Microbiological acceptance criteria, specifications of herbal drugs and herbal drug preparations in various pharmacopoeias: a global scenario.

PURPOSE: A pharmacopoeia is a compendium of guidelines and criteria for drug quality. It was established by a national or regional entity and has legal significance. This applies to administration of drugs in a particular nation or region. METHOD: In this study, the differences and similarities of microbiological acceptance criteria, specifications for microbial enumeration of herbal drugs and herbal drug preparations in 14 national and international pharmacopeias were investigated. RESULTS: It was found that 12 pharmacopeias have given separate microbial limits for total aerobic microbial count (TAMC) and total yeast and mold count (TYMC), and a list of specified microorganisms for which acceptance criteria are defined. However, similarities were noticed in Ph.Eur, Ph. Helv and, BP. Salmonella, and Escherichia coli are the most common pathogens specified for herbal preparations in which boiling water is added prior to use and for internal use in all Pharmacopoeias because they serve as indicators of potential contamination. CONCLUSION: From this study, it can be concluded that the differences in microbial limit tests and their acceptance criteria as specified in the various pharmacopoeias need to be harmonized. It will become a more convenient option for global drug manufacturers to import/export herbal drugs, and this would also eliminate the burden of performing various analytical methods and comply with different microbial acceptance criteria set by various pharmacopoeias. The comparative data obtained from this study will be used to develop strategies for revisions of pharmacopoeias in a harmonized manner with respect to microbiological acceptance criteria, specifications for microbial enumeration of herbal drugs and herbal drug preparations.

The Botanical Safety Consortium: A public-private partnership to enhance the botanical safety toolkit.

Botanical dietary supplement use is widespread and growing, therefore, ensuring the safety of botanical products is a public health priority. This commentary describes the mission and objectives of the Botanical Safety Consortium (BSC) - a public-private partnership aimed at enhancing the toolkit for conducting the safety evaluation of botanicals. This partnership is the result of a Memorandum of Understanding between the US FDA, the National Institute of Environmental Health Sciences, and the Health and Environmental Sciences Institute. The BSC serves as a global forum for scientists from government, academia, consumer health groups, industry, and non-profit organizations to work collaboratively on adapting and integrating new approach methodologies (NAMs) into routine botanical safety assessments. The objectives of the BSC are to: 1) engage with a group of global stakeholders to leverage scientific safety approaches; 2) establish appropriate levels of chemical characterization for botanicals as complex mixtures; 3) identify pragmatic, fit-for-purpose NAMs to evaluate botanical safety; 4) evaluate the application of these tools via comparison to the currently available safety information on selected botanicals; 5) and integrate these tools into a framework that can facilitate the evaluation of botanicals. Initially, the BSC is focused on oral exposure from dietary supplements, but this scope could be expanded in future phases of work. This commentary provides an overview of the structure, goals, and strategies of this initiative and insights regarding our first objectives, namely the selection and prioritization of botanicals based on putative toxicological properties.

A Graded Approach for Evaluating Health Claims about Plant-Based Food Supplements: Application of a Case Study Methodology.

The implementation of REGULATION (EC) No 1924/2006 has led to the formation of a list of health claims that can be used in food supplements (EU 432/2012). However, such supplements are often composed of plant preparations with claims omitted from this list. The peculiarity of plants is related to their long history of use, that could allow claims based on traditionally recognized health effects. In addition, the scientific literature has been enriched over the years through clinical studies that have assessed the bioavailability and efficacy of bioactive components, and investigated their mechanisms of action. Based on existing recognized models which aim to classify research according to the level of scientific evidence, Synadiet developed a three-grade model (A, B or C) for assessing plants health claims. In this paper, the applicability of the model is illustrated through an example for which a Grade B health claim attesting the possible contribution of red clover isoflavones to the improvement of blood lipid levels in postmenopausal women has been attributed. The model appears able to be easily extrapolated to claims pertaining to other plants. If adopted by consensus at European level, this model could initiate the implementation of a positive list of health claims on plant preparations.

PCR-based rapid diagnostic tools for the authentication of medicinal mistletoe species.

