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Bisbibenzyls are specialized metabolites found exclusively in liverworts, until recently; they represent chemical markers of liverworts. Their occurrence in vascular plants was noticed in 2007, when they were found in Primula veris subsp. macrocalyx from Russia. This report prompted us to chemically analyze the two most common Serbian Primula species, P. veris subsp. columnae and P. acaulis, in order to determine the presence of bisbibenzyls in them. Our study revealed nine structurally distinct bisbibenzyls (1-9), identified based on 1D and 2D NMR, IR, UV and HRESIMS data. Among them were five previously undescribed compounds (2-6). The remaining compounds found and previously described in the literature were: the bisbibenzyls riccardin C (1), isoperrottetin A (7), isoplagiochin E (8) and 11-O-demethylmarchantin I (9), as well as 4-hydroxyphenylmethylketone (10) and 4-hydroxy-3-methoxyphenylmethylketone (11). Riccardin C was the most dominant bisbibenzyl in both species studied. Previously, it was the first bisbibenzyl found in vascular plants (P. veris subsp. macrocalyx). An assessment of the cytotoxic activity of the isolated compounds against A549 lung cancer and healthy MRC5 cell lines was also the subject of our study. Compounds 6 and 9 exhibited significant cytotoxic activity expressed by IC 50 values of 12 μM, but the selectivity was not satisfactory., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Herein n-, iso- and anteiso-series of very-long-chained (VLC) alkanes (C 21 -C 35 ), fatty acid benzyl esters (FABEs; C 20 -C 32 ), and 2-alkanones (C 23 -C 35 ) were identified in the wax of Primula veris L. and P. acaulis (L.) L. (Primulaceae). For the very first time in a sample of natural origin, the presence of iso- and anteiso-VLC FABEs and 2-alkanones was unequivocally confirmed by synthetic work, derivatization, and NMR. It should be noted that the studied species produced unusually high amounts of branched wax constituents (e. g., >50 % of 2-alkanones were branched isomers). The domination of iso-isomers, probably biosynthesized from leucine-derived starters, is a unique feature in the Plant Kingdom. The plant organ distribution of these VLC compounds in P. acaulis samples (different habitats and phenological phases) pointed to their possible ecological value. This was supported by a eutectic behavior of binary blends of FABEs and alkanes, as well as by high UV-C absorption by FABEs., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

Our experiments may help to answer the question of whether cowslip ( Primula veris L.) is a rich source of bioactive substances that can be obtained by efficient extraction with potential use as a food additive. A hypothesis assumed that the type of solvent used for plant extraction and the individual morphological parts of Primula veris L. used for the preparation of herbal extracts will have key impacts on the efficiency of the extraction of bioactive compounds, and thus, the health-promoting quality of plant concentrates produced. Most analysis of such polyphenolic compound contents in extracts from Primula veris L. has been performed by using chromatography methods such as ultra-performance reverse-phase liquid chromatography (UPLC-PDA-MS/MS). Experiments demonstrated that the most effective extraction agent for fresh study material was water at 100 °C, whereas for dried material it was 70% ethanol. The richest sources of polyphenolic compounds were found in cowslip primrose flowers and leaves. The aqueous and ethanol extracts from Primula veris L. were characterized by a quantitatively rich profile of polyphenolic substances, and a high antioxidative potential. Selective extraction with the use of mild conditions and neutral solvents is the first step to obtaining preparations from cowslip primrose with a high content of bioactive substances.

The present study encompasses green synthesis of silver nanoparticles (AgNPs) using aqueous leaf extract of Arabian Primrose within 6 min of reaction at 60 °C, pH 7 and their characterisation using physico-chemical analytical techniques. UV-Visible spectroscopy elucidated the surface plasmon resonance around 420 nm. FESEM and TEM images revealed that AgNPs were spherical with average diameter 10-60 nm. XRD pattern confirmed their crystalline nature. The leaf extract rich in phenolics and flavonoids was subjected to GC-MS analysis that identified bioactive compounds helping in reduction and stabilisation of AgNPs. The synthesised AgNPs possessed high anti-oxidant potential against DPPH and H 2 O 2 radicals. Incidentally, the AgNPs acted as excellent nanocatalyst towards borohydride reduction and degradation of structurally different organic dyes. The AgNPs also exhibited selective colorimetric sensing of hazardous mercuric, ferric ions and ammonia. Such AgNPs were cytotoxic against HeLa cells (IC 50 7.18 µg/mL) and compatible towards normal L 20 B cells. These AgNPs showed effective anti-microbial activity against different human pathogens tested (bacterial and fungal). This is probably the first report of AgNPs synthesis using Arabian Primrose leaf extract showing strong anti-oxidant, catalytic, biosensing, anti-cancer and anti-microbial activities and find remarkable applications in medical, industrial and ecological sectors.

Background: The immune response is influenced by the administration of omega-3 polyunsaturated fatty acids (PUFA). Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE) are affected by PUFA. The combination of evening primrose/hemp seed oil (EPO/HSO) has essential fatty acids (EFAs) for human optimal health due to the favorable ratio of omega-6/omega-3 and antioxidantal properties. The study was conducted to evaluate the effects of EPO/HSO on improving the membrane fatty acids composition of spleen and blood cells and immunologic factors in compared to rapamycin (RAPA) in the EAE model., Methods and Materials: Chronic-EAE was induced by induction of MOG in C57BL/6J mice (female, age: 6-8 weeks, weight 18-21). Mice were assigned to 5 groups (6/group) to evaluate the therapeutic effects of EPO/HSO supplement in comparison with rapamycin: A group; EPO/HSO + RAPA, B group; RAPA, C group; EPO/HSO. Results were compared to two control groups (EAE and naive). The fatty acid profile of the spleen and blood cell membrane was evaluated. Real-time-polymerase chain reaction was used for the evaluate the genes expression levels of interleukin (IL) -4, IL-5, and IL-13 in lymphocytes. Also, IL-4 of serum was evaluated by enzyme-linked immunosorbent assay (ELISA)., Results: Our findings indicated that EPO/HSO therapy significantly increased the percentage of essential fatty acids in cell membrane of the spleen and blood. The relative expression of IL-4, IL-5, and IL-13 genes in lymphocytes and serum level of IL-4 was significantly increased in the HSO/EPO treated group versus other groups., Conclusion: These results point to potential therapeutic effects on the repair of the structure of cell membranes and suppression of inflammation by EPO/HSO in EAE., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this paper was to analyze selected physicochemical properties and the pro-health potential of wines produced in southeastern Poland, in the Subcarpathian region, and commercial Carlo Rossi wines enhanced with cowslip ( Primula veris L.). This study used ultra-performance reverse-phase liquid chromatography (UPLC)-PDA-MS/MS to perform most of the analysis, including the polyphenolic compounds and saponin content in wines enriched by Primula veris L. The initial anthocyanin content in Subcarpathian (Regional) red wine samples increased four times to the level of 1956.85 mg/L after a 10% addition of Primula veris L. flowers. For white wines, a five-fold increase in flavonol content was found in Subcarpathian (Regional) and wine samples, and an almost 25-fold increase in flavonol content was found in Carlo Rossi (Commercial) wine samples at the lowest (2.5%) Primula veris L. flower addition. Qualitative analysis of the regional white wines with a 10% Primula veris L. flower enhancement demonstrated the highest kaempferol content (197.75 mg/L) and a high quercetin content (31.35 mg/L). Thanks to wine enrichment in triterpenoid saponins and in polyphenolic compounds from Primula veris L. flowers, which are effectively extracted to wine under mild conditions, both white and red wines can constitute a highly pro-health component of diets, which is valuable in preventing chronic heart failure.

