Hepatotoxicity in Cattle Associated with Salvia reflexa Diterpenes, including 7-Hydroxyrhyacophiline, a New Seco-Clerodane Diterpene.

A case of baled alfalfa hay contaminated with multiple weeds induced hepatotoxicity and death in cattle. The hepatotoxic compounds were isolated by bioassay-guided fractionation using a mouse model and identified as salviarin, salvianduline D, rhyacophiline, and 7-hydroxyrhyacophiline. The structure of 7-hydroxyrhyacophiline has not been previously reported. All compounds were found to induce severe acute hepatic necrosis within 24-48 h after a single oral dosage (260-280 mg/kg). The identified diterpenes are known to be found among different Salvia species which led to finding dried plant parts of Salvia reflexa within bales of weedy hay and subsequently a population of S. reflexa was found along the field edges and irrigation ditch banks of the alfalfa hay field. It was thus determined that S. reflexa was responsible for the hepatotoxicity observed in cattle fed the contaminated hay.

Cytotoxic Effects of Salvinorin A, A Major Constituent of Salvia divinorum.

S. divinorum is a psychoactive plant that has been consumed as a recreational drug of abuse in the last years. Salvinorin A is its main constituent, and is responsible for the observed psychoactive effects. Both S. divinorum and salvinorin A have become controlled drugs in several countries, but they are not listed in the Schedules of the United Nations Drug Conventions. Regarding the effects of S. divinorum consumption, almost all studies are based on in vivo or on surveys, and there are no studies in vitro on its toxicity. Furthermore, all studies are focused on the acute toxicological effects of the plant. So, it is of utmost importance to further investigate the effects of S. divinorum and salvinorin A, particularly using in vitro models, after prolonged exposures. In this context, the present work evaluated the in vitro toxicity induced by S. divinorum or salvinorin A in six cell lines, through MTT assays and LC50 determination. Overall, results showed that both S. divinorum and salvinorin A are cytotoxic, dose- and time-dependent. Also, Hep G2 and Caco 2 (to a lesser extent) cells showed lower sensitivity to S. divinorum and salvinorin A when compared to the other studied cell lines. To our knowledge, this is the first work focused on the in vitro toxicity of S. divinorum and salvinorin A using a variety of cell lines, which are extensively described in literature and have been widely used in several in vitro studies.

From divination to madness: features of acute intoxication with Salvia use.

Salvia divinorum is a psychoactive botanical plant that is increasingly used for the 'legal' highs that it can produce. It is readily available for purchase on the Internet, and most abusers are unaware of the toxicity and abuse potential associated with its use. As the use of novel compounds among abusers is not uncommon, physicians need to increase their awareness and recognition of these new substances. Herein, we report a case of an acute presentation of Salvia intoxication.

From local to global-fifty years of research on Salvia divinorum.

ETHNOPHARMACOLOGICAL RELEVANCE: In 1962 ethnopharmacologists, Hofmann and Wasson, undertook an expedition to Oaxaca, Mexico. These two researchers were the first scientists to collect a flowering specimen of Salvia divinorum allowing the identification of this species. While the species' traditional use is confined to a very small region of Mexico, since Hofmann and Wasson's expedition 50 years ago, Salvia divinorum has become globally recognized for its main active constituent, the diterpene salvinorin A, which has a unique effect on human physiology. Salvinorin A is a kappa-opioid agonist and the first reported psychoactive diterpene. METHODS: This review concentrates on the investigation of Salvia divinorum over the last 50 years including ethnobotany, ethnopharmacology, taxonomy, systematics, genetics, chemistry and pharmacodynamic and pharmacokinetic research. For the purpose of this review, online search engines were used to find relevant research. Searches were conducted between October 2011 and September 2013 using the search term "Salvia divinorum". Papers were excluded if they described synthetic chemical synthesis of salvinorin A or analogues. RESULTS: Ethnobotanically there is a comprehensive body of research describing the traditional Mazatec use of the plant, however, the modern ethnobotanical use of this plant is not well documented. There are a limited number of botanical investigations into this plant and there are still several aspects of the botany of Salvia divinorum which need further investigation. One study has investigated the phylogenetic relationship of Salvia divinorum to other species in the genus. To date the main focus of chemistry research on Salvia divinorum has been salvinorin A, the main active compound in Salvia divinorum, and other related diterpenoids. Finally, the effects of salvinorin A, a KOR agonist, have primarily been investigated using animal models. CONCLUSIONS: As Salvia divinorum use increases worldwide, the emerging cultural use patterns will warrant more research. More botanical information is also needed to better understand this species, including germination, pollination vector and a better understanding of the endemic environment of Salvia divinorum. As well there is a gap in the genetic knowledge of this species and very little is known about its intra-species genetics. The terpenes in Salvia divinorum are very well documented, however, other classes of constituents in this species warrant further investigation and identification. To date, the majority of the pharmacology research on Salvia divinorum has focused on the effects of salvinorin A using animal models. Published human studies have not reported any harmful effects when salvinorin A is administered within the dose range of 0.375-21µg/kg but what are the implications when applied to a larger population? More data on the toxicology and safety of Salvia divinorum are needed before larger scale clinical trials of the potential therapeutic effects of Salvia divinorum and salvinorin A are undertaken.

