Differentiating Dyes: A Spectroscopic Investigation into the Composition of Scarlet Bloodroot (Haemodorum coccineum R.Br.) Rhizome.

Haemodorum coccineum, commonly known as scarlet bloodroot, is a plant native to New Guinea and the northern most parts of Australia. The highly coloured H. coccineum is used by communities in Larrakia country for dyeing garments and occasionally to treat snake bites. Previous studies into H. coccineum have focused on its taxonomic classification, with this being the first evaluation of the chemical composition of the plant. Haemodoraceae plants are reported to contain phenylphenalenones (PhPs), which are highly conjugated polycyclic oxygenated aromatic hydrocarbons. We report the characterisation of 20 compounds extracted from the rhizome of H. coccineum: four sugars and 16 compounds belonging to the PhP family. The compounds include five aglycones and seven glycosylated compounds, of which four contain malonate esters in their structures. Characterisation of these compounds was achieved through 1D and 2D NMR, MS analysis and comparison to the known phytochemistry of other species from the Haemodorum genus. Preliminary anti-microbial activity of the crude extract shows significant inhibition of the growth of both gram-positive and gram-negative bacteria, but no activity against Candida albicans.

Determination of Selected Isoquinoline Alkaloids from Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis Extracts by Liquid Chromatography and Their In Vitro and In Vivo Cytotoxic Activity against Human Cancer Cells.

The search for new substances with cytotoxic activity against various cancer cells, especially cells that are very resistant to currently used chemotherapeutic agents, such as melanoma cells, is a very important scientific aspect. We investigated the cytotoxic effect of Chelidonium majus, Mahonia aquifolium and Sanguinaria canadensis extracts obtained from different parts of these plants collected at various vegetation stages on FaDu, SCC-25, MCF-7, and MDA-MB-231 cancer cells. Almost all the tested extracts showed higher cytotoxicity against these cancer cells than the anticancer drug etoposide. The highest cytotoxicity against the FaDu, SCC-25, MCF-7 and MDA-MB-231 cancer cell lines was obtained for the Sanguinaria candensis extract collected before flowering. The cytotoxicity of extracts obtained from different parts of Chelidonium majus collected at various vegetation stages was also evaluated on melanoma cells (A375, G361 and SK-MEL-3). The highest cytotoxic activity against melanoma A375 cells was observed for the Chelidonium majus root extract, with an IC50 of 12.65 µg/mL. The same extract was the most cytotoxic against SK-MEL-3 cells (IC50 = 1.93 µg/mL), while the highest cytotoxic activity against G361 cells was observed after exposure to the extract obtained from the herb of the plant. The cytotoxic activity of Chelidonium majus extracts against melanoma cells was compared with the cytotoxicity of the following anticancer drugs: etoposide, cisplatin and hydroxyurea. In most cases, the IC50 values obtained for the anticancer drugs were higher than those obtained for the Chelidonium majus extracts. The most cytotoxic extract obtained from the root of Chelidonium majus was selected for in vivo cytotoxic activity investigations using a Danio rerio larvae xenograft model. The model was applied for the first time in the in vivo investigations of the extract's anticancer potential. The application of Danio rerio larvae xenografts in cancer research is advantageous because of the transparency and ease of compound administration, the small size and the short duration and low cost of the experiments. The results obtained in the xenograft model confirmed the great effect of the investigated extract on the number of cancer cells in a living organism. Our investigations show that the investigated plant extracts exhibit very high cytotoxic activity and can be recommended for further experiments in order to additionally confirm their potential use in the treatment of various human cancers.

Understanding of black salve toxicity by multi-compound cytotoxicity assays.

