HPLC/Fluorescence-Diode Array Detection for Rapid and Reliable Determination of Illegal Synthetic Drugs in Male Sexual Herbal and Honey Remedies: Comparative Study with UFLC-MS.

BACKGROUND: Nutraceuticals (NTCs), as honey and tablets with herbal extract are subjected to adulteration. OBJECTIVE: For NTCs claimed to enhance sexual performance, synthetic drugs (sildenafil, tadalafil, avanafil, vardenafil, and dapoxetine) are common adulterants, so they were selected to be simultaneously analyzed in the current study. Natural aphrodisiacs (icariin and yohimbine) are claimed to be present in many fake NTCs, so they were also included in the study. METHODS: In order to achieve the target of the current study, three liquid chromatographic methods with different unique detectors were developed and validated. RESULTS: High performance liquid chromatography (HPLC) with fluorescence detection enables rapid and reliable determination of natively fluorescent yohimbine, tadalafil vardenafil, and dapoxetine and it is the first report to analyze these compounds as adulterants in counterfeit NTC. Although the diode-array detector (DAD) enables the analysis of the seven adulterants, the fluorescence detector (FLD) shows better sensitivity and selectivity with lower LOQs and LODs. On the other hand, ultra-fast liquid chromatography-mass spectrometry (UFLC-MS) offers the advantages of peak identity confirmation, and it is of comparable sensitivity and selectivity to HPLC-FLD. CONCLUSION: One or more of these synthetic drugs were found in the analyzed NTCs while natural aphrodisiacs were absent. HIGHLIGHTS: Aphrodisiac nutraceuticals, NTCs, were analyzed for adulterants: five aphrodisiac synthetic drugs (adulterants) and two natural claimed aphrodisiacs. UFLC-MS and HPLC-DAD/FLD were compared for illicit NTCs analysis; all NTCs show the presence of synthetic aphrodisiacs and the absence of natural ones.

Simultaneous analysis of 23 illegal adulterated aphrodisiac chemical ingredients in health foods and Chinese traditional patent medicines by ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry.

This paper presents an application of ultra high-performance liquid-chromatography-quadrupole-TOF high resolution mass spectrometry (UHPLC-Q-TOF HRMS) for simultaneous analysis of 23 illegal adulterated aphrodisiac type chemical ingredients in health foods and Chinese Traditional Patent Medicines (CTPMs). The mass spectrometer was operated in Information Dependent Acquisition (IDA) mode, which provides crucial information for the elemental composition analysis, structure elucidation and quantitative analysis simultaneously. Quantitative analysis was performed using the peak areas of the precursor ions in the XICs. The method validation included assessment of selectivity, sensitivity, calibration curve, accuracy, precision, recovery, matrix effect and stability. The results show good linear relationship with the concentrations of the analytes over wide concentration ranges (e.g., 0.05-10 µg/g for sildenafil) as all the fitting coefficients of determination r2 are >0.9984. The detection limits (LODs) were in the range of 0.002-0.1 µg/g. The recoveries were able to reach 82.5-103.6%, while the matrix effects ranged from 87.7 to 109.3%. The intra- and inter-day accuracies were in the range of 82.3-113.8%, while the intra- and inter-day precision ranged from 0.4 to 13.6%. Among 40 batches of health foods and 32 batches of CTPMs (including 28 capsules, 32 tablets, 10 liquid and 2 pills) samples, 28 batches of heath foods were positive. The detected chemical ingredients involved sildenafil, tadalafil, aildenafil and sulfoaildenafil. This method can be used for the screening, identification and quantification of illegal adulterated aphrodisiac chemical ingredients in health foods and CTPMs. Moreover, the LC-Q-TOF MS is very useful to structural elucidation of unknown compound.

In vitro antioxidant potential of selected aphrodisiac medicinal plants.

