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Recently, the demand for improved brain function and concentration has increased in the dietary supplement market. However, to artificially enhance their pharmacological efficacy, dietary supplements may be illegally adulterated with unauthorised substances. Therefore, we developed a rapid and accurate method to simultaneously determine 11 nootropic substances using an ultra-high-performance liquid chromatography (UPLC) system equipped with a photodiode array (PDA) detector. In addition, sample preparation procedures were semi-optimised for various types of matrices, including solid (hard capsule, tablet, powder, and pill) and liquid (oil and extract) samples. The method was validated to determine the limit of detection (LOD), limit of quantification (LOQ), method detection limit (MDL), method quantitation limit (MQL), specificity, linearity, precision, accuracy, recovery, stability, and matrix effects. The validation results satisfied international validation guideline requirements. To test the applicability of the method, 55 real samples advertised as effective brain health, memory, and cognition supplements were analysed. Among the real samples, vinpocetine (2.483 and 7.296 µg/g), and kavain (69–44.056 µg/g) were detected. In addition, the detected compounds were confirmed by comparing their fragmentation patterns with those of the reference standards using liquid chromatography–quadrupole-time-of-flight mass spectrometry (LC-QTOF/MS). In conclusion, the UPLC-PDA method not only rapidly and accurately quantifies illegal nootropics but also enables the pre-emptive investigation and identification of 11 nootropic substances in illegal dietary supplements to protect public health.

Background Pyrrolizidine alkaloids (PAs) are naturally occurring plant toxins associated with potential hepatic and carcinogenic diseases in humans and animals. The concern over PAs has increased as the consumption of herbal medicines has increased. Objective This study aimed to develop and validate a sensitive analytical method to determine 28 PAs in five herbal medicines using liquid chromatography (LC)-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS). Additionally, this study identified and quantified the amount of PAs in 10 samples of each herbal medicine. Methods The pretreatment in the proposed LC-MS/MS analysis comprised solvent extraction using 0.05M H2SO4 in 50% methanol and clean-up step using an mixed-mode cationic exchange (MCX)-solid-phase extraction (SPE) cartridge. The PA contents in herbal medicines were measured by using the developed method. Results The proposed method had recoveries ranging from 72.5–123.7% for the Atractylodis Rhizoma Alba, 70.6–151.7% for Alba Chrysanthmi Flos, 80.6–130.9% for Leonuri Herba, 70.3–122.9% for Gastrodiae Rhizoma, and 67.1–106.9% for Glycyrrhizae Radix. Even though a few samples showed recoveries in unsatisfactory values, the proposed method indicated entirely sufficient recoveries and precision in most samples. In monitoring results, only Leonuri Herba contained two PAs, which indicated Retrorsine (4/10) of 84.7–120.9 μg/kg and Senkirkine (10/10) of 60.9–170.7 μg/kg. Conclusion The results obtained from this study demonstrate that the proposed method is fit for purpose to determine 28 PAs in herbal medicines. Therefore it could serve as a regulatory method capable of being used for controlling the risks of PAs in certain medicinal plants and dietary supplements. Highlights An LC-MS/MS method for the determination of 28 pyrrolizidine alkaloids in herbal medicines was developed and validated through this study. The proposed method is considered as an useful method for monitoring pyroolizidine alkaloids in herbal medicines.

Various synthetic dyes are artificially added to herbal medicines for the purpose of visual attraction. In order to monitor the illegal usage of synthetic dyes in herbal medication, a rapid and straightforward analysis method to determine synthetic dyes is required. The study aimed to develop and validate a high-performance liquid chromatography (HPLC) analysis to determine ten synthetic dyes in Hawthorn fruit, Cornus fruit, and Schisandra fruit. Ten synthetic dyes such as Tartrazine, Sunset yellow, Metanil yellow, Auramine O, Amaranth, Orange II, Acid red 73, Amaranth, New Coccine, Azorubine, and Erythrosine B, were extracted using 50 mM ammonium acetate in 70% MeOH, then separated by gradient elution with a mobile phase consisting of acetonitrile and 50 mM ammonium acetate in distilled water using a photodiode array detector (PDA) at 428 nm or 500 nm. In addition, this study established the LC-MS/MS method to confirm the existence of synthetic dyes in the positive sample solution. The HPLC analysis had good linearity (r2 &gt, 0.999). The recoveries of this method ranged from 74.6~132.1%, and the relative standard deviation (RSD) values were less than 6.9%. Most of the samples fulfilled the acceptance criteria of the AOAC guideline. This study demonstrates that the HPLC analysis can be applied to determine ten synthetic dyes in herbal medication.

