Simultaneous detection of sibutramine and phenolphthalein in a diet jelly health food product that caused health problems.

The Public Health Center in Chiba Prefecture, Japan, received a consultation from a resident of Chiba Prefecture who consumed a diet jelly health food product and experienced health problems. To investigate the cause of the health problems, we examined the two food products for the presence of pharmaceutical ingredients. A screening analysis using ultra-high-performance liquid chromatography with a photodiode array detector (UPLC-PDA) indicated the presence of sibutramine and phenolphthalein in the food product. Analysis using an ultra-high-performance liquid chromatography-quadrupole-Kingdon trap mass spectrometer (UHPLC-Q-Kingdon trap MS) confirmed the presence of sibutramine and phenolphthalein. Quantitative analysis using UPLC-PDA showed that sibutramine and phenolphthalein were present at 15 and 16 mg/bag and 2.4 and 2.6 mg/bag, respectively. According to the drug insert for sibutramine capsules in the United States, the recommended medicinal dose of sibutramine should not exceed 15 mg/day, and the amount ingested in the present case exceeded that value. The present study results indicated that ingestion of the jelly health food product may cause health problems.

Methods for quantifying phenolphthalein in slimming tea.

Because of increasing levels of obesity and the development of social network platforms, the online sale of slimming beverages has become very common. However, such beverages are often reported to contain hazardous drugs, such as phenolphthalein. In the present study, we established a rapid and sensitive immunochromatography gold-labeled monoclonal antibody (mAb) (IGM) method and a fluorescence-labeled mAb (IFM) method for determining phenolphthalein in ten types of slimming tea. The monoclonal antibodies and coating antigens were produced in our laboratory. Under optimal conditions, the cut-off limits were 250 ng mL-1 (IFM strip) and 500 ng mL-1 (IGM strip) in both 0.01 M phosphate-buffered saline (PBS) and samples of slimming tea. The mean recoveries were 96.2 to 104.7% for the IGM strip and 90.7 to 104.7% for the IFM strip. The data showed that the IFM strip was more sensitive than the IGM strip and that results could be generated within 10 min. Consequently, this novel technique represents a rapid and convenient method with which to detect phenolphthalein.

Exudate - phenolphthalein pH test for evaluation of validity in seeds of Libidibia ferrea.

Seed companies are looking for promising, quick and effective alternatives to determine the physiological quality of seeds. The objective of the current work was to study the efficiency of the exudate - phenolphthalein pH test to evaluate the seeds of two lots of Libidibia ferrea (Mart. ex Tul.) L. P. Queiroz var. ferrea. The statistical design for the the exudate - phenolphthalein pH test was completely randomized with four replicates of 50 seeds in a factorial design (2 x 5), two seed lots and five soaking periods (30, 60, 90, 120, and 150 minutes), respectively, using two constant temperatures (25 and 30°C). The percentage of viability and germination of the seeds did not differ in the temperatures of 25 and 30°C and in the soaking periods by the exudate - phenolphthalein pH test. Thus, it is recommended that the test be conducted for at least 30 minutes in distilled and deionized water at the constant temperature of 25 or 30°C to evaluate the vigor of the Libidibia ferrea.

A graphene tip coupled with liquid chromatography tandem mass spectrometry for the determination of four synthetic adulterants in slimming supplements.

Slimming supplements were popularly sold online driven by the increasement of obesity and the development of social networking platform. However, events of drug abuse in slimming supplements were also frequently reported. In this study, a graphene tip solid-phase extraction (Gtip SPE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established for determining fenfluramine, phenolphthalein, bumetanide, and sibutramine in slimming supplements. It was validated in terms of linearity (0.9985-0.9995), LOD (1.8ngmL-1), LOQ (5.6ngmL-1), intra-day precision (<5.1%), inter-day precision (<7.3%), and recovery (82.9-95.2%). Sibutramine is the most commonly used drug, which was detected in Bihais, Galong, and Aolist, with content 12.4, 3.6, 20.3mgg-1, respectively. Phenolphthalein was also found with content lower than 5.2mgg-1. The successful application of Gtip SPE and UPLC-MS/MS method indicated its advantage in analyzing low level of contaminates resulted from violation of regulation.

Colorimetric Detection of Thiophenol Based on a Phenolphthalein Derivative and Its Application as a Molecular Logic Gate.

