Survey on the Quality of the Top-Selling European and American Botanical Dietary Supplements Containing Boswellic Acids

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The Ishraq Program for out-of-school girls: From pilot to scale-up

In response to the needs of adolescent girls in Egypt for improved schooling, health, and livelihood prospects, Ishraq (sunrise)—a multidimensional program for 12–15-year-old out-of-school girls—was launched in 2001 by the Population Council in collaboration with CEDPA, Save the Children, Caritas, and local nongovernmental organizations. The program combined traditional tested program elements (literacy, life skills, nutrition) with more innovative ones (sports, financial education). It focused on building a multilayered platform to support and institutionalize the program by educating and mobilizing communities around issues of importance to adolescent girls; forging partnerships between international NGOs, government institutions, and local NGOs; and building capacities of local facilitators and partners to implement the program. This report identifies ways to continue expanding Ishraq, for which community demand remains high, including a new generation of girls’ programs to support graduates in exercising their rights and becoming active members of the community; and notes many important lessons on recruitment strategy, program structure and implementation, capacity-building, and partnerships

The Ishraq Program for out-of-school girls: From pilot to scale-up [Arabic]

In response to the needs of adolescent girls in Egypt for improved schooling, health, and livelihood prospects, Ishraq (sunrise)—a multidimensional program for 12–15-year-old out-of-school girls—was launched in 2001 by the Population Council in collaboration with CEDPA, Save the Children, Caritas, and local nongovernmental organizations. The program combined traditional tested program elements (literacy, life skills, nutrition) with more innovative ones (sports, financial education). It focused on building a multilayered platform to support and institutionalize the program by educating and mobilizing communities around issues of importance to adolescent girls; forging partnerships between international NGOs, government institutions, and local NGOs; and building capacities of local facilitators and partners to implement the program. This report identifies ways to continue expanding Ishraq, for which community demand remains high, including a new generation of girls’ programs to support graduates in exercising their rights and becoming active members of the community; and notes many important lessons on recruitment strategy, program structure and implementation, capacity-building, and partnerships

Reporting guidelines for medicinal plant extracts used in pharmacological and toxicological research: ConPhyMP

Every year, the number of studies that evaluate the pharmacological effects, (clinical) efficacy or the toxicity of medicinal plant extracts is constantly increasing, but the reporting quality remains unsatisfactory. One of the main reasons is that there is a lack of detailed reporting standards for guidance. In response to this long-standing challenge, a core group of nine experts with proficiency in phytochemical analysis, including editors-in-chief of leading specialist journals, and based in different research settings globally, developed the Consensus based reporting guidelines for Phytochemical Characterisation of Medicinal Plant extracts (ConPhyMP) through a multi-staged development process. This incorporated a) a global survey among medicinal plant researchers, b) a core group, who reviewed and developed the guidelines through a Delphi process, and c) an advisory group of 20 experts, including editors of leading journals and scientific societies in medicinal plants research, who provided feedback and sanctioned the final guidelines. The ConPhyMP guidelines comprise two tables with accompanying explanatory figures. The first table provides recommendations for reporting the starting material and its initial processing, and the second table presents recommendations for conducting and reporting the analytical methods for defining the chemical profile based on the type of extracts used in the research. The group hopes that the ConPhyMP will support authors as well as peer reviewers and editors assessing these studies for publication and assist the production of evidence-based guidance of studies utilising medicinal plant extracts

Best practice in the chemical characterisation of extracts used in pharmacological and toxicological researcht - the ConPhyMP-guidelines

