Rapid determination of propylthiouracil and methimazole by surface-enhanced Raman scattering based on sodium alginate-protected silver nanoparticles.

A rapid, ultrasensitive and selective method has been established to determine antithyroid drugs [propylthiouracil (PTU) and methimazole (MTZ)] by surface-enhanced Raman scattering (SERS). Silver nanoparticles (AgNPs) were synthesized by reduction of silver nitrate (AgNO3) with sodium alginate (SA) and were used as SERS substrates. Through optimization of the SERS conditions, including the volume of SA-protected AgNPs solution, pH of Britton-Robinson (BR) buffer solution and concentration of NaCl solution, linear responses were obtained for PTU and MTZ in the concentration ranges of 3.02 × 10-9-1.06 × 10-5 mol L-1 and 1.21 × 10-9-1.21 × 10-5 mol L-1, respectively. By the present method, the limits of detection (LODs) for the determination of PTU and MTZ were as low as 1.58 × 10-10 mol L-1 and 2.97 × 10-11 mol L-1. The method was successfully applied for the determination of PTU and MTZ in real samples. The recovery of PTU and MTZ from actual samples ranged between 97.20 and 104.80%, with relative standard deviations (RSDs) less than 3.63%.

Extrapolating In Vitro Screening Assay Data for Thyroperoxidase Inhibition to Predict Serum Thyroid Hormones in the Rat.

Thyroperoxidase (TPO) is an enzyme essential for thyroid hormone (TH) synthesis and a target site for a number of xenobiotics that disrupt TH homeostasis. An in vitro high-throughput screening assay for TPO inhibition, the Amplex UltraRed-TPO (AUR-TPO), has been used to screen the ToxCast chemical libraries for this action. Output from this assay would be most useful if it could be readily translated into an in vivo response, namely a reduction of TH in serum. To this end, the relationship between TPO inhibition in vitro and serum TH decreases was examined in rats exposed to 2 classic TPO inhibitors, propylthiouracil (PTU) and methimazole (MMI). Serum and gland PTU, MMI, and TH levels were quantified using tandem liquid chromatography mass spectrometry. Thyroperoxidase activity was determined in thyroid gland microsomes treated with PTU or MMI in vitro and ex vivo from thyroid gland microsomes prepared from exposed animals. A quantitative model was constructed by contrasting in vitro and ex vivo AUR-TPO results and the in vivo time-course and dose-response analysis. In vitro:ex vivo correlations of AUR-TPO outputs indicated that less than 30% inhibition of TPO in vitro was sufficient to reduce serum T4 by 20%, a degree of regulatory significance. Although further testing of model estimates using other TPO inhibitors is essential for verification of these initial findings, the results of this study provide a means to translate in vitro screening assay results into predictions of in vivo serum T4 changes to inform risk assessment.

Food Consumption Patterns and Body Composition in Children: Moderating Effects of Prop Taster Status.

This cross-sectional study determined whether 6-n-propylthiouracil (PROP) taster status moderates the relationship between food consumption patterns and body composition in children. Children were recruited (n = 342, 50% female, 8-10 y) from across New Zealand. Using a food frequency questionnaire, these food consumption patterns were derived: Processed Foods, Fruit and Vegetables, and Breakfast Foods. Body composition variables included: body fat (%), fat mass (kg), fat mass index (FMI, kg/m2), body mass index (kg/m2) and waist to height ratio (W:Ht). Following adjustment for confounders, Processed Foods were positively associated with %fat (p = 0.015), fat mass (p = 0.004) and FMI (p = 0.016). Taste test strips determined PROP status. For Breakfast Foods, there were small negative associations with all body composition variables (p ≤ 0.001 to 0.037). The population sample was also stratified by PROP taster status. For the non-tasters, there were small to moderate negative associations between Breakfast Foods and each body composition variable (p = 0.003-0.045) except W:Ht (p = 0.112), and these relationships were stronger for girls compared to boys. For the tasters, there were small to moderate positive associations between Processed Foods with %fat (p = 0.030), fat mass (p ≤ 0.001) and FMI (p = 0.014). In conclusion, sensitivity to bitterness may moderate the relationship between food consumption patterns and body composition in children.

