A simplex-optimized chromatographic separation of fourteen cosmetic preservatives: analysis of commercial products

An ion-interaction high-performance liquid chromatography (HPLC)-diode-array detection method is developed and optimized for the separation of typical antimicrobial agents used in cosmetics and hygiene products. The most used preservatives contain different molecular structures, different functionalities, and are characterized by different chemical properties. Organic acids, alkyl esters of benzoic acids, alkyl p-hydroxy benzoic acids (parabens), phenol derivatives, and carbanilides represent the most used preservatives, and are often present in multicomponent mixtures. In order to develop a multicomponent method to be used in quality control analysis, the ion-interaction reagent reversed-phase HPLC technique seems to be particularly suitable, because it allows for the simultaneous separation of acidic, basic, and neutral species. The experimental conditions of the method are developed by OVAT (one variable at a time) treatment and further optimized by a multivariate approach based on a Simplex algorithm that works on a desirability function targeted to maximize the resolution in a multicomponent mixture. The new method proposed that is able to simultaneously separate fourteen preservatives is applied in the analysis of commercial products.     

Comparing micellar electrokinetic chromatography and microemulsion electrokinetic chromatography for the analysis of preservatives in pharmaceutical and cosmetic products

In this study, separation and determination of nine preservatives ranging from hydrophilic to hydrophobic properties, which are commonly used as additives in various pharmaceutical and cosmetic products, by micellar electrokinetic chromatograpy (MEKC) and microemulsion electrokinetic chromatography (MEEKC) were compared. The effect of temperature, buffer pH, and concentration of surfactant on separation were examined. In MEKC, the separation resolution of preservatives improved markedly by changing the sodium dodecyl sulfate concentration. Temperature and pH of running buffers were used mainly to shorten the magnitude of separation time. However, in order to detect all preservatives in a single run in a MEEKC system, a microemulsion of higher pH was needed. The separation resolution was improved dramatically by changing temperature, and a higher concentration of SDS was necessary for maintaining a stable microemulsion solution, therefore the separation of the nine preservatives in MEEKC took longer than in MEKC. An optimum MEKC method for separation of the nine preservatives was obtained within 9.0 min with a running buffer of pH 9.0 containing 20 mM SDS at 25 degrees C. A separation with baseline resolution was also obtained within 16 min using a microemulsion of pH 9.5 which composed of SDS, 1-butanol, and octane, and a shorter capillary column at 34 degrees C. Finally, the developed MEKC and MEEKC methods determined successfully preservatives in various cosmetic and pharmaceutical products.     

Large volume sample stacking with EOF and sweeping in CE for determination of common preservatives in cosmetic products by chemometric experimental design

This study proposes a capillary electrophoresis method incorporating large volume sample stacking, EOF and sweeping for detection of common preservatives used in cosmetic products. The method was developed using chemometric experimental design (fractional factorial design and central composite design) to determine multiple separation variables by efficient steps. The samples were loaded by hydrodynamic injection (10 psi, 90 s), and separated by phosphate buffer (50 mM, pH 3) containing 30% methanol and 80 mM SDS at -20 kV. During method validation, calibration curves were found to be linear over a range of 5-100 μg/mL for butyl paraben and isobutyl paraben; 0.05-10 μg/mL for ethyl paraben; 0.2-50 μg/mL for dehydroacetic acid; 0.5-70 μg/mL for methyl paraben; 5-350 μg/mL for sorbic acid; 0.02-450 μg/mL for p-hydroxybenzoic acid and 0.05-10 μg/mL for salicylic acid and benzoic acid. The analytes were analysed simultaneously and their detection limits (S/N = 3) were down to 0.005-2 μg/mL. The analysis method was successfully used for detection of preservatives used in commercial cosmetics.     