BACKGROUND: Taxilli Herba (TH) and Visci Herba (VH), defined as the leaves and branches of the mistletoe species Taxillus chinensis and Viscum coloratum, respectively, are popular herbal medicines in East Asia. However, commercial TH and VH products are frequently adulterated with related inauthentic mistletoe species, posing efficacy and safety concerns. Accurate species identification of herbal medicinal products is a prerequisite for quality control, but traditional morphological identification methods are hampered by difficulties in discriminating among closely related species and in identifying the source materials in processed products. PURPOSE: This study aimed to develop sequence-characterized amplified region (SCAR) markers and a multiplex-SCAR assay for rapid and accurate identification of authentic TH and VH. METHODS: The matK region was sequenced in a total of 20 samples from five mistletoe species, namely T. chinensis and V. coloratum, and three species often found in adulterated herbal medicines, T. sutchuenensis, V. articulatum, and Macrosolen tricolor. Species-specific nucleotide polymorphisms were identified and short regions (21-22 bp) containing at least two species-specific nucleotides close to the 3' end were incorporated into SCAR primers that produced uniquely sized PCR amplicons for each species. The five SCAR primer sets were also combined into a multiplex-SCAR assay. RESULTS: The SCAR primers successfully generated amplicons of the expected size for each target species even with low-DNA templates or with templates containing DNA from multiple samples. No amplification was observed in non-target species. The SCAR markers and the multiplex-SCAR assay successfully identified commercial TH and VH products that were counterfeit or adulterated in both dried and processed products. CONCLUSION: This is the first report to illustrate discrimination of genuine medicinal mistletoe species with DNA-based marker assays, enabling rapid and accurate species identification. The SCAR assays developed in this study will facilitate the standardization of commercial mistletoe products.

Quality evaluation of rhubarb based on qualitative analysis of the HPLC fingerprint and UFLC-Q-TOF-MS/MS combined with quantitative analysis of eight anthraquinone glycosides by QAMS.

Rhubarb is one of the most ancient and important herbal medicines, but its current quality evaluation (QE) methods have some limitations. In this study, a new method was developed for the comprehensive QE of rhubarb. First, fingerprints of 28 batches of three species of rhubarb samples were determined by HPLC, the reference fingerprint was established and the common peaks were assigned. Second, the components of common peaks in the fingerprints were identified by ultrafast liquid chromatography quadrupole time-of-flight mass spectrometry. Finally, a method for the simultaneous determination of the contents of eight anthraquinone glycosides in rhubarb using quantitative analysis of multiple components by a single marker (QAMS) was established, and the contents of these eight components in 28 batches of rhubarb determined by QAMS and the external standard method were compared. The results showed that there were 31 common peaks in the rhubarb fingerprint. The components of these 31 common peaks were identified, and 20 of them were unambiguously confirmed by reference substances, including eight anthraquinone glycosides. The contents of eight anthraquinone glycosides in the 28 batches of rhubarb determined by QAMS and the external standard method were not significantly different. In conclusion, the method established in this study can be used for the comprehensive QE of rhubarb and can also provide a reference for the QE of other herbal medicines.

Efficacy of Aloe-Vera Use for Prevention of Chemotherapy-Induced Oral Mucositis in Children with Acute Lymphoblastic Leukemia: A Randomized Controlled Clinical Trial.

Oral mucositis can be caused by chemotherapy and can affect a patient's quality of life. Nowadays, to prevent chemotherapy-induced oral mucositis (CIOM) is a crucial point in palliative care centers. This trial aimed to assess the effectiveness of aloe-vera in that concept. The trial was accomplished at Hematology Department of Hospital of Children of Damascus University, Syria. Acute lymphoblastic leukemia (ALL) children were the population from which 26 children were enrolled in the study. They were aged between 3 and 6 years old and were randomly referred according to the intervention into two groups, Aloe-vera (AV) and sodium bicarbonate 5% (13 each). Spongeous sticks were used to help in applying the material on tongue, labial and buccal mucosa, lips, floor of the mouth, and hard palate. Two blinded external examiners evaluated oral mucosa weekly for up to 2 months using the World Health Organization grading scale. Mann-Whitney U test was used to analyze data. According to the observed findings, CIOM degrees were less severe in the aloe-vera group than in the sodium bicarbonate group. Statistically significant difference of occurrence of different CIOM degrees between groups was recorded in the 2nd, 3rd, 4th, and 7th weeks of follow-up period. Moreover, Mann-Whitney U test indicated that patients in the sodium bicarbonate group began CIOM sooner than those in the aloe-vera group with a statistically significant difference (p = .001). These findings show that topical application of aloe-vera solution is effective in the prevention of CIOM in ALL children.

Formulation, Standardization, and a Preliminary Before-After Clinical Trial on Effect of a Traditional Herbal Formula (Maqliasa) on Manifestations of Ulcerative Colitis.