Influences of Psychological Traits and PROP Taster Status on Familiarity with and Choice of Phenol-Rich Foods and Beverages.

Plant phenolics are powerful antioxidants and free radical scavengers that can contribute to the healthy functional properties of plant-based food and beverages. Thus, dietary behaviours rich in plant-based food and beverages are encouraged. However, it is well-known that the bitter taste and other low-appealing sensory properties that characterize vegetables and some other plant-based foods act as an innate barrier for their acceptance. The aim of this study was to investigate the influence of psychological traits and PROP status (the responsiveness to bitter taste of 6-n- propylthiouracil) on the choice of and familiarity with phenol-rich vegetables and beverages varying in recalled level of bitterness and astringency. Study 1 aimed at assessing the variations of the sensory properties of vegetable and coffee/tea items with two check-all-that-apply (CATA) questionnaires (n = 201 and n = 188 individuals, respectively). Study 2 aimed at investigating how sensitivity to punishment, to reward, and to disgust, food neophobia, private body consciousness, alexithymia, and PROP responsiveness affect choice and familiarity with phenol-rich foods (n = 1200 individuals). A Choice Index was calculated for vegetables (CV) and coffee/tea (CC) as a mean of the choices of the more bitter/astringent option of the pairs and four Familiarity Indices were computed for vegetables (FV) and coffee/tea (FC), higher (+) or lower (-) in bitterness and astringency. Subjects higher in food neophobia, sensitivity to punishment or sensitivity to disgust reported significantly lower choice indices than individuals lower in these traits, meaning that they systematically opted for the least bitter/astringent option within the pairs. Familiarity with vegetables was lower in individuals high in sensitivity to punishment, in food neophobia and in alexithymia, irrespective of their sensory properties. The Familiarity Index with coffee/tea characterized by higher bitterness and astringency was lower in individuals high in food neophobia, sensitivity to disgust, and alexithymia. No significant effect of PROP was found on any indices. The proposed approach based on product grouping according to differences in bitterness and astringency allowed the investigation of the role of individual differences in chemosensory perception and of psychological traits as modulators of phenol-rich foods preference and consumption.

Analysis of thyreostats in bovine feces using ultra high performance liquid chromatography with tandem mass spectrometry.

A method based on ultra-high performance liquid chromatography was developed and validated to detect six thyreostatic compounds: tapazole, thiouracil, methylthiouracil, dimethylthiouracil, propylthiouracil, and phenylthiouracil in faeces of bovine. Thyreostats were extracted from the matrix with a mixture of methanol and buffer (pH = 8). Next step was derivatization of analytes with 3-iodobenzylbromide. The liquid chromatographic separation of derivatives was obtained on a SB-C18 column (50 × 2.1 mm; 1.8 µm, Agilent) with gradient elution using a mobile phase consisting of acetonitrile/0.1% acetic acid within 7.5 min. The analysis was performed on a Shimadzu NEXERA X2 ultra-high performance liquid chromatograph with triple quadrupole MS 8050 instrument operating in positive electrospray ionization mode. Depending on the target compound, two or three diagnostic signals (selected reaction monitoring transitions) were monitored. The procedure was validated according to the Commission Decision 2002/657/EC. Recovery and repeatability met the performance criteria specified by this document for banned compounds. The recovery ranged from 97.5 to 110.5%, and repeatability did not exceed 14.1%. Decision limits and detection capabilities were below 10 µg/kg. The highest decision limits and detection capabilities concentrations were observed for phenylthiouracil of 3.48 and 6.96 µg/kg, respectively.

Validation of edible taste strips for assessing PROP taste perception.