BACKGROUND : Research on medicinal plants and extracts derived from them differs from studies performed with single compounds. Extracts obtained from plants, algae, fungi, lichens or animals pose some unique challenges: they are multicomponent mixtures of active, partially active and inactive substances, and the activity is often not exerted on a single target. Their composition varies depending on the method of preparation and the plant materials used. This complexity and variability impact the reproducibility and interpretation of pharmacological, toxicological and clinical research. OBJECTIVES : This project develops best practice guidelines to ensure reproducibility and accurate interpretations of studies using medicinal plant extracts. The focus is on herbal extracts used in pharmacological, toxicological, and clinical/intervention research. Specifically, the consensus-based statement focuses on defining requirements for: 1) Describing the plant material/herbal substances, herbal extracts and herbal medicinal products used in these studies, and 2) Conducting and reporting the phytochemical analysis of the plant extracts used in these studies in a reproducible and transparent way. THE PROCESS AND METHODS : We developed the guidelines through the following process: 1) The distinction between the three main types of extracts (extract types A, B, and C), initially conceptualised by the lead author (MH), led the development of the project as such; 2) A survey among researchers of medicinal plants to gather global perspectives, opportunities, and overarching challenges faced in characterising medicinal plant extracts under different laboratory infrastructures. The survey responses were central to developing the guidelines and were reviewed by the core group; 3) A core group of 9 experts met monthly to develop the guidelines through a Delphi process; and. 4) The final draft guidelines, endorsed by the core group, were also distributed for feedback and approval to an extended advisory group of 20 experts, including many journal editors. OUTCOME : The primary outcome is the “Consensus statement on the Phytochemical Characterisation of Medicinal Plant extracts“ (ConPhyMP) which defines the best practice for reporting the starting plant materials and the chemical methods recommended for defining the chemical compositions of the plant extracts used in such studies. The checklist is intended to be an orientation for authors in medicinal plant research as well as peer reviewers and editors assessing such research for publication.Willmar Schwabe GmbH & Co. KG, Germany.https://www.frontiersin.org/journals/pharmacologydm2022Paraclinical Science

Tunable polymeric mixed micellar nanoassemblies of Lutrol F127/Gelucire 44/14 for oral delivery of praziquantel: a promising nanovector against hymenolepis nana in experimentally-infected rats

Hymenolepiasis represents a parasitic infection of common prevalence in pediatrics with intimidating impacts, particularly amongst immunocompromised patients. The present work aimed to snowball the curative outcomes of the current mainstay of hymenolepiasis chemotherapy, praziquantel (PRZ), through assembly of polymeric mixed micelles (PMMs). Such innovative nano-cargo could consolidate PRZ hydrosolubility, extend its circulation time and eventually upraise its bioavailability, thus accomplishing a nanoparadigm for hymenolepiasis tackling at lower dose levels. For consummating this goal, PRZ-PMMs were tailored via thin-film hydration technique integrating a binary system of Lutrol F127 and Gelucire 44/14. Box-Behnken design was planned for optimizing the nanoformulation variables employing Design-Expert® software. Also, in Hymenolepis nana-infected rats, the pharmacodynamics of the optimal micellar formulation versus the analogous crude PRZ suspension were scrutinized on the 1st and 3rd days after administration of a single oral dose (12.5 or 25 mg/kg). Moreover, in vitro ovicidal activity of the monitored formulations was estimated utilizing Fuchsin vital stain. Furthermore, the in vivo pharmacokinetics were assessed in rats. The optimum PRZ-PMMs disclosed conciliation between thermodynamic and kinetic stability, high entrapment efficiency (86.29%), spherical nanosized morphology (15.18 nm), and controlled-release characteristics over 24 h (78.22%). 1H NMR studies verified PRZ assimilation within the micellar core. Additionally, the in vivo results highlighted a significant boosted efficacy of PRZ-PMMs manifested by fecal eggs output and worm burden reduction, which was clearly evident at the lesser PRZ dose, besides a reversed effect for the intestinal histological disruptions. At 50 µg/mL, PRZ-PMMs increased the percent of non-viable eggs to 100% versus 47% for crude PRZ, whilst shell destruction and loss of embryo were only clear with the applied nano-cargo. Moreover, superior bioavailability by 3.43-fold with elongated residence time was measured for PRZ-PMMs compared to PRZ suspension. Practically, our results unravel the potential of PRZ-PMMs as an oral promising tolerable lower dose nanoplatform for more competent PRZ mass chemotherapy