A bioactivity guided program exploring the interaction of phytochemicals in the entire plant Primula macrocalyx with the organic anion transporters (OAT1 and OAT3) and microorganisms led to the elucidation of ten known flavones (1-4, 6-10, 12) and two previously undescribed flavones (5, 11). The structures of the compounds were determined by extensive analysis of spectroscopic data, as well as by comparison with data from previous reports. Two known flavones (9, 12) are reported for the first time from the family Primulaceae. All compounds were evaluated for inhibition of OAT1 and OAT3. Six flavones (2, 3, 6-8, 12) showed potent inhibitory activity on OAT1, while seven flavones (2, 3, 6-9, 12) showed marked inhibitory activity on OAT3, with IC 50  ≤ 10.0 µM. Antimicrobial activities of crude fractions against sixteen microorganisms were tested to give a target yeast strain Candida rugosa for further evaluation of MICs on the isolates. Three flavones (7, 8, 12) showed marked antifungal activity with MIC < 2.0 µM. To our knowledge, this study is the first to evaluate these flavones as inhibitors of the OAT1 and OAT3, and as antifungal agents.

Background: High interest in chronic heart failure (CHF) is accounted for by its high incidence, poor prognosis, growing number of hospital admissions due to the heart failure relapse, and inadequate treatment. These facts necessitate a search for new pharmacological agents for the CHF correction. Herbal medicinal products appear to be very promising as they have a noticeable therapeutic effect and tend to be more harmless in comparison to the most of synthesized medications., Purpose: Our aim was to study the composition of the Primula veris L. solid herbal extract (PVSHE) and its effects on the myocardial contractile function in animals with experimental CHF., Study Design: The study design involved the identification of the raw material composition of the P. veris L. extract. For the experimental part of our research, we used the model of CHF to elucidate the cardioprotective properties of PVSHE., Methods: The active extract constituents were isolated by thin-layer chromatography and column chromatography; the extract components were identified by high-performance liquid chromatography, ultraviolet spectroscopy (UVS), and nuclear magnetic resonance spectroscopy (NMRS). To model CHF, L-isoproterenol at a dose of 2.5 mg/kg was intraperitoneally injected to the experimental rats twice a day for 21 days. Cardiac output was assessed with the loading test, adrenoreactivity test, and maximum isometric loading test; CHF markers adrenomedullin and copeptin were detected in blood plasma with ELISA kit for adrenomedullin and copeptin (Coud-Clone Corp., USA)., Results: P. veris L. solid herbal extract contains flavonoid aglycons (apigenin, quercetine, kaemferol), flavonoid glycosides (cinarozid, rutin, hyperozid), as well as polymethoxylated flavonoids acting as chemotaxonomic markers for the genus Primula (8-methoxy-flavone; 3',4'methylenedioxy-5'-methoxyflavone). The substance 3',4'methylenedioxy-5'-methoxyflavone has been isolated from the primrose herb for the first time. We showed that the PVSHE has a cardioprotective effect when it was administered at a dose of 30 mg/kg in the experimental CHF, as evidenced by a lower number of animal death, lower level of CHF markers in the blood plasma of the experimental animals, the higher increase in rate of myocardial contraction and relaxation, the higher level of left ventricular pressure (LVP) and of maximum intensity of structural performance (MISP), as compared to the control group., Conclusion: P. veris L. solid herbal extract contains flavonoid aglycons, flavonoid glycosides, and polymethoxylated flavonoids. The herbal agent increases the myocardial contractility in experimental CHF., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Experimental chronic heart failure (CHF), caused by administration of L-isoproterenol (2.5 mg/kg twice a day intraperitoneally for 21 days), promotes uncoupling of respiration and oxidative phosphorylation. The rate of mitochondrial oxygen consumption in the metabolic state V3 by Chance in animals with CHF decreased by 53.3% (p<0.05) with malate using (as an oxidation substrate feeding сomplex I of the electron transport chain (ETC)), by 70.6% (p<0.05) with succinate using (сomplex II substrate) and by 63.6% (p<0.05) when malate and succinate were added simultaneously. The respiratory control ratio significantly decreased 2.3 times for сomplex I, 2.5 for сomplex II, and 2.6 times for the simultaneous operation of two respiratory chain complexes in mitochondria of CHF rats compared to intact animals. Mitochondrial dysfunction in experimental CHF is evidently due to the development of oxidative stress. It was revealed that the content of malonic dialdehyde (MDA) in the group of rats with experimental CHF was higher by 54.7% (p<0.05), as compared with intact animals. The activity of superoxide dismutase (SOD) and catalase was lower by 17.5% (p<0.05), and by 18.4%, respectively than in the intact group. The dense extract from herba of Primula veris L. (DEHPV) 30 mg/kg limits the development of mitochondrial dysfunction in rats with experimental CHF, as evidenced by an increase in the role of V3 respiration for the first and second respiratory chain complexes in 1.7 (p<0.05) and 2.0 times (p<0.05), respectively, the ratio of respiratory control (RCR) - 1.7 times (p<0.05) for сomplex I and 2 times (p<0.05) for сomplex II compared with the negative control. The concentration of MDA was by 15.7% (p<0.05), lower and the activity of SOD was by 56.3% (p<0.05) higher.

