Cecropia pachystachya Trécul: a promising ingredient for skin-whitening cosmetics

Abstract Because it promotes the lightening of pigment spots, tyrosinase inhibition is one of the mechanisms of depigmenting cosmetic products. Considering the adverse effects produced by synthetic depigmenting actives, the search for new therapeutic options is desirable, and plant extracts are possible candidates for hyperpigmentation treatment. Glycolic extracts of Cecropia pachystachya Trécul are, therefore, the focus of this study. Its chemical characterization, antioxidant activity, tyrosinase inhibition, and cell viability were evaluated. Glycolic extracts were obtained by macerating the leaves of C. pachystachya in grain alcohol and glycerin or propylene glycol. Both had a similar chemical constitution, the glycerin being more efficient in concentrating phenolic compounds and flavonoids. Analyses by UHPLC-MS detected quinic acid, chlorogenic acid isomers, proanthocyanidin dimers type B and C, catechin/epicatechin, orientin/isoorientin, isoorientin 2"-O-xyloside, vitexin/isovitexin, and rutin. 5-O-caffeoylquinic acid was then quantified was then quantified, with predominance in the extract produced with propylene glycol. These extracts showed a high antioxidant capacity by the method of DPPH, ß-carotene, and nitric oxide. As for depigmenting activity, both extracts were able to inhibit tyrosinase. Cell viability assay also revealed that the extracts could safely be used in concentrations of ≤ 125 µg/mL. Thus, this study demonstrated for the first time that the glycolic extracts of C. pachystachya have promising chemical and biological characteristics for the development of a multifunctional cosmetic with antioxidant and tyrosinase-inhibition activities

Advanced Formulation Design for Topical Creams Assisted with Vibrational Spectroscopic Imaging.

Vibrational spectroscopic imaging has become useful analytical tools for quality control of drug products. In this study, we applied microscopic attenuated total reflection (ATR)-IR and confocal Raman microscopy to elucidate microscopic structure of creams and for the formulation design in the development of semi-solid drug products. The model creams were prepared with prednisolone (PRD) and fluconazole (FLC) as active pharmaceutical ingredients and oily solvents such as mineral oil (MO), isopropyl myristate (IPM), benzyl alcohol (BA) and diethyl sebacate (DES). As a result of microscopic ATR-IR imaging, several domains indicating oily internal phase were observed, which had absorption around 1732 and 1734 cm-1 derived from MO, IPM and DES. In addition, domains of BA around 1009 cm-1 were observed at the complemental or similar position in the formulation with MO or DES, respectively. These results suggested that the creams were oil-in-water type and the distribution of domains would reflect the compatibility of the solvents. The contents of PRD and BA were determined quantitatively in each layer after the intentional separation of the creams and the results agreed well with the imaging analysis. Whereas, confocal Raman imaging allowed to visualize the distribution of the components in depth direction as well as two-dimensional plane. In particular, the Raman imaging would ensure the coexistence of FLC and BA as oily phase in the cream. From these results, the feasibility of spectroscopic imaging techniques was successfully demonstrated for the formulation design of semi-solid dosage forms.

Scope and limitations of nuclear magnetic resonance techniques for characterisation and quantitation of vitamin D in complex mixtures.

BACKGROUND: The accurate determination of vitamin D in skin is of considerable importance in evaluating penetration of skin health products through the different layers of the skin. OBJECTIVE: We report on the characterisation and quantitation of vitamin D in an idealised sample and in complex mixtures which mimic that of a typical skin cream, using qNMR, 2D NMR and DOSY techniques. METHODS: The characterisation and quantitation conditions were acquired over several heterogeneous samples, allowing for analysis of how the dynamic range and complexity of the different sample mixtures affect the limits of detection (LOD) and limits of quantitation (LOQ) of vitamin D. NMR is of particular value to this task as it is non-destructive, uses a primary ratio method for quantification, and tolerates a wide variety of hydrophilic and hydrophobic components within a given matrix. RESULTS: In this investigation, we have attained a trueness level <10%, repeatability values of <1% and brought the limit of quantitation down to 100 nmol/L (≈limit of baseline range of vitamin D2 and D3 per litre seen in vivo), commenting on the limitations observed, such as peak overlap and sensitivity limits. CONCLUSIONS: Pure shift optimised sequences allow us to reduce peak overlapping, allowing further characterisation of individual compounds and the separation of complex mixtures using NMR.

