Development, physicochemical evaluation, and in vivo permeation studies of topical formulations containing 0.1% tacrolimus

Abstract The objective of this paper was to develop and evaluate two semi-solid pharmaceutical forms containing 0.1% tacrolimus: cream (CRT01) and gel (GLT01). For the evaluation of physicochemical stability, at times 0, 30, 60 and 90 days, at 23°C and at 40°C, High Performance Liquid Chromatography coupled with a Diode Array Detector (HPLC-DAD) was employed. This method was developed and validated for tacrolimus quantification. The occlusivity test and skin permeation assay were also performed, using an animal model (Wistar rats), and the CRT01 and GLT01 were compared to the 0.1% tacrolimus ointment (PFU01) obtained from the University Pharmacy, Federal University of Rio de Janeiro, Brazil. CRT01 and GLT01 presented a homogeneous aspect and consistency adequate for topical products, along with sensory characteristics above PFU01. They also presented adequate physicochemical stability for 90 days and a lower occlusive effect than PFU01 (p<0.05). CRT01 showed greater affinity for the skin when compared to PFU01 and GLT01, with low systemic absorption. The CRT01 semi-solid formulation was considered the most adequate one to treat patients with atopic dermatitis or other dermatologic inflammatory diseases, promoting rational use of tacrolimus

Mupirocin ointments: In vitro x In vivo bioequivalence evaluation

Abstract Bioequivalence (BE) assessment of topical drug products is a long-standing challenge. Agencies such as the Food and Drug Administration (FDA) and the European Medicines Agency (EMA) have published several drafts in recent years suggesting different approaches as alternative to evaluate the BE. A proposed Topical Classification System (TCS) has even been discussed. Given the above, the objective of this research was to use in vitro and in vivo BE approaches to evaluate Brazilian marketed mupirocin (MPC) ointments, previously classified as TCS class The in vitro permeation test (IVPT) was performed by applying formulations to pig skin by Franz cells. The in vivo methodology was dermatopharmacokinetic (DPK). These approaches (in vivo tape stripping and IVPT) demonstrated capability of distinguishing among different formulations, thus making them useful methodologies for BE evaluation.

Effect of Mesona chinensis polysaccharide on the pasting, rheological, and structural properties of tapioca starch varying in gelatinization temperatures.

The effects of Mesona chinensis polysaccharide (MCP) on the pasting, rheological properties, granule size, and water mobility of tapioca starch (TS) were investigated at different gelatinization temperatures (75 °C and 95 °C). The structures of tapioca starch-Mesona chinensis polysaccharide (TM) gels formed at different gelatinization temperatures were investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). Results showed that the peak, trough, and final viscosities of TM-95 mixtures were lower than that of TM-75 mixtures. Addition of MCP had a significant reduce (p < 0.05) on the granule size and transversal relaxation time of TM mixtures at the two gelatinization temperatures. Rheological analysis also showed that the addition of MCP increased the consistency indexes (K) and decreased the flow behavior indexes (n) of TM-95 and TM-75 gels. XRD results confirmed the diffraction peak of TM-95 gels became blunt and wider, and the diffraction peak at 17° and 23° of TM-75 gels could be observed after MCP added. In addition, the microstructures of TM-75 gels were more compact than that of TM-95 gels. These results can promote the development of TS-based products and application of MCP at different gelatinization temperatures.

Presence of Bisphenol A and Parabens in a Neonatal Intensive Care Unit: An Exploratory Study of Potential Sources of Exposure.

