Arbutin and hidroquinone cuantification in cowberry, bearberry and pear leaves

Universitatea de Medicină şi Farmacie „Carol Davila“, Facultatea de Farmacie, Bucureşti, RomâniaObiective. Pornind de la utilizarea tradiţională a frunzelor de păr în tratamentul infecţiilor urinare, lucrarea prezintă cercetările efectuate în scopul identificării dar şi determinării conţinutului de heterozide fenolice de tip C1 (arbutozidă) comparativ cu două surse vegetale (Vitis idaea folium şi Uvae ursi folium) întrebuinţate în terapeutică şi citate de literatură cu un conţinut apreciabil în aceşti derivaţi. Material şi metode. Materialul utilizat pentru cercetare a fost reprezentat de frunzele de păr (Pyrus communis folium, recoltate la maturitate din zona Olteniei), merişor (Vitis idaeae folium, achiziţionate sub formă de ceai monocomponent din reţeaua unităţilor farmaceutice din România) şi strugurii ursului (Uvae ursi folium, achiziţionat sub formă de ceai monocomponent dintr-o unitate farmaceutică din Serbia). Metoda folosită a fost analiza fitochimică urmărindu- se identificarea constituenţilor chimici activi în soluţiile extractive eterice, alcoolice şi apoase, identificarea hidrochinonei prin microsublimare şi determinarea conţinutului în arbutozidă şi hidrochinonă printr-o metodă volumetrică. Rezultate. Din punct de vedere chimic calitativ s-au identificat steroli, agliconi flavonici, saponozide, flavone, acizi polifenolici, poliholozide mixte de tip mucilag, taninuri, proantociani, intensitatea reacţiilor fiind dependentă de natura materiilor prime vegetale. Prin microsublimare s-a evidenţiat prezenţa cristalelor aciculare de hidrochinonă în cele trei materii prime vegetale; în prezenţa clorurii de fer (III), cristalele capătă un aspect penat. Cantitativ frunzele de strugurii ursului conţin 5,44% arbutozidă şi 2,44% hidrochinonă, cele de merişor 3,44% arbutozidă şi 1,39% hidrochinonă, iar cele de păr 1,22% arbutozidă şi 0,832% hidrochinonă. Concluzii. Pe baza cercetărilor efectuate se poate considera că frunzele recoltate de la specia indigenă Pyrus communis pot fi întrebuinţate singure sau asociate cu alte produse vegetale în tratamentul infecţiilor urinare