The present study aimed to evaluate the antioxidant activity of six selected aphrodisiac medicinal plants. Useful parts of the selected medicinal plants were collected and extracted in methanolic solvent. The antioxidant activity of selected plant extract was determined through different antioxidant assays, namely DPPH radical scavenging assay and ferric reducing antioxidant assay. Moreover, antioxidant compounds, like total phenolics and total flavonoids contents, were also determined. Results showed that Mucuna pruriens seed extract displayed high contents of phenolic compounds with total phenolic content of 683.15±4.28 mg GAE/g dry plant material while the least phenolic content was observed in Asparagus racemosus (195.5±3.02 mg GAE/g dry plant material). Highest total flavonoids content was found in Anacyclus pyrethrum roots (156.58±4.01 µg CE/g) and the least content was found in Asparagus racemosus roots. Among the studied plant extracts, the highest radical scavenging activity was shown by Mucuna pruriens seed extract (82.05±0.55%) and the least percent scavenging activity was observed in Tribulus terrestris extract (36.40±2.01%). Vitamin C was used as positive control for antioxidant assays showing 93.54±0.9% radical scavenging activity. The plant extract also exhibited a strong reducing potential against free radicals. Therefore, the present study concluded that all the studied medicinal plants possess varying concentrations of secondary active metabolites responsible for the antioxidant properties of the tested plant extracts.

Detection, identification and quantification by 1H NMR of adulterants in 150 herbal dietary supplements marketed for improving sexual performance.

One hundred and fifty dietary supplements (DS) marketed to increase sexual performance were analyzed. All these formulations were claimed to contain only natural compounds, plant extracts and/or vitamins. (1)H NMR spectroscopy was used for detecting the presence of adulterants and for their identification and quantification. Mass spectrometry was used as a complementary method for confirming the chemical structures. 61% of DS were adulterated with phosphodiesterase-5 inhibitors (PDE-5i) (27% with the PDE-5i medicines sildenafil, tadalafil and vardenafil, and 34% with their structurally modified analogues). Among them, 64% contained only one PDE-5i and 36% mixtures of two, three and even four. The amounts of PDE-5i medicines were higher than the maximum recommended dose in 25% of DS tainted with these drugs. Additional 5.5% DS included other drugs for the treatment of sexual dysfunction (yohimbine, flibanserin, phentolamine, dehydroepiandrosterone or testosterone). Some DS (2.5%) contained products (osthole, icariin) extracted from plants known to improve sexual performance. Only 31% of the samples could be considered as true herbal/natural products. A follow-up over time of several DS revealed that manufacturers make changes in the chemical composition of the formulations. Lack of quality or consistent manufacture (contamination possibly due to inadequate cleaning of the manufacturing chain, presence of impurities or degradation products, various compositions of a given DS with the same batch number, inadequate labelling) indicated poor manufacturing practices. In conclusion, this paper demonstrates the power of (1)H NMR spectroscopy as a first-line method for the detection of adulterated herbal/natural DS and the need for more effective quality control of purported herbal DS.

In search of the perfect aphrodisiac: parallel use of bitter tonics in West Africa and the Caribbean.