The genus Primula is the largest among the Primulaceae and is widespread mainly in the cold and temperate regions of the Northern Hemisphere. Since the beginning of the Twentieth century, several studies on the phytochemical composition of different species of Primula have been carried out. The main constituents examined were tissue and epicuticular flavonoids and saponins, which are of therapeutic significance. Only in recent years studies of the volatiles emitted by leaves and flowers have been carried out as well, but they are restricted to a small number of species. Only a few authors have documented the morphology and function of glandular trichomes in relation to the production of flavonoids and volatile organic compounds (VOCs). The use of Primula in folk medicine is described in the literature. Investigation of the biological and pharmacological activities of Primula are reported. This study aims at providing a collection of publications on the genus Primula along with a critical revision of literature data. It focuses on the possible taxonomic significance of the secondary metabolites and on their ecological role as attractors for pollinators and deterrents against herbivores and parasites, in order to build the base for further studies., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, the changes caused by variation of altitude to the essential oils (EOs), fatty acid methyl esters (FAMEs), and antimicrobial activities of Primula vulgaris Huds. subsp. vulgaris (Pvv) and P. vulgaris Huds. subsp. sibthorpii (Hoffmanns) W.W.Sm. & Forrest (Pvs)) grown in Turkey were investigated. Major fluctuations in the composition of Pvv and Pvs oils included methyl-4-methoxy salicylate (4.5-35.3%; Pvv and 3.2-37.2%; Pvs), (Z,Z,Z)-7,10,13- hexadecatrienal (5.1-21.8%; Pvv and 4.4-15.2%; Pvs ) and flavone (5.5-14.9%; Pvv and 1.6-18.0%; Pvs). Fatty acid profile (C60-CI6o) changes were noted in .Pvv and Pvs. Methyl hexadecanoate (2.4-9.3%) and methyl octadecanoate (1.0-4.7%) were present in all the FAME samples of the plants. The antimicrobial activity of the EOs of Pvv and Pvs were tested against nine bacterial species, which showed activity against Mycobacterium smegmatis with minimum inhibitory concentrations (MIC) varying from 8.5 to 59.2 pg/mL in all samples, respectively, depending on the altitude at which the oils were obtained.

In continuation of previous studies on glandular exudates of Primula, we analyzed eleven so far unstudied species and several populations for exudate composition. Unsubstituted flavone and unusually substituted flavones, normally predominant in Primula exudates, were not detected in all of the analyzed samples. Instead, some species exhibited regular substituted flavonoids, and in some cases, no flavonoids could be detected at all. The detection of a diterpene (1) in P. minima exudates is new to Primula. On basis of MS and NMR, 1 was structurally characterized as ent-kaur-16-en-19-oic acid. Comparative profiling of exudates as performed by HPLC and TLC against authentic markers indicated further the presence of the benzoquinone primin and derivatives in some exudates. Thus, exudates of newly studied species contrast markedly with those analyzed so far. The significance of observed exudate diversification is discussed in view of the phylogeny of derived lineages in European alpine regions.

Background: The application of alcohol-containing medicinal products to children has been a subject of discussion for many years. A safety threshold of 0.125‰ blood alcohol concentration following the administration of a single dose has been recommended by the European Medicines Agency.The aim of this clinical study was to prove the safety of administering a fixed combination of thyme herb and primrose root fluid extract (Bronchicum(®) Elixir) containing 4.9% ethanol., Methods: The herbal drug was administered for a period of 7-9 days to 16 children (ages 1-12 years) suffering from acute bronchitis for ≤ 48 h. After 3-5 days, a blood sample was taken 45 min (children ≥ 5 years: also 0 and 90 min) after application of the drug. The efficacy was assessed using the Bronchitis Severity Score. Global efficacy and tolerability were rated by the investigator and patients., Results: All measured blood ethanol concentrations were below the threshold (mean value after 45 min: 0.0029 ± 0.0057‰ and after 90 min: 0.0051 ± 0.0078‰). The Bronchitis Severity Score decreased from 6.6 ± 1.0 to 0.9 ± 1.6 points. Global efficacy was assessed as "very good" and "good" in 60% (investigator) and 80% (patients) of cases. Global tolerability was rated as "very good" and "good" in more than 90% of cases., Conclusion: In conclusion, oral administration of the drug containing 4.9% ethanol to children (age 1-12 years) demonstrated a favourable risk/benefit ratio of the drug., (© Georg Thieme Verlag KG Stuttgart · New York.)

Cough is one of the characteristic symptoms of upper respiratory tract infections (URI). Herbal medicines are often used in the treatment of the cough associated with infection and to accelerate recovery or support the immune system. An example of such products are extracts of thyme and primrose, and also their combination with thymol. Thymus vulgaris is a spasmolytic, antimicrobial, anti-inflammatory, immunomodulatory and antioxidant agent. The most important component responsible for the activity of thyme is thymol contained in the volatile thyme oil. Primrose is, most of all, a saponine agent with expectorant and secretolytic activity, showing also spasmolytic, anti-inflammatory and antimicrobial effects. Because of the additive effects of the mentioned extracts and their wide activity profile they are often used as a combination drug. The use of this combination was assessed in the URI with the effects of alleviation of cough and dyspnea, and shortening the length of the disease. The effectiveness of the drug containing the extracts of thyme and primrose with the addition of thymol (Bronchosol®) was comparable to synthetic ambroxol and its safety has been proved. Moreover, the in-vitro antibacterial and antifungal effects of this drug have been evaluated.

The aim of this study was to investigate the effect of different penetration enhancers, containing essential fatty acids (EFAs), on the transdermal delivery of flurbiprofen. Evening primrose oil (EPO), vitamin F, and Pheroid technology all contain fatty acids and were compared using a cream-based formulation. This selection was to ascertain whether EFAs solely, or EFAs in a Pheroid delivery system, would have a significant increase in the transdermal delivery of a compound. Membrane release studies were performed, and the results indicated the following rank order for flurbiprofen release from the different formulations: vitamin F >> control > EPO >> Pheroid. Topical skin delivery results indicated that flurbiprofen was present in the stratum corneum-epidermis and the epidermis-dermis. The average percentage flurbiprofen diffused to the receptor phase (representing human blood) indicated that the EPO formulation showed the highest average percentage diffused. The Pheroid formulation delivered the lowest concentration with a statistical significant difference (p < 0.05) compared with the control formulation (containing 1% flurbiprofen and no penetration enhancers). The control formulation presented the highest average flux, with the EPO formulation following the closest. It could, thus, be concluded that EPO is the most favorable chemical penetration enhancer when used in this formulation., (Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Three flavonol glycosides were isolated from the leaves of Primula sieboldii. They were identified as quercetin 3-O-β-[xylopyranosyl-(1-->2)-β- glucopyranosyl-(1-->6)-β-glucopyranoside] (1), kaempferol 3-O-β-[glucopyranosyl-(1-->2)-β-glucopyranosyl-(1-->6)-β-glucopyranoside] (2) and kaempferol 3- O-β-[xylopyranosyl-(1-->2)-β-glucopyranosyl-(1-->6)-β-glucopyranoside] (3). Their chemical structures were determined by UV, 1H and 13C NMR spectroscopy, LC-MS and acid hydrolysis. Compounds 1 and 3 are found in nature for the first time. They were also detected in the flowers, together with two anthocyanins, malvidin 3,5-di-O-glucoside and a minor petunidin dihexoside.