Development of a tandem thin-layer chromatography-high-performance liquid chromatography method for the identification and determination of corticosteroids in cosmetic products

A solid-phase extraction clean-up and and a liquid chromatographic method with ultraviolet detection were developed for the analysis of 51 corticosteroids in cosmetic samples in order to screen commercial samples for the presence of undeclared synthetic corticosteroids. A thin-layer chromatographic analysis was carried out on silica gel plates, using different eluants and detection reagents. When such a preliminary chromatographic separation gave some indications about the presence of steroid compounds, the methanol extracts from real samples were applied to a solid-phase extraction C₁₈ cartridge, and the analytes eluted with ethyl ether. The high-performance liquid chromatographic separation was then carried out for the identification and determination of the analytes using a Purospher RP-18 column, an isocratic or a gradient elution with a mixture acetonitrile-water and a photodiode-array detector. The accuracy of the method was determined by spiking experiments on home-made cosmetic samples. The analytical recoveries were satisfactory.     

Determination of methylisothiazolinone and methylchloroisothiazolinone in cosmetic products by ultra high performance liquid chromatography with tandem mass spectrometry

Isothiazolinone biocides are broad‐spectrum preservatives that are widely used in cosmetics, household, and industrial products. An increase in the number of cases of allergic contact dermatitis to isothiazolinone preservatives, namely, methylisothiazolinone and methylchloroisothiazolinone, have been recently noticed. The Food and Drug Administration relies on analytical methods to quantify levels of use of cosmetic ingredients and support enforcement action against products that are not in compliance with the law. In this study, an efficient ultra high performance liquid chromatography with tandem mass spectrometry method was developed and validated for the determination of methylisothiazolinone and methylchloroisothiazolinone in selected cosmetic products. The lower limit of quantitation was determined to be 0.1 μg/g for both preservatives. A survey of 24 cosmetic products was conducted and found concentrations of methylisothiazolinone and methylchloroisothiazolinone ranging from not quantified, or below the lower limit of quantitation, to 89.64 μg/g and not quantified to 10.31 μg/g, respectively.     

Analysis of multi-class preservatives in leave-on and rinse-off cosmetics by matrix solid-phase dispersion

Matrix solid-phase extraction has been successfully applied for the determination of multi-class preservatives in a wide variety of cosmetic samples including rinse-off and leave-on products. After extraction, derivatization with acetic anhydride, and gas chromatography–mass spectrometry analysis were performed. Optimization studies were done on real non-spiked and spiked leave-on and rinse-off cosmetic samples. The selection of the most suitable extraction conditions was made using statistical tools such as ANOVA, as well as factorial experimental designs. The final optimized conditions were common for both groups of cosmetics and included the dispersion of the sample with Florisil (1:4), and the elution of the MSPD column with 5 mL of hexane/acetone (1:1). After derivatization, the extract was analyzed without any further clean-up or concentration step. Accuracy, precision, linearity and detection limits were evaluated to assess the performance of the proposed method. The recovery studies on leave-on and rinse-off cosmetics gave satisfactory values (>78% for all analytes in all the samples) with an average relative standard deviation value of 4.2%. The quantification limits were well below those set by the international cosmetic regulations, making this multi-component analytical method suitable for routine control. The analysis of a broad range of cosmetics including body milk, moisturizing creams, anti-stretch marks creams, hand creams, deodorant, shampoos, liquid soaps, makeup, sun milk, hand soaps, among others, demonstrated the high use of most of the target preservatives, especially butylated hydroxytoluene, methylparaben, propylparaben, and butylparaben.     

Determination of 1,4-dioxane in cosmetic products by high-performance liquid chromatography

A rapid high-performance liquid chromatographic procedure has been developed for the assay of 1,4-dioxane in cosmetic products. After solid-phase extraction, using Bond Elut CN and Bond Elut C18 cartridges, samples were analysed directly on a LiChrospher CH-8 reversed-phase column with spectrophotometric detection at 200 nm and acetonitrile - water as eluent. Recovery of 1,4-dioxane from different cosmetic matrices was between 81.5 and 90.1% in the 30-90 microgram g(-1) range. The minimum quantifiable amount was 6.5 microgram g(-1). The method is simple, reproducible and specific and is suitable for routine analyses of commercial cosmetics.     