BACKGROUND: Black salve is a controversial complementary and alternative medicine (CAM) associated with skin toxicity and skin cancer treatment failures. Black salve formulations vary between manufacturers and contain a number of botanical and synthetic constituents. The skin cancer cytotoxicity of a number of these constituents has not been assessed to date. The alkaloids from the rhizomes of Sanguinaria canadensis, a key black salve ingredient, have had their single compound cytotoxicity assessed; however, whether they possess synergistic cytotoxicity with other compounds has not been studied and is of direct clinical relevance. This research aimed to improve our understanding of the skin cancer cytotoxicity of black salve constituents. METHODS: The cytotoxicity of individual and combination black salve constituents were assessed against the A375 melanoma and A431 squamous cell carcinoma cell lines. Cytotoxicity was determined using the Resazurin assay with fluorescence measured using a Tecan Infinite 200 Pro Microplate reader, compound cytotoxicity being compared to that of the topical cancer therapeutic agent, 5- fluouracil. Docetaxal was used as a positive control. Dunnetts p value was used to determine whether significant synergistic cytotoxicity was present. RESULTS: Sanguinarine was the most cytotoxic compound tested with a 24-hour IC50 of 2.1 µM against the A375 Melanoma cell line and 3.14 µM against the A431 SCC cell line. All black salve constituents showed greater cytotoxicity against the two skin cancer cell lines tested than the skin cancer therapeutic 5-Fluouracil with 24 hours of compound exposure. Chelerythrine and minor Quaternary Benzophenanthridine Alkaloids (QBAs) present in black salve, at concentrations not having a cytotoxic effect by themselves, boosted the cytotoxic effects of sanguinarine. This could be a synergistic rather than additive cytotoxic effect although the synergistic effect was cell line and concentration dependent. CONCLUSIONS: Black salve contains several cytotoxic compounds, a number of which have been found to possess synergistic cytotoxicity for the first time against skin cancer cell lines. In addition, these compounds together increase the overall cytotoxic effect. Assessing multi-compound cytotoxicity in herbal medicine can provide additional information about both their therapeutic and toxicity potential. As black salve is currently being used by patients, further cytotoxicity work should be undertaken to assess whether synergistic cytotoxicity exists when tested in normal skin cells.

Botanical Briefs: Bloodroot (Sanguinaria canadensis).

Bloodroot (Sanguinaria canadensis) is a plant that historically has been used in medicine for its antimicrobial, antihypertensive, anti-inflammatory, and antineoplastic properties. In dermatology, bloodroot has been utilized for its cytotoxic effects; it has been marketed as black salve as an anticancer treatment, but it does not come without notable toxicities. Unwanted cosmetic outcomes and even irreversible scarring and premalignant conditions have been reported. This article aims to bring awareness to both the therapeutic potential of S canadensis as well as the potential toxicities and risks associated with this North American plant.

Determination of Cytotoxic Activity of Sanguinaria canadensis Extracts against Human Melanoma Cells and Comparison of Their Cytotoxicity with Cytotoxicity of Some Anticancer Drugs.

Melanoma is an enormous global health burden, and should be effectively addressed with better therapeutic strategies. Therefore, new therapeutic agents are needed for the management of this disease. The aim of this study was the investigation of cytotoxic activity of some isoquinoline alkaloid standards and extracts obtained from Sanguinaria canadensis-collected before, during, and after flowering-against three different human melanoma cells (A375, G361, SK-MEL-3). The cytotoxicity of these extracts was not previously tested on these melanoma cell lines. Determination of alkaloid contents was performed by HPLC-DAD using Polar RP column and mobile phase containing acetonitrile, water, and 1-butyl-3-methylimidazolium tetrafluoroborate. The cytotoxicity of alkaloid standards was investigated by determination of cell viability and calculation of IC50 values. Significant differences were observed in the alkaloids content and cytotoxic activity of the extracts, depending on the season of collection of the plant material. In the Sanguinaria canadensis extracts high contents of sanguinarine (from 4.8543 to 9.5899 mg/g of dry plant material) and chelerythrine (from 42.7224 to 6.8722 mg/g of dry plant material) were found. For both of these alkaloids, very high cytotoxic activity against the tested cell lines were observed. The IC50 values were in the range of 0.11-0.54 µg/mL for sanguinarine and 0.14 to 0.46 µg/mL for chelerythrine. IC50 values obtained for Sanguinaria canadensis extracts against all tested cell lines were also very low (from 0.88 to 10.96 µg/mL). Cytotoxic activity of alkaloid standards and Sanguinaria canadensis extracts were compared with the cytotoxicity of anticancer drugs-etoposide, cisplatin, and hydroxyurea. In all cases except the one obtained for cisplatin against A375, which was similar to that obtained for Sanguinaria canadensis after flowering against the same cell line, IC50 values obtained for anticancer drugs were higher than the IC50 values obtained for sanguinarine, chelerythrine, and Sanguinaria canadensis extracts. Our results showed that Sanguinaria canadensis extracts and isoquinoline alkaloids, especially sanguinarine and chelerythrine, could be recommended for further in vivo experiments in order to confirm the possibility of their application in the treatment of human melanomas.

Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts.