This work investigated the epicuticular and tissue flavonoids, the volatiles and the glandular trichome structure of the leaves of four species of Primula L. that grow in the Italian Eastern Alps. Primula albenensis Banfi and Ferlinghetti, P. auricula L., P. farinosa L., P. halleri Gmelin produce farinose exudates that are deposited on the leaf surface as filamentous crystalloids. In addition to compounds already known, a new flavone, the 3,5-dihydroxyflavone, was isolated from the acetone extract of leaf farinas and three new flavonol glycosides, 3'-O-(β-galactopyranosyl)-2'-hydroxyflavone, isorhamnetin 3-O-α-rhamnopyranosyl-(1→3)-O-[α-rhamnopyranosyl-(1→6)]-O-β-galactopyranoside, quercetin 3-O-α-rhamnopyranosyl-(1→3)-O-[α-rhamnopyranosyl-(1→6)]-O-β-galactopyranoside, were isolated from the MeOH extract of the leaves. All the structures were elucidated on the basis of their (1)H and (13)C NMR data and 2D NMR techniques, as well as on HPLC-MS. The leaf-volatiles emitted by these Primula species were mainly sesquiterpene hydrocarbons, with the exception of P. albenensis, which produced almost exclusively a non-terpene derivative; P. halleri flowers were also examined and the volatiles emitted by the flower parts (corolla and calyx) were compared with the corresponding leaves., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Three flavonoids were isolated from the leaf MeOH extracts of Primula latifolia Lapeyr. and Primula vulgaris Hudson collected from Italian Alps: rutin (1) and kaempferol 3-neohesperidoside (2) from P. latifolia, and kaempferol 3-β-O-glucopyranosyl-(1 → 2) gentiobioside (3) from P. vulgaris. The structures were assigned on the basis of their (1)H and (13)C NMR data, including those derived from 2D NMR, as well as on HPLC-MS results. This article is the first to report on P. vulgaris tissue flavonoids after Harborne's study in 1968 and the first work ever on these compounds from P. latifolia.

Two orphan flavonoids containing an oxepin structure in ring A and named as Primcortusin (1) and 3'-OH-Primcortusin (2) were isolated from leaf exudates of Primula cortusoides, while P. glutinosa exudates yielded two dihydrochalcone derivatives (3,4). These novel structures have not been detected in other species of Primula so far. Chemical structures were elucidated by 2D NMR and mas spectrometry. The nature of compounds 1 and 2 is discussed, and ideas about their possible origin and that of unsubstituted flavone and other irregular substituted Primula flavones are presented.

Different parts of three plants (Primula auriculata, Fumaria vaillantii and Falcaria vulgaris) were extracted with three different solvents to yield 72 crude extracts. The phytochemical analysis (chemical screening, GC-MS) of three plants was investigated for their antioxidant and antibacterial activity using nine Gram-positive and Gram-negative bacteria. The principal antioxidant and antimicrobial components were determined using HPLC with UV detection. All extracts possessed antibacterial activity especially methanolic extracts from flowers of P. auriculata. The DPPH-radical scavenging assay exhibited high antioxidant activities in three plants (more than 80% at 50μg). The F. vulgaris showed high content of carvacrol (29.8%) as main component. The contents of carvacrol and fumaric acid in the methanolic-water extracts were 1119 and 1966mg/l respectively. Our results indicate that these plants would be able to promise sources of natural products with potential antibacterial and antioxidant activity., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Accumulation profiles of fatty acids and α-tocopherol were analysed at three ripening stages in the seed oils of Primula and Echium species. Total seed oils were increased considerably with maturation, while α-tocopherol contents decreased in both species. Increased levels of ALA and oleic acid, and decrease in linoleic and palmitic acids at late ripening stages of Primula sibthorpii, and slightly fluctuations of all examined fatty acids in Echium italicum were observed. Considerable amounts of α-tocopherol (27.4 mg/100 g), linoleic acid (42.76%) and ALA (25.46%) and GLA (4.11%) were detected in Ribes alpinum. Typical accumulation patterns of the examined parameters may be useful for species characterisation and biochemical monitoring of seed development in natural conditions.

In recent years, considerable attention has been paid to plants as potent natural drugs for their ameliorative roles against free-radical-mediated oxidative stress. Therefore, their interactions with cell membrane lipids and proteins, which generally serve as primary targets of lipid peroxidation, are of much interest. In the current investigation, in vitro and ex vivo studies are performed in order to estimate possible effects of different extracts of Primula heterochroma Stapf. on red blood cell membranes of rat erythrocytes using colorimetric methods. The results indicate that binding of the extracts to lipids and proteins of red blood cell membranes both significantly inhibits lipid peroxidation, and also increases red blood cell integrity against hemolysis. Moreover, a polyphenol extract, in particular, demonstrates notable antihemolytic activity in hydrogen peroxide-induced hemolysis model (IC(50) = 199.49 ± 9.1 μg ml(-1)).

A novel HPTLC-densitometric method was developed for separation and quantitation of primulasaponin I and II in different matrices. HPTLC silica gel 60 F254(S), 20 cm × 10 cm, plates with ethyl acetate:water:formic acid (5:1:1 v/v) as the mobile phase were used. Densitometric determinations were performed in reflectance mode at 540 nm after derivatization with vanillin reagent. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Primulasaponins were quantified in the range of 150-450 ng with RSD of repeatability and intermediate precision between 0.8 and 1.4% and accuracy within the acceptance limits. The method was tested on commercial herbal medicinal preparations claiming to contain primula root extract., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A combined study was carried out on the micromorphology and chemistry of glandular trichomes with focus on Primula vialii and P. vulgaris, respectively. Epifluorescence microscopy was applied to study the auto-fluorescent properties of flavonoids and their localization in glandular trichomes. Both species differed in the morphology of the glandular trichomes and in the exudate flavonoid composition. Leaf glands from P. vialii and from some species of subgen. Primula exhibited uniform glandular fluorescence, but notable differentiation was observed within a single leaf of P. vulgaris. Our observations indicate that exudate flavonoids are not transported from the tissue to the glandular hairs. Conversely, only the newly isolated glycoside 1 (kaempferol 3-O-(2"'-rhamnosyl)-robinobioside) was obtained from leaf tissue of P. vialii after removal of the exudate. Its structure was confirmed by NMR and mass spectrometry. This glycoside was not detected in tissue extracts of P. vulgaris after similar treatment. The observed chemical diversity is discussed, with focus on possible correlation with glandular structures and tissue differentiation in Primula, and also against other studied species. Aspects of biosynthesis in relation to tissue-specific flavonoid diversification are shortly addressed.

