Determination of Multiple Neurotransmitters through LC-MS/MS to Confirm the Therapeutic Effects of Althaea rosea Flower on TTX-Intoxicated Rats.

Tetrodotoxin (TTX) inhibits neurotransmission in animals, and there is no specific antidote. In clinical practice in China, Althaea rosea (A. rosea flower) extract has been used to treat TTX poisoning. In this work, the efficacy of the ethyl acetate fraction extract of A. rosea flower in treating TTX poisoning in rats was investigated. A high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to determine nine neurotransmitters in rat brain tissue, including γ-aminobutyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT), noradrenaline (NE), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 5-hydroxyindole-3-acetic acid (5-HIAA), epinephrine (E), and tyramine (Tyn). The detoxifying effect of A. rosea flower was verified by comparing the changes in neurotransmitters' content in brain tissue before and after poisoning in rats. The assay was performed in multiple reaction monitoring mode. The quantification method was performed by plotting an internal-standard working curve with good linearity (R2 > 0.9941) and sensitivity. Analyte recoveries were 94.04-107.53% (RSD < 4.21%). Results indicated that the levels of 5-HT, DA, E, and NE in the brains of TTX-intoxicated rats decreased, whereas the levels of GABA, Tyn, and 5-HIAA showed an opposite trend, and HVA and DOPAC were not detected. The levels of all seven neurotransmitters returned to normal after the gavage administration of ethyl acetate extract of A. rosea flower to prove that the ethyl acetate extract of A. rosea flower had a therapeutic effect on TTX poisoning. The work provided new ideas for studies on TTX detoxification.

Impact of copper treatment on phenylpropanoid biosynthesis in adventitious root culture of Althaea officinalis L.

Althaea officinalis has been widely used in various pharmaceutical applications. The biological effects and significance of phenylpropanoids in numerous industries are well studied. However, fulfilling consumer demand for these commercially important compounds is difficult. The effect of heavy-metal toxic influence on plants is primarily due to a strong and rapid suppression of growth processes, as well as the decline in activity of the photosynthetic apparatus, also associated with progressing senescence processes. Some of the secondary metabolite production was triggered by the application of heavy metals, but there was not a stress response. In the adventitious root culture of A. officinalis, copper-mediated phenylpropanoid biosynthesis has been investigated in both concentration-and duration-dependent manners. High-performance liquid chromatography (HPLC) analysis revealed a total of nine different phenolic compounds in response to different concentrations of copper chloride. In this study, high productivity of phenolic compounds was observed in the copper chloride treated-adventitious root culture of A. officianalis. In particular, a low concentration of copper chloride led to a significant accumulation of phenolic compounds under optimal conditions. Moreover, all genes responsible for phenylpropanoid biosynthesis may be sensitive to phenolic compound production following copper treatment. Especially, the highest change in transcript level was observed from AoANS at 6 h. According to our findings, treatment with copper chloride (0.5 mM) for 48 or 96 h can be an appropriate method to maximize phenylpropanoid levels in A. officinalis adventitious root culture.

Study of Superoxide Dismutase Activity in Long-Term Cultivated Artemisia and Althaea "hairy" Roots.

The aim of the study was to evaluate the long-term effect of Agrobacterium rhizogenes genes transfer on plant antioxidant system by the study of superoxide dismutase (SOD) activity in "hairy" roots of Artemisia and Althaea spp plants. PCR analyses revealed stability of the transformation and presence of bacterial rol B and rol C genes in the "hairy" roots after 4-6 years from the transformation event. SOD activity in the roots of untransformed in vitro cultivated plants used for the initiation of "hairy" roots growth was in the range of 45.8 ± 8.7 U/µg (Althaea officinalis) and 275 ± 97.1 U/µg (Artemisia ludoviciana). After a long-term in vitro cultivation more than half of tested "hairy" root lines (54%) showed a significant increase in SOD activity values compared to untransformed roots. The highest SOD activity values of "hairy" root lines (24-fold increase) were founded in A. officinalis (1105 ± 174 U/µg) and A. dracunculus (1356 ± 402 U/µg). The increase of the activity was found also in "hairy" roots of A. vulgaris (up to 375 ± 28.2 U/µg, sevenfold increase), A. ludoviciana (1001 ± 191 U/µg, 3.6-fold increase), and A. tilesii (438 ± 104 U/µg, 1.6-fold increase). The results of our study indicate that transformation by wild-type A. rhizogenes not harboring any foreign genes implementing in SOD activity regulation can often stably activate plant antioxidant enzyme system. This effect, observed in the "hairy" roots of five plant species in 4-6 years of the transformation event, obviously, should be taken into account in works aimed at creating transgenic plants by Agrobacterium-mediated transformation.

