Detection of pyrrolizidine alkaloids in German licensed herbal medicinal teas

BackgroundBecause of the hepatotoxic, mutagenic, and cancerogenic effects of pyrrolizidine alkaloids (PAs) the German Federal Institute for Risk Assessment (BfR) recommends not to exceed a daily PA intake of 0.007 µg/kg body weight (0.42 µg/60 kg adult). In a recent study conducted by the BfR, up to 5647 µg PA/kg dried herbal material were detected in tea products marketed as food.PurposeThe present study aimed at elucidating whether medicinal teas licensed or registered as medicinal products contain PAs as well.Study designOne hundred sixty-nine different commercially available medicinal teas, i.e. 19 nettle (Urtica dioica L.), 12 fennel (Foeniculum vulgare Mill.), 14 chamomile (Matricaria recutita L.), 11 melissa (Melissa officinalis L.) and 4 peppermint (Mentha piperita L.) teas as well as 109 tea mixtures were analyzed for the presence of 23 commercially available PAs.MethodLC/MS was used for the determination of the PAsResultsIn general, the total PA contents ranging 0–5668 µg/kg. Thirty percent of the tested single-ingredient tea products and 56.9% of the tested medicinal tea mixtures were found to contain PA concentrations above the limit of quantification (LOQ) of 10 µg/kg. In 11 medicinal teas PA contents >300 µg/kg dry herb were determined thus exceeding the recommended limit for PA intake by BfR. In addition three products of the investigated tea mixtures revealed extremely high PA contents of 4227, 5137, and 5668 µg/kg. Generally, single-ingredient tea products contained much less or even no detectable amounts of PAs when compared to the tea mixtures. PAs in the range between 13 and 1080 µg/kg were also detected in five analyzed aqueous herbal infusions of the medicinal tea mixture products with the highest PA content. Two out of the five investigated herbal infusions exceeded the recommended BfR limit for PA intake.ConclusionThis study demonstrates clearly that also medicinal teas licensed as medicinal products may partly contain high amounts of PAs exceeding current recommendations. For that reason manufacturers are advised to carry out more rigorous quality control tests devoted to the detection of PAs. This is very important to minimize PAs in medicinal teas accounting for possible additional exposure of the consumer to PAs from other food sources (e.g. honey).     

An investigation on the antimutagenic properties of South African herbal teas

The antimutagenic properties of South African herbal teas were investigated using the Salmonella typhimurium mutagenicity assay. Aqueous extracts of fermented and unfermented rooibos tea (Aspalathus linearis) and honeybush tea (Cyclopia intermedia) both possess antimutagenic activity against 2-acetylaminofluorene (2-AAF) and aflatoxin B(1) (AFB(1))-induced mutagenesis using tester strains TA98 and TA100 in the presence of metabolic activation. A far less inhibitory effect was noticed against the direct acting mutagens, methyl methanesulfonate (MMS), cumolhydroperoxide (CHP), and hydrogen peroxide (H(2)O(2)) using TA102, a strain designed to detect oxidative mutagens and carcinogens. Depending on the mutagen used, the unfermented tea exhibited the highest protective effect. A similar response regarding the protection against mutagenesis was obtained when utilising different variations of the double layer Salmonella assay. The double layer technique proved to be more effective to detect the protective effect of the different tea preparations against the direct acting mutagens. With respect to indirect mutagens, the highest protection was noticed when the carcinogen was metabolically activated in the presence of the tea extract as compared with when the tea extract was incubated in a separate layer with the bacteria. The current data suggest that two mechanisms seem to be involved in the antimutagenicity of the tea extracts towards carcinogens that require metabolic activation: (i) the tea components may interfere with cytochrome P450-mediated metabolism of these mutagens and (ii) the direct interaction between the tea constituents, presumably the polyphenolic compounds, with the promutagens and/or the active mutagenic metabolites. However, the mild and/or lack of protection and in some cases even enhancement of mutagenesis induced by direct acting or oxidative mutagens, provide new perspectives regarding the role of the polyphenolic compounds known to exhibit antioxidant properties, in the protection against mutagenesis in the Salmonella assay. The present study provides the first evidence on the antimutagenic activity of honeybush tea and further evidence on the antimutagenicity of rooibos tea.     