Contact transfer risk from fentanyl-contaminated RSDL® Kit.

The RSDL® (Reactive Skin Decontamination Lotion) Kit contains a lotion-impregnated sponge extensively studied for the removal or neutralization of chemical warfare agents from skin. Pilot investigation of efficacy with industrial threat compounds noted that synthetic opioid fentanyl citrate was removed by the RSDL Kit but not chemically inactivated by the lotion. This implies that after use the RSDL Kit will contain intact fentanyl, which may pose a dermal health hazard if the fentanyl is then transferred to skin after use without proper handling. This in vitro investigation studied the contaminated RSDL Kit using three different concentrations of fentanyl with a skin contact time of 15 min under direct interaction from passive contact, light touch, and leaning with one hand. It was demonstrated that the expected transfer of fentanyl from contaminated RSDL depends on 1) the concentration of fentanyl and 2) the area of the exposed surface. From a toxicological perspective, the contact risk of fentanyl under the conditions tested can be considered low but not absent. The present study determined that a contaminated RSDL Kit, used for removal of fentanyl, should be handled with proper care. Use of protective gloves in operational use and washing skin afterwards is advised to prevent undesired contamination.

Rapid and sensitive LC-MS/MS method for simultaneous quantification of capsaicin and dihydrocapsaicin in microdialysis samples following dermal application.

A bioanalytical LC-MS/MS method was developed and validated for the simultaneous quantification of capsaicin (CAPS) and dihydrocapsaicin (D-CAPS) in dermal microdialysis samples from rats. Capsaicinoids were separated by using a C18 column, with a mobile phase of water and acetonitrile, both with 0.1% of formic acid, eluted as a gradient. Compounds were detected by using an electrospray ionization source operating in the positive mode (ESI+) to monitor the m/z transitions of 306.1 > 137.0 for CAPS and 308.1 > 137.0 for D-CAPS. The method showed linearity in the concentration range of 0.5-100 ng/ml for CAPS and 0.25-100 ng/ml for D-CAPS, with coefficients of determination of ≥ 0.99. The inter- and intra-day precision, accuracy, and compound stability in different conditions were in accordance with the limits established by the US Food and Drug Administration guidelines. The recovery of the drugs by microdialysis were dependent on the flow rate, but independent of drug concentration. For CAPS, calibration of the in vitro microdialysis probes by dialysis and retrodialysis resulted in statistically similar drug recovery of 68.5% ± 5.9% and 77.8% ± 6.6%, respectively, at a flow rate of 0.5 µl/min. For D-CAPS, the recovery by dialysis was lower than by retrodialysis, at 51.4% ± 6.6% and 92.6% ± 2.4%, respectively. This difference was attributed to the binding of D-CAPS to the plastic tubing, which was experimentally evaluated and mathematically modeled. In vivo recoveries were 75.7% ± 6.3% for CAPS and 81.9% ± 1.5% for D-CAPS at the same flow rate. The analytical method showed high specificity, accuracy, and sensitivity, and suitability for dermatopharmacokinetic studies. These results will allow the determination of the actual free concentration of these drugs in dermatopharmacokinetic experiments, as shown in a pilot experiment with a commercial cream containing capsaicinoids.

Identification and characterization of unknown degradation impurities in beclomethasone dipropionate cream formulation using HPLC, ESI-MS and NMR.

