Determination of Cyanide and Cyanoglycosides in Sweetened Bean Paste by HPLC with Fluorescence Detection.

It is necessary to evaluate the efficiency of reduction for cyanide and cyanoglycosides during the manufacturing process from raw material beans to sweetened bean paste in a food hygiene control system from the viewpoint of food safety. Analytical methods for cyanide and cyanoglycoside determination in sweetened bean paste by HPLC with fluorescence detection were developed. In analysis of collection time of free cyanide in the free cyanide assay, the recovery was improved by extending the collection time, the recovery rate was >80% by 2 h. The accuracy, repeatability and intra-laboratory precision of the free cyanide assay were 82.3, 2.0, and 2.4%, respectively. The method for cyanoglycoside analysis was evaluated by 5 repeated spiked recovery experiments at a concentration of 10 ppm. The accuracy, repeatability and intra-laboratory precision of the cyanoglycoside method were 82.2, 1.9, and 3.4%, respectively. These analytical methods will enable the analysis of cyanide and cyanoglycosides in sweetened bean paste without using steam distillation method in the pretreatment.

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.

[Pharmaceutical Ingredients Detected in Dietary Supplements Purchased by Chiba Prefecture (Fiscal Years 2014-2021)].

In the pharmaceutical ingredients contamination testing of 702 commercial dietary supplement products, during fiscal years 2014-2021, 14 pharmaceutical ingredients, barrenwort, leaf axils of senna, and small leaf of senna were detected in 28 products. Screening and confirmation of the pharmaceutical ingredients in the products were performed by ultra-high performance liquid chromatography with photodiode array detector and ultra-high performance liquid chromatography-quadrupole-kingdon trap mass spectrometry, respectively. In particular, leaf axils and small leaf of senna were identified by stereomicroscopy and scanning electron microscopy. Furthermore, we found several pharmaceutical ingredients that exceeded the daily medicated dosage; therefore, it is important to prevent the distribution of such products to prevent the occurrence of health hazard. For that reason, it is necessary to continue the sample purchasing and testing systems to monitor the distribution of products containing pharmaceutical ingredients.

[The Configuration of Tadalafil and Tadalafil Analogues Found in Dietary Supplements].

This study determined the configuration of the isomers of tadalafil, nortadalafil, and homotadalafil in dietary supplements. The products purchased over the Internet studied included a honey product and a tablet, which contained tadalafil, and a candy, which contained nortadalafil and homotadalafil. Each of the pharmaceutical ingredients isolated from the products was measured with circular dichroism (CD).As a result, the CD spectrum of each isolated pharmaceutical ingredient was found to align with the standard CD spectrum of the 6R,12aR isomer, confirmed that each isolated tadalafil or tadalafil analogue included in a 6R,12aR isomer. According to a report, among the stereoisomers of tadalafil, the 6R,12aR isomers have the most potent inhibitory activities of phosphodiesterase-type-5. From the report, the potential strength of the inhibitory activity of the 6R,12aR isomers of nortadalafil and homotadalafil was suggested. Therefore, it seemed that the 6R,12aR isomer often used in the product.

[Simultaneous determination of statins in dietary supplements by ultra-performance liquid chromatography].

A method for the determination of 12 statins [atorvastatin (ATOR), cerivastatin (CERI), fluvastatin (FLU), lovastatin (LO), lovastatin acid (LOA), mevastatin (ME), mevastatin acid (MEA), pitavastatin (PITA), pravastatin (PRA), rosuvastatin (ROSU), simvastatin (SIM), and simvastatin acid (SIMA)] in dietary supplements by ultra-performance liquid chromatography (UPLC) has been developed. Statins were ultrasonically extracted with 50% (v/v) methanol. Clean-up was performed using an Oasis MAX mini-cartridge column with methanol and methanol containing 0.2% (v/v) phosphoric acid as an eluting solvent. UPLC separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm i.d. × 150 mm, 1.7 µm) with 0.2% (v/v) phosphoric acid aqueous solution-acetonitrile gradient. The method was validated for dietary supplements spiked with the 12 statins at the quantitation limits and 10 times the quantitation limits, and the recoveries of statins were between 89.2% and 100.9%. Relative standard deviation values of repeatability and intermediate precision were not more than 7%. The analytical method was applied to 24 commercial dietary supplements. LO and LOA were found at maximum concentrations of 4.85 mg/packet and 1.28 mg/capsule, respectively. Other statins were not detected. When a dietary supplement was consumed according to the directions on the package, the daily intake of LO was 6.74 mg. This could be dangerous to consumers because it exceeds one half of the lowest recommended daily dose of LO (10 mg).