A phenolphthalein-based colorimetric probe bearing a dinitrobenzene group is reported as a thiophenol (PhSH)-selective chemodosimeter. PhSH can react with chemodosimeter 1 to afford phenolphthalein. The addition of PhSH to the aqueous solution of 1 followed by a change in pH of the resulting solution to basic induces a selective color change from colorless to pink. Furthermore, using PhSH and base as inputs and color change of 1 by naked eye as an output, leads to the construction of an AND logic gate.

Detection of Synthetic Drugs as Adulterants in Natural and Herbal Slimming Products by LC-ESI-MS/MS with Polarity Switching.

The purpose of this study was to develop an LC-tandem MS method for the simultaneous detection of common synthetic drugs as adulterants in natural and herbal slimming products. Sixteen drugs belonging to a wide range of pharmaceutical classes were studied. Included in the list of drugs were anorexics, anxiolytics, antidepressants, diuretics, laxatives, and stimulants. The method used a C18 column (4.6 × 50 mm and 1.8 µm particle size). Separation of the drugs was achieved by gradient elution using 4 mM ammonium formate in water + 0.1% formic acid as the aqueous component and 4 mM ammonium formate in methanol + 0.1% formic as the organic component of the mobile phase. As not all of the analytes ionized in the positive mode, the mass spectrometer was operated in the electrospray ionization mode with polarity switching. The samples were extracted with methanol and the use of 50% acetonitrile in water and 50% methanol in water were investigated as diluents for injection into the LC-MS system. Utilizing both diluents, the validation parameters including accuracy, precision, LOD, and LOQ were assessed. The validation results and utilization of the method to analyze a variety of weight-loss supplements indicate that the two diluents give similar results and can be used interchangeably. This knowledge provides the user with the option of selecting either diluent for sample preparation depending on the sample matrix without having to revalidate the method. The method was applied to the analysis of weight-loss supplements available in local pharmacies, herbal pharmacies, and over the Internet.

Evaluation of vibrational spectroscopic methods to identify and quantify multiple adulterants in herbal medicines.

To counter the growth of herbal medicines adulterated with pharmaceuticals crossing borders, rapid, inexpensive and non-destructive analytical techniques, that can handle complex matrices, are required. Since mid-infrared (MIR), near infrared (NIR) and Raman spectroscopic techniques meet these criteria, their performance in identifying adulterants in seized weightloss herbal medicines is definitively determined. Initially a validated high pressure liquid chromatography methodology was used for reference identification and quantification of the adulterants sibutramine H2O·HCl, fenfluramine HCl and phenolphthalein. Of 38 products, only sibutramine and phenolphthalein were detected by HPLC. The spectroscopic measurements showed Raman was ill-suited due to sample burning and emission while NIR lacked adulterant selectivity. Conversely, MIR demonstrated apt identification performance, which manifested as spectrally meaningful separation based on the presence and type of adulterant during principal component analysis (test set validated). Partial least squares regression models were constructed from the MIR training sets for sibutramine and phenolphthalein - both models fitted the training set data well. Average test set prediction errors were 0.8% for sibutramine and 2.2% for phenolphthalein over the respective concentration ranges of 1.7-11.7% and 0.9-34.4%. MIR is apposite for the screening of anorectic and laxative adulterants and is the most viable technique for wider adulterant screening in herbal medicines.

Active pharmaceutical ingredients detected in herbal food supplements for weight loss sampled on the Dutch market.

Herbal food supplements claiming to reduce weight may contain active pharmacological ingredients (APIs) that can be used for the treatment of overweight and obesity. The aim of this study was to determine whether herbal food supplements for weight loss on the Dutch market contain APIs with weight loss properties. Herbal food supplements intended for weight loss (n = 50) were sampled from August 2004 to May 2013. An HPLC-DAD-MS/MS method was used to screen for the presence of the APIs in herbal supplements. In 24 samples the APIs sibutramine, desmethylsibutramine (DMS), didesmethylsibutramine (DDMS), rimonabant, sildenafil and/or the laxative phenolphthalein were identified 41 times. The presence of these APIs was, however, not stated on the label. The potential pharmacological effects of the detected APIs were estimated using data from reported effective doses of approved drugs. Use of 20 of the 24 herbal food supplements may result in potential pharmacological effects. Furthermore, risk assessment of phenolphthalein, a suspected carcinogen and found to be present in 10 supplements, based on the margin of exposure (MOE) approach, resulted in MOE values of 96-30,000. MOE values lower than 10,000 (96-220) were calculated for the daily intake levels of four out of these 10 supplements in which phenolphthalein was found. However, taking into account that weight loss preparations may be used for only a few weeks or months rather than during a lifetime, MOE values may be two to three orders of magnitude higher. The current study shows that the use of food supplements with sibutramine, DMS, DDMS and/or phenolphthalein could result in pharmacological effects.