BACKGROUND: Newborns in neonatal intensive care units (NICUs) are in contact with a variety of medical products whose production might include synthetic chemicals with hormonal activity. OBJECTIVES: Our aim was to assess the content of bisphenol A (BPA) and parabens (PBs) and the hormone-like activities of a subset of medical products commonly used in NICUs in prolonged intimate contact with NICU newborns. METHODS: Fifty-two NICU items were analyzed, determining the concentrations of BPA and PBs [methyl- (MeP), ethyl- (EtP), propyl- (PrP), and butylparaben (BuP)] and using the E-Screen and PALM-luciferase assays to measure the in vitro (anti-)estrogenic and (anti-)androgenic activity, respectively, of the extracts. Items found to have elevated BPA/PB content or hormone-like activities were further extracted using leaching methodologies. RESULTS: BPA was found in three-fifths and PBs in four-fifths of tested NICU items, and ∼25% and ∼10% of extracts evidenced estrogenic and anti-androgenic activity, respectively. The highest BPA content was found in the three-way stopcock (>7.000 ng/g), followed by patterned transparent film dressing, gastro-duodenal feeding tubes, sterile gloves, single-lumen umbilical catheters, and intravenous (IV) infusion extension sets (concentrations ranged from 100 to 700 ng/g BPA). A total PB concentration (∑PBs) >100 ng/g was observed in several items, including light therapy protection glasses, patterned transparent film dressing, winged IV catheters, IV infusion extension sets, and textile tape. The highest estrogenic activity [>450 pM estradiol equivalent (E2eq)] was found in small dummy nipples, three-way stopcocks, and patterned transparent film dressing and the highest anti-androgenic activity [>5 mM procymidone equivalent units per gram (Proceq/g)] in small dummy nipples and three-way stopcocks. DISCUSSION: According to these findings, neonates might be exposed to multiple sources of BPA and PBs in NICUs via inhalation, dermal, oral, and IV/parenteral routes. There is a need to address the future health implications for these extremely vulnerable patients and to adopt precautionary preventive measures as a matter of urgency. https://doi.org/10.1289/EHP5564.

Imaging Analysis Enables Differentiation of the Distribution of Pharmaceutical Ingredients in Tacrolimus Ointments.

We demonstrated the difference in the distribution state of pharmaceutical ingredients between tacrolimus (TCR) original ointment and six kinds of generic medicines. Two-dimensional imaging and depth analysis using attenuated total reflection Fourier transform infrared (ATR FT-IR) spectroscopy and confocal Raman microscopy were used, in addition to the evaluation of pharmaceutical properties, including spreading properties, rheological properties, and amount of solvent. The solvents, such as propylene carbonate and triacetin, in TCR ointments formed liquid droplets and dispersed in hydrocarbon oils. Waxes, white beeswax and beeswax, formed other domains. Confocal Raman microscopy could detect liquid droplet size without coalescence of that on germanium or glass surfaces. The combination of ATR FT-IR and confocal Raman imaging would be a powerful tool to reveal the size and shape of liquid droplets of pharmaceutical ingredients in semisolid formulations.

Development and Validation of Analytical Method for Simultaneous Estimation of Active Constituents in a Polyherbal Ointment by Gas Chromatography.

Background: The simultaneous, quantitative determination of all active ingredients present in the analgesic formulation (Dazzle ointment) requires an ideal and novel method by which these phytoconstituents can be separated with the highest resolution without any interference from one another. Objective: The present work was conducted to develop and validate a quantitative method for the simultaneous estimation of all five phytoconstituents present in a polyherbal analgesic ointment by GC. Methods: α-Pinene, 1,8-cineole, camphor, menthol, and methyl salicylate present in the ingredients of the ointment were analyzed and quantified by GC using a crosslinked 5% phenyl polydimethylsiloxane capillary column, nitrogen as a carrier gas, and a flame-ionization detector. Aniline was used as the internal standard. Method validation was also performed in order to demonstrate its selectivity, linearity, accuracy, precision, LOD, LOQ, and robustness. Results: The calibration curves of all five marker compounds showed good linear correlation coefficients (r² >0.998) within the tested ranges. The precision of the method was tested by carrying out intra- and interday analyses of the same sample. RSD values were observed to be <1.00%. The accuracy of the method, determined by performing recovery studies, was found to be between 99.25 and 101.39%. The developed method was also demonstrated to be robust (RSD <1.29%) by making small but deliberate variations in method parameters. Conclusions: The developed GC method is simple, precise, and accurate, it and can be used for the rapid quality control testing of the polyherbal formulation. Highlights: The developed GC method will assist in the standardization of polyherbal analgesic formulation consists of α-pinene, 1,8-cineole, camphor, menthol, and methyl salicylate as active constituents.