Best Practice in the chemical characterisation of extracts used in pharmacological and toxicological research—The ConPhyMP—Guidelines

The Advisory group on Consensus statement on the Phytochemical Characterisation of Medicinal Plant extracts (ConPhyMP) is a consortium of experts on Pharmacognosy and Phytochemistry open access articleBackground: Research onmedicinal plants and extracts derived fromthem differs from studies performed with single compounds. Extracts obtained from plants, algae, fungi, lichens or animals pose some unique challenges: they are multicomponent mixtures of active, partially active and inactive substances, and the activity is often not exerted on a single target. Their composition varies depending on the method of preparation and the plant materials used. This complexity and variability impact the reproducibility and interpretation of pharmacological, toxicological and clinical research. Objectives: This project develops best practice guidelines to ensure reproducibility and accurate interpretations of studies using medicinal plant extracts. The focus is on herbal extracts used in pharmacological, toxicological, and clinical/intervention research. Specifically, the consensus-based statement focuses on defining requirements for: 1) Describing the plant material/herbal substances, herbal extracts and herbal medicinal products used in these studies, and 2) Conducting and reporting the phytochemical analysis of the plant extracts used in these studies in a reproducible and transparent way. The process and methods: We developed the guidelines through the following process: 1) The distinction between the three main types of extracts (extract types A, B, and C), initially conceptualised by the lead author (MH), led the development of the project as such; 2) A survey among researchers of medicinal plants to gather global perspectives, opportunities, and overarching challenges faced in characterising medicinal plant extracts under different laboratory infrastructures. The survey responses were central to developing the guidelines and were reviewed by the core group; 3) A core group of 9 experts met monthly to develop the guidelines through a Delphi process; and. 4) The final draft guidelines, endorsed by the core group, were also distributed for feedback and approval to an extended advisory group of 20 experts, including many journal editors. Outcome: The primary outcome is the “Consensus statement on the Phytochemical Characterisation of Medicinal Plant extracts“ (ConPhyMP) which defines the best practice for reporting the starting plant materials and the chemical methods recommended for defining the chemical compositions of the plant extracts used in such studies. The checklist is intended to be an orientation for authors in medicinal plant research as well as peer reviewers and editors assessing such research for publicatio

Considerations to be taken when carrying out medicinal plant research : what we learn from an insight into the IC50 values, bioavailability and clinical efficacy of exemplary anti-inflammatory herbal components

Medicinal plants represent a big reservoir for discovering new drugs against all kinds of diseases including inflammation. In spite the large number of promising anti-inflammatory plant extracts and isolated components, research on medicinal plants proves to be very difficult. Based on that background this review aims to provide a summarized insight into the hitherto known pharmacologically active concentrations, bioavailability, and clinical efficacy of boswellic acids, curcumin, quercetin and resveratrol. These examples have in common that the achieved plasma concentrations were found to be often far below the determined IC50 values in vitro. On the other hand demonstrated therapeutic effects suggest a necessity of rethinking our pharmacokinetic understanding. In this light this review discusses the value of plasma levels as pharmacokinetic surrogates in comparison to the more informative value of tissue concentrations. Furthermore the need for new methodological approaches is addressed like the application of combinatorial approaches for identifying and pharmacokinetic investigations of active multi-components. Also the physiological relevance of exemplary in vitro assays and absorption studies in cell-line based models is discussed. All these topics should be ideally considered to avoid inaccurate predictions for the efficacy of herbal components in vivo and to unlock the “black box” of herbal mixtures