BACKGROUND: Ulcerative Colitis (UC) is a chronic condition that represents a group of intestinal disorders causing prolonged inflammation of the digestive tract. Nowadays, efforts to investigate new remedies have led to a committed movement toward the integration of traditional and complementary medicine into mainstream medicine. In Traditional Persian Medicine (TPM), Maqliasa is one of the most frequent gastrointestinal prescriptions which is claimed to be effective in both remission and recurrent phases of UC. OBJECTIVE: The aim of this pilot study was to examine the effect of Maqliasa on UC symptoms. METHODS: Through a non-randomized before-after uncontrolled clinical trial, 13 outpatients with active UC were enrolled in the study. They continued their conventional drug regimens plus Maqliasa capsules (2 capsules t.i.d.) for 28 days. Three visits were arranged for each patient- the first admission, day 14th admission and day 28th admission. The patients were evaluated by Lichtiger Colitis Activity Index. Wilcoxon signed-rank test was used for statistical analysis. RESULTS: The Lichtiger colitis activity index improved in the study group was compared to the corresponding baseline values: baseline score: 7.8 ± 0.5; after two weeks: 5.9 ± 0.6 (P=0.002); after four weeks: 3.6 ± 0.5 (P=0.001). CONCLUSIONS: This study showed the promising effect of Maqliasa in the treatment of active UC. However, due to some of the limitations of the study, conducting future high-quality randomized clinical trials would be crucial.

Rapid magnetic particles-based enzyme immunoassay for the quality control of Glycyrrhiza spp. based on glycyrrhizin content.

Glycyrrhizin (GC) is a triterpenoid saponin isolated from the roots of Glycyrrhiza spp., a medicinal plant that is present in 70% of Kampo prescriptions. Since the GC content in Glycyrrhiza spp. affects its various pharmacological activities, Glycyrrhiza spp. is prescribed to contain at least 2% of GC in the Japanese pharmacopoeia, and its quality control based on GC content is required. In this study, a magnetic particles-based enzyme immunoassay (MPs-EIA) was developed using specific monoclonal antibody against GC (MAb 2H2) for the detection of GC in Glycyrrhiza spp. In this system, the immunoreaction time using primary and secondary antibodies was reduced by taking advantage of the wide surface area of magnetic particles (MPs) conjugated with GC by N,N'­carbonyldiimidazole (CDI)-mediated method. Optimization of MPs-EIA revealed that total assay time (~2 h) was reduced to over half of that of conventional indirect competitive enzyme-linked immunosorbent assay (ELISA) (~5 h). In addition, the GC concentration was detectable within the range from 97.7 to 781 ng/mL, with a limit of detection of 71.4 ng/mL. A series of further validation analyses support the reliability and accuracy of the developed MPs-EIA for the detection of GC in Glycyrrhiza spp. Since the present MPs-EIA overcomes the disadvantage of ELISA in terms of rapidity, it provides a useful approach for the effective quality control of Glycyrrhiza spp., especially when handling multiple samples.

Quality Assessment of Sitopaladi Churna Using High-Performance Liquid Chromatography Coupled with Multivariate Analysis.

"Sitopaladi churna," a well-known formulation of Ayurveda, is prescribed to treat the disease like Bronchitis, Pneumonia, allergic conditions, viral infection of the respiratory tract and as a natural antioxidant. A novel method based on reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to photodiode array detector was established and validated for sitopaladi churna. Here, HPLC fingerprints data obtained for 28 samples including standard sample were then treated to chemometric analysis like principal component analysis and hierarchical clustering analysis for further analysis to evaluate the differences in market samples of sitopaladi churna. Additionally, one major marker compound, piperine was quantified and it also facilitated for relative retention time. The simulative mean spectrum was also generated. The validation results showed that the developed method was simple, precise and stable. Thus, the developed chromatographic method adjoined with multivariate analysis can be used as an efficient and practical approach for quality assessment of sitopaladi churna.

Extracting and Analyzing Pyrrolizidine Alkaloids in Medicinal Plants: A Review.

Pyrrolizidine alkaloids (PAs) are distributed in plant families of Asteraceae, Boraginaceae, and Fabaceae and serve in the chemical defense mechanism against herbivores. However, they became a matter of concern due to their toxicity associated with the high risk of intake within herbal preparations, e.g., phytopharmaceutical formulations, medicinal teas, or other plant-derived drug products. In 1992, the German Federal Ministry of Health established the first limits of PA content for fourteen medicinal plants. Because of the toxic effects of PAs, the Federal Institute of Risk Assessment (BfR) established more stringent limits in 2011, whereby a daily intake <0.007 µg/kg body weight was recommended and valid until 2018. A threefold higher limit was then advised by BfR. To address consumer safety, there is the need for more efficient extraction procedures along with robust, selective, and sensitive analytical methods to address these concerns. With the increased prevalence of, e.g., phytopharmaceutical formulations, this timely review comprehensively focuses on the most relevant extraction and analysis strategies for each of those fourteen plant genera. While a variety of extraction procedures has been reported, differences in PA content of up to 1110 ppm (0.11% (w/w)) were obtained dependent on the nature of the solvent and the applied extraction technique. It is evident that the efficient extraction of PAs requires further improvements or at least standardization of the extraction conditions. Comparing the various analytical techniques applied regarding selectivity and sensitivity, LC-MS methods appear most suited. This review shows that both standardized extraction and sensitive determination of PAs is required for achieving appropriate safety levels concerning public health in future.