Effect of topical marshmallow (Althaea officinalis) on atopic dermatitis in children: A pilot double-blind active-controlled clinical trial of an in-silico-analyzed phytomedicine.

Atopic dermatitis (AD) is a chronic relapsing eczematous skin disease, which primarily affects infants and young children. Due to the side effects of commonly used drugs for its treatment, the development of safer therapeutic strategies is needed. There are many reports on the topical use of marshmallow (Althaea officinalis) for a range of skin diseases in Persian medicine. The main aim of the present investigation was evaluating the efficacy of marshmallow in children with mild-to-moderate atopic dermatitis. Another aim of the study was screening the anti-allergic and anti-inflammatory potential of phytocomponents against target proteins, including TNF-alpha, IL6, and PDEs A, B, and D enzymes with PDB IDs: 2AZ5, 1P9M, 3I8V, 4KP6, and 1Y2K, respectively, along with their respective standard ligands using computational docking analysis. A pilot clinical trial was designed to investigate the safety and efficacy of Althaea officinalis in children with AD. The diagnosis of AD was made according to the criteria of Hanifin and Rajka. Children between 3 months and 12 years old were participated in this trial and randomly allocated into two parallel intervention and control groups. The intervention group used Althaea officinalis 1% ointment while the positive control group used Hydrocortisone 1% ointment twice a day for a week and after that, three times per week for a period of 3 weeks. The severity of AD was measured using the SCORAD score at the end of each assessment visits. A total number of 22 patients completed the study. A significant decrease of the SCORAD score was observed in both groups. At the end of the study, this score change, which indicates the improvement of the patients was significantly higher in the intervention group in comparison to the baseline (p-value = .015) and week 1 (p-value = .018). In the docking analysis of the study, 33 phytochemical compounds were identified, which were docked into the active site of IL6, TNF-alpha, and human PDE4 isoenzymes. Affinity toward the selected enzymes was significantly higher in glycosylated compounds. The results of this pilot study showed that the efficacy of Althaea officinalis 1% ointment in a decrease of disease severity is more than Hydrocortisone 1% in children with AD. However, further studies are needed to confirm this finding. Moreover, the docking analysis revealed that the inhibitory activity of compounds with free hydroxyl groups such as glycosylated compounds was better than others, probably due to the hydrogen bond interaction of hydroxyl groups of the ligands with the enzymes.

Additions of optimum water, spent mushroom compost and wood biochar to improve the growth performance of Althaea rosea in drought-prone coal-mined spoils.