Here today, gone tomorrow and back again? A review of herbal marijuana alternatives (K2, Spice), synthetic cathinones (bath salts), kratom, Salvia divinorum, methoxetamine, and piperazines.

Despite their widespread Internet availability and use, many of the new drugs of abuse remain unfamiliar to health care providers. The herbal marijuana alternatives, like K2 or Spice, are a group of herbal blends that contain a mixture of plant matter in addition to chemical grade synthetic cannabinoids. The synthetic cathinones, commonly called "bath salts," have resulted in nationwide emergency department visits for severe agitation, sympathomimetic toxicity, and death. Kratom, a plant product derived from Mitragyna speciosa Korth, has opioid-like effects, and has been used for the treatment of chronic pain and amelioration of opioid-withdrawal symptoms. Salvia divinorum is a hallucinogen with unique pharmacology that has therapeutic potential but has been banned in many states due to concerns regarding its psychiatric effects. Methoxetamine has recently become available via the Internet and is marked as "legal ketamine." Moreover, the piperazine derivatives, a class of amphetamine-like compounds that includes BZP and TMFPP, are making a resurgence as "legal Ecstasy." These psychoactives are available via the Internet, frequently legal, and often perceived as safe by the public. Unfortunately, these drugs often have adverse effects, which range from minimal to life-threatening. Health care providers must be familiar with these important new classes of drugs. This paper discusses the background, pharmacology, clinical effects, detection, and management of synthetic cannabinoid, synthetic cathinone, methoxetamine, and piperazine exposures.

Composition of three essential oils, and their mammalian cell toxicity and antimycobacterial activity against drug resistant-tuberculosis and nontuberculous mycobacteria strains.

Tuberculosis (TB) is the most ancient epidemic disease in the world and a serious opportunistic disease in HIV/AIDS patients. The increase in multidrug resistant Mycobacterium tuberculosis (MDR-TB, XDR-TB) demands the search for novel antimycobacterial drugs. Essential oils (EOs) have been widely used in medicine and some EOs and their major components have been shown to be active against M. tuberculosis. The aim of this work was to evaluate the antimycobacterial and cell toxicity activities of three EOs derived from Salvia aratocensis, Turnera diffusa and Lippia americana, aromatics plants collected in Colombia. The EOs were isolated by hydrodistillation and analyzed by GC/MS techniques. The EOs were tested against 15 Mycobacterium spp using a colorimetric macrodilution method and against mammalian Vero and THP-1 cells by MTT. The activity was expressed as minimal concentration in microg/mL that inhibits growth, and the concentration that is cytotoxic for 50 or 90% of the cells (CC50 and CC90). The major components were epi-alpha-cadinol (20.1%) and 1,10-di-epi-cubenol (14.2%) for Salvia aratocensis; drima-7,9(11)-diene (22.9%) and viridiflorene (6.6%) for Turnera diffusa; and germacrene D (15.4%) and trans-beta- caryophyllene (11.3%) for Lippia americana. The most active EO was obtained from S. aratocensis, with MIC values below 125 microg mL(-1) for M. tuberculosis Beijing genotype strains, and 200 to 500 microg mL(-1) for nontuberculous mycobacterial strains. The EOs were either partially or non toxic to Vero and THP-1 mammalian cells with CC50 values from 30 to > 100 microg mL(-1), and a CC90 > 100 microg mL(-1). The EOs obtained from the three aromatic Colombian plants are an important source of potential compounds against TB. Future studies using the major EO components are recommended.

Phytotoxic activity of Salvia x jamensis.

A study has been carried out on the surface exudate of Salvia x jamensis, which showed a significant phytotoxic activity against Papaver rhoeas L. and Avena sativa L.. Bioguided separation of the exudate yielded active fractions from which 3 beta-hydroxy-isopimaric acid (1), hautriwaic acid (2), betulinic acid (3), 7,8 beta-dihydrosalviacoccin (4), isopimaric acid (5), 14 alpha-hydroxy-isopimaric acid (7), 15,16-epoxy-7 alpha, 10 beta-dihydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide (8), cirsiliol (5,3',4'-trihydroxy-6,7-dimethoxyflavone, 9) and two new neoclerodane diterpenes (6 and 10) were isolated. The structures of 6 and 10 were identified as 15,16-epoxy-10 beta-hydroxy-clerod-3,13(16),14-trien-17,12;18,19-diolide and 15,16-epoxy-7 alpha,10-dihydroxy-clerod-2,13(16),14-trien-17,12;18,19-diolide respectively on the basis of spectroscopic data analysis. All compounds, but 7, 8 and 10, were active in inhibiting the germination of the tested species.