Context: The genus Primula (Primulaceae) has been used in traditional medicine to treat convulsion and microbial or viral infections., Objective: In the present study, we evaluate antioxidant, antihemolytic, and protective effects of flavonoid-rich fractions of endemic Primula heterochroma Stapf. (Primulaceae) against Fe(2+)-induced lipid peroxidation and oxidative stress in rat brain in vitro., Materials and Methods: Aerial parts of plant were defatted and extracted with 60% acetone. Then, 60% acetone extract was fractionated sequentially with n-hexane, ethyl acetate, and water. Antioxidant activity of fractions was evaluated by employing six different assays, i.e., 1,1-diphenyl-2-picryl hydrazyl (DPPH) and hydrogen peroxide scavenging, metal chelating and reducing power activities and hemoglobin-induced linoleic acid system and Fe(2+)-induced lipid peroxidation and oxidative stress in rat brain. Also, its antihemolytic activity was determined using 2,2'-azobis(2-amidinopropane) dihydrochloride-induced hemolysis in rat erythrocyts., Results: Among the flavonoid-rich fractions of Primula heterochroma, aqueous fraction demonstrated the most protective effect through decreasing brain thiobarbituric acid reactive substances (TBARS) levels at a dose 200 μg mL(-1) (40%, p < 0.001 versus iron group). Also, the aqueous fraction showed better activity in Fe(2+) chelating (89 ± 3.8 μg mL(-1)) and DPPH radical scavenging (394.4 ± 18.4 μg mL(-1)) models than other fractions. The probable protective mechanism of flavonoid-rich fractions may be attributed to their Fe(2+) chelating, DPPH radical scavenging and reducing power activities. Also, the n-hexane fraction demonstrated a higher protective effect in the hemoglobin-induced linoleic acid system and 2,2'-azobis(2-amidinopropane) dihydrochloride-induced hemolysis (67 ± 2.6 μg mL(-1))., Discussion and Conclusion: Results of this study demonstrate Primula heterochroma is a rich source of natural antioxidant compounds.

In continuation of previous work, exudate flavonoid profiles of 22 new accessions of the genus Primula L. aligned to different subgenera were studied for the first time. Profiling was done by comparative TLC and UV-HPLC against authentic marker compounds. Most of the studied species accumulated the typical Primula flavonoids, comprising derivatives of unsubstituted flavone, of 5,8- or 5,6-hydroxyflavones including partly highly oxygenated flavones, together with 2,2'-diOH-chalcone in their exudate. The profile of P. edelbergii belonging to subgen. Sphondylia differed largely from the majority of Primula species studied so far, showing accumulation tendencies similar to those observed earlier for the closely related genus Dionysia. The phylogenetic significance of this diversification is shortly addressed.

Covering: 1995 to June 2011. The macrocyclic bisbibenzyl family of natural products are commonly found in liverworts and other bryophytes, though the recent isolation of riccardin C from a primrose extract has demonstrated their existence in higher flowering plants. Each has a core comprising four aromatic rings and two ethano bridges, being derived in Nature from two molecules of lunularin. Sub-classes are distinguished by the connectivity between these lunularin units, while individual natural products are distinguished by the hydroxy- and/or alkoxy-substituents decorating the core structures. Further diversification results from halogenation and oxidation, which may lead to dimerization or the creation of additional rings. The review provides a timely update to a previous Natural Product Reports article by Keseru and Nógrádi (Nat. Prod. Rep. 1995, 12, 69-75) and largely focuses on new additions to the family and the strategies used to effect their chemical synthesis.

Sinupret(®), a herbal medicinal product made from Gentian root, Primula flower, Elder flower, Sorrel herb, and Verbena herb is frequently used in the treatment of acute and chronic rhinosinusitis and respiratory viral infections such as common cold. To date little is known about its potential antiviral activity. Therefore experiments have been performed to measure the antiviral activity of Sinupret(®) oral drops (hereinafter referred to as "oral drops") and Sinupret(®) dry extract (hereinafter referred to as "dry extract"), in vitro against a broad panel of both enveloped and non-enveloped human pathogenic RNA and DNA viruses known to cause infections of the upper respiratory tract: influenza A, Chile 1/83 (H1N1) virus (FluA), Porcine Influenza A/California/07/2009 (H1N1) virus (pFluA), parainfluenza type 3 virus (Para 3), respiratory syncytial virus, strain Long (RSV), human rhinovirus B subtype 14 (HRV 14), coxsackievirus subtype A9 (CA9), and adenovirus C subtype 5 (Adeno 5). Concentration-dependent antiviral activity (EC(50) between 13.8 and 124.8 μg/ml) of Sinupret(®) was observed against RNA as well as DNA viruses independent of a viral envelope. Remarkable antiviral activity was shown against Adeno 5, HRV 14 and RSV in which dry extract was significantly superior to oral drops. This could be ascertained with different assays as plaque-reduction assays in plaque forming units (PFU), the analyses of a cytopathogenic effect (CPE) and with enzyme immunoassays (ELISA) to determine the amount of newly synthesised virus. Our results demonstrate that Sinupret(®) shows a broad spectrum of antiviral activity in vitro against viruses commonly known to cause respiratory infections., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Combinations of thyme, cowslip and liquorice roots are often used in the treatment of upper respiratory tract infections. Therefore the volatile fraction of herbal teas prepared from thyme (Thymus vulgaris) and the effect of combining with cowslip (Primula veris/P. elatior) and liquorice roots (Glycyrrhiza glabra) on the volatiles were analyzed. Volatile compounds were isolated by hydrodistillation and solid phase extraction and analyzed by GC-MS. Thymol was also quantified by HPLC. The total amount of volatiles as well the thymol content was decreased with increasing proportions of cowslip or liquorice in the infusion extracts whereas the proportion of monoterpene hydrocarbons increased., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The vacuolar and epicuticular flavonoids and the volatiles of the leaves and parts of flower of P. spectabilis Tratt., an endemic species in the Italian Oriental Alps, were investigated. From a MeOH extract of the leaves two flavone glycosides, 8-C-β-glucopyranosylluteolin 7-O-α-arabinofuranoside (1) and 6-C-α-arabinofuranosylapigenin (2) were isolated, in addition to a flavone and three flavonols already known from species of Primula. From an EtOH extract of leaf exudates, 7,3',4'-tri-O-methylquercetin was obtained. The structures were elucidated on the basis of their 1D ¹H- and ¹³C NMR data and 2D NMR techniques, as well as of HPLC-MS. The volatiles emitted by the leaves were mainly constituted by non-terpene derivatives, followed by comparable proportions of hemiterpens, oxygenated monoterpenes and sesquiterpene hydrocarbons. In flowers, monoterpene hydrocarbons were the most represented chemical class followed by non-terpene derivatives. Different proportions of compounds were found when individual parts of flowers were examined separately; calyx produced a greater proportion (approx. 49.5%) of non-terpenes as its volatile metabolites. P. spectabilis has glandular trichomes in the hyaline margins of the epidermal depressions, distributed on the adaxial leaf blade. Glandular hairs were also present on the corolla. Correlations of phytochemical data with the morphological features of leaf, flower and glandular hair are discussed, and a hypothesis is proposed on the ecological roles of the flavonoids and volatile compounds on the general fitness of the species and cross-pollination strategies., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Primula macrophylla (Primulaceae) is reported as to be useful in asthma, restlessness, insomnia and fish poisoning. Antifungal and toxic activities of crude extract, fractions and a pure isolated compound exhibited statistically significant activities. Excellent antifungal activity was found in the crude extract, benzene and ethyl acetate fractions against T. longifusis and against M. canis with different MIC values. Antileishmanial activity (IC(50) = 50ug/mL) was observed as compared to standard drug Amphotericin B, and cytotoxic activity (LD(50) = 47.919microg/mL) was also found in the chloroform fraction. While pure compound 2-phenylchromone (Flavone) isolated from the chloroform fraction showed good activity (IC(50) = 25microg/mL) against Leishmania and cytotoxicity (LD(50) = 2.0116 microg/mL) in Brine Shrimp experiments. From antileishmanial and cytotoxic activity it can be concluded that 2-phenylchromone is the major compound responsible for these activities.