Ecosystem degradation as a result of coal mining is a common phenomenon in various regions of the world, especially in arid and semi-arid zones. The implementation of appropriate revegetation techniques can be considered crucial to restore these degraded areas. In this regard, the additions of spent mushroom compost (SMC) and wood biochar (WB) to infertile and degraded soils have been reported to enhance soil fertility and plant growth under water (W) deficit conditions. However, the combined application of W, SMC and WB to coal mine degraded soils, to promote Althaea rosea growth and facilitate subsequent restoration, has not been explored yet. Hence, in the current study a pot experiment was carried out by growing A. rosea on coal mine spoils to assess the influence of different doses of W, SMC and WB on its morpho-physiological and biochemical growth responses. The results indicated that several plant growth traits like plant height, root length and dry biomass significantly improved with moderate W-SMC-WB doses. In addition, the simultaneous application of W-SMC-WB caused a significant decrease in hydrogen peroxide (H2O2) (by 7-56%), superoxide anion (O2●‒) (by 14-51%), malondialdehyde (MDA) (by 23-46%) and proline (Pro) contents (by 23-66%), as well as an increase in relative water content (by 10-27%), membrane stability index (by 2-24%), net photosynthesis rate (by 40-99%), total chlorophylls (by 43-113%) and carotenoids (by 31-115%), as compared to the control treatment. The addition of SMC and WB under low-W regime enhanced leaf water use efficiency, and soluble sugar content, also boosting the activity of superoxide dismutase, catalase, peroxidase and ascorbate peroxidase in leaf tissues, thus reducing the oxidative stress, as proved by low levels of H2O2, O2●‒, MDA and Pro contents. Finest growth performance under optimum doses of W (60% field capacity), SMC (1.4%) and WB (0.8%) suggest that revegetation of A. rosea with the recommended W-SMC-WB doses would be a suitable and eco-friendly approach for ecological restoration in arid degraded areas.

Tolerance and distribution of cadmium in an ornamental species Althaea rosea Cavan.

The ornamental plants of Althaea rosea Cavan. were exposed to cadmium (Cd) at concentrations of 0, 5, 10, 50 and 100 mg·kg-1 to evaluate the potential of accumulation capacity and tolerance ability. The results showed that A. rosea was a Cd tolerance plant, and Cd accumulation was 4.57 mg·kg-1 in shoot and 9.43 mg·kg-1 in root at 100 mg·kg-1 Cd concentration. The high tolerance ability could be explained by the distribution characteristics and the defense mechanism. The accumulated Cd was allocated in root, older leaves, and subcellular level of the cell wall and supernatant soluble fraction to protect the physiological leaves from damage. The defense systems included the enzyme systems of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) that could effectively eliminate the excessive ROS, and the non-enzymes system of total soluble proteins and non-protein thiols (NP-SH) that played an important role in detoxification. Thus, A. rosea could be used as a potential species for phytoremediation in Cd contaminated areas and beautify the environment.

The evaluation ofefficacy and safety of Cough (EMA) granules used for upper respiratory disorders.

Ivy leaf is used for the treatment of respiratory diseases with the intensive mucus formation, respiratory infections, and irritating cough coming from the common cold. Conferring to clinical trials, the efficacy, and tolerability of ivy leaf is good. The main compounds accountable for biological activity are triterpene and saponins. Ivy leaves show convulsive/antispasmodic, anti-inflammatory, antimicrobial, analgesic, anthelmintic and anti-thrombin activity. Not only ivy but also marshmallow and mustard seeds are used for these indications. This study was conducted to evaluate the efficacy and safety of Cough (EMA; European Medicines Agency) granules used for upper respiratory disorders. This clinical trial was conducted on 150 patients, out of which 75received the Cough (EMA) granules and 75received the placebo. The age range of patients was 3 years to above 15 years. The sample paired t-test was applied to evaluate the significant level. Cough (EMA) granules were found effective in the treatment of cough, cold, and flu symptoms. The new treatment Cough (EMA) granules were safe and well tolerated in patient at given specific age group. The study recommends that Cough (EMA) granules can be used effectively in the treatment of upper respiratory tract infection.

Phytochemical Characterization of Low Molecular Weight Constituents from Marshmallow Roots (Althaea officinalis) and Inhibiting Effects of the Aqueous Extract on Human Hyaluronidase-1.