The present study focuses on identifying the degradation profile and pathways of unknown impurities from beclomethasone dipropionate (BDP) topical cream formulation reported under accelerated stability conditions. Six degradation impurities were observed during the accelerated stability testing of BDP topical cream formulation, and these thermally labile degradation impurities were primarily identified using a simple, effective and mass compatible isocratic reversed-phase high-performance liquid chromatography with ultraviolet detection method. The degradation impurities found in this sample were of very low concentration levels, thus the concentration of these impurities in the sample was enriched by mimicking the thermal degradation conditions to structurally elucidate the unknown impurities. These BDP thermal degradation impurities were isolated using preparative liquid chromatography and followed by pre-concentration using rota-vapour. Further, the collected thermal degradation impurities were characterized using ESI-MS, and the major impurity was identified using 1H and C13 NMR spectroscopy, and DEPT technique. Plausible degradation pathway and mechanism of each impurity from BDP has been proposed based on the obtained mass and NMR spectral data. Thus, the present method is simple and suitable to be applied towards BDP assay in various formulations, and also to investigate the thermal stability and degradation kinetics of the final drug product.

Effective tool for assessment of the quality of barrier creams - relationships between rheological, textural and sensory properties.

The efficiency of barier creams (BC) in the prevention of occupational skin diseases is closely related to their mechanical, rheological but also sensory properties. The measurement procedure we elaborated, which simulates the spreadability conditions on the skin and evaluation whether the structure reconstruction occurs (hysteresis loop test, shear rate dependence of normal force), allows the assessment of the effectivness of the BC in terms of mechanical resistance and its adhesion to the skin surface. In this thesis an effort was made to define the impact of the human factor - the product application on skin - on the efficiency of medical devices for cutaneous use. Creams' performance such as the spreadability or the feeling on the skin during and after application, which mostly determine users willingness to use them systematically and rigorously, have been linked to parameters obtained during the structure analysis and to rheological properties. Moreover an attempt has been made to correlate the values of basic textural properties with rheological parameters determined by viscoelasticity and classic flow analysis. Instrumental analysis of selected BC products demonstrated a good correlation with organoleptic tests carried out on probands. The applicability of our tool for quality evaluation of BC has been confirmed.

In Vivo Determination of Moisturizers Efficacy on Human Skin Hydration by Confocal Raman Spectroscopy.

This research work deals with in vivo testing of the efficacy of commercial moisturizer products on the hydration of human skin, as there are various in vitro and ex vivo studies questioning their activity. Confocal Raman spectroscopy was used for this purpose of assessing the efficacy of moisturizers on skin hydration mainly owing to its simple, non-invasive, non-destructive, timesaving, and cost-effective nature. Water content and natural moisturizing factor (NMF) of stratum corneum were analyzed and compared using this method at high wavenumber (2500-4000 cm-1) and fingerprint (400-1800 cm-1) spectral regions, respectively, as these two parameters are correlated to skin hydration. Four commercial moisturizer products of different brands were tested on volar forearm region of healthy human female volunteers. This study was conducted for a period of 30 days with 0, 7, and 30 days as time points of analysis. The results of this study clearly indicate that not all the moisturizer products hydrate the skin to the expected levels, and this extent of skin hydration varies with duration of application of these products.

[Determination of 42 glucocorticoids in infant eczema products by auto-solid phase extraction-ultra high performance liquid chromatography-tandem mass spectrometry].

A method was developed for the determination of 42 glucocorticoids in infant eczema products by auto-solid phase extraction-high performance liquid chromatography-tandem mass spectrometry (ASPE-UHPLC-MS/MS).The analytes were extracted by acetonitrile after dispersing in saturated sodium chloride solution.Macromolecules matrix were precipitated by adding potassium and zinc acetate, and then purified by HLB SPE column.The chromatographic separations were performed on a DIKMA EndeavorsilTM C18 column (100 mm×2.1 mm, 1.7 µm).Acetonitrile and 0.1%(v/v) formic acid aqueous solution were used as the mobile phases.Electrospray ionization (ESI) source was operated in the positive mode using multiple reaction monitoring (MRM) scanning mode.The results were quantified by external standard method.The correlation coefficients of linear calibration curves were greater than 0.998 in the corresponding mass concentration ranges.The average recoveries at three spiked levels ranged from 78.7% to 101.5%, and the relative standard deviations (RSDs) were 2.0%-8.8%(n=6).The LODs (S/N ≥ 3) and LOQs were 0.002-0.031 mg/kg and 0.0007-0.103 mg/kg, respectively.The method is simple, rapid, sensitive, and reliable, and can be used for the detection of glucocorticoids in infant eczema products.