Several new accessions of the genus Primula and of the closely related Cortusa matthioli have been studied for their exudate flavonoid profiles. Unsubstituted flavone, 5-hydroxy-, 2'-hydroxyflavone and 5,2'-dihydroxyflavone were found as main components. Several other rather unusual substitution patterns were also found. 8-O-Substituted flavones appear to be accumulated more often than 6-O-substituted products. Chalcones with corresponding substitution patterns were of scattered occurrence, while flavanones could so far not be detected in exudates of Primula species. The distribution of externally accumulated flavonoids, including literature data, is related to current taxonomic views on the infrageneric grouping of Primula and related taxa. Organ-specific accumulation, biosynthetic and chemosystematic aspects are briefly addressed.

Objective: To study the chemical constituents of Primula sikkmensis Hook., Methods: The chemical constituents of the plant were isolated and purified by column chromatography and their structures were elucidated by spectroscopic analysis., Results: Seven compounds were isolated and identified as kaempferol, quercetin, Prunetin, hexacosanol, Apigenin, beta-sitosterol, quercetin-3-O-beta-D-glucoside, respectively., Conclusion: Compounds I--VII are obtained for the first time from this plant.

The taxonomy of alpine Primula species has long been in dispute because of high morphologic variability and several hybridisations. In Primula species, the trichome height and the colour of hair-tips are usually indicated as diacritic characters, but in our experience this is not adequate. The present study, focused on Primula auricula, Primula daonensis and Primula hirsuta, therefore proposes the use of other morphologic trichome parameters (size and dimensional ratio of stalk, neck and gland head). Phytochemical investigations about the flavonoid composition (epicuticular and vacuolar) of leaves, as taxonomic markers, have also been performed. We report the isolation and identification of two new flavonol glycosides, isorhamnetin 3-O-(2,6-di-O-beta-D-glucopyranosyl-beta-D-glucopyranoside) (1) and kaempferol 3-O-(2-O-alpha-L-rhamnopyranosyl-6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside) (2) and of eight known flavonoids. Size and dimensional ratio of the three trichome elements (stalk, neck and glandular head) are typical for each species analysed. The flavonoid profile well characterise the entities under study. Three different profiles have been obtained with both vacuolar and epicuticular flavonoids. The morphologic and phytochemical markers proposed in this work seem to be parameters which significatively discriminate the species under study.

Study Objective: The objective of the study was to assess the efficacy and tolerability of a fixed combination of dry extracts of thyme herb and primrose root (thyme-primrose combination) and matched placebo in patients suffering from acute bronchitis with productive cough., Methods: In a double-blind, placebo-controlled, multicentre Phase IV study, 361 outpatients with acute bronchitis and > or = 10 coughing fits during the day, onset of bronchial mucus production with impaired ability to cough up at a maximum of 2 days prior to recruitment, and a Bronchitis Severity Score (BSS) > or = 5 score points were randomly assigned to an 11-day treatment (1 tablet three times daily) with either thyme-primrose combination (Bronchipret TP FCT; N = 183) or placebo (N = 178). After the baseline examination (Visit 1 = Day 0), 2 control examinations were scheduled (Visit 2 = Day 4; Visit 3 = Day 10/end of treatment). The efficacy of the study treatment on acute bronchitis was evaluated by the patient's daily counting of coughing fits during the daytime (manual counter), assessment of acute bronchitis related symptoms and by the investigator's assessment of the most important symptoms of acute bronchitis using the BSS. Evaluation of tolerability was based upon adverse event (AE) monitoring, measurement of vital signs as well as the patient's and investigator's global judgement of tolerability at study end. Primary outcome was the change in frequency of coughing fits during daytime on days 7-9 according to patient's accurate daily recording with a manual counter and documentation in the diary. Treatment effects were analysed by analysis of variance (ANOVA) adjusted for centre effects. Due to significant deviation from the "preconditions" of the ANOVA, the Mann-Whitney-Wilcoxon test (stratified by centre) was carried out additionally., Results: The mean reduction in coughing fits on days 7 to 9 relative to baseline (primary endpoint) was 67.1% under thyme-primrose combination compared to 51.3% under placebo (p < 0.0001). In the thyme-primrose combination group, a 50% reduction in coughing fits from baseline was reached about 2 days earlier compared to the placebo group. The symptoms of acute bronchitis (BSS) improved rapidly in both groups, but regression was faster and the responder rates compared to placebo were higher at Visit 2 (77.5% vs 60.1%; p = 0.0006) and Visit 3 (92.9% vs 75.8%; p < 0.0001) under the treatment of thyme-primrose combination. Treatment was well tolerated with no difference in the frequency or severity of AE between thyme-primrose combination and placebo groups. Severe or serious AE were not reported., Conclusion: Oral treatment of acute bronchitis with thyme-primrose combination for about 11 days was superior to placebo in terms of efficacy. The treatment was safe and well tolerated.

