A new sildenafil analogue, cyclopentynafil (1) and a new tadalafil analogue, N-octylnortadalafil (2) were isolated from a dietary supplement illegally marketed for erectile dysfunction. The structures of the sildenafil and tadalafil analogues were elucidated by using HPLC-photodiode array (PDA), LC-MS, high-resolution MS, NMR and circular dichroism (CD). These compounds were determined to be 5- In this paper, we report the analysis and structural elucidation of a new sildenafil analogue, cyclopentynafil and a new tadalafil analogue, N-octylnortadalafil, that were isolated from a dietary supplement illegally marketed for erectile dysfunction. ExperimentalChemicals and Reagents HPLC-grade acetonitrile and all other reagents (analytical grade) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan).Sample The examined product was purchased as a processed food composed mainly of walnuts through the Internet and was composed of four pieces of ivory tablets (400 mg). The product properties were as follows: product name; Highperwalnup 2, producing company; Art Creation Co., Ltd., sales company; Ogawa Planning Co., Ltd., date of purchase; December 28, 2007.Preparation of Sample Solution One tablet was finely powdered, and 100 mg of the powder was ultrasonically extracted in 10 ml of 70% methanol for 15 min. The extract was centrifuged at 1700ϫg. The supernatant was filtered through a 0.45 mm filter. The filtrate was used for HPLC, and a portion of it was diluted 10-fold with methanol for liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis.HPLC Analysis HPLC analysis was performed using a JASCO PU-2089 apparatus equipped with a photodiode array (PDA) detector model MD-2015 (JASCO Corporation, Tokyo, Japan). The sample solution was separated by using a TSK-GEL ODS-80Ts column (150ϫ4.6 mm i.d., 5 mm, Tosoh Co., Tokyo, Japan). The mobile phase was an acetonitrile/water/phosphoric acid (100 : 900 : 1) mixture solution containing 5 mmol/l sodium hexanesulfonate (eluent A) and an acetonitrile/water/phosphoric acid (900 : 100 : 1) mixture solution containing 5 mmol/l sodium hexanesulfonate (eluent B). The gradient elution was started at 90% eluent A, and was linearly decreased to 55% A in 25 min and to 10% A in 44-49 min. The flow rate of the mobile phase was set at 1.0 ml/min, and the injection volume was 20 ml. The column temperature was maintained at 40°C. The PDA detection wavelength was set from ultraviolet (UV) 200 to 400 nm, and max-plot chromatographic monitoring was performed (200-400 nm).LC-ESI-MS Analysis LC-ESI-MS analysis was performed using a Waters alliance 2695 separation module and ZQ mass spectrometer (Waters Corporation, Milford, MA, U.S.A.). The sample solution was separated by using an Atlantis dC18 column (150ϫ2.1 mm i.d., 3 mm, Waters Corporation). The mobile phase was 0.1% formic acid aqueous solution (eluent A) and acetonitrile containing 0.1% formic acid (eluent B). The gradient elution began at 95% eluent A, and linearly decreased to 80% A in 15 min and ...

A tadalafil analogue was detected in a dietary supplement marketed for tonic e#ect, along with hydroxyhomosildenafil and aminotadalafil. The tadalafil analogue was isolated by preparative thin layer chromatography (TLC) and its structure was elucidated using high-performance liquid chromatography (HPLC), liquid chromatography electrospray ionization-mass spectrometry (LC-ESI-MS), Fourier transform-ion cyclotron resonance-mass spectrometry (FT-ICR-MS) and nuclear magnetic resonance (NMR) spectroscopy. The compound was determined to be methyl-1-(1,3-benzodioxol-5-yl)-2-(chloroacetyl)-2,3,4,9-tetrahydro-1H-pyrido [3,4-b]indole-3-carboxylate. This is the first report of detection of this compound in a dietary supplement.

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 solutionacetonitrile 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 .

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.

A rapid and simple method for determination of theophylline, theobromine and ca#eine in dietary supplements containing guarana by ultra-performance liquid chromatography (UPLC) has been developed. Theophylline, theobromine and ca#eine were extracted from finely powdered samples with water in a boiling water bath for 20 min, then the extracts were filtered and the filtrates were subjected to UPLC. Liquid samples were diluted with water and filteres, and the filtrates were subjected to UPLC. UPLC separation was performed on an AQUITY UPLC BEH C 18 column (2.1 mm i.d.ῒ50 mm, 1.7 mm, Waters) with 10 mmol/L ammonium acetate bu#er (pH 4.0)ῌacetonitrile gradient and eluates were monitored at 275 nm. .4ΐ, respectively. The quantitation limits of theophylline, theobromine and ca#eine were 10 mg/g (seed, seed powder, tablet and capsule) and 2.0 mg/mL (liquid) each. When this analytical method was applied to commercial dietary supplements, theophylline, theobromine and ca#eine were found at concentrations of 4.45 mg/tablet, 5.48 mg/tablet, 139 mg/tablet, respectively. Taking 4 tablets of this product according to the directions on the package could be dangerous to consumers because of possible overdosing of these ingredients.