Simultaneous determination of eleven preservatives in cosmetics by micellar electrokinetic chromatography

A new method for the simultaneous quantitation of 11 preservatives-imidazolidinyl urea, benzyl alcohol, dehydroacetic acid, sorbic acid, phenoxyethanol, benzoic acid, salicylic acid, and four parabens (methyl, ethyl, propyl, and butyl)-in cosmetics by micellar electrokinetic capillary chromatography was established and validated. The separation was performed using an uncoated fused-silica capillary (50 pm id x 60.2 cm, effective length 50 cm) with a running buffer consisting of 15 mmol/L sodium tetraborate, 60 mmol/L boric acid, and 100 mmol/L sodium dodecyl sulfate. A 1:10 dilution of the running buffer was used as the sample buffer to extract the cosmetic samples. The key factors, such as the concentration and pH of the running and sample buffers, which influence quantitative analysis of the above 11 preservatives in cosmetic samples, were investigated in detail. The linear ranges of the calibration curves for imidazolidinyl urea and the other 10 preservatives were 50-1000 and 10-200 mg/L, respectively. The correlation coefficients of the standard curves were all higher than 0.999. The recoveries at the concentrations studied ranged from 93.0 to 102.7%. RSDs were all less than 5%. The new method with simple sample pretreatment met the needs for routine analysis of the 11 preservatives in cosmetics.     

Optimization by experimental design and artificial neural networks of the ion-interaction reversed-phase liquid chromatographic separation of twenty cosmetic preservatives

Particular attention are recently receiving antimicrobial agents added as preservatives in hygiene and cosmetics commercial products, since some of them are suspected to be harmful to the human health. The preservatives used belong to different classes of chemical species and are generally used in their mixtures. Multi-component methods able to simultaneously determinate species with different chemical structure are therefore highly required in quality control analysis. This paper presents an ion interaction RP-HPLC method for the simultaneous separation of the 20 typical antimicrobial agents most used in cosmetics and hygiene products, that are: benzoic acid, salicylic acid, 4-hydroxybenzoic acid, methyl-, ethyl-, propyl-, butyl-, benzyl-benzoate, methyl-, ethyl-, propyl-, butyl-, benzyl-paraben, o-phenyl-phenol, 4-chloro-m-cresol, triclocarban, dehydroacetic acid, bronopol, sodium pyrithione and chlorhexidine. For the development of the method and the optimization of the chromatographic conditions, an experimental design was planned and models were built by the use of artificial neural network to correlate the retention time of each analyte to the variables and their interactions. The neuronal models developed showed good predictive ability and were used, by a grid search algorithm, to optimize the chromatographic conditions for the separation of the mixture.     

Simultaneous determination of multiclass preservatives including isothiazolinones and benzophenone‐type UV filters in household and personal care products by micellar electrokinetic chromatography

In this work, a simple and reliable micellar electrokinetic chromatography method for the separation and quantification of 14 preservatives, including isothiazolinones, and two benzophenone‐type UV filters in household, cosmetic and personal care products was developed. The selected priority compounds are widely used as ingredients in many personal care products, and are included in the European Regulation concerning cosmetic products. The electrophoretic separation parameters were optimized by means of a modified chromatographic response function in combination with an experimental design, namely a central composite design. After optimization of experimental conditions, the BGE selected for the separation of the targets consisted of 60 mM SDS, 18 mM sodium tetraborate, pH 9.4 and 10% v/v methanol. The MEKC method was checked in terms of linearity, LODs and quantification, repeatability, intermediate precision, and accuracy, providing appropriate values (i.e. R² ≥ 0.992, repeatability RSD values ?9%, and accuracy 90–115%). Applicability of the validated method was successfully assessed by quantifying preservatives and UV filters in commercial consumer products.     