Urinary concentrations of morphine after the administration of herbal teas containing Papaveris fructus in relation to doping analysis

A quantitative method for the analysis of morphine in human urine in the concentration range between 0.25 and 2 microg/ml is described and validated. Morphine was determined after enzymatic hydrolysis of the urine. After liquid-liquid extraction with dichloromethane-methanol (9:1) at pH 9.5, morphine was derivatized with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) and analyzed with GC-MS (full scan). The limit of quantification of the method was 0.25 microg/ml. Two Papaveris fructus containing herbal teas were administered to five male volunteers and urine samples were taken quantitatively during the first 12 h after the administration. The morphine concentration in the tea was 10.4 and 31.5 microg/ml, respectively. Morphine was detected in the urine of all volunteers by 1 h after drinking the tea. Maximum morphine concentrations, 4.3 and 7.4 microg/ml, respectively, were obtained 4-6 h after administration. Doping positive urine samples were delivered for 1-9 h.     

Comparative analysis of the volatile fraction of fruit juice from different Citrus species

The volatile composition of fruit from four Citrus varieties (Powell Navel orange, Clemenules mandarine, and Fortune mandarine and Chandler pummelo) covering four different species has been studied. Over one hundred compounds were profiled after HS-SPME-GC-MS analysis, including 27 esters, 23 aldehydes, 21 alcohols, 13 monoterpene hydrocarbons, 10 ketones, 5 sesquiterpene hydrocarbons, 4 monoterpene cyclic ethers, 4 furans, and 2 aromatic hydrocarbons, which were all confirmed with standards. The differences in the volatile profile among juices of these varieties were essentially quantitative and only a few compounds were found exclusively in a single variety, mainly in Chandler. The volatile profile however was able to differentiate all four varieties and revealed complex interactions between them including the participation in the same biosynthetic pathway. Some compounds (6 esters, 2 ketones, 1 furan and 2 aromatic hydrocarbons) had never been reported earlier in Citrus juices. This volatile profiling platform for Citrus juice by HS-SPME-GC-MS and the interrelationship detected among the volatiles can be used as a roadmap for future breeding or biotechnological applications.     

Characterization of volatile fraction of typical Irpinian wines fermented with a new starter yeast

Non-Saccharomyces yeasts are microorganisms that play an important role in the fermentation dynamics, compositions and flavour of wine. The aromatic compounds responsible for varietal aroma in wine are mainly terpenes, of which the most important group are the monoterpenes because of their volatility and odour if present in a free form. In fact, some terpenyl-glycosides do not contribute to the aroma unless they are hydrolysed. The glycosylated form of terpenes can be converted by hydrolysis with β-glycosidases produced by yeasts during the winemaking process, into aromatic compounds. In this study we utilized a non-Saccharomyces yeast, with a high extra-cellular glycosidase activity, isolated from grapes of cultivars typical of Irpinia region. This strain, identified as a Rhodotorula mucillaginosa (strain WLR12), was used to carry out an experimental winemaking process and the results were compared with those obtained with a commercial yeast starter. Chemical and sensorial analysis demonstrated that the wines produced with WLR12 strain had a more floral aroma and some sweet and ripened fruit notes compared to those obtained with commercial yeast. The data also showed an increasing of the free terpenes fraction that, however, did not significatively modify the bouquet of the wines.     

The volatile microbiome

The first detailed temporal study of the human microbiome shows that individual body habitats exhibit surprising variation over time yet maintain distinguishable community structures.     