Extract RE was obtained from the roots of Althaea officinalis in a yield of 8.1%, related to the dried plant material, by extraction with MeOH-H2O (1:1), followed by precipitation with EtOH to remove high molecular weight constituents. Phytochemical investigation of RE revealed the presence of N-phenylpropenoyl-l-amino acid amides 1-5, 8% glycine betaine 6, about 9% total amino acids with proline as the main compound, and about 61% mono- and oligomeric carbohydrates with sucrose as the main compound. Further fractionation revealed the presence of a hypolaetin diglycoside (12) and four hypolaetin glycosides (7-9 and 11) with O-sulfocarbohydrate moieties; additionally, 4'-O-methylisoscutellarein-8-O-ß-d-(3″-O-sulfo)glucuronopyranoside (10) and the diglycosylated coumarin haploperoside D (13) were identified. The hypolaetin-O-sulfoglycosides 7-10 are new natural products. RE inhibited the enzymatic activity of surface-displayed human hyaluronidase-1 on Escherichia coli F470 cells with an IC50 of 7.7 mg/mL. RE downregulated mRNA expression of hyal-1 in HaCaT keratinocytes at 125 and 250 µg/mL, respectively. These data contribute to a deeper phytochemical understanding of marshmallow root extracts and to the positive influence of extracts used for therapy of irritated and inflamed buccal tissue and cough.

In vivo screening and evaluation of four herbs against MRSA infections.

BACKGROUND: Recently, we reported high in vitro antibacterial efficacy of Althaea officinalis, Ziziphus jujuba, Cordia latifolia and Thymus vulgaris out of a total 21 plants against wide range of bacteria including MRSA. This study was therefore, designed to confirm efficacy of these four herbs against MRSA in an animal model. METHODS: A pilot study was conducted to establish the dose of S. aureus (KY698020) required to induce clinical infection. Afterword, in main trial, efficacy of aforementioned plant extracts on the course of sore throat was checked by evaluating general health, gross lesion score, bacterial load and hematology in mice. RESULTS: Pilot study revealed that 40 µl dose of 107 CFU/ml could induce infection which persist upto 08 days post infection. Mice treated with T. vulgaris and Z. jujuba showed reduction in gross lesion score of both heart and lungs. Treatment with only some plants could significantly decrease bacterial load of throat (T. vulgaris) heart, blood and joint (C. latifolia, and T. vulagris). Hematological indicators confirmed in vivo control of MRSA infection in all treatment groups except A. officinalis. CONCLUSION: This is first report confirming in vivo anti-MRSA potential of C. latifolia and T. vulgaris and highlight the need to explore bioactive constituents of these plants. Moreover, previously reported in vitro antibacterial efficiency of A. officinalis could not be validated in current study.

Role of Ni-tolerant Bacillus spp. and Althea rosea L. in the phytoremediation of Ni-contaminated soils.

In our current study, four nickel-tolerant (Ni-tolerant) bacterial species viz, Bacillus thuringiensis 002, Bacillus fortis 162, Bacillus subtilis 174, and Bacillus farraginis 354, were screened using Ni-contaminated media. The screened microbes exhibited positive results for synthesis of indole acetic acid (IAA), siderophore production, and phosphate solubilization. The effects of these screened microbes on Ni mobility in the soil, root elongation, plant biomass, and Ni uptake in Althea rosea plants grown in Ni-contaminated soil (200 mg Ni kg-1) were evaluated. Significantly higher value for water-extractable Ni (38 mg kg-1) was observed in case of Ni-amended soils inoculated with B. subtilis 174. Similarly, B. thuringiensis 002, B. fortis 162, and B. subtilis 174 significantly enhanced growth and Ni uptake in A. rosea. The Ni uptake in the shoots and roots of B. subtilis 174-inoculated plants enhanced up to 1.7 and 1.6-fold, respectively, as compared to that in the un-inoculated control. Bacterial inoculation also significantly improved the root and shoot biomass of treated plants. The current study presents a novel approach for bacteria-assisted phytoremediation of Ni-contaminated areas.

Immune-Stimulatory Effects of Althaea rosea Flower Extracts through the MAPK Signaling Pathway in RAW264.7 Cells.