ETHNOPHARMACOLOGICAL RELEVANCE: Enslaved Africans in the Americas had to reinvent their medicinal flora in an unknown environment by adhering to plants that came with them, learning from Amerindians and Europeans, using their Old World knowledge and trial and error to find substitutes for their homeland herbs. This process has left few written records, and little research has been done on transatlantic plant use. We used the composition of aphrodisiac mixtures across the black Atlantic to discuss the adaptation of herbal medicine by African diaspora in the New World. Since Africans are considered relatively recent migrants in America, their healing flora is often said to consist largely of pantropical and cultivated species, with few native trees. Therefore, we expected Caribbean recipes to be dominated by taxa that occur in both continents, poor in forest species and rich in weeds and domesticated exotics. MATERIALS AND METHODS: To test this hypothesis, we compared botanical ingredients of 35 African and 117 Caribbean mixtures, using Dentrended Correspondence Analysis, Cluster Analysis, Indicator Species Analysis and Mann-Whitney U tests. RESULTS: Very few of the 324 ingredients were used on both continents. A slightly higher overlap on generic and family level showed that Africans did search for taxa that were botanically related to African ones, but largely selected new, unrelated plants with similar taste, appearance or pharmacological properties. Recipes from the forested Guianas contained more New World, wild and forest species than those from deforested Caribbean islands. We recorded few 'transatlantic genera' and weeds never dominated the recipes, so we rejected our hypothesis. CONCLUSIONS: The popularity of bitter tonics in the Caribbean suggests an African heritage, but the inclusion of Neotropical species and vernacular names of plants and mixtures indicate Amerindian and European influence. We show that enslaved Africans have reinvented their herbal medicine wherever they were put to work, using the knowledge and flora that was available to them with great creativity and flexibility. Our analysis reveals how transplanted humans adapt their traditional medical practises in a new environment.

Analysis of herbal dietary supplements for sexual performance enhancement: first characterization of propoxyphenyl-thiohydroxyhomosildenafil and identification of sildenafil, thiosildenafil, phentolamine and tetrahydropalmatine as adulterants.

Nine herbal dietary supplements intended to be beverages for enhancing sexual performance were analyzed before their possible launch on the market. Four of them contained a sildenafil analog reported for the first time as an adulterant. After isolation and characterization using NMR, MS, IR and UV, this analog was named propoxyphenyl-thiohydroxyhomosildenafil as the ethoxy chain on the phenyl ring of the already known analog thiohydroxyhomosildenafil was replaced by a propoxy moiety. One formulation was tainted with thiosildenafil, another unapproved PDE-5 inhibitor. Sildenafil along with the natural alkaloid tetrahydropalmatine that has no documented effect for enhancing erectile dysfunction were identified in two formulations. Another formulation was adulterated with phentolamine, a drug that is not approved for boosting male sexual performance when taken orally. The last formulation containing osthole, a bioactive natural coumarine improving sexual dysfunction, is most probably truly natural.

Screening of Indian aphrodisiac ayurvedic/herbal healthcare products for adulteration with sildenafil, tadalafil and/or vardenafil using LC/PDA and extracted ion LC-MS/TOF.

Ayurvedic/herbal healthcare products are considered safe under the impression that they are derived from natural products. But recently, there have been several reports worldwide on the adulteration of synthetic PDE-5 inhibitors in aphrodisiac herbal formulations. Therefore, the objective of the present study was to explore the presence of synthetic PDE-5 inhibitors (sildenafil, tadalafil and/or vardenafil) in ayurvedic/herbal healthcare products sold in Indian market for aphrodisiac/related uses. In total, 85 herbal formulations (HFs) were included in the study. The formulations were extracted with methanol and subjected to centrifugation. The supernatant was analysed by HPLC and LC-MS/TOF. Early detection of the presence of sildenafil, tadalafil and vardenafil in the herbal samples was done by the study of extracted ion mass chromatograms at the m/z values of respective parent ions, and two prominent fragments of each. In case of sildenafil and tadalafil, adulteration was also detected by comparing the relative retention times (RR(T)) and UV spectra. Further substantiation was done through comparison of accurate mass spectra with those of the two available standards. Of the 85 HFs tested, only one was eventually found to be adulterated with sildenafil. The extent of adulterant in this sample was determined to the therapeutic dose in the formulation. The study thus indicates emergence of the problem of adulteration of Indian herbal products with PDE-5 inhibitors.

Use of a hydrolytic procedure and spectrometric methods in the structure elucidation of a thiocarbonyl analogue of sildenafil detected as an adulterant in an over-the-counter herbal aphrodisiac.