Bioanalytik von Ginsenosiden und Boswelliasäuren

Heute gewinnen pflanzliche Arzneimittel im Zeichen einer verstärkten Hinwendung zu natürlichen, relativ nebenwirkungsarmen Medikamenten zunehmend an Bedeutung, so auch die über Jahrtausende hinweg traditionell angewandte Ginsengwurzel (Panax ginseng) und der Indische Weihrauch (Boswellia serrata). Neben der Struktur- und Extraktanalytik konzentriert sich die Forschung in zunehmenden Maße darauf, die zahlreichen pharmakologisch untersuchten und klinisch beobachteten Wirkungen dieser beiden Arzneipflanzen einzelnen Inhaltsstoffen zuzuordnen. Bei der Ginsengwurzel gestaltet sich dies jedoch besonders schwierig. Dies ist begründet durch die große Anzahl strukturell ähnlicher Ginsenoside sowie durch deren unterschiedlicher Metabolismus. Zwar ist aus In-vitro-Experimenten bekannt, daß die Degradation über die stufenweise Deglukosylierung stattfindet, allerdings ist bislang nicht geklärt, ob intakte Ginsenoside überhaupt resorbiert werden und welche der vielen in vitro ermittelten Degradationsprodukte tatsächlich den systemischen Kreislauf erreichen. Anders als bei Ginseng, kann die therapeutische Wirkung des Indischen Weihrauches wohl definierten Inhaltsstoffen, AKBA und KBA, zugeschrieben werden. Dennoch mangelt es hier an verlässlichen pharmakokinetischen Daten, da bislang keine validierte bioanalytische Methode zur Verfügung stand. Im Rahmen der Bioanalytik von Ginsenosiden und Boswellliasäuren kamen in der vorliegenden Arbeit die Nano-ESI-MS(n)-Technik sowie die HPLC-Analytik zur Anwendung. Bereits bei der Strukturanalyse von Ginsenosiden erwies sich die Kombination der Nano-ESI-Technik mit MS(n)-Experimenten in einer Quadrupol-Ionenfalle als besonders vorteilhaft. Im Vergleich zu konventionellem ESI bietet Nano-ESI nicht nur den Vorzug, mit kleinsten Substanzmengen lange Messungen durchführen zu können, sondern auch den Vorteil einer effektiveren, weniger diskreminierenden Ionisation. Sowohl die hohe Sensitivität der Nano-Elektrosprayionisierung bei der Analyse von glykosidischen Verbindungen als auch die umfangreichen Strukturinformationen infolge mehrerer aufeinanderfolgender stoßinduzierter Fragmentierungen machen diese Technik zu einer attraktiven und effizienten Methode zur Analyse von Ginsenosiden. In MS(n)-Experimenten äußert sich das charakteristische Fragmentierungsverhalten der Ginsenoside in der sequentiellen Abspaltung der Zuckereinheiten in sukkzessiven Fragmentierungsschritten. Mit dieser Methode konnten die Zuckerketten an verschiedenen Positionen des Protopanaxadiol- bzw. Protopanaxatriolaglykons der Ginsenoside identifiziert, die glykosidischen Verknüpfungen innerhalb der einzelnen Zuckereinheiten durch spezifische Ringfragmente bestimmt und die genaue(n) Verknüpfungsposition(en) enzymatisch eingeführter Galaktose(n) lokalisiert werden. Bisher wurden allerdings Nano-ESI-MS/MS bzw. MS(n)-Untersuchungen primär an wässrigen oder organischen Lösungen von isolierten / aufgereinigten Stoffen oder Stoffgemischen durchgeführt. In dieser Arbeit wurde erstmals diese Technik in der Bioanalytik zur Identifizierung von Ginsenosiden und deren Degradationsprodukte in Humanplasma und -urin angewandt. Obwohl die optimale Analytkonzentration für die Nano-Elektrosprayionisierung im allgemeinen bei 10-5M liegt, ist es gelungen, die Ginsenoside anhand ihrer spezifischen Fragmentionen im MS/MS Modus bis zu einer Konzentration von 2 ng/mL (10-8M) in biologischen Matrices nachzuweisen. Auch wenn die Molekülionenpeaks bei diesen geringen Konzentrationen im Rauschen untergehen, können die Ginsenoside durch selektive Isolierung der gewünschten Vorläuferionen und deren anschließende Fragmentierung in der Quadrupol-Ionenfalle auf der Basis der Detektion spezifischer Fragmente identifiziert werden. Da bei der Fragmentierung der gleichen Vorläuferionenmasse in Leerplasma bzw. Urin keine charakteristischen Fragmentionen gebildet werden, handelt es sich somit um einen spezifischen Nachweis der Ginsenoside in biologischen Matrices. Vor dem Hintergrund dieser vielversprechenden Vorversuche wurde eine Pilotstudie zum qualitativen Screening von Ginsenosiden und deren Metaboliten in Humanplasma und –urin durchgeführt. Es konnte gezeigt werden, daß Protopanaxatriol-Ginsenoside sowohl im Magen hydrolysiert als auch intestinal degradiert werden. Schon in den ersten Stunden nach der einmaligen oralen