Pyrrolizidine alkaloids (PAs) that are plant toxin naturally produced for protection against herbivores in some plant families. They are associated with the potential hepatotoxic and carcinogenic diseases serious hepatic disease in humans and animals. As the concern of human health risk posed by exposure to PAs has been gradually increased, precise and reliable analysis is required for monitoring PAs. The present study developed a new and simple pretreatment using 50% MeOH (methanol) for quantification analysis of the PAs contained with high content in the herbal medicines. Another pretreatment method using cation-ion exchange solid-phase extraction (MCX-SPE) was employed for determining most of the PAs that are not contained in the herbal medicines. That is, the proposed LC–MS/MS method coupled with MCX-SPE extraction and 50% MeOH extraction method was developed. And to evaluate the reliability of its application for Farfarae Flos and Lithospermi Radix, a validation study was conducted. In addition, monitory study was performed with ten samples in each herbal medicine. As a result, the proposed method had good linearity with r2 ≥ 0.997. Also, the recoveries indicated to be in the ranges of 70.4–118.0% for the Farfarae Flos, 70.2–119.7% for the Lithospermi Radix. In two herbal medicines, the intra-day precision was revealed to satisfy the reference criteria in most of the PAs. In monitoring results, most of the PAs were not contained in two herbal medicines, whereas a part of PAs revealed to have high concentration in Farfarae Flos and Lithospermi Radix. The proposed method is considered as a simple and reliable method to quantify 28 PAs present in two herbal medicines. Especially, the simple MeOH extraction method seems to be available for quantification analysis of certain PAs in herbal medicines with high content.

Recently, prohibited glucocorticoids (GCs) have been detected in commercial cosmetic products; they have been illegally added to enhance certain pharmacological efficacy and to make illegal profits. Therefore, the health of consumers has been exposed to the adverse effects of these tampered products. In this study, 95 samples are related to skincare, such as lotions, creams, solutions, gels and powders, with deceptive advertisements in the online or offline markets and their compositions were determined. From these samples, 43 GCs (beclomethasone, betamethasone, dexamethasone, budesonide, hydrocortisone, prednisolone, etc.) were simultaneously quantified by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The limit of quantification (LOQ, 1.0–90.0 ng mL−1), recoveries (spiked in cream samples, 84.61–118.34%; spiked in solution samples, 82.51–116.05%) and linearities (spiked in all samples, R2 > 0.99 at 100.0–2000.0 ng mL−1) were measured for method validation. The precisions (RSD) for intra-day and inter-day analysis were less than 14.19% and 12.82%. Also accuracies were 80.24–119.90% and 80.08–117.13%, respectively. The RSD of stability was ≤14.25%. Three of the 43 compounds were detected in the cream and solution forms, betamethasone at 24.2% was the most frequently used adulterant, followed by triamcinolone acetonide (8.4%) and clobetasol propionate (3.2%). The measured concentration ranges for betamethasone, clobetasol propionate, and triamcinolone acetonide were 29.2–331.0 μg g−1, 66.0–119.0 μg g−1, and 15.2–28.6 μg g−1, respectively. Therefore, with respect to public health and safety, the information obtained from this research should be available to the public.

Two synthetic cannabinoid analogues were detected using high-performance liquid chromatography (HPLC)–diode array detector, and gas chromatography–time-of-flight-mass spectrometry during the inspection of illegal products in an airmail package. The analogues were separated by semi-preparative HPLC, and their structures were determined by performing liquid chromatography–high-resolution-mass spectrometry, infrared analysis, and nuclear magnetic resonance spectroscopy. Compound 1 was MDMB(N)-Bz-F, which has been reported previously. Compound 2 was elucidated as adamantan-1-yl 1-pentyl-1H-indazole-3-carboxylate (APINAC), in which the amide group of APINACA was replaced with an ester group. Because there has been no chemical or pharmacological data about this compound until now, this is the first report of its detection in illegal products.