Evaluación del extracto fluido de Pluchea carolinensis (Jacq) G Don. por el método de clases de toxicidad en ratas / Evaluation of fluid extract from Pluchea carolinensis (jacq) G Don by the toxicity class method in rats

Introducción: la salvia de playa, Pluchea carolinensis (Jacq) G Don., ha sido utilizada en la medicina tradicional para eliminar fiebres, digestiones lentas, dolores de riñones, de cabeza, ronqueras. Objetivos: clasificar al extracto fluido de salvia comenzando por el nivel de dosis 2 000 mg/kg, según el método de clase de toxicidad aguda. MÉTODOS: se evaluó el extracto fluido de hojas secas de salvia de playa mediante el ensayo de las clases de toxicidad, que permite clasificar la sustancia en un rango de toxicidad. Se utilizaron 6 hembras de la sublínea Cenp:SPRD, procedentes del CENPALAB. Se les administró una dosis única de 2 000 mg/kg de peso corporal del extracto fluido de salvia de playa por vía oral, previo ayuno. El período de observación fue de 14 d, en el cual se monitorearon las condiciones ambientales diariamente, la aparición de signos de toxicidad y muerte, así como el peso corporal en los días 0, 7 y 14 del ensayo. Resultados: los animales alcanzaron 100 por ciento de supervivencia. No se observaron signos de toxicidad, tras la administración de la sustancia ensayo en la dosis máxima de 2 000 mg/kg. En la evaluación anatomopatológica no se observaron alteraciones macroscópicas en la superficie externa de los animales y en ninguna de sus cavidades, órganos y tejidos. Conclusiones: de acuerdo con el método de toxicidad de clases la sustancia no es clasificada, DL50 es mayor que 2 000 mg/kg(AU)

Introduction: beach salvia, Pluchea carolinensis (Jacq)G Don has been used for long in traditional medicines to abate fevers, slow digestion, kidney pains, headache, and hoarseness. Objectives: to classify fluid extract from beach salvia according to the acute toxicity class method, by starting with a 2000 mg/kg dose. Methods: the fluid extract from beach salvia dry leaves was evaluated through the toxicity class test that allows classifying the substance into a range of toxicity. Six Cenp:SPRD subline female rats from CENPALAB (Center of lab animal production) were used. They were orally administered a single dose of 2000 mg of beach salvia fluid extract per kg of body weight on fasting. The observation period was 14 days to monitor daily environmental conditions, the occurrence of toxicity signs and death as well as the body weight at 0th,7th and 14th day. Results: the six rats survived. No sign of toxicity was observed after the administration of the extract at 2000 mg/kg dose. In the anatomopathological assessment, no macroscopic alterations were seen in the external surface or in the cavities, organs and tissues of the animals. Conclusions: according to this toxicity class method, the substance is not classified since LD50 is higher than 2 000 mg/kg(AU)

Evaluación del extracto fluido de Pluchea carolinensis (Jacq) G Don. por el método de clases de toxicidad en ratas / Evaluation of fluid extract from Pluchea carolinensis (jacq) G Don by the toxicity class method in rats

Introducción: la salvia de playa, Pluchea carolinensis (Jacq) G Don., ha sido utilizada en la medicina tradicional para eliminar fiebres, digestiones lentas, dolores de riñones, de cabeza, ronqueras. Objetivos: clasificar al extracto fluido de salvia comenzando por el nivel de dosis 2 000 mg/kg, según el método de clase de toxicidad aguda. MÉTODOS: se evaluó el extracto fluido de hojas secas de salvia de playa mediante el ensayo de las clases de toxicidad, que permite clasificar la sustancia en un rango de toxicidad. Se utilizaron 6 hembras de la sublínea Cenp:SPRD, procedentes del CENPALAB. Se les administró una dosis única de 2 000 mg/kg de peso corporal del extracto fluido de salvia de playa por vía oral, previo ayuno. El período de observación fue de 14 d, en el cual se monitorearon las condiciones ambientales diariamente, la aparición de signos de toxicidad y muerte, así como el peso corporal en los días 0, 7 y 14 del ensayo. Resultados: los animales alcanzaron 100 por ciento de supervivencia. No se observaron signos de toxicidad, tras la administración de la sustancia ensayo en la dosis máxima de 2 000 mg/kg. En la evaluación anatomopatológica no se observaron alteraciones macroscópicas en la superficie externa de los animales y en ninguna de sus cavidades, órganos y tejidos. Conclusiones: de acuerdo con el método de toxicidad de clases la sustancia no es clasificada, DL50 es mayor que 2 000 mg/kg.