Direct bioautography hyphenated to direct analysis in real time mass spectrometry: Chromatographic separation, bioassay and mass spectra, all in the same sample run.

Mass spectra were recorded directly in situ the bioautogram, i.e., in the presence of microorganisms, bioassay medium and substrate reagent. The desorption-based direct analysis in real time mass spectrometry (DART-MS) was applied immediately after direct bioautography (DB). It turned out to be an advantageous combination, as it offered the possibility of a straightforward mass spectrometric detection of bioactive analytes within the bioautogram, and at the same time, it was discriminating microorganism cells and highly polar bioassay medium ingredients which could otherwise stress the MS system. DB-DART-MS was investigated for bioactive compounds in cosmetics using the Bacillus subtilis and Aliivibrio fischeri bioassays for detection of Gram-positive and Gram-negative antimicrobials, respectively, and the planar yeast estrogen screen for detection of estrogen-effective compounds. The influences of the three different bioassay matrices on the analyte response and DB-DART-MS performance on different layers were studied on the example of parabens in hand creams. It was shown that with increasing culture medium complexity, the ion suppression increased. As proof-of-principle, the mass spectrometric quantification at the nanogram level in situ the bioautogram was verified by comparison to HPTLC-DART-MS. The total paraben contents of hand creams 1 and 2 were 0.17-0.20% and 0.30-0.34%, respectively, depending on the method used (DB-DART-MS with two different bioassays or HPTLC-DART-MS as well as on RPW or NP plate). In contrast to the current practice of applying the sample twice and subjecting one track to the bioassay and another to MS, the introduced hyphenation DB-DART-MS is straightforward and highly efficient.

A phase II study to evaluate the safety and efficacy of topical 3% amphotericin B cream (Anfoleish) for the treatment of uncomplicated cutaneous leishmaniasis in Colombia.

BACKGROUND: Pentavalent antimonials (Sb5) are the first-line drugs for treating cutaneous leishmaniasis in Colombia; however, given problems with toxicity, compliance, availability, and cost, it is imperative to look for better therapeutic options. Intravenous amphotericin B (AmB) has been used extensively to treat visceral leishmaniasis; however, evidence on its topical use for cutaneous leishmaniasis is limited. Anfoleish is a topical formulation based on 3% AmB, which was developed following GMP standards by HUMAX and PECET. Anfoleish was shown to be safe and efficacious in animal model and in an open label study in CL patients. Hereafter we show the results of the first controlled and randomized study assessing the safety and efficacy of Anfoleish administered topically, two or three times per day for 28 days, for the treatment of non-complicated cutaneous leishmaniasis in Colombia. METHODS: An open-label, randomized, non-comparative phase Ib/II clinical trial was performed. Adult volunteers with a parasitologically confirmed diagnosis of cutaneous leishmaniasis were randomly allocated to receive Anfoleish cream either 3 (TID group) or 2 (BID group) times per day for 4 weeks. RESULTS: 80 out of 105 subjects screened were included in the study. In intention to treat analysis, final cure was observed in 13 (32.5%) out of 40 subjects (IC 95% = 20.1-48) and in 12 (30%) out of 40 subjects (IC 95% = 18.1-45.5) in the BID and TID group respectively. In the per protocol analysis, cure rates were 39.4% (n = 13) (IC 95% = 24.7-56.3) and 35.3% (n = 12) (IC 95% = 21.5-52.1) in the BID and TID groups respectively. Anfoleish proved to be safe, and the few adverse events reported were local, around the area of application of the cream, and of mild intensity. CONCLUSION: Anfoleish showed to be a safe and well-tolerated intervention. Its efficacy results however do not support at this time continuing with its clinical development or recommending it for the treatment of CL. Additional, studies to improve its current formulation are needed before thinking in conducting additional studies in patients. TRIAL REGISTRATION: Registered in clinicaltrials.gov NCT01845727.