Einnahme von Ginsana G115 Kapseln wurden die Hydrolyseprodukte G-Rh1 und hydratisiertes G-Rh1 in Humanplasma detektiert. Die schnelle Resorption dieser beiden Verbindungen aus dem oberen Gastrointestinaltrakt deutet auf die Hydrolyse des Ginsenosides Rg1 im Magen hin. Das spätere Auftreten eines weiteren monoglukosylierten Protopanaxatriols, des Degradationsproduktes G-F1, liefert den ersten In-Vivo-Hinweis auf einen intestinalen Metabolismus von Protopanaxatriol-Ginsenoside. Im Gegensatz zu den Protopanaxatriolginsenosiden passieren die Protopanaxadiol-Ginsenoside den Magen unverändert, wie aus der Abwesenheit jeglicher Protopanaxadiol-Degradationsprodukte im Plasma und Urin in den ersten Stunden nach der Applikation geschlossen werden kann. Erst im unteren Gastrointestinaltrakt werden Protopanaxadiol-Ginsenoside durch intestinale Bakterien zu „Compound-K“ abgebaut und anschließend resorbiert. Der Nachweis von Ginsenosid Rb1 im Plasma, sowie weiterer Ginsenoside im Urin eines Probanden zeigt, daß auch Ginsenoside in ihrer intakten Form resorbiert werden können. Dennoch sind weitere Studien hierzu notwendig, um zu klären, ob die Resorption intakter Ginsenoside die Regel oder eher eine Ausnahme darstellt. Mit der Identifizierung des Hydrolyseproduktes G-Rh1, der Degradationsprodukte G-F1 sowie „Compound-K“ in Humanplasma und -urin konnte schließlich die Frage geklärt werden, welche der zahlreichen in vitro bestimmten Degradationsprodukte tatsächlich den systemischen Kreislauf erreichen. Damit ist die Basis für eine spätere Quantifizierung dieser Verbindungen nach ihrer Isolierung bzw. Herstellung und Charakterisierung als Referenzsubstanzen geschaffen. Außerdem können diese neuen Erkenntnisse helfen, pharmakologische Ergebnisse aus In-vitro-Versuchen mit In-vivo-Daten besser zu korrelieren, da sie erste Hinweise geben, welche der in vitro getesteten Substanzen für die in vivo beobachteten Effekte verantwortlich sein könnten. Bisher wurde Nano-ESI-MS(n) in der Bioanalytik pflanzlicher Arzneistoffe nicht eingesetzt. In dieser Arbeit wurde diese Technik erstmals zum Screening von Ginsenosiden und deren Degradationsprodukte in Humanplasma und –urin benutzt. Darüber hinaus konnte gezeigt werden, daß Nano-ESI-MS(n) auch eine empfindliche und sensitive Methode zur Identifizierung und zum Nachweis anderer medizinisch relevanter glykosidischer Verbindungen in biologischen Matrices darstellt, woraus sich neue Perspektiven für den Einsatz dieser Technik in der Metabolitenforschung sowie in der Bioanalytik pflanzlicher Xenobiotika ergeben. In den letzten Jahren rückte auch der Indische Weihrauch immer mehr in den Mittelpunkt des wissenschaftlichen sowie therapeutischen Interesses. Aufgrund der selektiven Hemmung der 5-Lipoxygenase gewinnen AKBA und KBA, als neue entzündungshemmende Verbindungen, zunehmend an Bedeutung. Während die quantitative Analytik der Boswelliasäuren in Extrakten und in verschiedenen Fertigarzneimitteln erhebliche Fortschritte erzielen konnte, fehlten auf dem Gebiet der Bioanalytik bislang validierte analytische Methoden zur Durchführung pharmakokinetischer Studien. Vor diesem Hintergrund wurde eine HPLC-Methode zur Bestimmung von KBA in Humanplasma entwickelt und validiert. Die Methode ist durch eine einfache Probenvorbereitung gekennzeichnet, die sich auf eine Festphasenextraktion der KBA aus der komplex zusammengesetzten biologischen Matrix beschränkt. Im Anschluß an die chromatographische Trennung auf einer RP-C18 Säule erfolgt die Quantifizierung der KBA mittels UV-Detektion bei 250 nm. Um eine adäquate Bestimmung der KBA in Humanplasma zu gewährleisten, wurde eine umfassende Validierung durchgeführt. Alle ermittelten Validierungsparameter, wie Spezifität, Linearität, Präzision, Richtigkeit, Reproduzierbarkeit, untere Quantifizierungsgrenze und Stabilität lagen innerhalb der vorgegebenen Grenzen. Damit erfüllt die entwickelte HPLC-Methode die allgemein gültigen Anforderungen an die Validierung bioanalytischer Methoden. Mit der Erstellung einer Plasmakonzentrations-Zeit-Kurve im Rahmen einer ersten Pilotstudie konnte diese Methode zudem ihre praktische Anwendbarkeit unter Beweis stellen. Somit steht für zukünftige pharmakokinetische Studien eine validierte HPLC-Analytik zur Bestimmung von KBA, einer der Hauptwirkstoffe des Indischen Weihrauches, zur Verfügung, die sich durch hohe Spezifität, Reproduzierbarkeit