Althaea rosea (Linn.) is a medicinal plant from China and Korea that has been traditionally used to control inflammation, to stop bedwetting and as a mouthwash in cases of bleeding gums. Its flowers are employed medicinally for their emollient, demulcent and diuretic properties, which make them useful in chest complaints. Furthermore, a flower extract decoction is used to improve blood circulation, for the treatment of constipation, dysmenorrhoea, haemorrhages, etc. However, the possible mechanisms of the immune-stimulatory effect remains to be elucidated. Therefore, we investigated the role of Althaea rosea flower (ARF) extracts in the immune-stimulatory effect of macrophages and the underlying mechanisms of action. ARF water extract (ARFW) could dose-dependently increase NO production and cytokines (IL-6 and TNF-α). We also found that ARFW significantly increased the expression of iNOS and COX-2 proteins in RAW264.7 cells. Consistent with these results, MAPK protein (JNK, ERK, p38) expression levels were induced after treatment with ARFW. Additionally, ARFW showed a marked increase in the phosphorylation level of IκBα and subsequent IκBα degradation allowing NF-κB nuclear translocation. These results suggest that the immune-stimulatory effect of A. rosea flower extracts is mediated through the translocation of NF-κB p65 subunit into the nucleus from the cytoplasm and subsequent activation of pro-inflammatory cytokines (IL-6 and TNF-α) and other mediators (iNOS and COX-2), which occurs mainly through MAPK signalling pathway. Thus, we suggest that ARFW could be considered as a potential therapeutic agent useful in the development of immune-stimulatory compounds.

To evaluate the efficacy and safety of CofNovex plus (EMA) syrup.

The cough and cold are very widespread conditions and a common purpose for advice in general practice. Utmost often the sign and symptoms of cough are produced by acute viral airway infection and the course is frequently benign. But it can be converted into bacterial super-infection and can cause acute bronchitis. Herbal medicines are used to treat symptoms of the cough and cold, and among these medicines Ivy leaf is used to treat mucous discharge and irritation in throat due to the cough and cold. In addition to synthetic substances such as acetylcysteine, carbocisteins, ambroxol and bromhexine, herbal medicines contain saponins, which are used in these indications. Not just Ivy, but also the marshmallow and mustard seeds used for these indications. This clinical trial was conducted in 220 patients, in which 110 receive the CofNovex plus European Medicines Agency (EMA) syrup and 110 receive the placebo. The age range of patients was 3 years to above 15 years. The sample paired t-test was applied to evaluate the significant level. CofNovex plus (EMA) syrup was very effective in treating cough and cold symptoms. The new treatment CofNovex plus (EMA) syrup was safe and well tolerated in patient at given specific age group.

Eating marshmallows reduces ileostomy output: a randomized crossover trial.

AIM: Anecdotally, many ostomates believe that eating marshmallows can reduce ileostomy effluent. There is a plausible mechanism for this, as the gelatine contained in marshmallows may thicken small bowel fluid, but there is currently no evidence that this is effective. METHOD: This was a randomized crossover trial. Adult patients with well-established ileostomies were included. Ileostomy output was measured for 1 week during which three marshmallows were consumed three times daily, and for one control week where marshmallows were not eaten. There was a 2-day washout period. Patients were randomly allocated to whether the control or intervention week occurred first. In addition, a questionnaire was administered regarding patient's subjective experience of their ileostomy function. RESULTS: Thirty-one participants were recruited; 28 completed the study. There was a median reduction in ileostomy output volume of 75 ml per day during the study period (P = 0.0054, 95% confidence interval 23.4-678.3) compared with the control week. Twenty of 28 subjects (71%) experienced a reduction in their ileostomy output, two had no change and six reported an increase. During the study period, participants reported fewer ileostomy bag changes (median five per day vs six in the control period, P = 0.0255). Twenty of 28 (71%) reported that the ileostomy effluent was thicker during the study week (P = 0.023). Overall 19 (68%) participants stated they would use marshmallows in the future if they wanted to reduce or thicken their ileostomy output. CONCLUSION: Eating marshmallows leads to a small but statistically significant reduction in ileostomy output.

Comparison between three different saliva substitutes in patients with hyposalivation.