[Construction and application of modern research paradigm of new foreign introducing traditional Chinese medicine].

With the development of society, more and more foreign natural medicines have entered China, but they are faced with the problem that they are not given with the traditional Chinese medicine(TCM) properties when they are used with other TCM. When we carried out the research on introducing foreign natural plant resources to TCM, gradually, a new research model and field with original characteristics of TCM--the study of new foreign introducing TCM had been formed. In the process of it, we explored and summarized research rules to form the research paradigm, which will be conducive to the standardization and scientization of new foreign introducing TCM research. For the research of foreign introducing TCM, we analyzed the research background, reviewed the application history, sorted out the research status, elaborated the concept and summarized the research achievements. On this basis, we put forward the concept and system of the modern research paradigm of new foreign introducing TCM, studied and clarified the core elements, properties and research principles of this paradigm, and summarized the research contents and methods of new foreign introducing TCM. We interpreted the foundation of paradigm construction from multiple perspectives. The paradigm was the practical application of theoretical innovation of TCM. Under the guidance of it, more and more varieties of new foreign introducing TCM will be studied, the study will be more standardized, the conclusions will be more scientific and reliable, which will inspire and guide more researchers to focus on the research of new foreign introducing TCM, and the paradigm itself will be constantly improved. The construction and application of the paradigm will certainly accelerate the innovative research of foreign introducing TCM and play a historic role in promoting the enrichment of traditional Chinese medicine varieties.

Mycotoxin Contamination Concerns of Herbs and Medicinal Plants.

Plants and medicinal herbs that are available on the market do not always meet quality and safety standards. One particular concern is the risk of contamination with mycotoxins. Aflatoxins and ochratoxin A are the most frequently described mycotoxins in herbal products and have repeatedly been reported to occur at concentrations which exceed regulatory levels set by the European Union (EU). Possible solutions include enforcing existing limits, and for the new materials, establishing tighter limits and mandate the growth of medicinal plants in EU member countries under more strict conditions.

Microbial contamination in herbal medicines: a serious health hazard to elderly consumers.

INTRODUCTION: The use of herbal medicine is on the rise worldwide, and safety issues associated with herbal medicines may have an exacerbated impact in elderly because this population has an increased susceptibility and sensitivity to health complications due to the aging process. METHODS: This cross-sectional study was carried out at a primary health care unit in the city of Macapa, Brazil. The herbal medicines used and the sociodemographic characteristics of 123 voluntarily consenting participants were collected using a structured questionnaire. A total of 132 herbal medicines with oral or topical administration were donated by the elderly for microbial analysis before consumption, and 18 water samples used in the preparation of homemade herbal medicines were collected. Bacterial and fungal counts and identification of bacterial pathogens (Escherichia coli, Salmonella spp., Pseudomonas aeruginosa and Staphylococcus aureus) were performed according to the regulations of the Brazilian Pharmacopoeia and World Health Organization. Water analysis for the detection of coliforms and E. coli was carried out using Colilert® according to the manufacturer's instructions and the techniques established by Standard Methods. RESULTS: Of the study participants, 78.8% were women. Bacterial growth was observed in samples from 51.5% of study and 35.6% had fungal growth. A total of 31.8% of the herbal medicine samples exceeded the safety limits (CFU/g ≤ 105), including 16.7% of the homemade herbal medicines and 15.1% of the commercial herbal medicines. It was also found that 31.0% of the samples exceeded the safety limit for fungal growth. The microorganisms most commonly isolated from the herbal medicines were S. aureus (49.2%), followed by Salmonella spp. (34.8%), E. coli (25.8%), and P. aeruginosa (14.4%). Of water samples analyzed, 77.8% were positive for total coliforms (1 ml) and in 66.7% water samples E. coli was detected (1 ml), making them unfit for consumption. CONCLUSIONS: The use of homemade and commercial herbal medicines is a major risk to the health of elderly who use these therapies due to the lack of microbial quality standards. We observed levels of viable bacteria and fungi that were above safety limits; in addition, we were able to isolate pathogenic bacteria from these herbal medicines.