance of the CA/COL ratio for the antioxidant activity of rosemary extracts was also analysed. The presence of COL in rosemary extracts increased the antioxidant activity with an optimal CA/COL ratio of 2.5-3.0. Olive extract and grape seed extract seem to be very promising additives for use as technological antioxidants.

Unexpectedly high concentrations of 2,4-di-tert-butylphenol in human urine.

Synthetic phenolic antioxidants (SPAs) have received increasing attention due to the reports of toxicity and environmental contamination. Nevertheless, limited information was available on human burdens of these SPAs, with the exception of 2,6-di-tert-butyl-4-methylphenol (BHT). In our study, BHT as well as six other SPAs were analyzed in human urine samples from United States donors. Three SPA congeners were detected in human urine: BHT, 2,4-di-tert-butylphenol (DBP), and 3-tert-butyl-4-hydroxyanisole (BHA). BHT, which is the congener received most concerns, was detected at low concentrations [geometric mean (GM): 0.06 ng/mL], whereas four of its metabolites were detected at relatively high concentrations (GM: 1.68 ng/mL). Surprisingly, DBP was detected at extremely high concentrations (GM: 18.3 ng/mL). The concentrations of DBP (GM: 25.8 ng/mL), BHT (0.853 ng/mL), and metabolites (GM: 10.5 ng/mL) increased significantly after the urine samples were hydrolyzed by ß-glucuronidase (p < 0.01), indicating the prevalence of the conjugated forms of SPAs and their metabolites in human urine. DBP, which has previously received little attention, was the predominant congener, contributing 88.2% and 63.6% to total target concentrations in the urine samples before and after ß-glucuronidase hydrolysis, respectively. Thus, previous studies have vastly underestimated the burdens of SPAs to humans. To our knowledge, this is the first study revealing the presence of DBP in human urine.

A stir foam composed of graphene oxide, poly(ethylene glycol) and natural latex for the extraction of preservatives and antioxidant.

A stir foam composed of graphene oxide, poly(ethylene glycol) and natural latex (GO-PEG-NL) was prepared for use in micro-solid phase extraction sorbent of preservative agents and antioxidants from cosmetic products. The extracted analytes were quantified by GC-MS. Under the optimized conditions, the calibration plots are linear in the concentration ranges between 5.0 µg·L-1 to 1.0 mg·L-1 for benzoic acid, of 10.0 µg·L-1 to 1.0 mg·L-1 for 2-methyl-3-isothiazolinone (MI), and between 1.0 µg·L-1 and 1.0 mg·L-1 for both 3-tert-butyl-4-hydroxyanisole (BHA) and 2,6-di-tert-butyl-p-hydroxytoluene (BHT). The LODs are 1.0 µg·L-1 for benzoic acid, 5.0 µg·L-1 for MI and 0.5 µg·L-1 for both BHA and BHT. The stir-foam can be easily prepared, is inexpensive and well reproducible (RSDs <3%, for n = 6). It can be re-used for up to 12 times after which extraction efficiency has dropped to 90%. The method was successfully applied to the determination of preservatives and antioxidants in cosmetic samples. Recoveries from spiked samples ranged between 94.5 ± 2.1% and 99.8 ± 1.8%. Graphical abstract A stir foam was prepared from graphene oxide, poly(ethylene glycol) and natural latex (GO-PEG-NL) and is shown to be a most viable sorbent for the microextraction of trace amounts of preservative agents and antioxidants from cosmetic products.

Migration Study of Butylated Hydroxytoluene and Irganox 1010 from Polypropylene Treated with Severe Processing Conditions.

Safety concerns have emerged over the increased use of polypropylene (PP) in food-packaging markets. Some antioxidants in PP can migrate to foods and cause undesirable effects in humans. In this study, migration behaviors of butylated hydroxytoluene (BHT) and Irganox 1010 (I-1010) in PP sheets were determined according to the US FDA migration test conditions. In particular, we tested the effects of severe conditions of food processing and storage, such as autoclave heating (sterilization at about 121 °C), microwave radiation (700 W), and deep freezing (-30 °C) on migration of antioxidants. Migrant concentrations were higher in 95% ethanol as lipid food simulant, because of the hydrophobic nature of both PP and antioxidants. Autoclave heating treatment increased migrant concentrations compared with other processing conditions. Moreover, increased migrant concentrations by deep freezing condition were attributed to the brittleness of PP at freezing temperature. Regardless of processing conditions, BHT which has a lower molecular weight, migrated faster than I-1010. PRACTICAL APPLICATION: The antioxidants with hydrophobic nature such as butylated hydroxytoluene (BHT) and Irganox 1010 (I-1010) in polypropylene sheets would be migrated to foods, which is an important issue for industrial production food packaging materials. Migration behavior was promoted by severe processing conditions such as autoclave heating, microwave radiation, freezing, and especially autoclave heating treatment led the highest migration among them. Therefore, control of chemical additive migration from polypropylene food packaging is needed for safe food processing.