In vitro release testing method development for ophthalmic ointments.

It is essential as well as challenging to develop a reliable in vitro release testing method for determining whether differences in release profiles exist between qualitatively and quantitatively equivalent ophthalmic ointment formulations. There is a lack of regulatory guidance on in vitro release testing methods for ophthalmic formulations. Three different in vitro release testing methods 1) USP apparatus 4 with semisolid adapters; 2) USP apparatus 2 with enhancer cells; and 3) Franz diffusion cells were investigated. Qualitatively and quantitatively equivalent ointments were prepared via hot melting and simple mixing methods using four different sources of excipients (i.e. white petrolatum). The ointment formulations were characterized for content uniformity, particle size, and rheological parameters. All the formulations showed adequate content uniformity and similar particle size. The ointments prepared via the hot melting processes showed higher rheological parameters, as did the ointments prepared using 'white' petrolatum that exhibited a yellowish color. The three in vitro release testing methods were compared and evaluated for reproducibility, discriminatory capability, and correlation with the rheological parameters. Compared with the compendial methods, the non-compendial method (Franz diffusion cells) showed poorer reproducibility. All three methods possessed the ability to discriminate between the ophthalmic ointments with manufacturing differences. However, the USP apparatus 4 method displayed the largest margin of discrimination between the release profiles of the different ophthalmic ointments. In addition, the in vitro release rate obtained using the USP apparatus 4 method showed the strongest logarithmic linear correlation with the rheological parameters (Power law consistency index (K value) and crossover modulus) compared to the other two methods.

The palliative efficacy of modified Mohs paste for controlling canine and feline malignant skin wounds.

In veterinary medicine, the management of malignant skin wounds is highly challenging. We conducted a study on seven case animals (four dogs and three cats) which presented with malignant skin wounds. All seven animals had signs and symptoms which were controlled following treatment with a modified Mohs paste. Upon obtaining informed consent from their owners, the animals requiring management of malignant wounds were enrolled in this study. The modified Mohs paste was prepared by mixing zinc chloride, zinc oxide starch powder, glycerin, and distilled water. The modified Mohs paste was topically applied to and left to remain on the malignant wounds for one hour, under controlled conditions. Once the paste was removed, the wounds were irrigated with a solution of sterile saline. At the first examination, the wounds of each animal were observed for signs of exudate, malodor, and bleeding. In every case, visible improvement was observed immediately after the modified Mohs paste treatment. Specifically, the size of the malignant wounds, and the number of times the dressing gauze required changing, significantly decreased (p < 0.05 and p < 0.01, respectively). The open malignant skin wounds caused by mammary gland tumors disappeared in two cases. The Mohs paste has been shown to be a viable option for the palliative treatment in canine and feline malignant skin wound management.

Development of a Single Ion Pair HPLC Method for Analysis of Terbinafine, Ofloxacin, Ornidazole, Clobetasol, and Two Preservatives in a Cream Formulation: Application to In Vitro Drug Release in Topical Simulated Media-Phosphate Buffer Through Rat Skin.