Distribution of various illegal or counterfeit drugs of seven approved erectile dysfunction drugs and their analogues has been increased, causing health problems such as cardiovascular disorder, tachycardia, headache, or vision disturbance. We used liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) to determine the erectile dysfunction drugs and their analogues in various counterfeit drugs. Eleven erectile dysfunction drugs and their analogues were detected, with sildenafil and its analogues being the most counterfeited compounds (73.8 %), followed by tadalafil and its analogues (25.4 %). The limits of detection (LOD) and the limits of quantitation (LOQ) of liquid-type and solid-type negative samples ranged from 0.1 to 3.3 ng/mL or ng/g and from 0.3 to 10.0 ng/mL or ng/g, respectively. The recoveries ranged from 84.3 to 112.3 % and 83.2 to 110.2 %, respectively. The contents of sildenafil and tadalafil in the various counterfeit drugs ranged from 21.0 to 947.5 mg/g and from 0.2 to 170.2 mg/g, respectively.

A new tadalafil analogue was identified along with homotadalafil during routine screening of an adulterated dietary supplement using HPLC-DAD. The UV spectrum of this analogue was almost identical with that of tadalafil. This compound was isolated from the supplement by using semi-preparative HPLC and its structure was subsequently elucidated by performing Q-TOF/MS/MS and NMR spectroscopic experiments. The spectral data indicate that this tadalafil analogue is a dimeric compound that consists of an ethylamino group and two pretadalafil moieties. NOE experiments and comparison with (1)H NMR spectra of tadalafil and trans-tadalafil suggested the trans-relationship between the substituents on piperidine rings in the pretadalafil moieties.

Recently, the number of the cases in which weight loss products have been sold with illegal adulterants has increased, as awareness of the problems of obesity grows. In this study, we developed simultaneous analysis methods to rapidly and accurately identify ingredients illegally mixed with foods and dietary supplements. Twenty-three anti-obesity drugs in foods and dietary supplements were determined by developed and validated UPLC and LC-MS/MS methods. The UPLC method were validated for the LOD and LOQ in the ranges 0.05-3.0 and 0.2-10.0 μg/mL, respectively. The determination coefficient was over 0.999, precision was6.2 %, and the accuracy was 80.8-103.9 %. The mean recoveries ranged from 80.3 to 109.3 % and RSD of stability was less than 2.1 %. The determination of the 23 anti-obesity drugs was accomplished by electrospray ionization LC-MS/MS using multiple reaction monitoring (MRM). The LODs and LOQs were in the ranges 0.03-7.5 and 0.09-30.0 ng/mL, respectively. The LC-MS/MS method was validated for linearity (r(2) 0.99), precision (RSD 10.7 %), accuracy (94.1-109.1 %), recovery (80.5-113.5 %), and the RSD of stability was7.8 %. Using the newly developed and validated method, 193 samples were tested, and 55 were found to be adulterated.

The objective of this study was to determine the presence of corticosteroids in illegal herbal medicines using ultra-high-performance liquid chromatography-tandem mass spectrometry. We collected 212 herbal medicine samples that were advertised as being effective for treatment of joint pain and bone aches. Samples were from the Korean commercial market during a span of four years (2010-2013), and the method was validated. The limits of quantification ranged from 0.47 to 15.0 ng/mL, and recoveries ranged from 80.6% to 119.5%. The intra- and interday precision ranged from 0.18% to 8.82% and from 0.09% to 8.96%, respectively. Among the samples, three samples (1.4%) were identified as adulterants. Dexamethasone was the only compound detected in the adulterated products. As the corticosteroid-adulteration of herbal medicines may become a major problem and lead to side effects, the continued development of screening procedures for herbal medicines is critical.

Phosphodiesterase type 5 inhibitors (PDE-5 inhibitors) are used in the treatment of erectile dysfunction. In recent years, a number of reports have been conducted on dietary supplements contaminated with PDE-5 analogues. In this study, 58 analogues of PDE-5 inhibitors were sorted into five groups: tadalafil, sildenafil, hongdenafil, vardenafil, and other analogues. These analogues were then evaluated using a liquid chromatographyquadrupole-time of flight mass spectrometry (LC-QTOF-MS) electrospray ionization mass method. Each compound has a unique fragmentation ion, which can be easily analyzed qualitatively. The fragmentation pathways of the analogues were elucidated based on the QTOF-MS and MS/MS data. Common ions were confirmed for each group by analyzing the structural characteristics and fragmentation pathways. Specifically, common ions were observed at m/z 169.08 and 135.04 (tadalafil analogues), m/z 311.15 and 283.12 (sildenafil analogues and hongdenafil analogues), and m/z 312.16 and 151.09 (vardenafil analogues). The advantage of this method is that the structure of unknown components can be determined by interpreting the product ions. Hence, the developed method can be used for the identification of unknown compounds. Fragmentation pathways may also aid in the detection and identification of PDE-5 inhibitor analogues.