A novel delivery method is described that incorporates taste stimuli into edible strips for determining n-propylthiouracil (PROP) taster status. Edible strips that contained 400 or 600 nanomoles of PROP were prepared for psychophysical studies. Using these strips, we measured taste intensity, taste hedonics, and taste quality responses in a sample of healthy volunteers (n = 118). Participants were also asked to assess a single NaCl strip, a quinine strip, 3 NaCl solutions, and 3 PROP solutions. All psychophysical data were subsequently analyzed as a function of TAS2R38 genotype. The use of PROP strips for distinguishing between individuals with at least 1 PAV allele and individuals with other genotypes was assessed and compared with the use of PROP solutions for making this same distinction. For the 2 PROP strips and PROP solutions, individuals who expressed at least 1 PAV allele could perceive the bitter taste of PROP. Individuals who expressed 2 AVI alleles responded similarly to 400 nanomole PROP strips and blank strips. Furthermore, individuals with 2 AVI alleles responded to 0.032 and 0.32 mM PROP solutions at intensities that were similar to water, though intensity ratings to 3.2 mM PROP solution exceeded water. In general, those with at least 1 PAV allele rated the bitter taste of PROP as unpleasant in both delivery methods (strips or solutions). Psychophysical data from PROP strips and solutions were consistent with TAS2R38 genotype. These results support the validity of edible taste strips as a method for assessing PROP taste perception in humans.

Offering "dip" promotes intake of a moderately-liked raw vegetable among preschoolers with genetic sensitivity to bitterness.

BACKGROUND: Evidence-based strategies for promoting vegetable consumption among children are limited. OBJECTIVE: To determine the effects of providing a palatable "dip" along with repeated exposure to a raw vegetable on preschoolers' liking and intake. PARTICIPANTS: One hundred fifty-two predominately Hispanic preschool-aged children studied in Head Start classrooms in 2008. DESIGN: A between-subjects, quasiexperimental design was used. A moderately-liked raw vegetable (broccoli) was offered twice weekly at afternoon snacks for 7 weeks. Classrooms were randomized to receive broccoli in one of four conditions differing in the provision of dip. Bitter taste sensitivity was assessed using 6-n-propylthiouracil. INTERVENTION: Broccoli was provided in four conditions: with regular salad dressing as a dip, with a light (reduced energy/fat) version of the dressing as a dip, mixed with the regular dressing as a sauce, or plain (without dressing). MAIN OUTCOME MEASURES: Mean broccoli intake during 7 weeks of exposure and broccoli liking following exposure. STATISTICAL ANALYSES: Descriptive statistics were generated. Multilevel models for repeated measures tested effects of condition and bitter sensitivity on mean broccoli intake during exposure and on pre- and post-exposure liking while adjusting for classroom effects and potential covariates. RESULTS: The majority of Hispanic preschoolers (70%) showed sensitivity to the bitter taste of 6-n-propylthiouracil. Children's broccoli liking increased following exposure but did not vary by dip condition or bitter sensitivity. Bitter-sensitive children, however, ate 80% more broccoli with dressing than when served plain (P<0.001); effects did vary based on whether regular or light dressing was provided as a dip or sauce. Dip did not promote broccoli intake among bitter-insensitive children. CONCLUSIONS: Providing dip­regular, light, or as a sauce­increased raw broccoli intake among bitter-sensitive Hispanic preschoolers. Findings suggest that offering low-fat dips can promote vegetable intake among some children who are sensitive to bitter tastes.

Optimization of carrier-mediated three-phase hollow fiber microextraction combined with HPLC-UV for determination of propylthiouracil in biological samples.

Carrier-mediated three-phase hollow fiber microextraction combined with high-performance liquid chromatography-ultra violet detection (HPLC-UV) was applied for the extraction and determination of propylthiouracil in biological samples. Propylthiouracil (PTU) was extracted from 7.5 mL of the basic solution (the source phase) with pH 12 into an organic phase (n-octanol containing 6% (w/v) of Aliquat 336 as the carrier) impregnated in the pores of a hollow fiber, and finally was back extracted into 24 µL of the acidic solution located inside the lumen of the hollow fiber (the receiving phase). The extraction was performed through the gradient of counter ion from the source to the receiving phase. The effects of different variables on the extraction efficiency were studied simultaneously using an experimental design. A half-fractional factorial design was employed for screening to determine the variables significantly affecting the extraction efficiency. Then, the factors with significant effect were optimized using a central composite design (CCD) and the response surface equations were developed. The optimal experimental conditions obtained from this statistical evaluation included: source phase, pH 12; temperature, 25°C; extraction time, 40 min; counter ion concentration, 2 mol L(-1) of NaClO(4); organic solvent 6% of Aliquat in octanol and without salt addition in the source phase. Under the optimized conditions, the preconcentration factors were between 125 and 198 and also the limit of detections (LODs) ranged from 0.1 µg L(-1) to 0.4 µg L(-1) in different biological samples. The calibration curve was linear (r(2)=0.998) in the concentration range of 0.5-1000 µg L(-1). Finally, the feasibility of the proposed method was successfully confirmed by extraction and determination of PTU in human plasma and urine as well as the bovine milk and meat samples in microgram per liter, and suitable results were obtained (RSDs<6.3%).