Present work reports an HPLC method with UV detection for quantification of terbinafine, ofloxacin, ornidazole, and clobetasol in a cream formulation along with two preservatives methyl and propyl paraben. The chromatographic separation and quantification was achieved by an octyl bonded column and a gradient elution program involving an ion-pairing reagent, hexanesulfonic acid (0.2%, pH modified to 2.7 using orthophosphoric acid) and acetonitrile. The method was simple and devoid of buffer salts and therefore advantageous for system and column life. The three step gradient program was initiated with 30% (v/v) acetonitrile for the first 5 min and ramped linearly to 60% in the next 7 min. The mobile phase remained constant for the next 11 min and then concluded at 30% (v/v) of acetonitrile. Flow rate throughout was 0.8 mL/min, and all the signals were monitored at 243 nm. The method was applied for assay of a cream formulation and its in vitro permeation studies to determine the penetration profile of the four drugs and two preservatives. A marketed cream formulation was selected for the permeation study, which was carried out using a diffusion cell consisting of topical simulated media, phosphate buffer (pH=6.8) solution containing 1% sodium lauryl sulfate as a receiver medium.

Quantitative analysis of α-mangostin in hydrophilic ointment using near-infrared spectroscopy.

The objective of this research was to quantify the α-mangostin content in mangosteen pericarp (MP) ointment as a colloidal dispersion using near-infrared (NIR) spectroscopy. Various concentrations of MP (IP and EP) ointments containing both internal and external pericarps were prepared and the NIR spectra of these ointments were measured. The NIR spectrum of each ointment was correlated with α-mangostin concentration by partial least square (PLS) regression. Validation of the models was performed and their predictive ability was also investigated. The equation and R(2) value for the prediction of α-mangostin concentration in IP ointment were y=0.9843x+0.4441 and 0.9730 and those in EP ointment were y=0.9569x+0.1142 and 0.9136, respectively. The biases of the IP and EP ointment models were 0.23 and 0.00, respectively. The results showed that NIR could be a useful tool for the quality control of herbal medicine in hydrophilic ointment without any sample preparation. It could predict α-mangostin content in hydrophilic ointment at very low concentration with sufficient accuracy.

Strength and ultrasonic properties of cemented paste backfill.

This paper presents the strength (UCS) and ultrasonic pulse velocity (UPV) properties of cemented paste backfill (CPB) produced from two different mill tailings (Tailings T1 and T2). A total of 240 CPB samples with diameter×height of 5 × 10 cm and 10 × 20 cm prepared at different binder dosages (5-7 wt.%) and water-to-cement ratios (3.97-5.10) were subjected to the UPV and UCS tests at 7, 14, 28 and 56-days of curing periods. UCS and UPV of CPB samples increased with increasing the binder dosage and reducing the w/c ratio irrespective of the sample size and tailings type. CPB samples with a diameter × height of 5 × 10 cm were observed to produce consistently higher (up to 1.69-fold) UCSs than those of 10 × 20 cm CPB samples at all binder dosages and w/c ratios. However, at the corresponding binder dosages and w/c ratios, the maximum variation of UPV between the CPB samples of 5 × 10 cm and 10 × 20 cm was only 7.45%. Using the method of least squares regression, the UCS values were correlated with the UPV values for CPB samples of 10 × 20 cm in size. A linear relation with a high correlation coefficient appeared to exist between the UCS and UPV for CPB samples. These findings suggest that the UPV is essentially independent of the sample size. In this regard, the UPV test can be suitably exploited for the rapid estimation of the strength and quality of CPB samples even using small samples with concomitant benefits of reducing sample size.

Spectrometric analysis of mercury content in 549 skin-lightening products: is mercury toxicity a hidden global health hazard?

BACKGROUND: Cosmetic skin lightening is practiced worldwide. Mercury is a well-documented melanotoxin added to some lightening products. However, mercury can cause many dermatologic, renal, and neurologic problems. The Food and Drug Administration limits the amount of mercury in cosmetic products to trace amounts, 1 ppm. OBJECTIVE: The objective of this study was to quantitatively evaluate a large international sample of lightening products for mercury content, focusing on products available to US consumers either online or in stores. METHODS: A total of 549 skin-lightening products, manufactured in 32 countries, were purchased online in the United States, Taiwan, and Japan and in stores in the United States, China, Taiwan, Thailand, Japan, and Sri Lanka. Cosmetics were screened for mercury content above 200 ppm using a low-cost portable x-ray fluorescence spectrometer. RESULTS: Of the 549 tested products, 6.0% (n = 33) contained mercury above 1000 ppm. In all, 45% of mercury-containing samples contained mercury in excess of 10,000 ppm. Of lightening products purchased in the United States, 3.3% were found to contain mercury in excess of 1000 ppm. LIMITATIONS: Our study did not evaluate creams for other melanosuppressive ingredients. Only 1 sample of each product was tested. CONCLUSION: Our study confirms the national and global presence of mercury in skin-lightening products.