und Präzision auszeichnet. Verlässliche Daten zur Pharmakokinetik am Menschen sind heute von besonderer Bedeutung, da sie helfen, die Dosierung zu optimieren, die biopharmazeutischen Eigenschaften von Präparaten zu verbessern und die Sicherheit bei der Anwendung zu erhöhen. Insbesondere im Hinblick auf bereits festgestellte Interaktionen von pflanzlichen Arzneimitteln mit Medikamenten reicht es nicht mehr aus, wenn sich pflanzliche Arzneimittel, wie beispielsweise Ginseng und Indischer Weihrauch, nur auf ihre langjährigen tradierten Anwendungserfahrungen berufen. So wird auch bei Phytopharmaka in verstärktem Maße eine wissenschaftliche Absicherung durch bioanalytische Forschung erwartet. Mit der Identifizierung der Ginsenoside und deren Degradationsprodukte im Menschen, der erstmaligen Anwendung von Nano-ESI-MS(n) in der Metabolitenforschung und der Entwicklung einer validierten bioanalytischen Methode zur Bestimmung der 11-Keto-Beta-Boswelliasäure in Humanplasma wurde diesen Erfordernissen Rechnung getragen.In terms of the widespread use of herbal remedies Panax ginseng and Boswellia serrata are gaining more and more scientific and therapeutic interest. Inspite of recent advances in the structural elucidation and analysis of their active ingredients in extracts as well as in pharmaceutical preparations bioanalytical studies are still sparse. So even though the metabolism of ginseng has been thoroughly studied in animals and in vitro using acids, enzymes and intestinal bacteria, knowledge concerning the metabolic fate of ginsenosides, considered to be the active ingredients of Panax ginseng, in humans is generally lacking. Thus it remains to be clarified if any of the various protopanaxatriol and protopanaxadiol ginsenosides can be absorbed from the human GIT and which of the previously in vitro determined metabolites reach the systemic circulation. Knowing about these active components is actually of great importance to understanding the clinical effects, preventing herb-drug interactions and optimizing the biopharmaceutical properties of ginseng preparations. Therefore the attention with regard to ginseng is focused on its metabolic pathway in humans. For the purpose of screening human plasma and urine for the presence of any ginsenosides and possible metabolites nano-ESI-MS(n), a sensetive and specific mass spectrometric method, was used. Nano-ESI in combination with collision induced dissociation (CID) in an ion trap has already revealed to be an efficient and rapid method for the structural analysis of ginsenosides. Due to the sequential elimination of both sugar moieties ginsenosides show a characteristic fragmentation pattern in MS(n) experiments. The high sensitivity of nano-ESI and the capability of carrying out several consecutive fragmentations in the ion trap allowed the identification of the carbohydrate moieties at different positions of the protopanaxadiol and protopanaxatriol aglycone, the linkage position within the sugar moieties and the exact point of attachment of enzymatically introduced galactose(s) in ginsenoside derivatives. Although the optimum analyte concentration for nano-ESI is generally known to be a 10-5 molar solution, ginsenosides could be detected in biological matrices up to 2 ng per mL (10-8M) by means of their characteristic fragment ions in the MS/MS modus. Of course the molecular ions of the ginsenosides in such low concentrations are not distinguishable from the background noise anymore, nevertheless they can be identified by the isolation of the corresponding precursor ion followed by its fragmentation in the ion trap. Furthermore the identification of the ginsenosides on the basis of their characteristic fragmentation ions in the MS/MS modus offers a substantial specificity advantage, since fragmentation of the same precursor ion mass in blank plasma or urine doesn´t yield these fragmentation ions. Within the scope of a pilot study the metabolic pathway of ginseng could be elucidated in humans after the oral adminstration of Ginsana G115, a commercial ginseng preparation. Both plasma and urine data were in good agreement with each other. The rapid absorption of the monoglucosylated protopanaxatriol Rh1 and hydrated Rh1, both hydrolysis products of ginsenoside Rg1, in the first hours after the oral administration from the upper part of the digestive tract indicates that ginsenoside