Estudio de estabilidad de tabletas de propiltiouracilo 50 mg / Study of the 50 mg Propylthiouracil tablets stability

Se desarrolló el estudio de estabilidad de las tabletas de propiltiouracilo 50 mg y se determinó su fecha de vencimiento. Este estudio se realizó por los métodos de vida de estante y de estabilidad acelerada mediante cromatografía líquida de alta eficiencia, validados en el Centro de Investigación y Desarrollo de Medicamentos. El estudio de vida de estante se desarrolló por un periodo de 24 meses a temperatura ambiente; mientras que el estudio de estabilidad acelerada se efectuó sometiendo el producto a la influencia de la luz, la humedad y la temperatura; se realizó el anàlisis durante 3 meses, para los 2 primeros y durante 6 meses para el estudio de la temperatura. La formulación de propiltiouracilo tabletas 50 mg cumplió con las especificaciones de calidad descritas en la farmacopea. Los resultados del estudio de estabilidad por vida de estante después de transcurridos los 24 meses indicaron que el producto mantenía los paràmetros que determinan su calidad durante ese tiempo, y en los estudios acelerados no se observó degradación significativa del producto. Se estableció 2 años como fecha de vencimiento en las condiciones señaladas.

Autors developed a stability study of 50 mg Propylthiouracil tablets and determination of its expiry date. This study was conducted by fixed life methods and of accelerated stability by high-performance liquid chromatography, validated in Drugs Research and Development Center. Fixed life study was conducted during 24 months at room temperature; whereas the accelerated stability study was conducted exposing the product to light influence, humidity and temperature; during 3 months a analysis was performed for the two first ones and over 6 months in the case of temperature study. Propylthiouracil formula (50 mg tablets) fulfilled the quality specifications described in Pharmacopeia. Results of stability study by fixed life after 24 monhts showed that thr product maintain the parameter determining its quality during this period, and in the accelerted studies there was not a significant degradation of product. Two years was the expity date established in above mentioned conditions.

Validación del método analítico para el control de la calidad y estudio de estabilidad de propiltiouracilo 50 mg / Analytical method validation for quality control and the study of the 50 mg propylthiouracil stability

Se desarrolló y validó un método analítico por cromatografía líquida de alta resolución, para el control de la calidad y los estudios de estabilidad del propiltiouracilo 50 mg, tabletas. El método se basó en la separación del principio activo a través una columna cromatográfica Lichrospher 100 RP-18 RP-18 (5 µm) (250 x 4 mm), con detección ultravioleta a 272 nm, para lo cual se empleó una fase móvil compuesta por una mezcla desgasificada de solución amortiguadora fosfato de potasio monobásico 0,025 M a pH= 4,6, y acetonitrilo en una proporción de 80:20, con una velocidad de flujo de 0,5 mL/min. El método analítico resultó lineal, preciso, específico y exacto en el intervalo de concentraciones estudiadas.

A high-performance liquid chromatography analytical method was developed and validated for the quality control and stability studies of 50 mg Propylthiouracil tablets. Method is based in active principle separation through a 100 RP-18 RP-18 (5 µm) (250 x 4 mm) Lichrospher chromatography with UV detection to 272 nm, using a mobile phase composed by a ungaseous mixture of a 0.025 M buffer solution-monobasic potassium phosphate to pH= 4,6 ad acetonitrile in a 80:20 ratio with a flux speed of 0,5 mL/min. Analytical method was linear, precise, specific and exact in the study concentrations interval.