Gentianae radix – from tradition to modern phytotherapy

Universitatea de Medicină şi Farmacie „Carol Davila“, Facultatea de Farmacie, Bucureşti, RomâniaObiective: În flora României sunt cunoscute 14 specii ale genului Gentiana, răspândite în zonele montane. O serie de preparate pe bază de rădăcină de ghinţură (Gentianae radix) sunt recomandate în fitoterapie, sau utilizate în mod tradiţional, în anorexie, gastrite hipoacide, disfuncţie biliară, helmintiaze. Lucrarea de faţă îşi propune analiza unei materii vegetale colectată din Munţii Iezer-Păpuşa în vederea stabilirii identităţii acesteia, precum şi caracterizarea fitochimică a 2 extracte vegetale – macerat apos, macerat hidroalcolic (forme farmaceutice de administrare folosite în mod tradiţional în zona Muscel), pentru verificarea corectitudinii posologiei. Material şi metode: Materia primă vegetală studiată a constituit-o o probă de organ subteran colectată din flora spontană (Munţii Iezer-Păpuşa), presupusă a fi rădăcină de ghinţură. Determinarea identităţii s-a realizat pe baza datelor din literatura de specialitate şi a rezultatelor următoarelor teste farmacognostice: examen macroscopic (stabilirea caracterelor morfologice şi organoleptice); examen microscopic (evidenţierea caracterelor anatomice, pe preparate clarificate cu soluţie de cloral hidrat 800 g/L); examen chimic calitativ (identificarea unor principii active, prin reacţii chimice specifice efectuate în soluţii extractive eterică, alcoolică şi apoasă); microsublimare (identificarea compuşilor xantonici, pe baza reacţiei cu hidroxid de sodiu); analiză cromatografică (stabilirea amprentei cromatografice CSS, folosind condiţiile experimentale descrise în monografia ,,Gentianae radix “ din Farmacopeea Europeană ediţia a 7-a); indice de amăreală (stabilirea organoleptică a gradului de amăreală). Extractele vegetale s-au obţinut prin macerare în apă, respectiv în alcool 32o, timp de 14 zile, conform unor reţete tradiţionale aplicate de localnicii din zona Muscel (judeţul Argeş). Acestea au fost caracterizate chimic calitativ (identificarea principiilor active prin reacţii specifice) şi cantitativ (determinarea extractivului, folosind metoda descrisă în Farmacopeea Europeană ediţia a 7-a, capitolul 2.8.16.). Ca referinţe (pentru comparare) s-au folosit probe de ,,Ceai de ghinţură (Gentianae radix)” şi ,,Tinctură de ghinţură”, procurate din reţeaua de distribuţie farmaceutică. Rezultate: Caracterele morfologice şi anatomice, principiile active identificate şi amprenta cromatografică CSS au dovedit că proba analizată este Genţianae radix, provenită de la Gentiana asclepiadea. Comparativ cu referinţa, proba analizată are un conţinut mai mare de principii active (intensitate mai mare a reacţiilor de identificare, gust amar superior). În cele 3 soluţii extractive (macerat apos, macerat hidroalcolic, tinctură de referinţă) s-au identificat oze şi alţi compuşi reducători, flavone, tanin, acizi polifenolcarboxilici şi suplimentar, mucilagii în maceratul apos. S-au determinat următoarele valori pentru totalul de substanţe solubile (extractiv): maceratul apos – 0,49%; maceratul hidroalcoolic – 1,01%; tinctură – 4,48%. Concluzii: Proba analizată este Gentianae radix, provenită de la Gentiana asclepiadea şi este superioară calitativ probei ,,Ceai de ghinţură (Gentianae radix)”, procurată din reţeaua de distribuţie farmaceutică. Pe baza conţinutului de total substanţe extrase, considerăm că se poate administra o lingură macerat hidroalcolic sau 2 linguri macerat apos (obţinute după reţeta tradiţională)

Traditional Medicinal Plants—A Possible Source of Antibacterial Activity on Respiratory Diseases Induced by Chlamydia pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae and Moraxella catarrhalis

Background. Nowadays, phytotherapy offers viable solutions in managing respiratory infections, disorders known for considerable incidence in both children and adults. In a context in which more and more people are turning to phytotherapy, finding new remedies is a topical goal of researchers in health and related fields. This paper aims to identify those traditional medicinal plants that show potentially antibacterial effects against four Gram-negative germs (Chlamydia pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae, and Moraxella catarrhalis), which are considered to have high involvement in respiratory infections. Furthermore, a comparison with Romanian folk medicines was performed. Methods. An extensive review of books and databases was undertaken to identify vegetal species of interest in the context of the topic. Results. Some traditional Romanian species (such as Mentha × piperita, Thymus vulgaris, Pinus sylvestris, Allium sativum, Allium cepa, Ocimum basilicum, and Lavandulaangustifolia) were identified and compared with the plants and preparations confirmed as having antibacterial effects against specific germs. Conclusions. The antibacterial effects of some traditionally used Romanian medicinal plants are poorly investigated, and deserve further attention

Dinamica acumulării compuşilor lipofili în frunzele de Urtica dioica L.

University of Medicine and Pharmacy „Carol Davila“, Faculty of Pharmacy, Bucharest, RomaniaObjective: phytochemical characterization of Urtica dioica L. leaves and the selection of the raw materials with the highest content of sterols and carotenoids, in order to obtain pharmacologically active extracts. Material and methods. The leaves were harvested for a period of 7 months (March – September) from Racari Village, Dambovita County, Romania, in 2010. The batches were codified as B1 (March), B2 (April), B3 (May), B4 (June), B5 (July), B6 (August), B7 (September). In order to confirm the identity of the leaves, macroscopic, microscopic and qualitative exams were undertaken. The sterols (free and glycosidic sterols) were assessed spectrophotometrically, based on the formation of dehydration products with multiple conjugated double bonds in the presence of concentrated sulfuric acid and ferric chloride (catalyst). The carotenoids content were determined, before and after their release from esteric forms, by assessing the absorbance at λ = 460 nm. Results. The morphological and anatomical aspects concur with scientific literature. The leaves are thin and complete, with a triangular and pubescent lamina (L=0.5 to 10 cm, l=0.3 to 5 cm).The margin is serate and the apex is acuminate. The upper surface is dark green and the lower surface is light green. In all batches, the microscopic exam shows the following specific anatomical elements: subulate hairs and stinging hairs, secretory trichomes with bicellular heads and unicellular stalks, cluster-crystals of calcium oxalate. The active principles identified in the qualitative examination are: sterols, carotenoids, coumarines, flavonoids (except batch B4), polyphenolcarboxilic acids, tannins, proanthocyanidins, mucilages. The carotenoids determination indicates that the highest content of free and esteric forms are found in leaves harvested in June and July, respectively. Glycosidic sterols are prevailing. The leaves content of glycosidic sterols is increases with maturity of the plant. The highest content of free sterols was determined in samples harvested in May. Conclusions. The leaves of Urtica dioca L. represent a complex source of active principles. The selected batch was B3. The selection of the batch was based on the total amount of carotenoids and sterols