[Simultaneous determination of 25 illegally added drugs in diet health foods by ultra-high performance liquid chromatography].

An analytical method using ultra-high performance liquid chromatography (UPLC) was developed for qualitative and quantitative analysis of 25 illegally added drugs in diet health foods. The diet food samples were extracted using 40 mL methanol by sonication. After centrifugation, the supernatants were separated on a Waters HSS T3 column with gradient elution at a flow rate of 0.3 mL/min, coupling with diode array detection (DAD) in wavelength range from 200 nm to 400 nm. The binary mobile phase was acetonitrile and 10 mmol/L ammonium acetate solution (containing 0.1% formic acid). The correlation coefficient of standard curve for each drug in linearity range was not less than 0. 997, as well as the recoveries of all the drugs in diet health foods were 70.7%-104% with the relative standard deviations (RSDs) of 0.132%-5.03% at three spiked levels. Seventeen diet food samples were tested, in which phenolphthalein was found in three samples and emodin was found in one sample. The method is specific, easy, quick, and suitable for confirmation of the 25 illegally added drugs in diet health foods.

[Detection of phenolphthalein added illegally into anti-obesity and healthcare food by TLC-FTIR].

OBJECTIVE: To establish a TLC-FTIR method for detection of western medicine phenolphthalein added illegally into anti-obesity and healthcare food. METHODS: The sample was extracted with anhydrous alcohol. The stationary phase was the GF254 aluminium alloy silica gel plates (10 cm x 20 cm) while the developing solvent was acetic ether: petroleum ether (60-90): methanol = 10: 6: 1. The sample volume was 2 microl. After primary screening by the UV lamp 254nm and self-made 2% NaOH test paper, the preparative technique of TLC was used to separate the component. Then the component was detected by FTIR and compared with the FTIR spectrogram of standard substance. RESULTS: Five of the ten samples contained phenolphthalein with the same testing results by using the TLC scanning and HPLC. CONCLUSION: The established method is accurate and reliable and can be used for detection of phenolphthalein illegally added into the products.

Rapid screening of illicit additives in weight loss dietary supplements with desorption corona beam ionisation (DCBI) mass spectrometry.

Desorption corona beam ionisation (DCBI), the relatively novel ambient mass spectrometry (MS) technique, was utilised to screen for illicit additives in weight-loss food. The five usually abused chemicals - fenfluramine, N-di-desmethyl sibutramine, N-mono-desmethyl sibutramine, sibutramine and phenolphthalein - were detected with the proposed DCBI-MS method. Fast single-sample and high-throughput analysis was demonstrated. Semi-quantification was accomplished based on peak areas in the ion chromatograms. Four illicit additives were identified and semi-quantified in commercial samples. As there was no tedious sample pre-treatment compared with conventional HPLC methods, high-throughput analysis was achieved with DCBI. The results proved that DCBI-MS is a powerful tool for the rapid screening of illicit additives in weight-loss dietary supplements.

Study on detection of drugs in slimming health foods using GC-MS/MS.

The determination of five drugs, fenfluramine (FEN), N-nitrosofenfluramine (NFE), sibutramine (SIB), mazindol (MAZ) and phenolphthalein (PHP), was studied in slimming health foods using GC-MS/MS. These drugs have been detected at high rates, especially in slimming health foods. Prolonged or excessive consumption of non-approved or unauthorized pharmaceuticals may cause serious adverse health consequences. In this study, samples were extracted with methanol and ultrasonication. Analyses were performed by GC-MS/MS, using established MS/MS parameters in the electron ionization (EI) mode and chemical ionization (CI) mode. In the EI mode, the recoveries of five drugs from several types of slimming health foods such as tablets, capsules and tea-bags spiked at 1 µg/mg (except PHP, spiked at 4 µg/mg) were in the range of 85.0-110.7% and 100 µg/mg (except PHP, spiked at 200 µg/mg) were 94.9-102.9%, respectively. In the CI mode, good recoveries of 80.3-102.2% (spiked at low concentration) and 92.8-103.2% (spiked at high concentration) were also obtained. We evaluated the present method using four slimming health foods, in which drugs had previously been detected. The results were similar to the previous results. These findings indicate that the present procedure for evaluating five drugs in slimming health foods by means of GC-MS/MS is useful.