Validación del método analítico para el control de la calidad y estudio de estabilidad de propiltiouracilo 50 mg / Analytical method validation for quality control and the study of the 50 mg Propylthiouracil stability

Se desarrolló y validó un método analítico por cromatografía líquida de alta resolución, para el control de la calidad y los estudios de estabilidad del propiltiouracilo 50 mg, tabletas. El método se basó en la separación del principio activo a través una columna cromatográfica Lichrospher 100 RP-18 RP-18 (5 µm) (250 x 4 mm), con detección ultravioleta a 272 nm, para lo cual se empleó una fase móvil compuesta por una mezcla desgasificada de solución amortiguadora fosfato de potasio monobásico 0,025 M a pH= 4,6, y acetonitrilo en una proporción de 80:20, con una velocidad de flujo de 0,5 mL/min. El método analítico resultó lineal, preciso, específico y exacto en el intervalo de concentraciones estudiadas(AU)

A high-performance liquid chromatography analytical method was developed and validated for the quality control and stability studies of 50 mg Propylthiouracil tablets. Method is based in active principle separation through a 100 RP-18 RP-18 (5 µm) (250 x 4 mm) Lichrospher chromatography with UV detection to 272 nm, using a mobile phase composed by a ungaseous mixture of a 0.025 M buffer solution-monobasic potassium phosphate to pH= 4,6 ad acetonitrile in a 80:20 ratio with a flux speed of 0,5 mL/min. Analytical method was linear, precise, specific and exact in the study concentrations interval(AU)

Estudio de estabilidad de tabletas de propiltiouracilo 50 mg / Study of the 50 mg Propylthiouracil tablets stability

Se desarrolló el estudio de estabilidad de las tabletas de propiltiouracilo 50 mg y se determinó su fecha de vencimiento. Este estudio se realizó por los métodos de vida de estante y de estabilidad acelerada mediante cromatografía líquida de alta eficiencia, validados en el Centro de Investigación y Desarrollo de Medicamentos. El estudio de vida de estante se desarrolló por un periodo de 24 meses a temperatura ambiente; mientras que el estudio de estabilidad acelerada se efectuó sometiendo el producto a la influencia de la luz, la humedad y la temperatura; se realizó el anàlisis durante 3 meses, para los 2 primeros y durante 6 meses para el estudio de la temperatura. La formulación de propiltiouracilo tabletas 50 mg cumplió con las especificaciones de calidad descritas en la farmacopea. Los resultados del estudio de estabilidad por vida de estante después de transcurridos los 24 meses indicaron que el producto mantenía los paràmetros que determinan su calidad durante ese tiempo, y en los estudios acelerados no se observó degradación significativa del producto. Se estableció 2 años como fecha de vencimiento en las condiciones señaladas(AU)

Autors developed a stability study of 50 mg Propylthiouracil tablets and determination of its expiry date. This study was conducted by fixed life methods and of accelerated stability by high-performance liquid chromatography, validated in Drugs Research and Development Center. Fixed life study was conducted during 24 months at room temperature; whereas the accelerated stability study was conducted exposing the product to light influence, humidity and temperature; during 3 months a analysis was performed for the two first ones and over 6 months in the case of temperature study. Propylthiouracil formula (50 mg tablets) fulfilled the quality specifications described in Pharmacopeia. Results of stability study by fixed life after 24 monhts showed that thr product maintain the parameter determining its quality during this period, and in the accelerted studies there was not a significant degradation of product. Two years was the expity date established in above mentioned conditions(AU)

Pharmacologic treatment of hyperthyroidism during lactation.

QUESTION: I have a patient who has hyperthyroidism due to Graves disease. She was taking methimazole but discontinued when she found out she was pregnant. She is currently close to delivery and might require antithyroid therapy in the postpartum period. Can methimazole cross into human milk, and is breastfeeding safe for her infant? ANSWER: The exposure of infants to methimazole or propylthiouracil through breast milk is minimal and not clinically significant. Women with hyperthyroidism using methimazole or propylthiouracil should not be discouraged from breastfeeding, as the benefits of breastfeeding largely outweigh the theoretical minimal risks.

Determination of propylthiouracil in pharmaceutical formulation by high-performance liquid-chromatography with a post-column iodine-azide reaction as a detection system.