Simultaneous analysis of antioxidants and preservatives in cosmetics by supercritical fluid extraction combined with liquid chromatography-mass spectrometry

This study evaluated supercritical fluid extraction (SFE) combined with liquid chromatography-mass spectrometry (LC-MS) to determine trace preservatives and antioxidants including methylparaben (MP), ethylparaben (EP), propylparaben (PP), butylparaben (BP), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), alpha-tocopherol (alpha-t) and alpha-tocopherol acetate (alpha-ta) in cosmetic products. A supercritical fluid extraction procedure was used to isolate four paraben preservatives and four antioxidants from the cosmetic matrix before quantitative analysis. The optimum extraction condition was performed with static extraction for 5 min, then dynamic extraction for 20 min by using carbon dioxide supercritical fluid at 14,000 kPa and 65 degrees C. Methanol was used as collection solvent and the sea sand was chosen as a filling material. The analytes were separated on a C18 reversed-phase column using methanol-water as mobile phase and quantified by measuring its mass spectrometry. The linearity range is from 10 to 20,000 ng/g with RSD values below 18%. Detection limits are achieved at the level of 4.7-142 ng/g. It was successfully applied to the determination of paraben preservatives and antioxidants in cosmetics without tedious pretreatment.     

Determination of methyldibromoglutaronitrile in cosmetic products by high-performance liquid chromatography with electrochemical detection. Method validation

An increased frequency of contact allergy to methyldibromoglutaronitrile (MDBGN), a commonly used preservative in cosmetics and other consumer products, has been reported in recent years. A high-performance liquid chromatography (HPLC) method for the determination of MDBGN in cosmetic products has been validated in the present study. The identification is performed by reductive electrochemical detection of the bromine present in the molecule. The method is suitable for compliance testing of cosmetic products as well as for the research to support clinical and epidemiological studies.     

Determination of parabens in cosmetic products using multi-walled carbon nanotubes as solid phase extraction sorbent and corona-charged aerosol detection system

The potential of carbon nanotubes for the solid phase extraction of parabens in cosmetic products and the detection using a corona-charged aerosol detector (C-CAD) is presented in this work. The analytical procedure is based on a conventional solid phase extraction step for which 20 mg of multi-walled carbon nanotubes were packed in a 3-mL commercial SPE cartridge. Methylparaben, ethylparaben, propylparaben and butylparaben were thus isolated and preconcentrated from the pre-treated samples and subsequently separated on a RP-C18 column using acetonitrile:water, 50:50 (v/v) as mobile phase. The analytical signals for the individual parabens were obtained using C-CAD. The experimental variables affecting the extraction procedure and the instrumental detection have been deeply studied. Limits of detection were in the range of 0.5–2.1 mg L⁻¹, while the linear range was extended up to 400 mg L⁻¹. The average precision of the method varied between 3.3–3.8% (repeatability) and 4.3–7.6% (reproducibility). Finally, the optimized procedure was applied to the determination of the target preservatives in a variety of cosmetic products with satisfactory results.     

Occurrence of 1,4-dioxane in cosmetic raw materials and finished cosmetic products.

Surveys of cosmetic raw materials and finished products for the presence of the carcinogen 1,4-dioxane have been conducted by the U.S. Food and Drug Administration since 1979. Analytical methods are described for the determination of 1,4-dioxane in ethoxylated cosmetic raw materials and cosmetic finished products. 1,4-Dioxane was isolated by azeotropic atmospheric distillation and determined by gas chromatography using n-butanol as an internal standard. A solid-phase extraction procedure based on a previously published method for the determination of 1,4-dioxane in cosmetic finished products was also used. 1,4-Dioxane was found in ethoxylated raw materials at levels up to 1410 ppm, and at levels up to 279 ppm in cosmetic finished products. Levels of 1,4-dioxane in excess of 85 ppm in children's shampoos indicate that continued monitoring of raw materials and finished products is warranted.     