Anabolic–androgenic steroids (AASs) have been illegally used in counterfeit drugs to improve the performance of athletes. In addition, AASs have been used for cosmetic purpose by non-athletes. To determine the presence of 26 AASs, an analysis method using ultra-liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) was developed and validated. The validated method was applied to 19 counterfeit drugs collected from the Internet and off-line markets during 2014. Nearly 50% (9/19) of the samples contained one of these 26 AASs. In addition, the concentration ranges of the AASs ranged from 0.09 to 119,228.57 mg/kg in the suspected samples. The determined AASs primarily consisted of testosterone and testosterone 17-propionate (26%) followed by boldenone (21%). These results indicate the adulteration of over-the-counter counterfeit drugs, and the continuous monitoring of counterfeit drugs or dubious dietary supplements containing anabolic steroids is warranted.

The roots of Pueraria mirifica were extracted with70% aqueous ethyl alcohol and partitioned into ethyl acetate(EtOAc), n-butyl alcohol (BuOH), and H 2 O fractions, successively.From the EtOAc fraction, four norsesquiterpenes were isolatedthrough the repeated silica gel, octadecyl silica gel and SephadexLH-20 column chromatographies. On the basis of physicochemicaland spectroscopic data including nuclear magnetic resonance(NMR), mass spectrometry, and infrared spectroscopy, thechemical structures were identified as megastigm-5-en-3,9-diol(1), linarionoside B (2), 3,5,6,9-tetrahydroxymegastigm-7-ene (3)and 3,4,9-trihydroxymegastigma-5,7-diene (4). Especially, theconfiguration of the anomer hydroxyl group was determined as âfrom the coupling constants of the anomer proton (J =8.0Hz) inthe 1 H-NMR spectrum. These compounds were isolated for thefirst time from the roots of P. mirifica in this study.Keywords 3,4,9-trihydroxy-megastigma-5,7-diene · 3,5,6,9-tetrahydroxy megastigm-7-ene · linarionoside B · megastigm-5-en-3,9-diol · norsesquiterpene · Pueraria mirifica · White Kwao Krua서론태국에서 예로부터 약용으로 이용되는 칡(Kwao Krua)으로는 RedKwao Krua (Butea superba), Black Kwao Krua (Mucunacollettii) 및 White Kwao Krua (Pueraria mirifica)가 가장 많이 알려져 있다. P. mirifica는 갱년기여성에 대하여 에스트로겐대체요법으로 사용되고 있으며, Red Kwao Krua (B. superba)와 Black Kwao Krua (M. collettii)는 남성의 성기능 증진 및발기부전 예방 및 치료에 사용되고있다(Suntara, 1931). 최근 위의 3종의 태국칡이 건강기능식품, 화장품 등의 제품으로 우리나라에 허가없이 수입, 유통되고 있다. 제품 중에는 그 효능에대해 허위, 과장되어 광고되거나 안전성이 확인되지 않은 성분이 혼입되어 안전사고 발생의 우려가 증가하고 있다. 따라서 태국칡의 유효성분에 대한 과학적 효능검증과 아울러 지표성분을확보함으로서 시료의 정확한 동정과 표준화된 원료의 규명연구가 시급한 실정이다.P. mirifica (White Kwao Krua)는of Korea 콩과(Favaceae) 칡속의 덩굴식물이며, 주로 태국 북부의 첸마이 지방에서 자생한다. 잎은손바닥 모양이며 잎자루에서 3개의 잎이 난다. 2−3월에는 푸른빛을 띄는 보라색의 꽃이This is an Open Access article distributed under the terms of the Creative 잔가지의 끝에 30개의 무리를 이루며피고, 4월에는 깍지 안에 3−5개의 씨가 생겨난다(Malaivijitnond,2012). P. mirifica로부터는 현재까지 daidzein, genistein, daidzin,genistin, puerarin, puerarin-6"-monoacetate, mirificin과 같은