Rg1 has been decomposed in the stomach. The later appearance of another monoglucosylated protopanaxatriol, metabolite F1, represents an in vivo evidence, that intestinal metabolism of protopanaxatriol ginsenosides also takes place in humans. As can be concluded from the absence of any metabolites of the protopanaxadiol group in the first hours following drug administration the protopanaxadiol ginsenosides are hardly decomposed in the gastric juice. Instead, the prolonged time needed for the appearance of compound-K, the main metabolite of the protopanaxadiol ginsenosides, shows that their absorption takes place in the lower part of the intestine after being hydrolysed by intestinal bacteria. The detection of ginsenoside Rb1 indicates that ginsenosides can also be absorbed in their intact form. However further studies are necessary to clarify whether the absorption of intact ginsenosides is the rule or rather an exception. In summary it was proven by the above results that two metabolites of the protopanaxatriol group, namely Rh1 and F1, in addition to compound-K, the metabolite of the protopanaxadiol ginsenosides are absorbed in humans. This insight into the biotransformation of ginseng in humans gives an idea of the metabolites actually reaching the systemic circulation and thus facilitates further quantification studies. In addition it helps better correlating pharmacological results from in vitro experiments with in vivo data, since it provides hints, which of the in vitro tested substances may be responsible for the effects observed in vivo. Within the framework of this dissertation nano-ESI-MS(n) was first used in the bioanalysis of herbal remedies to identify ginsenosides and their metabolites in human plasma and urine. Moreover it has been shown that nano-ESI-MS(n) can also be applied as a sensitive method for the identification of other glycosidic substances of medicinal relevance in biological matrices, opening thus another field of application for this technique in the metabolic research of herbal remedies. Also Boswellia serrata became the subject of intensive research in the last decades. In contrary to most herbal remedies, where the active constituents are not yet known, the active principles of Boswellia serrata have been identified as the boswellic acids. Being selective inhibitors of the 5-Lipoxygenase pathway 3-O-acetyl-11-keto-beta-boswellic acid (AKBA) and 11-keto-beta-boswellic acid (KBA) are gaining more and more importance as novel antiinflammatory compounds. Inspite of their therapeutic importance no validated analytical method existed up till now for the quantitative determination of boswellic acids in human matrices. Therefore a specific reversed phase high-performance liquid chromatographic (HPLC) method has been developed and validated for the determination of KBA in human plasma. This method is characterized by a simple sample preparation procedure, which is restricted on the introduction of a single solid-phase extraction step prior to the HPLC analysis of KBA. The method has been shown to provide good sensitivity, accuracy and precision. A validation procedure was performed according to current guidelines for bioanalytical method validation. All results obtained during the validation fulfill the requirements and recommendations generally accepted for bioanalytical studies. Furthermore the clinical suitability of this method was verified by monitoring the plasma concentration of KBA in a preliminary pilot study. In conclusion this method is suitable for the determination of KBA in future pharmacokinetic studies. As these studies can help to evaluate the optimal dose and to improve biopharmaceutical properties they are very essential in order to employ Boswellia serrata under the best conditions of efficacy and safety. Espescially with respect of recently observed drug-herb-interactions it is not sufficient anymore if herbal remedies just refer to their long traditional use. Therefore the demand for scientific confirmation of efficacy and safety of herbal remedies in humans is constantly increasing. With the elucidation of the metabolic pathway of ginsenosides in humans using nano- ESI-MS(n) for the first time in the metabolic research and developing a validated bioanalytical method for the determination of 11-keto-beta-boswellic acid in human plasma first steps have been made in this direction