色谱柱选择对23种防腐剂测定结果的影响

以化妆品中23种防腐剂检测方法为例,探讨色谱柱选择对液相色谱方法测定结果的影响。参照《化妆品安全技术规范》甲基异噻唑啉酮等23个组分的检验方法,在2台不同的高效液相色谱仪上用15款不同品牌、型号的C₁₈色谱柱检测23种防腐剂,计算色谱峰的理论塔板数和分离度,对23种组分的分离效果进行分析,并应用USP (United States Pharmacopeia)数据库和PQRI (Product Quality Research Institute)数据库等2种等效色谱柱选择方法,对不同色谱柱的分离效果及等效性进行评价和预测。实验结果表明,15款色谱柱对23种防腐剂的分离效果差异显著,仅有2款色谱柱可以实现23种组分的完全分离。USP和PQRI数据库中2种等效色谱柱选择方法均无法预测出合适的等效色谱柱,对23种防腐剂的液相色谱分析参考价值均较小。色谱柱是影响23种防腐剂液相色谱法测定结果准确性的关键因素,有关实验室在应用该方法时,应考虑色谱柱选择性差异。化妆品基质复杂,如何在现有研究成果的基础上,开发色谱柱的筛选和预测评价体系,进而指导实际样品的分离是下一步研究的重点、难点。建议有关部门在制修订检测方法时,注重色谱柱的耐用性考察,完善系统适应性指标,细化色谱柱分类和增加描述信息,指导色谱柱的合理选择,从而规避由于色谱柱使用过程中选择依据缺失而导致测定结果不准确的风险。     

Determination of 19 Preservatives in Various Matrices by High-Performance Liquid Chromatography

A simple and sensitive high-performance liquid chromatography (HPLC) method was developed for the determination of 19 preservatives in cosmetic matrices. The composition of the mobile phase was optimized as a gradient to achieve a lower detection limit when compared to previously validated methods, and sample preparation conditions were investigated to optimize separation of the 19 preservatives. A C18 column was used with methanol, 0.05 mol/L ammonium acetate buffer, and water as the mobile phase under gradient elution conditions. Preservatives in cosmetics were extracted with 70% methanol using an ultrasonicator, after which they were analyzed with an HPLC-photodiode array detector. All preservatives were separated within 55 min. The recoveries ranged from 94.9% to 102.8%, with relative standard deviations of less than 3.2% and no correlation coefficients lower than 0.9986. Additionally, the developed method has a low detection limit, which makes it possible to analyze trace levels of compounds in various cosmetic and ingredient matrices.     

Determination of sunscreen agents in cosmetic products by supercritical fluid extraction and high-performance liquid chromatography

A rapid supercritical fluid extraction (SFE) procedure for the isolation of five of the most common sunscreen agents (2-ethylhexyl-p-dimethylaminobenzoate, 2-hydroxy-4-methoxybenzophenone, 2-ethylhexyl-p-methoxycinnamate, 4-methylbenzylidene camphor and 4-tert.-butyl-4′-methoxydibenzoylmethane) from cosmetic products is described. Investigation of the factors affecting the extraction efficiency in SFE indicated that sunscreen recoveries were affected mainly by the supercritical CO₂ pressure and by the trapping method. The sunscreens were analyzed by reversed-phase high-performance liquid chromatography after a 10-min extraction of the cosmetic product with CO₂ at 250 bar and 40°C, using sequential glass surface and C₁₈ sorbent as collection system. A quantitative comparison of SFE with a liquid extraction procedure was performed on commercial cosmetics. The SFE method yielded recoveries higher than 94.8% compared with conventional liquid extraction and exhibited a precision better than 5.3% relative standard deviation. Moreover, SFE minimized sample handling, reduced the consumption of harmful solvents and afforded a more effective purification of the cosmetic matrices.     