The adulteration of foods and dietary supplements with steroids has been well attested and has the potential to be dangerous owing to various possible side-effects. Therefore, detecting the presence of steroids in various health food products has become increasingly important. The purpose of this study was to monitor illegally adulterated health food products by applying multiple reaction monitoring techniques to tandem liquid chromatography-mass spectrometry (LC-MS/MS). Various food and supplement samples advertised for the treatment of arthritis, bone ache and joint pain were collected over a 4-year period (2010-13) from local and online Korean sources. The method was validated based on limits of quantification of 0.5-15.0 ng g(-1) and recoveries in spiked solid samples of 81-119%. Approximately 30% of the tested samples were identified as having been illicitly adulterated. Six compounds were observed overall, including dexamethasone (45.1%), cotrisone-21-aceteate and prednisone-21-acetate (16.2%), and betamethasone (14.4%), and found in some samples in high concentrations.

The purpose of this study was to develop and validate an ultra-performance liquid chromatography method for simultaneous analysis of 20 antihistamines (illegal additives) in dietary supplements. The limits of detection and quantitation of the method ranged from 1.5 to 2.5 µg/mL and from 20.0 to 50.0 µg/mL, respectively. The determination coefficient was >0.999, precisions were 0.2-5.1% (intra-day) and 0.1-8.8% (inter-day), and accuracies were 84.5-111.2% (intra-day) and 91.9-112.0% (inter-day). The mean recoveries of 20 targeted compounds from dietary supplements ranged from 75.4 to 119.3%. The relative standard deviations were

Commercially available non-opioid analgesics such as acetaminophen and non-steroidal anti-inflammatory drugs (NSAIDs) have been used to adulterate some foods and dietary supplements. Considering the rapid growth of the dietary supplement market, it is essential to analyse various analgesics used for adulteration over a time period. Acetaminophen and 16 NSAIDs used to adulterate food and dietary supplements were simultaneously determined by LC-MS/MS. The method was validated by determining the coefficient of determinations, limit of quantification and recovery, and samples were analysed for the determination of analgesics. Consequently, acetaminophen, diclofenac, ibuprofen, indomethacin, naproxen and piroxicam were detected in 53 samples (n = 214). Ibuprofen was the most commonly used adulterant, which was detected in a wide concentration range (1.06-233.40 mg g(-1)) and was present in about one-third of the adulterated samples. Various types of samples, in particular pills and capsules (73.6% of the total positive samples), were found to be adulterated with non-opioid analgesics. Samples containing high concentrations of analgesics can have a deleterious effect on human health, and thus the continued monitoring of adulterated food and dietary supplements is essential to maintain a healthy life.

Because of the rapid growth in dietary supplement availability and public concern for weight control, the investigation of foods and various dietary supplements illegally adulterated with weight loss compounds has become increasingly important. A total of 29 weight loss compounds, including sennoside, sibutramine, ephedrine and their analogues, found to be adulterated in foods and dietary supplements were simultaneously examined by LC-MS/MS. The 188 samples were collected between 2009 and 2012 in South Korea, and method validation was performed to determine the adulterants to the weight loss compounds. LODs, LOQs and linearity ranged from 0.03 to 7.5 ng ml⁻¹, from 0.08 to 30.00 ng ml⁻¹, and from 0.990 to 0.999, respectively. The results showed that nine weight loss compounds, namely bisacodyl, desmethylsibutramine, didesmethylsibutramine, ephedrine, fluoxetine, pseudoephedrine, sennoside A, sennoside B and sibutramine, were detected in 62 of all collected samples and were found in order of frequency as follows: sibutramine, 25.7%; sennoside A, 22.9%; sennoside B, 20.0%; fluoxetine, 8.6%; desmethylsibutramine, 7.1%; bisacodyl, ephedrine, and pseudoephedrine, 4.3%; and didesmethylsibutramine, 2.9%. Sibutramine, which was the most frequently found adulterant, ranged in levels from 0.03 to 132.40 mg g⁻¹ (2010), from 0.88 to 76.2 mg g⁻¹ (2011), and from 0.07 to 0.24 mg g⁻¹ (2012). Although the concentrations of most compounds ranged widely, some compounds such as bisacodyl and fluoxetine were found at high concentrations in several samples.

