IntroductionAflatoxin B1 (AFB1) is a toxic low‐molecular‐weight secondary metabolite of Aspergillus flavus and A. parasiticus. AFB1 was classified as a Group I carcinogen by the World Health Organisation for Research on Cancer in 1993. AFB1 is an unavoidable natural contaminant of some herbal medicine, able to cause serious health issues for humans consuming the related medicine.ObjectiveTherefore, this study aimed to develop an efficient fluorescence polarisation immunoassay (FPIA) and a rapid, low‐cost, and easy‐to‐use membrane‐based flow‐through immunoassay (MBA) for determination of AFB1 in herbal medicine Origanum vulgare L., Rubus idaeus L., Urtica dioica L. and Sorbus aucuparia L.ResultsA cut‐off level of the developed MBA was 0.8 ppb. Validation of the developed test was performed with blank and spiked samples. Using three naturally contaminated or three artificially spiked samples. The FPIA showed a linear working range of 8.6 to 64 ppb, and a half maximal inhibitory concentration (IC50) of 24 ppb.ConclusionThe results were in good correlation with the enzymelinked immunosorbent assay (ELISA) results (the IC50 0.1 ppb). Both the sample preparation and analysis are simple, cost‐effective and easy to perform on‐site in non‐laboratory environments. Liquid chromatography with tandem mass spectrometry (LC‐MS/MS) was used as a confirmatory technique.

The roots of licorice (Glycyrrhiza glabra L.) have been widely used in traditional and officinal medicines for the treatment of different diseases. Natural deep eutectic solvents (NADES) have become popular for the extraction of active principles from medicinal plants. However, the ability of NADES to co-extract trace elements during the isolation of target active compounds is rarely investigated. The aim of this study was to analyze the content of trace elements in acid-based NADES extracts from the roots of G. glabra and the health risks associated with them. In this study, we have tested for the first time the ability of several acid-based NADES to co-extract glycyrrhizic acid (GA) and trace elements from the roots of G. glabra. GA has been identified as the dominant phytochemical in G. glabra NADES extracts (0.145–0.495 mg/g). Due to the close pKa of lactic acid and GA, the yield of GA in lactic acid-based NADES was higher in comparison with other tested NADES. The yield of GA in NADES3-NADES5 was statistically significant and surpassed the yield of GA in water. The recovery of all elements (except Li) by all tested NADES was low (less than 6%). According to an ANOVA test, the hydrogen bond donor type plays a decisive role in the extraction of elements. A strong positive correlation between the recovery of GA and MPI was noted. The metal pollution index, hazard quotient, hazard index, and chronic daily intake were calculated and suggest that all tested NADES extracts of G. glabra roots were nontoxic and possess no health risk for both ingestion and topical application.

Herbal medicinal products (HMPs) are widely used in medical practice due to their availability, ease of use, and relatively safe pharmacological profile. However, medicinal plants are capable of accumulating heavy metals and arsenic which can have toxic effect on the human body when found in HMPs. The aim of the study was to summarise and analyse requirements of the Russian and foreign pharmacopoeias for the limits of heavy metals and arsenic in HMPs. National and regional pharmacopoeias have limits for the content of the major toxic elements (lead, cadmium, mercury, chromium, and arsenic) in HMPs. The study showed that the Indian and Japanese pharmacopoeias include only semi-quantitative methods for determination of heavy metals and arsenic, while the Russian and Chinese pharmacopoeias allow for the use of both quantitative and semi-quantitative methods. It was demonstrated that the limits for heavy metals and arsenic are the same for herbal substances and HMPs. The development of consistent approaches to determination of heavy metals and arsenic content, a systematic transition to quantitative methods of analysis, and establishment of individual limits for toxic elements in different HMP dosage forms, will make it possible to achieve the so-called consistent harmonisation, ensure reliable assessment of the content of heavy metals and arsenic, and minimize the risk of their entering human body with HMPs.

The aim of the investigation was to study in experiment total toxicity and immunological safety of a novel domestic therapeutic vaccine against recurrent respiratory papillomatosis and anogenital condylomatosis, the vaccine injected intramuscularly.Materials and Methods. we studied acute toxicity of the vaccine by the following parameters: clinical presentation of intoxication, median lethal dose size, the change of body weight of the surviving animals; chronic toxicity -by dynamics of general condition of the animals, body weight, hematological and biochemical indices of peripheral blood, functional status of central nervous system, cardiovascular system, kidneys, as well as pathomorphological changes of viscera.Allergenicity was studied by systemic and active cutaneous anaphylactic tests. we assessed immunotoxicity by direct a hemagglutination assay and a delayed-type hypersensitivity test, as well as by neutrophilic activity using luminal-dependent chemiluminescence. Proliferative activity of В-and Т-lymphocytes to lipopolysaccharides and concanavalin А (ConА) was estimated in a direct immunofluorescence test using immunocytochemical assay with anti-Ki-67 monoclonal antibodies.Results. Mean lethal doses (lD 50 ) were not reached in white outbread rats injected intramuscularly by the tested vaccine at a maximum possible single dose (25 ml/kg). Multi-dose administration to white outbread rats and chinchilla rabbits at doses of 0.043; 0.43; 0.86 ml/kg and 0.023; 0.23; 0.4 ml/kg respectively, did not cause significant damage of functional status of basal organs and systems of experimental animals.The findings of systemic anaphylaxis and active cutaneous anaphylactic response to the vaccine in albino guinea pigs at the doses of 0.033 and 0.33 ml/kg intramuscularly indicated the vaccine to exhibit no allergenic properties.Intramuscular vaccine injected to first filial hybrid mice (СВА×С57Bl/6)f 1 at the doses of 0.084; 2.5 and 25 ml/kg showed neither significant increase or decrease of inflammatory response intensity relating to reference values. The vaccine at the doses of 0.084 and 2.5 ml/kg did not cause any changes of vaccination titer (IgG), phagocytic activity, spontaneous or induced blastic transformation of lymphocytes compared to the control group. The vaccine administered at the dose of 25 ml/kg significantly reduced the antibody formation level relating to the controls, as well as produced a suppressive effect on spontaneous and induced phagocytic activity resulting in the significant reduction of ConА-induced proliferative activity of Т-lymphocytes by the number of Ki-67 positive cells compared to the control group.Conclusion. The animal study of different types of general toxicity of the vaccine against recurrent respiratory papillomatosis and anogenital condylomatosis, the vaccine injected intramuscularly, showed the vaccine to be low toxic and immunologically safe in the dose range from 0.033 to 2.5 ml/kg.