不同发酵茶中优势真菌的分离鉴定及产消化酶活性比较

目的 分离鉴定云南普洱茶、广州小青柑、广西六堡茶和湖南熙茯茶中的主要真菌,并探讨其产消化酶活性。方法 采用平板涂布法分离不同发酵茶中的真菌,通过其ITS序列进行分子鉴定;利用平板透明圈法测定不同真菌的产淀粉酶、蛋白酶、纤维素酶和脂肪酶活性。结果 从4种发酵茶中共分离出8种真菌,其中,在云南普洱茶中分离出2种真菌,分别为篮状菌（Talaromyces. sp.）和T. variabilis;广州小青柑中分离出3种真菌,分别为阿曲霉（Aspergillus amstelodami）、烟曲霉（A. fumigatus）和绳状篮状菌（T. funiculosus）;广西六堡茶中分离出2种真菌,分别为黑曲霉（A. niger）和冠突曲霉（A. cristatus）;湖南熙茯茶中仅分离出谢瓦曲霉（A.chevalieri）。其中Talaromyces. sp.、T. variabilis、A. niger、A.amstelodami、A. fumigatus、T. funiculosus有产纤维素酶活性,Talaromyces. sp.、T. variabilis、A. cristatus、A. fumigatus、T. funiculosus有产淀粉酶活性,Talaromyces. sp.、T. variabilis、A. niger、A. fumigatus、T. funiculosus有产蛋白酶活性,A. niger有产脂肪酶活性。在8种真菌中,T. variabilis、烟曲霉和绳状篮状菌可同时产纤维素酶、淀粉酶和蛋白酶;黑曲霉能同时产纤维素酶、蛋白酶和脂肪酶;冠突曲霉仅有产淀粉酶活性;阿曲霉仅有产纤维素酶活性。通过比较不同真菌的产酶活性表明,烟曲霉的产蛋白酶活性能力最强;Talaromyces sp.的产纤维酶活性能力最强;Talaromyces. sp.和绳状篮状菌的产淀粉酶活性最强。结论 发酵茶的真菌以曲霉菌属Aspergillus、散囊菌属Eurotium和篮状菌属Talaromyces为主,多数真菌具有产消化酶活性,这些酶可能在茶叶发酵过程中起重要作用,并影响发酵茶叶的品质。     

Separation of catechin compounds from different teas

Catechin compounds from Korean and Chinese green tea, and pu-erh, Indian black, Longjing, Tieguanyin, Bamboo, Jasmine, Oolong, Flower, Red teas, as potential anticancer and antioxidant components, were target material in this work. After extracting the green tea with water at 50 degrees C for 4 h, the extract was partitioned with water/chloroform, which was best suited to remove caffeine impurity from the extract. Further, the resulting extract was partitioned with water/ethyl acetate to deeply purify the five catechin compounds epigallocatechin, (+) catechin, epicatechin, epigallocatechin gallate and epicatechin gallate. The extracted samples were analyzed by reversed-phase high performance liquid chromatography. The mobile phase applied was the binary system of A (water/acetic acid, 100/0.1 vol%) and B (acetonitrile/acetic acid 100/0.1 vol%) from 90:10 to 70:30 (A:B vol%) in a linear gradient over 30 min time. The amount of catechin compounds extracted from Chinese green tea was 114.65% higher than from the Korean green tea. Comparing various tea sorts, the green teas contained more than 1.7 times of the five catechin compounds contained in other teas.     

Chemical composition of Crepis foetida L. and C. rubra L. volatile constituents and evaluation of the in vitro anti-inflammatory activity of salicylaldehyde rich volatile fraction

The genus Crepis L. (Asteraceae) comprises more than 200 currently recognized species. Several studies have been conducted on non-volatile phytochemicals of Crepis spp., featuring mainly sesquiterpene lactones and phenolic derivatives. Nevertheless, no report has been made on assessing the volatile constituents of the genus. Therefore, the present study is the first report to the chemical composition of the volatile constituents of two odoriferous Crepis spp., namely C. foetida L. and C. rubra L. Flowers and stems with leaves volatiles were analysed separately by gas chromatography coupled with mass spectrometry. In total, 37 volatile compounds were detected. Salicylaldehyde, carvacrol and aliphatic hydrocarbons are the main components of both C. foetida fractions, however C. rubra flowers are characterized by the abundance of β-sitosterol and eicosanoic acid, while the stems-leaves volatiles revealed to be more complex with hydrocarbons as main constituents.In addition, we investigated the salicylaldehyde rich volatile fraction for its in vitro activity on TNF-α induced ICAM-1 expression.     

The excellence of turnip mitochondrial fraction

Several mitochondrial fractions were screened for suitability in practical experiments designed for students, with the following issues in mind: avoiding the use of animals; minimal expenditure of labour and time; high enzyme activities; accessible instruments and low-cost materials. Turnips and potato tubers were identified as the best materials from which to extract purified mitochondria according to these criteria, with high respiratory activities and integrity maintained during five consecutive days. Mitochondrial respiration was assayed for succinate, exogenous NADH, malate/pyruvate and alpha-ketoglutarate oxidations with excellent results. At the fifth day, the respiratory control was still about 3, the integrity of the outer mitochondrial membrane maintained at values higher than 80%, and the enzymes retained more than 55% of the initial activities.     