Recent data have shown an association between polyunsaturated fatty acid and depression. This study examined the effect of the supplementation with n-6 fatty acid on the behavior of rats treated with imipramine and submitted to the Forced Swimming Test (FST). Non-supplemented imipramine-treated rats presented a significant reduction of immobility time in the FST whereas n-6 fatty acid-supplemented rats showed a significantly higher immobility time. Imipramine significantly increased norepinephrine plasma concentrations in the two groups. These results show that the diet supplemented with n-6 fatty acid altered the behavior of the animals in the FST, inhibiting the imipramine effect.

This paper describes the first liquid chromatographic method suitable for the simultaneous determination of bioactive compounds, saponins and phenolic glycosides, present in Primula elatior and Primula veris, including the NMR data of primulaverin and primeverin. Optimum separations were obtained with a Synergi 4mum Fusion RP 80A column, using 0.025% TFA in water and 5% acetonitrile in methanol as mobile phase. Saponins were detected by evaporative light scattering detection (ELSD), whereas the phenolic glycosides were monitored by UV at 210 nm. The method was validated for repeatability (sigma(rel) or=97.1%) and sensitivity (LOD

In a single-blind, randomized, bi-centric, prospective study, the non-inferiority of a fixed combination of thyme fluid extract and primrose root fluid extract (Bronchicum Elixir S, fluid test medication) was evaluated by comparison to a fixed combination of thyme fluid extract and primrose root tincture (Bronchcium Tropfen, drops test medication). The patients took either 6 x 5 ml of the fluid test medication (fluid group) or 5 x 1 ml of the drops test medication (drops group) daily. 189 outpatients (121 women, 68 men) suffering from acute, not previously treated bronchitis, lasting for less than 48 h, were randomized and treated with either fluid (94 patients: 66 women, 28 men) or drops (95 patients: 55 women, 40 men) over a time period of 7-9 days. 71 patients were excluded from the per-protocol (PP) collective because of violations regarding examination time points and/or intake of the study medication. The primary outcome criterion was to demonstrate the non-inferiority of the Score (BSS) at the end of the study compared to baseline. In the fluid group, the BSS decreased from 11.0 +/- 5.0 points at baseline to 2.6 +/- 4.6 (76%) at study end compared to a decrease from 11.0 +/- 4.8 points at baseline to 2.5 +/- 4.2 (77.1%) at study end in the drops group (Intention-to-treat (ITT) -analysis). The decrease of the BSS in both groups was highly significant (p < or = 10(-3)), but there was no difference between the two groups. Differences between the study sites were noticed regarding the baseline BSS, which were twice as high at study site 2 compared to study site 1 (probably due to the different way the patients were recruited). However, a statistically significant intergroup difference was not observed at any time point. At the end of the study, 52.1% of the patients of the fluid group were symptom free and 53.7% of the patients from the drops group were symptom free as compared by the ITT-analysis (secondary outcome criterion). For both parameters, the PP-analysis support the non-inferiority of the fluid compared to the drops. The global therapeutic efficacy of the fluid as well as of the drops was rated as being "very good" or "good" by 80% of the patients and clinical investigators. The tolerability was very good in both groups; neither serious adverse events nor clinically relevant findings in the safety parameters were observed. A total of 10 adverse events occurred, 5 in the fluid group and 5 in the drops group. Five of these adverse events (2 in the fluid group and 3 in the drops group) were considered to be possibly or probably related to the intake of the study medication. Neither serious nor unknown adverse drug reactions were observed. One drop-out occurred during the study, because of ineffectiveness of the study medication. In the global safety assessment, the tolerability of both medications was rated by about 90% of the patients and by clinical investigators as "good" or "very good". The study demonstrated that the fixed combination of thyme fluid extract and primrose root extract and the combination of thyme fluid extract and primrose root tincture were well tolerated and showed comparable results regarding their efficacy, e.g. decrease of the bronchitis symptoms (primary outcome criterion) and in relief of symptoms (secondary outcome criterion). The results of the study confirm the non-inferiority of the fluid, a combination of thyme fluid extract and primrose root extract when compared to the drops, a combination of thyme fluid extract and primrose root tincture.

Ten lipophilic flavones were isolated from the leaves of Primula veris from field cultivation - the newly described 3'-hydroxy-4',5'-dimethoxyflavone and 3'-methoxy-4',5'-methylenedioxyflavone, the previously known from chemical synthesis 3',4'-dimethoxyflavone, 2',5'-dimethoxyflavone, and also flavone, 2'-hydroxyflavone, 2'-methoxyflavone, 3'-methoxyflavone, 3',4',5'-trimethoxyflavone and 5,6,2',6'-tetramethoxyflavone (zapotin) which were previously known from plants. The same flavones were found in the leaves of P. veris obtained by in vitro propagation. The structural assignments were derived from (1)H NMR, (13)C NMR, EIMS and UV spectral data and the influence of B-ring oxygen substituents on the C-2, C-3 and H-3 NMR resonances in flavones unsubstituted in the A ring is taken into consideration.

Floral traits such as odor, color, and morphology are important pollinator attractants. Variation in floral traits may influence floral constancy, the tendency of pollinating insects to visit flowers of only one type. We investigated for the first time variation in odor between floral morphs in heterostylous species. We analyzed inter- and intraspecific odor variation in the "pin" and "thrum" floral morphs of sympatric Primula elatior and P. farinosa (Primulaceae). Floral volatiles were sampled with headspace sorption. Quantitative analysis and chemical identification were performed by gas chromatography coupled to mass spectrometry. The species produced different floral bouquets. P. elatior emitted mostly limonene with small amounts of alpha-pinene, myrcene, and sabinene. P. farinosa produced benzaldehyde, 4-oxoisophorone (2,6,6-trimethyl-2-cyclohexene-1,4-dione), benzyl alcohol, and benzyl acetate. These interspecific differences may play a role in promoting floral constancy and maintaining species integrity. Conversely, no differences were detected between the scents of pin and thrum morphs within each species. Heterostyly relies on pollinators visiting both floral morphs. There may be stabilizing selection against divergences in traits that may cause pollinators to develop floral constancy to only one of the floral morphs.