Simultaneous determination of tert-butylhydroquinone, propyl gallate, and butylated hydroxyanisole by flow-injection analysis with multiple-pulse amperometric detection.

We report the first amperometric method for the simultaneous determination of tert-butylhydroquinone (tBHQ), propyl gallate (PG), and butylated hydroxyanisole (BHA) using flow injection analysis coupled to multiple-pulse amperometry. A sequence of potential pulses was selected in order to detect tBHQ, PG, and BHA separately in a single injection step at a glassy carbon electrode without the need of a preliminary separation. A mixture of methanol and 0.040M Britton-Robinson buffer was used both as a carrier solution and for dilution of analyzed solutions before injection. The method is precise (RSD < 5%, n = 10), fast (a frequency of 140 injections h-1), provides sufficiently low quantification limits (2.51, 1.45, and 0.85µmolL-1 for tBHQ, PG, and BHA, respectively) and can be easily applied without high demands on instrumentation. As a practical application, the determination of these antioxidants contained in commercial chewing gum samples was carried out by applying a simple extraction procedure.

Simultaneous analysis of tert-butylhydroquinone, tert-butylquinone, butylated hydroxytoluene, 2-tert-butyl-4-hydroxyanisole, 3-tert-butyl-4-hydroxyanisole, α-tocopherol, γ-tocopherol, and δ-tocopherol in edible oils by normal-phase high performance liquid chromatography.

A normal-phase high performance liquid chromatography method for the simultaneous determination of tert-butylhydroquinone, tert-butylquinone, butylated hydroxytoluene, 2-tert-butyl-4-hydroxyanisole, 3-tert-butyl-4-hydroxyanisole, α-tocopherol, γ-tocopherol, and δ-tocopherol in edible oils was investigated. A silica column was used to separate the analytes with the gradient elution. An ultraviolet-visible detector was set at dual wavelengths mode (280 and 310nm). The column temperature was 30°C. The analytes were directly extracted with methanol. Results showed that the normal-phase high performance liquid chromatography method performed well with wide liner ranges (0.10∼500.00µg/mL, R2>0.9998), low limits of detection and quantitation (below 0.40 and 1.21µg/mL, respectively), and good recoveries (81.38∼102.34% in soybean oils and 83.03∼100.79% in lard, respectively). The reduction of tert-butylquinone caused by the reverse-phase high performance liquid chromatography during the injection was avoided with the current normal-phase method. The two isomers of butylated hydroxyanisole can also be separated with good resolution.

Optimization and Validation of a Fast UPLC Method for Simultaneous Determination of Hydroquinone, Kojic Acid, Octinoxate, Avobenzone, BHA, and BHT.

A previously published HPLC method for the simultaneous determination of six major components (hydroquinone, kojic acid, octinoxate, avobenzone, butylated hydroxyanisole, and butylated hydroxytoluene) in a skin-whitening cream was transferred and optimized to an ultra-performance LC system. Separation was achieved in a ZORBAX SB-Phenyl Rapid-Resolution High Throughput column (2.1 × 100 mm, 1.8 µm), using a mobile phase consisting of water with 0.1% acetic acid and acetonitrile at a flow rate of 0.7 mL/min. The column was maintained at 40°C, and detection was carried out at 230 nm using a diode-array detector. These chromatographic conditions allow the separation of the six compounds in 3 min instead of 14 min. The extraction procedure was optimized, reducing the time and demonstrating its suitability. The method was validated according to International Conference on Harmonization guidelines, with respect to specificity, precision, accuracy, and linearity. Selectivity was found to be satisfactory. Linear regression analysis data for all compounds showed a good linear relationship, with r2 > 0.999 in the concentration range of 50-120% of the label claim for each compound. The RSD for precision and accuracy of the method was found to be less than 2% for all compounds. Comparison of system performance with the previously published HPLC method was made with respect to analysis time, efficacy, and resolution. The proposed method is faster and consumes less solvent and was applied in the determination of six major compounds in batches of skin-whitening cream manufactured during the validation process.

Determination of synthetic phenolic antioxidants in edible oils using microvial insert large volume injection gas-chromatography.

Three synthetic phenolic antioxidants, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ), were determined in different edible vegetable oil samples. The analyses were carried out by gas chromatography-mass spectrometry (GC-MS) using microvial insert large volume injection (LVI). Several parameters affecting this sample introduction step, such as temperatures, times and gas flows, were optimised. Quantification was carried out by the matrix-matched calibration method using carvacrol as internal standard, providing quantification limits between 0.08 and 0.10 ng g(-1), depending on the compound. The three phenolic compounds were detected in several of the samples, BHT being the most frequently found. Recovery assays for oil samples spiked at two concentration levels, 2.5 and 10 ng g(-1), provided recoveries in the 86-115% range.