Isoquinoline alkaloids may have a wide range of pharmacological activities. Some of them have acetylcholinesterase activity inhibition. Nowadays, neurodegenerative disorders such as Alzheimer's disease have become a serious public health problem. Searching for new effective compounds with inhibited acetylcholinesterase activity is one of the most significant challenges of modern scientific research. The aim of this study was the in vitro investigation of acetylcholinesterase activity inhibition of extracts obtained from Sanguinaria canadensis collected before, during and after flowering. The acetylcholinesterase activity inhibition of these extracts has not been previously tested. The aim was also to quantify selected alkaloids in the investigated extracts by high performance liquid chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). The acetylcholinesterase activity inhibition of the tested plant extracts and respective alkaloid standards were examined using high performance liquid chromatography with diode-array detector (HPLC-DAD) for the quantification of 5-thio-2-nitro-benzoic acid, which is the product of the reaction between the thiocholine (product of the hydrolysis of acetylthiocholine reaction) with Ellman reagent. The application of the HPLC method allowed for elimination of absorption of interfering components, for example, alkaloids such as sanguinarine and berberine. It is revealed that the HPLC method can be successfully used for the evaluation of the acetylcholinesterase inhibitory activity in samples such as plant extracts, especially those containing colored components adsorbing at wavelength in the range 405-412 nm. The acetylcholinesterase inhibition activity synergy of pairs of alkaloid standards and mixture of all investigated alkaloids was also determined. Most investigated alkaloids and all Sanguinaria canadensis extracts exhibited very high acetylcholinesterase activity inhibition. IC50 values obtained for alkaloid standards were from 0.36 for berberine to 23.13 µg/mL for protopine and from 61.24 to 89.14 µg/mL for Sanguinaria canadensis extracts. Our investigations demonstrated that these plant extracts can be recommended for further in vivo experiments to confirm their acetylcholinesterase activity inhibition.

Dermatologic uses of bloodroot: a review and reappraisal.

Bloodroot (Sanguinaria canadensis) is a plant, native to North America, containing bioactive compounds that interrupt biological processes. It has been around for centuries and is known for its medicinal properties. Today, naturopathic remedies are becoming more and more popular, especially for skin ailments. There are an alarming number of online vendors marketing their bloodroot-containing products as cures for skin cancer without any scientific evidence supporting such claims. Clinical data concerning the efficacy of bloodroot primarily come from case studies with unfavorable outcomes involving patients who self-treated with bloodroot-containing black salves. However, recent preclinical studies have concluded that sanguinarine, the active component of bloodroot, shows positive evidence of being an efficacious treatment for skin cancers at micromolar doses. This article reviews the mechanism of action of bloodroot as a skin cancer treatment, its misuse in clinical dermatology, and the FDA's stance on products containing bloodroot that are marketed and sold to laypersons. Members of the public should be made aware of the dangers of self-treating with bloodroot-containing products through effective communication and education by clinicians.

Review of Common Alternative Herbal "Remedies" for Skin Cancer.

BACKGROUND: Alternative herbal remedies for skin cancer are commonly found on the Internet. Many websites contain inaccurate or false information regarding side effects and efficacy. OBJECTIVE: To review the evidence behind several commonly advertised herbal remedies that claim to cure skin cancer: black salve, eggplant, frankincense, cannabis, black raspberry, milk thistle, St. John's wort, and turmeric. METHODS: A PubMed search was performed using the common and scientific names of frequently advertised herbal remedies along with the terms "nonmelanoma skin cancer," or "basal cell carcinoma" or "squamous cell carcinoma," or "melanoma." RESULTS: Some preclinical studies have shown positive evidence that these substances can induce apoptosis in skin cancer, but clinical studies proving efficacy are either insufficient, nonexistent, or show negative evidence. Botanicals that were excluded are those that do not have published studies of their efficacy as skin cancer treatments. CONCLUSION: Online advertising may tempt patients to use botanical agents while citing efficacy found in preclinical studies. However, many agents lack strong clinical evidence of efficacy. Dermatologists must be aware of common herbal alternatives for skin cancer treatment to maintain effective patient communication and education.

Black salve treatment of skin cancer: a review.

The use of complementary and alternative medicines in Australia has grown significantly. Much of this growth is due to their ease of accessibility from online vendors, often marketed with claims that are not scientifically tested. Black salve is a topical escharotic compound containing the active component sanguinarine, derived from the bloodroot plant. It has been advertised as a natural treatment for skin cancer. This article reviews the current state of black salve as an alternative skin cancer treatment, discussing its distribution and regulation, and provides a summary of clinical and laboratory studies. Clinical trials in this area are lacking, with most clinical data in the form of case reports demonstrating suboptimal therapeutic and cosmetic outcomes associated with its use. However, in vitro studies of sanguinarine suggest it causes indiscriminate destruction of healthy and cancerous tissue at doses higher than 5 µM, limiting its practical utility. It is vital that members of the public are aware of the potential effects and toxicity of commercial salve products.

Carcinogenic potential of sanguinarine, a phytochemical used in 'therapeutic' black salve and mouthwash.