[Simultaneous content determination of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin in ZJHX rubber paste by double wavelength HPLC].

Notoginsenoside R1, ginsenoside Rg1, Re, Rb, and dracorhodin from ZJHX rubber paste were analyzed simultaneously by HPLC, with acetonitrile-water as the mobile phase for gradient elution at a flow rate of 1.0 mL x min(-1). The column temperature was 35 degrees C and the sample size was 10 microL. The detection wavelength was set at 203 nm for ginsenoside and 440 nm for dracorhodin, respectively. The results showed that all of notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and dracorhodin could be were separated well by baseline, with the linear ranges of 0.251-5.020 microg (R2 = 0.999 8), 0.520-10.400 microg (R2 = 0.999 9), 0.251-5.010 microg (R2 = 0.999 7), 0.505-10.100 microg (R2 = 0.999 8) and 0.160-3.270 microg (R2 = 0.999 9), respectively. Each component showed a good linear relationship, with the average recoveries ranging from 99.39% to 100.5%. The established method was so simple, accurate and highly reproducible that it could be used for quality control of ZJHX rubber paste.

Properties and analytical applications of the self-assembled complex {peroxidase-chitosan}.

A novel promising approach to the improvement of analytical properties of horseradish peroxidase based on its inclusion into self-assembled structures of chitosan is discussed. It is shown that the reasonable choice of a polyelectrolyte, a detailed investigation of its interaction with the enzyme and the conditions of the {peroxidase-polyelectrolyte} complex formation allow for stabilizing the biocatalyst in aqueous and aqueous-organic media without a substantial loss in its activity and developing corresponding analytical procedures and biosensors. The latter provides highly selective determination of a number of organic compounds and sensitive determination of heavy metal ions that becomes possible due to the specific interactions of the analytes with the polymer matrix. Besides, the application of the proposed analytical systems and biosensors provides the expansion of the range of the compounds, and poorly water soluble and slowly oxidized substrates of peroxidase as well, which could be determined and real samples which could be analyzed by enzymatic methods. Analytical performance of the developed spectrophotometric indicator procedures and biosensors based on the self-assembled complex {peroxidase-chitosan} is demonstrated in the determination of metal ions (Hg(II), Cd(II), and Pb(II)), phenothiazines (promazine, chloropromazine, and trifluoroperazine), phenolic compounds (phenol, hydroquinone, catechol, pyrogallol, quercetin, rutin, and esculetin), organic peroxides (tert-butyl peroxide, 2-butanone peroxide, and benzoyl peroxide) in various samples, including water-insoluble matrices.

[Simultaneous determination of six antibiotics in disinfection products by high performance liquid chromatography tandem mass spectrometry].

OBJECTIVE: To develop a method for determination of the Metronidazole, Chlortetracycline hydrochloride, Oxytetracycline dihydrate, Minocycline hydrochloride, Erythromycin and Tetracycline hydrochloride in disinfection products by high performance liquid chromatography tandem mass spectrometry(LC-MS/MS). METHODS: Samples were extracted by methanol and filtered through 0.45 microm PTFE membrane filter, then analyzed by LC-MS/MS using Waters Symmetry C18 (2.1 mm x 150 mm, 3.5 microm) column in positive ion scan mode. The mobile phase was 5 mmol/L ammonium acetate, methanol and acetonitrile. RESULTS: The linear range was 0-2000 ng/ml and the correlation coefficients were more than 0.998, the average recoveries ranged from 74.7% to 114% with the relative standard deviations between 1.6%-20.2%. The method was successfully used to detect the content of antibiotics in 115 disinfection products. CONCLUSION: The method is simple, sensitive, selective and suitable for the analysis of residual content of antibiotics in cream formulations of disinfection products.