Traditional Medicinal Plants—A Possible Source of Antibacterial Activity on Respiratory Diseases Induced by <i>Chlamydia pneumoniae</i>, <i>Haemophilus influenzae</i>, <i>Klebsiella pneumoniae</i> and <i>Moraxella catarrhalis</i>

Background. Nowadays, phytotherapy offers viable solutions in managing respiratory infections, disorders known for considerable incidence in both children and adults. In a context in which more and more people are turning to phytotherapy, finding new remedies is a topical goal of researchers in health and related fields. This paper aims to identify those traditional medicinal plants that show potentially antibacterial effects against four Gram-negative germs (Chlamydia pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae, and Moraxella catarrhalis), which are considered to have high involvement in respiratory infections. Furthermore, a comparison with Romanian folk medicines was performed. Methods. An extensive review of books and databases was undertaken to identify vegetal species of interest in the context of the topic. Results. Some traditional Romanian species (such as Mentha × piperita, Thymus vulgaris, Pinus sylvestris, Allium sativum, Allium cepa, Ocimum basilicum, and Lavandulaangustifolia) were identified and compared with the plants and preparations confirmed as having antibacterial effects against specific germs. Conclusions. The antibacterial effects of some traditionally used Romanian medicinal plants are poorly investigated, and deserve further attention

Wound Healing and Anti-Inflammatory Effects of a Newly Developed Ointment Containing Jujube Leaves Extract

Ziziphus jujuba Mill. (jujube) is a well-known medicinal plant with pronounced wound healing properties. The present study aimed to establish the chemical composition of the lyophilized ethanolic extract from Romanian Ziziphus jujuba leaves and to evaluate the healing and anti-inflammatory properties of a newly developed lipophilic ointment containing 10% dried jujube leaves extract. The ultra-High-Performance Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry method was used, and 47 compounds were detected, among them the novel epicatechin and caffeic acid. The extract contains significant amounts of rutin (29.836 mg/g), quercetin (15.180 mg/g) and chlorogenic acid (350.96 µg/g). The lipophilic ointment has a slightly tolerable pH, between 5.41–5.42, and proved to be non-toxic in acute dermal irritation tests on New Zealand albino rabbits and after repeated administration on Wistar rats. The ointment also has a healing activity comparable to Cicatrizin (a pharmaceutical marketed product) on Wistar rats and a moderate anti-inflammatory action compared to the control group, but statistically insignificant compared to indomethacin in the rat-induced inflammation test by intraplantar administration of kaolin. The healing and anti-inflammatory properties of the tested ointment are due to phenolic acids and flavonoids content, less because of minor components as apocynin, scopoletin, and isofraxidin

The Polyphenolic Profile and Antioxidant Activity of Five Vegetal Extracts with Hepatoprotective Potential