A sildenafil-related compound was detected in an herbal dietary supplement marketed as an aphrodisiac. The compound was identified as an analogue of sildenafil in which the carbonyl group in the pyrimidine ring of sildenafil was substituted with a thiocarbonyl group, and the methyl group on the piperazine ring was substituted with a hydroxyethyl group. Based on this structure, the compound was named thiohydroxyhomosildenafil. The structure of the compound was established using HPLCIMS, UV spectrometry, electrospray ionization-MS/MS, NMR spectrometry, and a hydrolytic process. One key product of hydrolysis was 1-(2-hydroxyethyl)-piperazine; the identification of this product defined the amine portion of the compound. Another key product of hydrolysis was hydroxyhomosildenafil, generated by hydrolysis of the thiocarbonyl group to a carbonyl group (C = S --> C = O). Hydroxyhomosildenafil was detected as a minor component in the dietary supplement.

The effect of saffron, Crocus sativus stigma, extract and its constituents, safranal and crocin on sexual behaviors in normal male rats.

In this study, the aphrodisiac activities of Crocus sativus stigma aqueous extract and its constituents, safranal and crocin, were evaluated in male rats. The aqueous extract (80, 160 and 320mg/kg body wt.), crocin (100, 200 and 400mg/kg body wt.), safranal (0.1, 0.2 and 0.4ml/kg), sildenafil (60mg/kg body wt., as a positive control) and saline were administered intraperitoneally to male rats. Mounting frequency (MF), intromission frequency (IF), erection frequency (EF), mount latency (ML), intromission latency (IL) and ejaculation latency (EL) were the factors evaluated during the sexual behavior study. Crocin, at all doses, and the extract, especially at doses 160 and 320mg/kg body wt., increased MF, IF and EF behaviors and reduced EL, IL and ML parameters. Safranal did not show aphrodisiac effects. The present study reveals an aphrodisiac activity of saffron aqueous extract and its constituent crocin.

Pociones sexuales. Relación entre alcohol, drogas y sexo / Sex potions. Relationships between alcohol, drugs and sex

A lo largo de la historia, el consumo de drogas y de alcohol ha estado íntimamente ligado a la conducta sexual. A pesar de ello, sin embargo, casi todos los esfuerzos por mejorar la salud sexual y reducir los niveles de consumo de drogas y alcohol se ocupan de ambos problemas como si fueran independientes. Con una disponibilidad potencialmente mayor que nunca de sustancias que tienen efectos relacionados con el sexo, analizar los vínculos entre sexo y consumo de drogas se ha convertido en un factor crítico para tratar ambos problemas. En el presente estudio nos ocupamos de las relaciones entre alcohol, drogas y conducta sexual, incluyendo el consumo de afrodisíacos y facilitadores sexuales, de cómo las sustancias están cada vez más vinculadas a la violencia sexual, y de cómo el propio sexo puede ser un medio para obtener drogas. Analizamos cómo aquellos que consumen drogas y alcohol son sexualmente más activos, tienen más posibilidades de practicar un sexo poco seguro y, por ese motivo, mayor riesgo de enfermedades de transmisión sexual o de embarazos no deseados. Vemos cómo los servicios de asistencia para el consumo de drogas y la salud sexual pueden beneficiarse de un acercamiento entre ambos problemas. Finalmente, en el área de la prevención, estudiamos cómo los servicios de salud sexual pueden utilizar las imágenes relacionadas con la droga para dirigirse a quienes corren el riesgo de contraer enfermedades de transmisión sexual, y cómo esas iniciativas cada vez mayores en la prevención de drogas deben desafiar la imagen sexual de muchas substancias a fin de reducir su atractivo (AU)