Evaluation of a short stability-indicating HPLC method for diclofenac sodium gels.

A fast and reproducible high performance liquid chromatography method has been developed for the determination of diclofenac sodium and its degradation products in commercial and in in-house produced ointments. The method employs a RP-LiChrospher select B (C8) column with a mobile phase containing methanol/water (63:37, v/v) and detection at 220 nm. This rapid and simple HPLC assay was used for QA/QC of large scale in-house produced diclofenac gel. The validation protocol was designed following international guidelines, e. g. ICH Q2(R1). Selectivity tests also included the separation of synthesis related by-products like 1-(2,6-dichlorphenyl)indoline-2-one (impurity A) and indoline-2-one (impurity E), and in addition selectivity with regard to several photodegradation products produced by both UV and simulated sunlight irradiation has been shown.

Pyrolysis-GC/MS for the identification of macromolecular components in historical recipes.

Analytical pyrolysis with thermally assisted hydrolysis and methylation was employed to investigate ancient ointments collected from Spanish vessels coming from the sixteenth century pharmacies. The ointments were reproduced on the basis of historical recipes and characterization was made in comparison with real samples. Characteristic markers indicate the presence of beeswax, of animal and plant lipids, and of natural resins. Analyses of old samples are consistent with the modern reproductions and with the analysis of raw materials. Multivariate data analysis was used to discriminate between the different types of lipidic materials, also in connection with their relative amount in the samples.

Phthalates in baby skin care products.

BACKGROUND: The systemic toxicity of phthalates has been extensively reported. Although rarely sensitizing, phthalates have been implicated in promoting the development of both atopy and contact dermatitis in animal models. Dermal absorption of phthalates may contribute to overall chemical burden. Infants may be particularly susceptible to chemical exposures. Baby skin care products may be a significant source of phthalate exposure. OBJECTIVE: We measured the phthalate content of 30 skin care products intended for babies and children. METHODS: Nineteen leave-on and 11 wash-off baby skin care products were analyzed for 17 unique phthalates by an independent laboratory using standard gas chromatographic mass spectrometry. RESULTS: Of 30 products tested, four had phthalate levels above the reporting limit (0.1-0.5 ppm); of these, only one had levels above 1 ppm (44 ppm). There was no statistical significance of phthalate detection in leave-on versus wash-off products (p = .578). CONCLUSION: The majority (26 of 30) of the baby skin care products analyzed did not have detectable phthalate levels. Four products had detectable phthalate levels. In baby skin care products, levels of the 17 phthalates tested are low overall, but occasional products may contain higher phthalate levels. Monitoring products to ensure safety standards are met may be warranted.

Determination of retinyl palmitate in ointment by HPLC with diode array detection.

A simple and rapid HPLC with diode array detection method was developed for the determination of retinyl palmitate present together with other active substances in an ointment. Chromatographic separation was performed on 100 RP-18 Lichrospher column of particle size 5 microm. The mobile phase was methanol:water (98:2, v/v) and flow rate was 2.0 mL/min in isocratic mode. Samples were analyzed for 30 min. Spectophotometric detection was conducted at 325 nm. Under these conditions, the method featured high sensitivity, good precision and comparability of results as proven by the method validation and statistical analysis of the results. The limits of detection and determination were 0.4317 mg/100 mL and 1.3081 mg/100 mL, respectively, recovery values were measured at three levels 80%, 100% and 120% and yielded 101.05%, 101.34% and 100.43%, respectively. The linearity range was checked from 2 mg/100 mL to 10 mg/100 mL. The precision and inter-day precision of the method was expressed by relative standard deviation value and did not exceed 1.68%.