Analysis of adulterated herbal medicines and dietary supplements marketed for weight loss by DOSY 1H-NMR.

Twenty herbal medicines or dietary supplements marketed as natural slimming products were analysed by diffusion ordered spectroscopy (DOSY) 1H-nuclear magnetic resonance (NMR) and DOSY-COSY 1H-NMR. The method allows analysis of the whole sample with the detection of both active and inactive ingredients in these complex matrices. Among the 20 formulations analysed, two were strictly herbal and four had a composition corresponding to declared ingredients on the packaging or the leaflet. The others were all adulterated. Eight formulations contain sibutramine alone at doses ranging from 4.4 to 30.5 mg/capsule. Five formulations contain sibutramine (from 5.0 to 19.6 mg/capsule or tablet) in combination with phenolphthalein (from 4.4 to 66.1 mg/capsule), and the last formulation was adulterated with synephrine (19.5 mg/capsule). Quantification of the actives was carried out with 1H-NMR. Several other compounds were also characterized including methylsynephrine, vitaberin, sugars, vitamins, etc. DOSY NMR is thus proposed as a useful tool for detection of unexpected adulteration.

Validación de técnicas para detección de sangre, sangre humana y grupo sanguíneo ABO en diferentes soportes y condiciones con fines forenses / Validation of techniques for detection of blood, human blood and sanguineous group ABO under different supports and conditions with forensic purposes

Las manchas encontradas en la escena del crimen se han constituido como elemento esencial en la resolución de investigaciones judiciales. Establecer su origen y grupo sanguíneo aporta información útil en el proceso de inclusión o exclusión de sospechosos o víctimas. En este estudio se evaluaron pruebas presuntivas de Piramidón, Luminol y Fenolftaleina con el fin de determinar su sensibilidad e interferencia frente a diferentes condiciones de soporte, temperatura, tiempo y ambiente. Además se evaluaron las mismas condiciones para los test de Takayama, Inhibición de la aglutinación y Absorción-elución que determinan el origen de la sangre y tipifican su grupo sanguíneo. Tanto las pruebas presuntivas como las confirmatorias no se ven afectadas en su mayoría al variar los soportes, la temperatura o las condiciones ambientales bajo las cuales fueron almacenadas. Respecto a la sensibilidad se encontró que en las pruebas presuntivas es alta en contraposición a las confirmatorias que requieren de muestras que no hayan sido sometidas a diluciones o lavados

Blood stains found at the scene of the crime have become an essential element when solving judicial investigations. To establish its origin and sanguineous group contributes to useful information in the inclusion process or exclusion of suspects or victims. In our study, we examined the Pyramidone, Luminol, and Phenolphthalein presumptive tests in order to establish their sensitivity (with different degrees of interference) and in different support, temperature, time, and storage conditions. The same was carried out to evaluate the agglutination inhibition, absorption-elution and Takayama tests, also used to establish blood origin and type group. In general terms, both the presumptive and the confirmatory tests were not affected in the different supports, temperature, time, and storage conditions we used. As far as sensitivity is concerned, we found that the presumptive tests remained high, while confirmatory tests required samples which had not been subject to dilutions or washings

[Identification of corrected retention indices of ten phenols in reversed-phase high performance liquid chromatography].

Corrected retention indices in RP-HPLC of ten phenols based on 2-keto-alkanes were examined on three ODS-silica columns. The RI values for phenols showed large variations. However, when the correction was applied, the variations diminished significantly. It is clear that the correction procedure improves the variation of phenols' RI values obtained from different ODS columns. The procedure is recommended as a method of standardization of HPLC data.

['Weight reduction preparations' from Thailand: unexpected complication for the traveller]. / 'Afslankmiddelen' uit Thailand: onverwachte reizigerszorg na terugkeer.

Two series of Thai 'slimming agents' purchased apparently without a medical consultation or prescription (one directly in Thailand and the other one indirectly in the Netherlands) were submitted for chemical analysis. Fenfluramine and diazepam were present in both series. One series also yielded phenolphtalein and vitamin B substances, while sennosides were found in the other series. Apparently travellers may unwittingly take along potent medicines when they return from far countries.