OBJECTIVE: The aim of the present study was to compare the efficiency of oral spray based on thermal spring water (Buccotherm®) versus commercial saliva substitute (Xeros®) and marshmallow root on the quality of life in patients with hyposalivation. MATERIALS AND METHODS: A total of 60 patients with unstimulated salivary flow rate <0.2 ml/min were randomized into three groups. In the first group, 30 patients were using Buccotherm®; in the second group, 15 patients were using Xeros®; and in the third group, 15 patients were using marshmallow root. Therapy lasted for 2 weeks; everyday, patients used one of the products four times a day. Quality of life was measured by the Croatian version of Oral Health Impact Profile 14 questionnaire, and visual analog scale was used to determine the intensity of dry mouth before and after therapy. Statistical analysis was performed by Wilcoxon signed-rank test and Kruskal-Wallis test. Standardized effect size was calculated for OHIP following treatment. RESULTS: Buccotherm® has shown the biggest effect on quality of life in patients with hyposalivation. Intensity of dry mouth was lower after the applied therapy whatever substitute patients used. CONCLUSIONS: We recommend the use of all three saliva substitutes for decreasing the intensity of dry mouth symptoms as well as improvement in the quality of life. CLINICAL RELEVANCE: Although all tested agents showed beneficial effect in alleviating hyposalivation symptoms, it seems that Buccotherm® was superior to Xeros® and marshmallow root.

[Study on composition, antibiotic activity and antioxidant activity of volatile oils from uyghur medicine Althaea rosea].

Althaea rosea is a type of mallow plant. Its dry flowers are one of common herb in Uyghur medicines and recorded to have several efficacies such as external application for detumescence, moistening lung and arresting cough, sweating and relieving asthma, diminishing swelling and promoting eruption, soothing the nerves and strengthening heart. However, there are only fewer studies on effective components of A. rosea and no literature about its volatile oil and pharmacological activity. In this paper, the volatile oil of A. rosea was obtained by using the chemical distillation and extraction method. The individual chemical components were separated from the volatile oil and identified by the Gas Chromatograph-Mass Spectrometer technique (GC-MS). The antioxidant activity against free radicals was detected by the'ultraviolet and visible spectrophotometer method. The antibiotic activity was detected by the filter paper diffusion method. The experimental results showed nearly 70 compounds in the volatile oil, with complex chemical components. With a low content, most of the compounds were aromatic and aliphatic compounds and their derivatives. A. rosea had a better antibiotic activity for common microorganisms, with a wide antibacterial spectrum. According to the results, the volatile oil of A. rosea will have a good application value in medicine, food and cosmetic industries, which provided a scientific basis for the development of natural A. rosea resources.

Unconventional myosin ID is expressed in myelinating oligodendrocytes.

Myelin is a dynamic multilamellar structure that ensheathes axons and is crucial for normal neuronal function. In the central nervous system (CNS), myelin is produced by oligodendrocytes that wrap many layers of plasma membrane around axons. The dynamic membrane trafficking system, which relies on motor proteins, is required for myelin formation and maintenance. Previously, we found that myosin ID (Myo1d), a class I myosin, is enriched in the rat CNS myelin fraction. Myo1d is an unconventional myosin and has been shown to be involved in membrane trafficking in the recycling pathway in an epithelial cell line. Western blotting revealed that Myo1d expression begins early in myelinogenesis and continues to increase into adulthood. The localization of Myo1d in CNS myelin has not been reported, and the function of Myo1d in vivo remains unknown. To demonstrate the expression of Myo1d in CNS myelin and to begin to explore the function of Myo1d in myelination, we produced a new antibody against Myo1d that has a high titer and specificity for rat Myo1d. By using this antibody, we demonstrated that Myo1d is expressed in rat CNS myelin and is especially abundant in abaxonal and adaxonal regions (the outer and inner cytoplasm-containing loops, respectively), but that expression is low in peripheral nervous system myelin. In culture, Myo1d was expressed in mature rat oligodendrocytes. Furthermore, an increase in expression of Myo1d during maturation of CNS white matter (cerebellum and corpus callosum) was demonstrated by histological analysis. These results suggest that Myo1d may be involved in the formation and/or maintenance of CNS myelin.

Beverage and culture. "Zhourat", a multivariate analysis of the globalization of a herbal tea from the Middle East.