A reliable and environmentally-friendly liquid-chromatographic method for multi-class determination of fat-soluble UV filters in cosmetic products

An environmentally-friendly analytical method for the simultaneous determination of 15 fat-soluble ultraviolet (UV) filters currently authorized by the European Union regulation on cosmetic products has been developed. The determination was performed by liquid chromatography with UV spectrophotometric detection. Different parameters, such as type of column, oven temperature, mobile phase composition and flow rate were studied. The best chromatographic separation was obtained under the following conditions: C18 column set at 60 °C and gradient ethanol:water (containing 1% formic acid and 20 mM of 2-hydroxypropyl-β-cyclodextrin) as mobile phase pumped at 1 mL min⁻¹. 2-Hydroxypropyl-β-cyclodextrin was added as mobile phase modifier to achieve the complete resolution of some of the chromatographic peaks. The 15 target compounds were separated in less than 30 min. The method was satisfactorily validated by analyzing three laboratory-made cosmetic samples besides of eleven commercially available cosmetic products containing different combination of the target UV filters. Good accordance of the found levels compared with those of the laboratory-made samples and those of the commercial samples (when available) was achieved. Moreover, excellent recoveries (97–104%) and good intra-day and inter-day precision values at different concentration levels, besides limits of detection values below the μg mL⁻¹ level, were obtained. These good analytical features, as well as their environmentally-friendly characteristics, make the presented method suitable not only for routine analysis in cosmetics industries, but also as candidate reference method for sunscreen analysis.     

Application of response function methodology for the simultaneous determination of potential fragrance allergens and preservatives in personal care products using micellar electrokinetic chromatography

A micellar electrokinetic chromatography method was developed for determination of 15 suspected fragrance allergens and preservatives. The target compounds are widely used as ingredients in many personal care products, and all of them are included in the European Regulation concerning cosmetic products. The method was optimized by using a central composite experimental design and response surface methodology. A modified chromatographic response function was defined to weigh the terms in the response function adequately. After optimization of experimental conditions, a background electrolyte of 100 mM sodium dodecyl sulphate and 24 mM sodium tetraborate and pH 9.0 was selected for the separation of the analytes. The developed methodology was evaluated in terms of linearity, limits of detection and quantification, precision and accuracy, showing appropriate values (i.e., R ²?=?≥0.99 and accuracy of 89–115 %). Finally, applicability of the micellar electrokinetic chromatography method was assessed by successfully quantifying fragrance allergens and preservatives in commercial personal care products. The most commonly found analyte was linalool (48.3 % of samples) followed by benzoic acid (37.6 %). All samples contained at least one of the target compounds, thus confirming the ubiquity of fragrance allergens and preservatives in personal care products.

Fast and sensitive high performance liquid chromatography analysis of cosmetic creams for hydroquinone, phenol and six preservatives

A fast and sensitive HPLC method for analysis of cosmetic creams for hydroquinone, phenol and six preservatives has been developed. The influence of sample preparation conditions and the composition of the mobile phase and elution mode were investigated to optimize the separation of the eight studied components. Final conditions were 60% methanol and 40% water (v/v) extraction of the cosmetic creams. A C18 column (100 mm × 2.1 mm) was used as the separation column and the mobile phase consisted of methanol and 0.05 mol/L ammonium formate in water (pH=3.0) with gradient elution. The results showed that complete separation of the eight studied components was achieved within 10 min, the linear ranges were 1.0-200 μg/mL for phenol, 0.1-150 μg/mL for sorbic acid, 2.0-200 μg/mL for benzoic acid, 0.5-200 μg/mL for hydroquinone, methyl paraben, ethyl paraben and propyl paraben, butyl paraben, and good linear correlation coefficient (≥0.9997) were obtained, the detection limit was in the range of 0.05-1.0 μg/mL, the average recovery was between 86.5% and 116.3%, and the relative standard deviation (RSD) was less than 5.0% (n=6). The method is easy, fast and sensitive, it can be employed to analyze component residues in cosmetic creams especially in a quality control setting.