Oxidative stress is among the major triggers for many important human functional disorders, which often lead to various metabolic or tissue diseases. The aim of the study is to obtain five standardized vegetal extracts (Cynarae extractum&mdash;CE, Rosmarini extractum&mdash;RE, Taraxaci extractum&mdash;TE, Cichorii extractum&mdash;CHE, and Agrimoniae extractum&mdash;AE) that contain active principles with an essential role in protecting liver cells against free radicals and quantify their antioxidant actions. The compounds of therapeutic interest from the analyzed extracts were identified and quantified using the UHPLC&ndash;HRMS/MS technique. Thus, the resulting identified compounds were 28 compounds in CE, 48 compounds in RE, 39 compounds in TE, 43 compounds in CHE, and 31 compounds in AE. These compounds belong to the class of flavonoids, isoflavones, phenolic acids and dicarboxylic acids, depsides, diterpenes, triterpenes, sesquiterpenes, proanthocyanidins, or coumarin derivatives. From the major polyphenolic compounds quantified in all the extracts analyzed by UHPLC&ndash;HRMS/MS, considerable amounts have been found for chlorogenic acid (619.8 &micro;g/g extract for TE&ndash;2032.4 &micro;g/g extract for AE), rutoside (105.1 &micro;g/g extract for RE&ndash;1724.7 &micro;g/g extract for AE), kaempferol (243 &micro;g/g extract for CHE&ndash;2028.4 &micro;g/g extract for CE), and for naringenin (383 &micro;g/g extract for CHE&ndash;1375.8 &micro;g/g extract for AE). The quantitative chemical analysis showed the highest content of total phenolic acids for AE (24.1528 &plusmn; 1.1936 g chlorogenic acid/100 g dry extract), the highest concentration of flavones for RE (6.0847 &plusmn; 0.3025 g rutoside/100 g dry extract), and the richest extract in total polyphenols with 31.7017 &plusmn; 1.2211 g tannic acid equivalent/100 g dry extract for AE. Several methods (DPPH, ABTS, and FRAP) have been used to determine the in vitro total antioxidant activity of the extracts to evaluate their free radical scavenging ability, influenced by the identified compounds. As a result, the correlation between the content of the polyphenolic compounds and the antioxidant effect of the extracts has been demonstrated. Statistically significant differences were found when comparing the antiradical capacity within the study groups. Although all the analyzed extracts showed good IC50 values, which may explain their antihepatotoxic effects, the highest antioxidant activity was obtained for Agrimoniae extractum (IC50ABTS = 0.0147 mg/mL) and the lowest antioxidant activity was obtained for Cynarae extractum (IC50ABTS = 0.1588 mg/mL). Furthermore, the hepatoprotective potential was evaluated in silico by predicting the interactions between the determined phytochemicals and key molecular targets relevant to liver disease pathophysiology. Finally, the evaluation of the pharmacognostic and phytochemical properties of the studied extracts validates their use as adjuvants in phytotherapy, as they reduce oxidative stress and toxin accumulation and thus exert a hepatoprotective effect at the cellular level

The Influence of Phytosociological Cultivation and Fertilization on Polyphenolic Content of Menthae and Melissae folium and Evaluation of Antioxidant Properties through In Vitro and In Silico Methods

Since medicinal plants are widely used in treating various diseases, phytoconstituents enrichment strategies are of high interest for plant growers. First of all, we investigated the impact of phytosociological cultivation on polyphenolic content (total flavonoids&mdash;TFL, and total polyphenols&mdash;TPC) of peppermint (Mentha piperita L.) and lemon balm (Melissa officinalis L.) leaves, using spectrophotometric methods. Secondly, the influence of chemical (NPK) and organic (BIO) fertilization on polyphenolic content and plant material quality was also assessed. Dry extracts were obtained from harvested leaves using hydroethanolic extraction solvents for further qualitative and quantitative assessment of phytoconstituents by FT-ICR MS and UHPLC-MS. Furthermore, the antioxidant activity of leaf extracts was determined in vitro using DPPH, ABTS and FRAP methods. Molecular docking simulations were employed to further evaluate the antioxidant potential of obtained extracts, predicting the interactions of identified phytochemicals with sirtuins. The concentration of polyphenols was higher in the plant material harvested from the phytosociological culture. Moreover, the use of BIO fertilizer led to the biosynthesis of a higher content of polyphenols. Higher amounts of phytochemicals, such as caffeic acid, were determined in extracts obtained from phytosociological crops. The antioxidant activity was dependent on polyphenols concentration, more potent inhibition values being observed for the extracts obtained from the phytosociological batches. Molecular docking studies and MM/PBSA calculations revealed that the obtained extracts have the potential to directly activate sirtuins 1, 5 and 6 through several polyphenolic compounds, such as rosmarinic acid, thus complementing the free radical scavenging activity with the potential stimulation of endogenous antioxidant defense mechanisms. In conclusion, growing medicinal plants in phytosociological cultures treated with biofertilizers can have a positive impact on plant material quality, concentration in active constituents and biological activity