Buchu (Agathosma betulina and A. crenulata): rightfully forgotten or underutilized?

Today, the term buchu refers to the two species in commerce, Agathosma betulina (P.J.Bergius) Pillans and Agathosma crenulata (L.) Pillans (Rutaceae). Its traditional use in urinary tract infections and related ailments made it a popular remedy, specifically in the US, in 19th century, but with the advent of antibiotics it became largely obsolete. Recent focus is on technological use and on the essential oil for use in the perfume and food-flavouring industry. A review of the scarce pharmacological research revealed moderate antimicrobial activity for a leaf extract but not the essential oil of both species in the MIC assay. In the 5-lipoxygenase (5-LO) assay the essential oil of both species revealed IC50 values of 50.37 ± 1.87 μg/ml and 59.15 ± 7.44 μg/ml, respectively. In another study 98% inhibitory activity was determined for 250 μg/ml of an ethanolic extract of A. betulina on cyclooxygenase (COX)-1 and a 25% inhibitory activity on COX-2. Analgesic activity of an ethanolic extract of A. betulina was shown in mice. Moderate antioxidant activity was determined for methanol:dichlormethane extracts of A. betulina and A. crenulata and an aqueous extract of A. betulina showed a Trolox equivalent antioxidant capacity (TEAC) of 11.8 µM Trolox. Recent in vitro studies with a commercial aqueous extract of buchu revealed increased uptake of glucose added to 3T3-L1 cell line, significant inhibition of the respiratory burst of neutrophils and monocytes, reduction in the expression of adhesion molecules and inhibition of the release of IL-6 and TNF-α. In diabetic rats the ingestion of aqueous buchu extract completely normalized the glucose level and in rats receiving a high fat diet the consumption of aqueous buchu extract resulted in less weight gain and less intraperitoneal fat gain as well as reduction of elevated blood pressure to normal associated with cardioprotective effects. Limitations in the hitherto conducted research lie in the undisclosed composition of the buchu extracts used and the difficulty in extrapolating data from animal studies to humans. Health claims for buchu products need to be substantiated by randomized, double-blind and placebo-controlled studies. Only then can they be promoted for their true therapeutic potential