A high-performance liquid chromatographic method with a post-column iodine-azide reaction has been chosen and tested for validity in quantitative determination of propylthiouracil in tablets. A mobile phase with a flow rate of 1.4 ml/min was conducted in the form of isocratic chromatography on a C18 column with acetonitrile-water-sodium azide solution (2.5%; pH 5.5) 24:26:50 (v/v/v). Unreacted iodine from post-column iodine-azide induced by reaction was monitored with visible detection at lambda=350 nm. The method proved both its linearity within the range of 8-100 nM (r2>0.9988) and satisfactory results of inter-day precision (RSD<4.2%) and accuracy (recovery>91%). The limits of detection (DDL) and quantification (DQL) reached the levels of 5 and 8 nM, respectively. The validation of the method comprised also its specificity. The results obtained proved the suitability and appropriateness of the suggested method for intended use.

Consumer liking of refined and whole wheat breads.

Preference for refined bread is often cited as a reason for the relatively low consumption of whole wheat bread; only a few studies, however, have examined consumer preferences between refined and whole wheat breads, and the results of these studies are inconclusive. Our objective was to determine if refined wheat bread is preferred to whole wheat bread. We hypothesized that people would prefer refined wheat bread. We conducted a taste test with 89 people. They rated their liking of 9 different breads chosen to represent several comparisons between equivalent refined and whole wheat breads. The participants also rated the intensity of 6-n-propylthiouracil (PROP) and completed a questionnaire about their bread preferences and purchasing habits. We classified the participants by their bread preference and their PROP taster status, and then examined the liking patterns of these subgroups. People preferred refined bread to whole wheat bread when both were made using equivalent ingredients and procedures. They liked the commercial samples of refined and whole wheat breads equally well. When people were classified by their bread preference, those who preferred refined bread liked the refined bread better in all comparisons. PROP nontasters liked all refined and whole wheat breads equally. Sensory preferences are a barrier to whole wheat bread consumption, but ingredient or processing modifications can improve liking of whole wheat bread to the level of refined bread.

1,3-Dibromo-5,5-dimethylhydantoin (DBH) as oxidant and precipitant for drug identification according to PH. EUR Analytical methods of pharmacopoeias with DBH in respect of environmental and economical concern, Part 15(1).

Elemental bromine serves as oxidant for the identification of propylthiouracil, 2-thiouracil and sulphur according to PH. EUR. 2002. Phenol is identified according to PH. EUR. 2002 with bromine water as the sparingly water-soluble 2,4,4,6-tetrabromo-2,5-cyclohexadiene-1-one. These tests can be performed better using 1,3-dibromo-5,5-dimethylhydantoin (DBH).

Determination of propylthiouracil using 1,3-dibromo-5,5-dimethylhydantoin (DBH): analytical methods of pharmacopeias with DBH in respect to environmental and economical concern: part 9.

USP 2000 [The United States Pharmacopeia, Rockville USA, 24th ed., 2000, pp. 1436] and PH. EUR. 1997 [European Pharmacopoeia, third ed., Council of Europe, Strasbourg, 1997, p. 1401] determine propylthiouracil using neutralization titration, whereby 0.1 M silver nitrate and twice boiling is necessary. With the application of 1,3-dibromo-5,5-dimethylhydantoin (DBH), the assay of propylthiouracil can be performed easily, faster and friendlier to environment. A mean deviation of 0.03% and a relative standard deviation of 0.3% are obtained. 5-Bromo-6-propyluracil is formed, when propylthiouracil is determined with DBH.

Development and validation of a high-performance liquid chromatographic method for the analysis of propylthiouracil in pharmaceuticals.

A simple, rapid, and stability-indicating high-performance liquid chromatographic (HPLC) method was developed and validated for the assay of propylthiouracil (PTU). The method was used to quantify PTU in topical formulations and in tablets. Excellent linearity was observed between PTU concentration and the peak area (R2= 0.999). The limit of detection was 1 ng, and the limit of quantitation was 1.2 ng. The method proved to be selective. Selectivity was validated by subjecting a stock solution of PTU to acidic, basic, and oxidative degradations. The peaks of the degradation products did not interfere with the peak of PTU. Excipients present in the dosage forms did not interfere with the analysis, and the recovery of PTU from each dosage form was quantitative.

Iodine-131 elimination from breast milk: a case report.

This case report describes the management of a breastfeeding mother who had been given radioactive iodine and technetium for diagnosis of thyroid disease. The mother requested to submit weekly milk samples for monitoring of radioactivity. Once activity fell below measurable counts, the mother resumed lactation.