Introduction: beach salvia, Pluchea carolinensis (Jacq)G Don has been used for long in traditional medicines to abate fevers, slow digestion, kidney pains, headache, and hoarseness. Objectives: to classify fluid extract from beach salvia according to the acute toxicity class method, by starting with a 2000 mg/kg dose. Methods: the fluid extract from beach salvia dry leaves was evaluated through the toxicity class test that allows classifying the substance into a range of toxicity. Six Cenp:SPRD subline female rats from CENPALAB (Center of lab animal production) were used. They were orally administered a single dose of 2000 mg of beach salvia fluid extract per kg of body weight on fasting. The observation period was 14 days to monitor daily environmental conditions, the occurrence of toxicity signs and death as well as the body weight at 0th,7th and 14th day. Results: the six rats survived. No sign of toxicity was observed after the administration of the extract at 2000 mg/kg dose. In the anatomopathological assessment, no macroscopic alterations were seen in the external surface or in the cavities, organs and tissues of the animals. Conclusions: according to this toxicity class method, the substance is not classified since LD50 is higher than 2 000 mg/kg.

DNA internal standard for the quantitative determination of hallucinogenic plants in plant mixtures.

Here, we show a new, simple, and rapid SYBR Green-based Real-Time PCR assay for the quantification of hallucinogenic plants in plant mixtures. As a test plant, Salvia divinorum Epling & Játiva-M., a perennial herb belonging to the Lamiaceae family able to induce hallucinations, changes in perception, or other psychologically induced changes with similar potency as LSD, was used. The method was tested on seven mixtures 100/0%, 80/20%, 60/40%, 40/60%, 20/80%, 10/90%, 0/100% (w/w) S. divinorum versus a non-hallucinogenic plant, Salvia officinalis. Total DNA was extracted from samples and quantified by Real-Time PCR. Arabidopsis thaliana genomic DNA was added, as internal standard, at the beginning of each extraction. A new formula for the interpretation of Real-Time PCR data, based on the relative quantification of DNA extracted from mixture versus a reference DNA extracted from a known amount of pure S. divinorum, was developed. The results of this work show an almost perfect correspondence between Real-Time PCR-calculated weight and the weight estimated by an analytical weighted method, proving the effectiveness of this method for the quantitative analysis of a given species in a plant mixture.

Psychopharmacology of the hallucinogenic sage Salvia divinorum.

At present, the Mexican mint Salvia divinorum is an unregulated hallucinogen. This has resulted in various on-line botanical companies advertising and selling S. divinorum as a legal alternative to other regulated plant hallucinogens. It is predictable that its misuse will increase rapidly. The active ingredient in S. divinorum is the neoclerodane diterpene, salvinorin A (1a), which has been shown to be a kappa agonist both in vitro and in vivo. This review will cover the current state of research into the psychopharmacology of S. divinorum.

The in vitro pharmacological activities and a chemical investigation of three South African Salvia species.

Salvia species (sage) are well known in folk medicine throughout the world. In South Africa sage is used against fever and digestive disorders. Three closely related South African species (Salvia stenophylla, Salvia repens and Salvia runcinata) were investigated for their anti-oxidant (DPPH assay); anti-inflammatory (5-lipoxygenase and cyclo-oxygenase assays); antimalarial (tritiated hypoxanthine incorporation assay); antimicrobial (disc diffusion and micro-dilution assays) properties and toxicity profile (tetrazolium-based assay). The solvent extracts exhibited anti-oxidant, antimalarial and antibacterial and poor anti-inflammatory properties. The essential oils exhibited anti-inflammatory and antimalarial properties, but displayed poor anti-oxidant and antimicrobial activity. The extract of Salviastenophylla and the essential oil of Salvia runcinata displayed the highest toxicity profile. Overall, Salvia runcinata displayed the most favorable activity of all three taxa tested with an IC(50) value of 6.09 (anti-oxidant); 29.05 (antimalarial) and 22.82 microg/ml (anti-inflammatory). Analytical procedures (GC-MS and HPLC-UV) were employed to generate chromatographic profiles for the essential oils and solvent extracts respectively. The HPLC analysis revealed the presence of rosmarinic acid in all three taxa while carnosic acid was only present in Salvia repens and Salvia stenophylla. The GC-MS analysis showed that oils were qualitatively and quantitatively variable. beta-Caryophyllene was present in large amounts in all three taxa. Other components present include camphor, alpha-pinene and alpha-bisabolol. The results of the in vitro pharmacological activities provide a scientific basis to validate the use of these Salvia species in traditional medicine in South Africa.