Enantiomeric determination of DOPA in dietary supplements containing Mucuna pruriens by liquid chromatography/mass spectrometry.

We developed a simple and rapid liquid chromatography/mass spectrometry (LC/MS) method for the enantiomeric determination of DOPA in dietary supplements containing Mucuna pruriens. L- and D-DOPA were ultrasonically extracted with 1% formic acid aqueous solution. The isolated extracts were analyzed by LC/MS using a Crownpak CR (-) column at 30℃. The mass spectrometer was operated in the positive mode of electrospray ionization, and the mobile phase was aqueous formic acid (pH 2.0). L-DOPA-ring-d3 was used as an internal standard. The method was validated for a dietary supplement spiked with L- and D-DOPA at 50 and 500 µg/g, respectively, and the recoveries of the DOPA enantiomers were between 97.5% and 101.3%. Relative standard deviation values of repeatability and intermediate precision were less than 7%. The method was applied to 14 dietary supplements. L-DOPA was detected in these supplements in the range of 0.88-12.8 mg/unit. D-DOPA was not detected.

[Current situation and issues for analyzing illegal drug products].

Thirty-two psychotropic substances (31 compounds and one plant) have been controlled as designated substances (Shitei-yakubutsu) in Japan by the Pharmaceutical Affairs Law since April 2007. Although the trafficking of these drugs has decreased because of this regulation, new designer drugs (synthetic cannabinoids and cathinones) have appeared, one after the other. As of October 2011, 40 compounds had been newly added to this category. Analytical methods have become more complicated due to this increase in the number of designated substances. Moreover, many reference substances for such designated substances and other new designer drugs are not commercially available. For the reasons stated above, a lot of time and effort is required to analyze the illegal drug products available on the market.

Binding of spermine and ifenprodil to a purified, soluble regulatory domain of the N-methyl-D-aspartate receptor.

The binding of spermine and ifenprodil to the amino terminal regulatory (R) domain of the N-methyl-D-aspartate receptor was studied using purified regulatory domains of the NR1, NR2A and NR2B subunits, termed NR1-R, NR2A-R and NR2B-R. The R domains were over-expressed in Escherichia coli and purified to near homogeneity. The K(d) values for binding of [(14)C]spermine to NR1-R, NR2A-R and NR2B-R were 19, 140, and 33 microM, respectively. [(3)H]Ifenprodil bound to NR1-R (K(d), 0.18 microM) and NR2B-R (K(d), 0.21 microM), but not to NR2A-R at the concentrations tested (0.1-0.8 microM). These K(d) values were confirmed by circular dichroism measurements. The K(d) values reflected their effective concentrations at intact NR1/NR2A and NR1/NR2B receptors. The results suggest that effects of spermine and ifenprodil on NMDA receptors occur through binding to the regulatory domains of the NR1, NR2A and NR2B subunits. The binding capacity of spermine or ifenprodil to a mixture of NR1-R and NR2A-R or NR1-R and NR2B-R was additive with that of each individual R domain. Binding of spermine to NR1-R and NR2B-R was not inhibited by ifenprodil and vice versa, indicating that the binding sites for spermine and ifenprodil on NR1-R and NR2B-R are distinct.