Stability of Mixed Preparations Consisting of Commercial Moisturizing Creams with an Ointment Base Investigated by Magnetic Resonance Imaging.

A moisturizing cream mixed with a steroid ointment is frequently prescribed to patients suffering from atopic dermatitis. However, there is a concern that the mixing operation causes destabilization. The present study was performed to investigate the stability of such preparations closely using magnetic resonance imaging (MRI). As sample preparations, five commercial moisturizing creams that are popular in Japan were mixed with an ointment base, a white petrolatum, at a volume ratio of 1 : 1. The mixed preparations were stored at 60°C to accelerate the destabilization processes. Subsequently, the phase separations induced by the storage test were monitored using MRI. Using advanced MR technologies including spin-spin relaxation time (T2) mapping and MR spectroscopy, we successfully characterized the phase-separation behavior of the test samples. For most samples, phase separations developed by the bleeding of liquid oil components. From a sample consisting of an oil-in-water-type cream, Urepearl Cream 10%, a distinct phase-separation mode was observed, which was initiated by the aqueous component separating from the bottom part of the sample. The resultant phase separation was the most distinct among the test samples. To investigate the phase separation quantitatively and objectively, we conducted a histogram analysis on the acquired T2 maps. The water-in-oil type creams were found to be much more stable after mixing with ointment base than those of oil-in-water type creams. This finding strongly supported the validity of the mixing operation traditionally conducted in pharmacies.

Mercury contamination in facial skin lightening creams and its health risks to user.

This study aims to determine concentrations of mercury in facial skin lightening cream according to different price categories (category I:

Full spectrum and selected spectrum based multivariate calibration methods for simultaneous determination of betamethasone dipropionate, clotrimazole and benzyl alcohol: Development, validation and application on commercial dosage form.

Five different chemometric methods were developed for the simultaneous determination of betamethasone dipropionate (BMD), clotrimazole (CT) and benzyl alcohol (BA) in their combined dosage form (Lotriderm® cream). The applied methods included three full spectrum based chemometric techniques; namely principal component regression (PCR), Partial Least Squares (PLS) and Artificial Neural Networks (ANN), while the other two methods were PLS and ANN preceded by genetic algorithm procedure (GA-PLS and GA-ANN) as a wavelength selection procedure. A multilevel multifactor experimental design was adopted for proper construction of the models. A validation set composed of 12 mixtures containing different ratios of the three analytes was used to evaluate the predictive power of the suggested models. All the proposed methods except ANN, were successfully applied for the analysis of their pharmaceutical formulation (Lotriderm® cream). Results demonstrated the efficiency of the four methods as quantitative tool for analysis of the three analytes without prior separation procedures and without any interference from the co-formulated excipient. Additionally, the work highlighted the effect of GA on increasing the predictive power of PLS and ANN models.

Opinion of the Scientific Committee on Consumer safety (SCCS) - Opinion on the safety of the use of deoxyarbutin in cosmetic products.

CONCLUSION OF THE OPINION: Although on the basis of the provided scientific data the use of deoxyarbutin as such can be considered safe for consumers in cosmetic products in a concentration up to 3% in face creams, hydroquinone will be formed at levels which raise concerns with regard to the safety of such products during life-cycle of the product (e.g. storage conditions and stability under in-use conditions). Therefore, the overall conclusion of the SCCS is that the use of deoxyarbutin up to 3% in face creams is not safe.

Opinion of the Scientific Committee on Consumer safety (SCCS)--Opinion on the safety of the use of α-arbutin in cosmetic products.