In a double-blind, randomized, placebo-controlled, multicenter, prospective study, the clinical efficacy and tolerability of a fixed combination of thyme fluid extract and primose root tincture (Bronchicum Tropfen) was investigated at a dosage of 30 drops (1 ml), taken orally five times daily. 150 outpatients (97 women, 53 men) suffering from acute, not previously treated bronchitis, lasting for less than 48 h, were randomized and treated with either verum (75 patients: 45 women, 30 men) or placebo (75 patients: 52 women, 23 men) over a time period of 7-9 days. 17 patients were excluded from the per-protocol (PP) collective because of either withdrawal from the trial (n = 2) or violations regarding examination time points and/or intake of the study medication (n = 15). The primary outcome criterion for efficacy assessment was the decrease of the Bronchitis Severity Score (BSS) at the end of the study compared to baseline. In the verum group, the BSS decreased from 12.0 +/- 4.4 points at baseline to 1.0 +/- 2.1 at study end compared to a decrease from 11.7 +/- 4.3 points at baseline to 6.5 +/- 4.8 at study end in the placebo group. The inter-group difference of 5.8 points was highly significant (p < or = 10(-3)) in favour of the verum medication [Intention-to-treat (ITT) analysis]. At the end of the study, significantly more patients were symptom free in the verum group (58.7%) than in the placebo group (5.3%) as compared by the ITT analysis (secondary outcome criterion). For both parameters, the PP analysis showed comparable results. The results for the concomitant variables of efficacy support the high superiority of the verum medication compared to placebo. The therapeutic effect was more pronounced the stronger the severity of the acute bronchitis was (as proofed by a stratified evaluation based on severity grade). The tolerability was very good in both groups; neither serious adverse events nor clinically relevant findings in the safety parameters were observed. A total of 7 adverse events occurred, 2 in the verum group and 5 in the placebo group. One of the two adverse events in the verum group was considered to be possibly related to the intake of the study medication. Neither serious nor unknown adverse drug reactions were observed. Two drop-outs occurred during the study related to adverse events, both in the placebo group. In the global safety assessment, the tolerability of both medications was rated as "good" or "very good" by more than 90% of the patients and physicians. In the present study, the fixed combination of thyme fluid extract and primrose root tincture was well tolerated and resulted in a clinically relevant and more pronounced decrease of the bronchitis symptoms (primary outcome criterion) and in shortening the duration of acute bronchitis (secondary outcome criterion) when compared to placebo.

The biological effects of epicuticular substances in farinose exudates accumulated on inflorescence shafts and calyces of Primula denticulata on human acute myeloid leukemia cells (HL-60) were analyzed. The crude material possessed little antioxidative capacity but strong cytostatic properties. Some of its known components (5-hydroxyflavone, 2'-hydroxyflavone, 5,2'-dihydroxyflavone, and 5,8-dihydroxyflavone) were further tested to identify the biologically active compounds. The effects of these flavones on cell cycle progression, mitochondrial membrane potential, and reactive oxygen species have been investigated by flow cytometry. The flavonol quercetin was included in the study as reference compound because of its known cytostatic properties and its activity as radical scavenger. Compared to quercetin the flavones induced little apoptosis (up to 40 microM), but despite their low toxicity, the Primula flavonoids possessed strong cytostatic properties even at low concentrations. The cell cycle distribution showed a characteristic time-dependent shift, giving evidence of a generally short-lived effect of the test compounds in the exposed cells. The antioxidative properties quantified according to two different methods correlated with the number of hydroxyl groups. Whereas quercetin strongly affected the mitochondrial membrane potential, none of the Primula flavones showed a comparable effect.

Pharmaceutical herbal preparations have been used for the treatment of respiratory diseases for a long time. Alongside with the well known herbs, there are many of crude drugs to be investigated and applied in therapeutics. Respiratory diseases are mainly treated by chemical medicines and antibiotics. The use of herbal preparations is limited and they are usually used as supplemental ones. Application of modern research techniques, the new data on biochemical processes in plants, chemical compounds and information on their value in therapy were obtained. The pharmacological effect of the compounds could be foreseen from their chemical structure. The development of rational technology of pharmaceutical preparations follows the identification of active constituents in pharmaceutical herbs. The proper use of pharmaceutical herbs supplies the organism with the natural substances, thus preventing various diseases. The primrose dried raw material for use in production of pharmaceutical preparations has been analyzed and its quality was evaluated. The dry primrose extract was produced and tested, and later it was used in development of pharmaceutical preparations. The liquid primrose extract and two compositions of primrose syrup were developed. The stability of experimental preparations was evaluated.

The chemical composition of the essential oils of five natural populations of P. ovalifolia from central and southwest China and their interpopulation variability were first analyzed by using GC-MS. Twenty-two essential oil compounds were obtained, in which eighteen ones were identified and characterized representing 95%-96% of the oil composition. Three main chemotypes, i.e., the methyl-acetyl-hydroquinone-rich, hydroquinone-rich, and acetyl-hydroquinone-rich chemotypes, were then differentiated, corresponding to the three groups obtained from the cluster analysis based on the essential oil composition percentages. Genetic variations among the five populations were also investigated using the Inter-Simple Sequence Repeats (ISSR) markers. Finally, the Mantel test showed that there was a significant correlation between two distance matrices based on the chemical compounds of essential oils and ISSR markers, confirming the congruence of interpopulation relationships in the P. ovalifolia revealed by the chemical and molecular markers

Primula obconica was introduced to Europe from Hubei, China in 1880, and has been cultivated worldwide as one of popular ornamental plants. The volatile oil of wild P. obconica collected from its original place, Yichang, Hubei was first investigated. A total of 43 compounds constituting 93.49% of the oil were identified by using GC and GC-MS. The major compounds were methyl 2,4-dihydroxy-5-methyl benzoate (30.41%), methyl 2,6-dihydroxy-4-methyl benzoate (29.27%), and hypnone (8.92%) etc. In comparison with the published data of some European cultivars, the native P. obconica seems to be allergen-free due to absence of primin and miconidin.

Essential oil compounds such as found in thyme extract are established for the therapy of chronic and acute bronchitis. Various pharmacodynamic activities for thyme extract and the essential thyme oil, respectively, have been demonstrated in vitro, but availability of these compounds in the respective target organs has not been proven. Thus, investigation of absorption, distribution, metabolism, and excretion are necessary to provide the link between in vitro effects and in vivo studies. To determine the systemic availability and the pharmacokinetics of thymol after oral application to humans, a clinical trial was carried out in 12 healthy volunteers. Each subject received a single dose of a Bronchipret TP tablet, which is equivalent to 1.08 mg thymol. No thymol could be detected in plasma or urine. However, the metabolites thymol sulfate and thymol glucuronide were found in urine and identified by LC-MS/MS. Plasma and urine samples were analyzed after enzymatic hydrolysis of the metabolites by headspace solid-phase microextraction prior to GC analysis and flame ionization detection. Thymol sulfate, but not thymol glucuronide, was detectable in plasma. Peak plasma concentrations were 93.1+/-24.5 ng ml(-1) and were reached after 2.0+/-0.8 hours. The mean terminal elimination half-life was 10.2 hours. Thymol sulfate was detectable up to 41 hours after administration. Urinary excretion could be followed over 24 hours. The amount of both thymol sulfate and glucuronide excreted in 24-hour urine was 16.2%+/-4.5% of the dose.