South Korea’s Ministry of Food and Drug Safety (MFDS) has been developing programs for the inspection and accreditation of food sanitation inspection institutions. Food sanitation inspection institutions such as MFDS regional offices, the Research Institute of Public Health and Environment and authorized private service providers in South Korea must participate in proficiency testing (PT) programs to comply with the Food Sanitation Act and MFDS Notification No. 2012-112. As the PT provider, the MFDS annually plans various microbiological and chemical PT programs for foods, cosmetics, and pharmaceutical products in accordance with ISO/IEC 17043. The aim of this project was to evaluate the performance of microbiological PT programs to ensure the quality of their routine test results. The test materials used were freeze-dried BioBalls from BTF Pty Ltd. Homogeneity and stability were investigated to assess the adequacy of the selected test materials. This project also contains data from inter-laboratory comparisons organized by MFDS in 2011 and 2012. More than 50 laboratories attended the PT program and submitted their results. Laboratory results were rated with z-scores according to the international standard ISO 13528. The results from 2011 and 2012 revealed that all participating laboratories had similar levels of proficiency. Most of the participants received a rating of “Satisfactory.” Moreover, the percentage of participants who received a rating of “Unsatisfactory” decreased from 3.5 % in 2011 to 2.0 % in 2012.

More than 46 phosphodiesterase type 5 (PDE5) inhibitor analogues have been found to be present as illegal adulterants in various forms of health food products (powder, tablet, capsule, etc.), thereby placing the health of consumers at risk through product intake. In this study, 164 samples advertised to be effective at enhancing male sexual performance were collected over a 4-year period (2009-2012) from the Korean on-line or off-line market and screened. An LC-MS/MS method was employed to screen for the presence of 48 compounds including sildenafil, tadalafil, vardenafil and their analogues. Method validation established LOQs (0.30-10.00 ng ml(-1) or ng g(-1)) and recoveries (spiked in liquid sample, 84-112%; spiked in solid sample, 83-110%). Most of the illicit products screened were adulterated with 14 of the PDE5 derivatives under examination, including considerable amounts of sildenafil and tadalafil; of the 48 compounds, tadalafil was the most frequent adulterant (42.6%), followed by sildenafil (27.9%). Specifically, tadalafil concentration ranges (mg g(-1)) in the samples collected over the 4-year period were determined as follows: 2.91-52.20 (2009), 4.50-108.10 (2010), 0.37-101.40 (2011), and 0.08-138.69 mg g(-1) (2012). The concentration ranges (mg g(-1)) of sildenafil were also at high levels: 4.90-117.96 (2009), 1.30-369.93 (2010), 0.03-241.77 (2011), and 18.34-297.91 mg g(-1) (2012). The results of screening for PDE5 inhibitor pharmaceuticals as adulterants in illicit health food products are of great significance with respect to the protection of public health and consumer safety.

A propoxyphenyl-linked thiohomosildenafil analogue, one of the sildenafil analogues, was found in an herbal product. It was isolated by semi-preparative high-performance liquid chromatography (HPLC). The structure was established based on a comparison of chromatographic and spectroscopic behaviour with other sildenafil analogues using HPLC with diode array detection, quadrupole time-of-flight mass spectrometry (Q-TOF/MS), and nuclear magnetic resonance (NMR) spectroscopy. The HPLC analysis showed separation from known sildenafil analogues with a similar chromatographic retention time. An [M + H](+) ion at m/z 519.22 was detected by mass spectrometry corresponding to an empirical formula of C24H34N6O3S2. The structure was similar to that of thiohomosildenafil, except that the ethoxy group attached to the phenyl ring was substituted for a propoxy group. It was assigned as 5-[2-propoxy-5-(4-ethylpiperazin-4-ylsulfonyl)phenyl]-3-methyl-1-n-propyl-4,5,dihydro-1H-pyrazole[7,1,d]pyrimidin-4-thione and named as propoxyphenyl-thiohomosildenafil because the structure was considerably similar to thiohomosildenafil.

This paper describes the development and validation of simultaneous analytical method for 38 compounds, sildenafil, tadalafil, vardenafil and their analogues in illicit erectile dysfunction (ED) products by the liquid chromatography–electrospray ionization-tandem mass method (LC–ESI-MS/MS). Chromatographic separation was performed on a C18 reversed-phase column using a gradient of solvent A: aqueous 2 mM ammonium formate solution and solvent B: acetonitrile (ACN). All components were monitored under time-scheduled multiple reaction monitoring (MRM) mode. The limits of detection (LOD) ranged from 0.004 ng/ml to 0.455 ng/ml and the limits of quantification (LOQ) ranged from 0.012 ng/ml to 1.5 ng/ml. Calibration curves were linear with correlation coefficients over 0.9991. Mean recoveries ranged from 73.6% to 111.3%, and relative standard deviation (RSD) was less than 10%. The intra- and inter-day precision ranged from 0.2% to 16.3% and from 0.2% to 17.0%, respectively. The proposed method was applied to investigate the 52 illicit ED products.