Black salves are escharotic skin cancer therapies in clinical use since the mid 19th century. Sanguinaria canadensis, a major ingredient of black salve formulations, contains a number of bioactive phytochemicals including the alkaloid sanguinarine. Despite its prolonged history of clinical use, conflicting experimental results have prevented the carcinogenic potential of sanguinarine from being definitively determined. Sanguinarine has a molecular structure similar to known polyaromatic hydrocarbon carcinogens and is a DNA intercalator. Sanguinarine also generates oxidative and endoplasmic reticulum stress resulting in the unfolded protein response and the formation of 8-hydroxyguanine genetic lesions. Sanguinarine has been the subject of contradictory in vitro and in vivo genotoxicity and murine carcinogenesis test results that have delayed its carcinogenic classification. Despite this, epidemiological studies have linked mouthwash that contains sanguinarine with the development of oral leukoplakia. Sanguinarine is also proposed as an aetiological agent in gallbladder carcinoma. This literature review investigates the carcinogenic potential of sanguinarine. Reasons for contradictory genotoxicity and carcinogenesis results are explored, knowledge gaps identified and a strategy for determining the carcinogenic potential of sanguinarine especialy relating to black salve are discussed. As patients continue to apply black salve, especially to skin regions suffering from field cancerization and skin malignancies, an understanding of the genotoxic and carcinogenic potential of sanguinarine is of urgent clinical relevance.

Caracterización y estudio fitoquímico de Justicia secunda valh (Sanguinaria, Singamochilla, Insulina) / Characterization and phytochemical study of Justicia secunda valh (sanguinaria, singamochilla, insulina)

Introducción: las hojas de Justicia secunda Vahl. son ampliamente utilizadas en las poblaciones rurales de la costa ecuatoriana. Los pobladores la utilizan con fines medicinales sin que se hayan comprobado las propiedades que se le atribuye. El presente estudio se realizó con los extractos hidroalcohólico, alcohólico y etéreo de las hojas. Objetivos: caracterizar la droga cruda y varios extractos de J. secunda para determinar la presencia de metabolitos o principios activos con potencial medicinal y de esta manera analizar la posible utilidad en la elaboración de productos farmacéuticos. Métodos: se analizaron parámetros físico-químicos de la droga cruda, las hojas se recolectaron en la noche, se secaron hasta peso constante en una estufa a 30 °C ± 1 °C por 5 días y posteriormente se trituraron con un mortero, se pesaron y se obtuvieron los extractos por maceración con diversos solventes orgánicos; como etanol al 90 por ciento, éter dietílico y una solución hidroalcohólica al 50 por ciento v/v como menstruos. En el análisis fitoquímico se utilizaron técnicas cualitativas para la determinación de los principales metabolitos presentes en los extractos así como técnicas gravimétricas para determinación de cenizas totales, sólidos totales y porciento de humedad. Resultados: se constató que el contenido de cenizas totales y la humedad residual halladas en la droga cruda se encuentra dentro de los límites establecidos en la USP 37 para artículos de origen botánico. Además, se comprobó la presencia de alcaloides, cumarinas y glucósidos fenólicos en el extracto hidroalcohólico; taninos, glucósidos fenólicos y triterpenos y/o esteroides en el extracto alcohólico y alcaloides y triterpenos y/o esteroides en el extracto etéreo. Conclusión: la gran variedad de componentes químicos presentes en las hojas de J. secunda, le confieren un alto potencial para su uso medicinal como antiséptico, antiinflamatorio, antioxidante, entre otros(AU)

Introduction: Justicia secunda Vahl. leaves are widely used by rural populations from the Ecuadorian coast for medicinal purposes, but the properties attributed to it have not been verified. A study was conducted of hydroalcoholic, alcoholic and etheric leaf extracts. Objectives: Characterize the crude drug and several extracts of J. secunda to determine the presence of metabolites or active principles with medicinal potential, and analyze their possible usefulness to produce pharmaceutical products. Methods: An analysis was conducted of the physicochemical parameters of the crude drug. The leaves were collected at night and dried to constant weight in a stove at 30 °C ± 1 °C for 5 days. They were then crushed in a mortar and weighed. The extracts were obtained by maceration in various organic solvents, such as 90 percent ethanol, diethyl ether and a 50 percent v/v hydroalcoholic solution. For phytochemical analysis, qualitative techniques were applied to determine the main metabolites present in the extracts. Gravimetric techniques were used for determination of total ashes, total solids and percentage of humidity. Results: It was found that content of total ashes and residual humidity in the crude drug are within the limits established by USP 37 for items of botanical origin. Additionally, alkaloids, coumarins and phenolic glucosides were found to be present in the hydroalcoholic extract; tannins, phenolic glucosides and triterpenes and/or steroids in the alcoholic extract; and alkaloids and triterpenes and/or steroids in the etheric extract. Conclusion: The great variety of chemical components present in the leaves of J. secunda grants them high potential for medicinal use as antiseptic, antiinflammatory and antioxidant, among others(AU)

Sanguinaria canadensis: Traditional Medicine, Phytochemical Composition, Biological Activities and Current Uses.