The "Zhourat" herbal tea consists of a blend of wild flowers, herbs, leaves and fruits and is a typical beverage of Lebanon and Syria. We aim to evaluate cultural significance of "Zhourat", to determine cultural standards for its formulation including key ingredients and to determine acceptable variability levels in terms of number of ingredients and their relative proportions, in summary what is "Zhourat" and what is not "Zhourat" from an ethnobotanical perspective. For this purpose we develop a novel methodology to describe and analyse patterns of variation of traditional multi-ingredient herbal formulations, beverages and teas and to identify key ingredients, which are characteristics of a particular culture and region and to interpret health claims for the mixture. Factor analysis and hierarchical clustering techniques were used to display similarities between samples whereas salience index was used to determine the main ingredients which could help to distinguish a standard traditional blend from a global market-addressed formulation. The study revealed 77 main ingredients belonging to 71 different species of vascular plants. In spite of the "Zhourat's" highly variable content, the salience analysis resulted in a determined set of key botanical components including Rosa x damascena Herrm., Althaea damascena Mouterde, Matricaria chamomilla L., Aloysia citrodora Palau, Zea mays L. and Elaeagnus angustifolia L. The major health claims for "Zhourat" as digestive, sedative and for respiratory problems are culturally coherent with the analysis of the traditional medicinal properties uses of its ingredients.

Quality control of herbs: determination of amino acids in Althaea officinalis, Matricaria chamomilla and Taraxacum officinale.

Analysis of raw materials and final products need reliable methods for the standardization of natural product drugs. Legal guideline also emphasizes on the qualitative and quantitative analyses of the plant constituents in an herbal product. In this study, thin layer chromatography (TLC) and amino acid analyzer was used for the determination of amino acids in plant extracts. Samples for this study were standards and aqueous extracts from Althaea officinalis, Matricaria chamomilla and Taraxacum officinale. Different amino acids in the extracts were detected through TLC. An automatic amino acid analyzer was used for the quantification of amino acids in the plant extracts under study.

The comprehensive strategy in the human health risk assessment of total chromium impurities in cough syrups with Marshmallow Root (Althaea officinalis) available in Polish pharmacies: regulatory aspects and special emphasis on Cr(VI) mode of action.

Chromium, which can currently only be considered pharmacologically active and not an essential element, is a very intriguing elemental impurity in final pharmaceutical products, especially traditional herbal medicinal products. This kind of traditional herbal medicinal product with Marshmallow root (Althaea officinalis L., radix) registered in the EU is widely used among the European population. The aim of this article is to propose a double regulatory strategy in assessing the human health risk of total chromium impurities in cough syrups with Marshmallow Root (Althaea officinalis) available in Polish pharmacies. We applied the strategy based on the requirements of the ICH Q3D (R1) guideline for the assessment of Cr impurities in final traditional herbal medicinal products with Marshmallow Root registered in the EU. Furthermore, we applied the strategy based on the concept of margin of exposure (MoE) considering Cr(VI) genotoxicity mode of action (MOA) and based on BMD10 for Cr(VI) as a point of departure (PoD). The total Cr content was in the range: 1.12-9.61 µg/L (in comparison with the ICH Q3D R1 guidelines: 1100 µg/g). Total Cr levels in a single dose were relatively high compared to raw results, but were not a threat to patients. Comparison of estimated results with oral PDE value for Cr in final drugs suggested by the ICH Q3D R1 guideline (10,700 µg/day) show that all the products analyzed were below this value (the highest result was 278.40 ng/day). Despite conservative assumptions, the MoE values obtained for Cr in daily dose for each Marshmallow Root cough syrup were above 10,000; therefore, exposure to Cr would not cause a health risk for specific population groups (children and adults). It can be summarized that each of the phytopharmaceuticals analysed with Marshmallow root available in Polish pharmacies does not represent a health hazard to patients. We confirm the safety of Cr impurities by applying a double regulatory strategy without the application of an expansive and demanding HPLC-ICP-MS technique for Cr speciation.