The primary purpose of this study was to develop and validate a method based on UPLC with UV and UPLC-MS-MS for the simultaneous analysis of different cannabinoids and synthetic cannabinoids in food as well as in herbal and dietary supplements. The limits of detection and quantitation of the method ranged from 0.1 to 0.3 and 0.3 to 0.9 μg/mL by UPLC with UV, respectively. The coefficient of determination was0.999; the intra- and interday precision of the method were 0.1-3.7 and 0.9-4.1%, respectively. The intra- and interday accuracy were 94.8-103.1 and 98.3-100.9%, respectively. The mean recoveries of nine cannabinoids obtained from tablet samples ranged from 81.1 to 105.4%. The mean extraction recoveries of nine target cannabinoids obtained from various types of samples (tablets, capsules, powders, liquids, cookies and candies) ranged from 82.26 to 112.40%. The relative standard deviation (RSD) of the stability of the prepared sample solutions was1.80%. Identification and quantification of the nine cannabinoids were accomplished by ion spray UPLC-MS-MS using multiple reaction monitoring. The UPLC-MS-MS method was validated for linearity (R(2)0.99); the precision was 0.1-4.0% (intraday) and 0.1-2.8% (interday), and the accuracy was 98.0-103.5% (intraday) and 97.1-103.2% (interday). The mean extraction recoveries of six types of samples were 82.2-114.5% and the RSD of stability was6.54%, complying with the established international guidelines. The results indicated that the method can be used for rapid and accurate screening of cannabinoids present in food.

Department of Life Science & Biotechnology, Korea University, KoreaReceived October 9, 2011, Accepted October 21, 2011Key Words : Isomer, Amino acid, HPLC, LC/TOFMSThe discovery of molecular asymmetry by Pasteur in 1848established the foundation for our understanding of mole-cular chirality. Recently, the interest in safe food andpharmaceuticals has been increasing owing to increase ofelderly population with longer average life expectancyaccording to improvements in income levels and develop-ment of medical technology. Isomers are compounds withthe same molecular formula but different dimensional struc-tures and dissimilar properties.

In order to rapidly screen for the virulence factor that produces pathogenic Escherichia coli in food, we have developed multiplex polymerase chain reaction (PCR) assays. The multiplex PCR assays detect 4 pathogenic genes of enterohaemorrhagic E. coli (EHEC), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), and enteroinvasive E. coli (EIEC). This allows for the generation of specific fragments 150, 179, 218, 401, 465, 584, and 881 bp for VT1, ST, LT, 16S rRNA, inV, VT2, and eaeA genes, respectively. The detection limit of 3 log CFU/mL for eaeA, LT, VT1, VT2, 4 log CFU/mL for inV, 6 log CFU/mL for ST by single PCR, while 5 log CFU/mL for VT1, VT2, 6 log CFU/mL for eaeA, LT, 7 log CFU/mL for ST, inV by multiplex PCR. This optimized detection method of pathogenic E. coli can be used as supportive data to revise the microbiological analytical manuals for the Korean Food Code.

The purpose of this study is to develop LC-MS-MS and LC-Q-Orbitrap/MS method for the analysis of the components of Pueraria mirifica, which are illegal additives in dietary supplements. Blank samples and samples spiked with miroestrol and isomiroestrol were used for the initial development and validation studies. Specificity, linearity, limit of quantification (LOQ), limit of detection (LOD), accuracy, precision, recovery and stability were employed as the validation parameters. The LODs of miroestrol and isomiroestrol were found to be 4.17 and 0.84 ng/mL, respectively, whereas their LOQs were 12.50 and 2.52 ng/mL, respectively. The determination coefficient was over 0.999, intra- and inter-day precisions were 0.8-6.9 and 1.9-9.8%, respectively, and intra and inter-day accuracies were 82.1-103.7 and 85.0-109.7%, respectively. The mean recoveries of the targeted compounds from the dietary supplements ranged from 86.9 to 108.9%. The relative standard deviations (RSDs) for recovery were

A new compound, miroestrol-3-O-β-D-glucopyranoside, was isolated from the roots of Pueraria mirifica (white kwao krua). The structure of the compound was established on the basis of NMR, ESI-MS, and IR spectroscopic data.