Sanguinaria canadensis, also known as bloodroot, is a traditional medicine used by Native Americans to treat a diverse range of clinical conditions. The plants rhizome contains several alkaloids that individually target multiple molecular processes. These bioactive compounds, mechanistically correlate with the plant's history of ethnobotanical use. Despite their identification over 50 years ago, the alkaloids of S. canadensis have not been developed into successful therapeutic agents. Instead, they have been associated with clinical toxicities ranging from mouthwash induced leukoplakia to cancer salve necrosis and treatment failure. This review explores the historical use of S. canadensis, the molecular actions of the benzophenanthridine and protopin alkaloids it contains, and explores natural alkaloid variation as a possible rationale for the inconsistent efficacy and toxicities encountered by S. canadensis therapies. Current veterinary and medicinal uses of the plant are studied with an assessment of obstacles to the pharmaceutical development of S. canadensis alkaloid based therapeutics.

Black salve and bloodroot extract in dermatologic conditions.

Due to extensive advertising of black salve's effectiveness in "curing" skin cancers and healing other skin conditions, many patients are turning to self-treating with black salve. Although black salve has not been proven to have anticancer properties, application of black salve has been shown to cause damage to healthy tissue and the need for further treatment. We describe a 35-year-old woman whose one-time application of black salve to a healing biopsy site resulted in skin erosion and formation of a dermatitic plaque with subsequent scarring. Dermatologists and other health professionals need to be aware of this increasingly popular product to be able to better inform and treat their patients.

Bloodroot associated eschar.

We present a case of a 60-year-old woman who, after using a blemish cream purchased at a health store, presented to our clinic with a large eschar on the right neck. The active ingredient in this cream was found to be Sanguinaria canadensis, a known escharotic. This substance is popularly marketed as a natural remedy for a host of skin diseases such as blemishes, moles, warts, skin tags, basal cell carcinomas, squamous cell carcinomas, actinic keratoses, and even melanoma. The patient was treated with topical corticosteroids but is still left with a scar from the healed eschar. Dermatologists must be aware of the increasing popularity of "natural" medicinal skin cancer therapies, such as bloodroot-containing "cancer salves" and "herbal cures," and emphasize to their patients that conventional excision, micrographic surgery, electrodessication, and/or curettage are highly successful treatments for most skin cancers with little morbidity.

Owners' perception of the efficacy of Newmarket bloodroot ointment in treating equine sarcoids.

A retrospective questionnaire-based survey was used to determine the perceived efficacy of Newmarket bloodroot ointment in treating equine sarcoids. In 49 horses with 74 sarcoids, 64 sarcoids responded either completely (n = 49) or partially (n = 15) while 10 did not respond or worsened. Sarcoids < 2 cm responded better to treatment (P < 0.001) than did larger sarcoids.

Perception des propriétaires sur l'efficacité de l'onguent à la sanguinaire de Newmarket pour traiter les sarcoïdes équines. Un sondage rétrospectif réalisé par questionnaire a été utilisé pour déterminer l'efficacité perçue de l'onguent à la sanguinaire de Newmarket pour traiter les sarcoïdes équines. Chez 49 chevaux atteints de 74 sarcoïdes, 64 sarcoïdes ont soit répondu complètement (n = 49) ou partiellement (n = 15) tandis que 10 n'ont pas répondu ou se sont aggravées. Les sarcoïdes de < 2 cm ont mieux répondu au traitement (P < 0,001) que les sarcoïdes plus grandes.(Traduit par Isabelle Vallières).

A review of topical corrosive black salve.

Black salve is a compound derived from various inert ingredients, but it can be transformed into a corrosive ointment by the addition of bloodroot (Sanguinaria canadensis) or zinc chloride. Black salve products have been advertised as a natural remedy for many ailments, ranging from bee stings to skin cancer. This article reviews the current literature surrounding this compound, which in its corrosive form can be dangerous for use without medical supervision. Patients should be educated about the lack of objective evidence supporting the clinical efficacy of black salve as a skin cancer treatment, as well as the possible cosmetic defects resulting from tissue necrosis secondary to the effects of bloodroot and zinc chloride.