CONCLUSION OF THE OPINION: The SCCS considers the use of α-Arbutin safe for consumers in cosmetic products in a concentration up to 2% in face creams and up to 0.5% in body lotions. A potential combined use of α-Arbutin and other hydroquinone releasing substances in cosmetic products has not been evaluated in this Opinion.

Liquid chromatography/tandem mass spectrometry for simultaneous determination of undeclared corticosteroids in cosmetic creams.

RATIONALE: Undeclared corticosteroids in creams intended for frequent use might cause serious side-effects, especially in children. In order to prevent this or find the cause, it was essential to develop a method for quick detection and quantification of low levels of corticosteroids. METHODS: Eleven corticosteroids were used in this study: prednisolone, methylprednisolone, prednisolone-21-acetate, fluocinolone acetonide, fluocinolone acetonide-21-acetate, hydrocortisone-21-acetate, dexamethasone, betamethasone, betamethasone dipropionate, clobetasol propionate and triamcinolone. Separation was achieved via liquid chromatography (LC), and mass spectrometric analysis was conducted by electrospray ionization triple-quadrupole mass spectrometry (MS/MS) in the multiple reaction monitoring mode using corticosterone as internal standard. RESULTS: Good separation by using a gradient-elution LC/MS/MS method with run time of 25 min enabled the use of a segmented detection method and consecutive decrease in detection limits. The proposed method has been validated in the linearity range of 10-1000 ng/mL with coefficients of determination higher than 0.990. The method has shown to have very low limits of quantification (0.75-3 ng/mL) with satisfactory precision and accuracy for each of the corticosteroids. CONCLUSIONS: An LC/MS/MS method for the rapid and simultaneous determination of low levels of eleven topical corticosteroids in creams was developed, optimized and validated. The proposed method can be used for testing of different products indicated for the treatment of atopic dermatitis, including "natural products", and "herbal creams" with "miraculous effects".

Safety evaluation of metal exposure from commonly used moisturizing and skin-lightening creams in Nigeria.

The concentrations of ten metals (Cd, Pb, Ni, Cr, Cu, Co, Fe, Mn, Zn and Al) were measured in some commonly used moisturizing and skin-lightening creams in Nigeria with a view to providing information on the risk of exposure to metals from the use of these products. The metal concentrations in these products were measured by atomic absorption spectrometry after acid digestion of the samples. The measured concentrations of metals in the skin moisturizing creams ranged from <0.15 to 6.3 µg/g Cd, <0.02 to 17.5 µg/g Cu, 2.25 to 6.25 µg/g Cr, <0.25 to 124.3 µg/g Al, 0.2 to 7.3 µg/g Pb, <0.03 to 10.7 µg/g Ni, 17.3 to 372.0 µg/g Zn, <0.02 to 1.0 µg/g Co, 17.75 to 28.8 µg/g Mn, <0.1 to 89.8 µg/g Fe while the concentrations of metals in the skin-lightening products ranged from <0.15 to 16.5 µg/g Cd, <0.02 to 10.0 µg/g Cu, 4.25 to 8.0 µg/g Cr, <0.25 to 128.0 µg/g Al, 0.5 to 4.5 µg/g Pb, <0.03 to 1.65 µg/g Ni, 24.7 to 267.5 µg/g Zn, <0.02 to 2.5 µg/g for Co, 19.3 to 31.8 µg/g Mn, 9.5 to 211.63 µg/g Fe. In a significant number (>93%) of the samples investigated the concentrations of Pb, Cd, Ni and Co were below the specified limit, or the maximal limit for impurities in colour additives in cosmetics for external use. However, Cr was found at concentrations above the allergenic limit of 1 µg/g. The results also showed that skin-lightening creams contained higher concentrations of the studied metals than the moisturizing creams, except for Ni, which indicates that persons who uses skin-lightening creams in preference to moisturizing ones, are exposed to higher concentrations of metals.