A number of 188 food and dietary supplement samples were collected from 2009 to the first half of 2013 in Korean online and offline stores. A method to identify phosphodiesterase-5 (PDE-5) inhibitors and their analogues using liquid chromatography-electrospray ionisation-mass spectrometry/mass spectrometry (LC-ESI-MS/MS) was validated. Limit of detection and limit of quantitation of liquid-type and solid-type negative samples ranged from 0.05 to 3.33 ng/mL or ng/g and from 0.15 to 10.00 ng/mL or ng/g, respectively. Recoveries ranged from 83% to 112%. Nineteen PDE-5 inhibitors and their analogues were detected, with tadalafil group compounds being the most frequently observed (53.0%), followed by the sildenafil group (42.5%). Tadalafil concentrations ranged from 0.08 to 138.69 mg/g. Compounds were most frequently detected in capsules (in 40 of 80 adulterated samples). To protect public health and food safety, appropriate monitoring of PDE-5 inhibitors and their analogues in foods and dietary supplements is recommended.

A tadalafil analogue was detected in an herbal product by high performance liquid chromatography-diode array detector (HPLC-DAD) with a similar chromatographic retention time to tadalafil. The compounds were separated using semi-preparative HPLC. The structure of the detected tadalafil analogue was elucidated by LC-quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) and nuclear magnetic resonance (NMR) spectroscopy. A positive ion at m / z 404.1644 was detected by LC-Q-TOF/MS, corresponding to a molecular formula of C 23 H 22 N 3 O 4 . This unknown compound was identified as an analogue of tadalafil containing an additional methylene group and named homotadalafil. Homotadalafil was detected in 10 of 91 herbal products at concentrations of 0.058 mg g −1 to 8.735 mg g −1 .

A suspected sibutramine analogue was detected in a slimming functional food by an ultra performance liquid chromatography-electrospray ionisation-time of flight mass spectrometry (UPLC-ESI-TOF/MS) method. The ultraviolet (UV) spectrum of this suspected compound showed close similarity to that of sibutramine. The sample was extracted with 70% MeOH and isolated by semi-preparative column chromatography. The structure of this compound was identified by spectroscopic analyses (nuclear magnetic resonance [NMR] technique, mass and tandem mass etc.). The structure of the unknown compound was demonstrated to be [(±)-dimethyl-1-[1-(3,4-dichlorophenyl)cyclobutyl]-N,N,3-trimethylbutan-1-amine (molecular formula C17H25NCl2) and named as chloro-sibutramine. Compared with sibutramine, it has one more chlorine atom than the 3-cholorophenyl group so was switched to 3,4-dichlorophenyl. Until now, chloro-sibutramine was isolated for the first time from the undeclared ingredient included in dietary supplements. Although the safety of chloro-sibutramine is unknown, there is a potential health risk to consumers because of a similar skeleton to sibutramine. For public health, this sibutramine analogue has been included in the inspection list of illegal adulterants in Korea.

In this study, a method for discriminating natural mozzarella cheese from cheese substitutes, using fatty acid profiles, phytosterol contents, and statistical comparison, was developed. A total of 27 cheeses were evaluated: eight natural mozzarella cheeses (NMCs), four imitation mozzarella cheeses (IMCs), 12 processed cheeses (PCs) and three mixed cheeses (MCs) composed of NMCs and IMCs. The fatty acid composition of the NMC class was distinct from those of the IMC and MC classes, but statistically similar (p

A new tadalafil analogue, acetaminotadalafil, was detected by HPLC in a bulk powder that is being used as an ingredient formanufacturing dietary supplements. The analogue was isolated by semi-preparative HPLC. A chemical structure of the new compound was elucidated by HPLC, LC-quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS), nuclear magnetic resonance (NMR), infrared (IR) and circular dichroism (CD) spectroscopy. The compound was identified as an acetylatedcompound of aminotadalafil. The structure of the previous unknown compound was confirmed as (6R,12aR)-2-acetamino-6-(1,3-benzodioxol-5-yl)-2,3,6,7,12,12a-hexahydro-pyrazino[1',2':1,6]pyrido[3,4-b]indole-1,4-dione and named as acetaminotadalafil.