Throughout history drug and alcohol use has been intimately linked with sexual behaviour. Despite this however most attempts to improve sexual health and reduce levels of drug and alcohol use treat them as separate issues. With the availability of substances that have sex related effects potentially greater than ever, examining the links between sex and substance use has become a critical factor in addressing both issues. Here we discuss the relationships between alcohol, drugs and sexual behaviour, including the use of drugs as aphrodisiacs and sexual facilitators, how substances are increasingly being implicated in sexual assault, and how sex itself can be a means of obtaining drugs. We examine how those who use alcohol and drugs are more sexually active, more likely to practise unsafe sex, and hence at greater risk of sexually transmitted infections and unwanted pregnancy. We consider how treatment services for substance use and sexual health may benefit from an understanding of each other issues. Finally in the area of prevention, we discuss how sexual health services might utilise drug related imagery to target those at risk of sexual transmitted infections and how those developing drug prevention initiatives must challenge the sexual image of many substances in order to reduce their appeal (AU)

Qualitative and quantitative determination of yohimbine in authentic yohimbe bark and in commercial aphrodisiacs by HPLC-UV-API/ MS methods.

The development and validation of a rapid qualitative and quantitative method based on an HPLC-UV-MS technique with atmospheric pressure chemical ionisation and electrospray ionisation for the analysis of yohimbine in a number of commercial aphrodisiac products is reported. HPLC with multiple-stage mass spectrometry experiments allowed the identification of the target compound and increased the selectivity of complex analyses such as those involved with multi-botanical preparations. The precision and the robustness of the method were improved by the use of two internal standards: codeine for UV detection and deuterium-labelled yohimbine for MS detection. Twenty commercial aphrodisiac preparations were analysed and the amount of yohimbine measured and expressed as the maximal dose per day suggested on product labels ranged from 1.32 to 23.16 mg.

Efficacy of digoxin specific Fab fragments (Digibind) in the treatment of toad venom poisoning.

Chan Su, a traditional Chinese medication, and Love Stone, a topical aphrodisiac, are both made from dried venom of the toad bufo bufo gargarizans and contain bufalin, cinobufotalin, cinobufagin, and other cardioactive steroids of the bufadienolide class. Deaths have occurred following ingestion of these products and the clinical course resembles digoxin toxicity. The purpose of this study was to determine the efficacy of digoxin specific Fab fragments in treating Chan Su poisoning. An ethanolic extract was prepared from Chan Su. Digoxin specific Fab fragments were reconstituted in normal saline to a concentration of 80 mg/ml. An approximate LD90 dose was determined in preliminary experiments. Mice were then randomly divided into a treatment group of 15 mice and a control group of 30 mice. The treatment group was pretreated with 20 ml/kg of digoxin specific Fab fragment solution by intraperitoneal injection at t = 0, followed by 10 ml/kg of digoxin specific Fab fragments intraperitoneal at t = 30 min. The control group was pretreated with equal volumes of intraperitoneal normal saline at the same times. Immediately following the 30 min injection, both groups were given the estimated LD90 dose of Chan Su extract by subcutaneous injection. An endpoint of survival at 6 h was chosen after preliminary results showed that all deaths occurred in the first 4 h. All 30 of the control mice had seizures followed by death compared to 11 seizures and 7deaths in the 15 treatment mice. These results were statistically significant by Fisher's exact test (p = 0.00003 for mortality and p = 0.009 for seizures). Digoxin specific Fab fragments are effective in the treatment of Chan Su poisoning in mice and may be effective for poisoning by other cardioactive steroids of the bufadienolide class.

Treatment of toad venom poisoning with digoxin-specific Fab fragments.

Toxicity from toad venom poisoning is similar to digoxin toxicity and carries a high mortality rate. We report on six previously healthy men who developed vomiting and bradycardia after ingesting a purported topical aphrodisiac. Each patient had positive apparent digoxin levels and the first four patients died of cardiac dysrhythmias. The last two patients recovered following treatment with digoxin Fab fragments. We analyzed samples of the purported aphrodisiac and found that it was identical to Chan Su, a Chinese medication made from toad venom. To our knowledge, this is the first reported use of digoxin Fab fragments to treat toad venom poisoning.