光肩星天牛幼虫虫粪挥发物成分分析

天牛幼虫排出的虫粪所释放的挥发性物质成为天敌搜寻天牛所在微栖境的重要化学信息物质.本试验采用固相微萃取技术,提取光肩星天牛Anoplophora glabripennis(Motschulsky)取食垂柳和金丝柳后产生虫粪的挥发性物质,利用气相色谱-质谱联用仪(GC-MS)进行鉴定.结果表明,虫粪挥发性物质主要为萜烯类物质.两种虫粪挥发物中含共有物质19种,其中à-蒎烯的含量最高,该物质在垂柳虫粪挥发物中相对含量为51.60%,在金丝柳虫粪中的相对含量为28.84%.在金丝柳虫粪中检测到的挥发物种类稍多于垂柳虫粪中检测到的种类.     

An herbal preparation ameliorates heart failure with preserved ejection fraction by alleviating microvascular endothelial inflammation and activating NO-cGMP-PKG pathway

BackgroundHeart failure with preserved ejection fraction (HFpEF) is a heterogeneous disease presenting a substantial challenge to clinicians. Currently, there is no safe and efficacious HFpEF treatment. In this study, we reported a standardized herbal medicinal product, QiShenYiQi (QSYQ), that can be used in the treatment of HFpEF.MethodsHFpEF mice were established by infusing a combination of N^ω-nitro-L-arginine methyl ester (L-NAME) and feeding them a high-fat diet for 14 weeks. In the 10th week, the HFpEF mice were given dapagliflozin or QSYQ via oral gavage for four weeks. The blood pressure, echocardiography, hemodynamics, leukocyte infiltration, and oxidative stress in HFpEF mice were evaluated. Besides, inflammatory factors, endothelial adhesion factors, and endothelial-mesenchymal transformation (EndMT) markers were investigated.ResultsQSYQ significantly attenuated concentric cardiac remodeling while improving diastolic function and left ventricular compliance in HFpEF mice. QSYQ also inhibited inflammation and immunocyte recruitment during HFpEF. The infiltration of CD8⁺, CD4⁺ T cells, and CD11b/c⁺ monocytes was substantially mitigated in the myocardium of QSYQ-treated mice. TNF-α, MCP-1, NF-κB, and NLRP3 levels also reduced after QSYQ treatment. Furthermore, QSYQ significantly reversed the elevated expression of endothelial adhesion factors and EndMT occurrence. These effects of QSYQ were demonstrated by the activation of NO-cGMP-PKG pathway and reduction of eNOS uncoupling in the HFpEF heart.ConclusionThese results provide novel evidence that QSYQ treatment improves HFpEF by inhibiting microvascular endothelial inflammation and activating NO-cGMP-PKG pathway.     

7种广西产甜茶抗氧化活性的比较

通过清除二苯代苦味酰基(1,1-diphenyl-2-picrylhydrazyl,DPPH)法、铁离子还原测定(ferric reducing antioxidant power,FRAP)法、亚铁离子螯合(ferrous ion-chelating,FIC)法、β-胡萝卜素-亚油酸法等4种体外抗氧化活性测定方法,首次比较了3种茶科属茶叶和7种广西产甜茶水提物的抗氧化活性,同时用Folin-Ciocalteus法测定了其总多酚的含量。研究结果表明,多穗柯甜茶和藤茶的总多酚含量高于其他供试品。藤茶具有很好的清除DPPH·的能力,高于绿茶及阳性对照水溶性维生素E(Trolox)和人工合成抗氧化剂二丁基羟基甲苯(butylated hydroxytoluene,BHT)。甜叶菊茶和悬钩甜茶的金属离子螯合能力均高于绿茶。其中,黄杞和牛白藤两种植物的抗氧化活性为首次报道。由此可见,广西产甜茶均具有较高的开发利用价值,是治疗和防治慢性代谢性疾病的重要研究对象。对广西产甜茶的研究对于了解和抢救某些少数民族的历史文化具有重要意义,为别样茶的抗氧化研究奠定了基础,值得进一步探索和开发。