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Hypericum perforatum--chemical profiling and quantitative results of St. John's Wort products by an improved high-performance liquid chromatography method.

The analysis of flavonoids, naphthodianthrones, and the phloroglucinol derivative hyperforin in H. perforatum is described in this article. In a 35-min HPLC run nine major compounds could be identified and baseline separated in the methanolic plant extracts. For an optimum separation the mobile phase consisted of 10 mM ammonium acetate buffer (pH 5.0) and an acetonitrile/methanol mixture; a Synergi MAX-RP 80 A column (C-12 material) was used as stationary phase. Detection was performed at 270 nm, and the identity of the compounds was confirmed in an LC-MS experiment. Commercial St. John's Wort products were analyzed and qualitative and quantitative results are discussed.

Calibration↗

Mammalian neurohormones: potential significance in reproductive physiology of St. John's wort (Hypericum perforatum L.)?

Melatonin and serotonin are indoleamine neurohormones that function as photoperiod signals in many species and have recently been found in St. John's wort, a medicinal plant used in the treatment of depression. There is no known role for melatonin in higher plants but melatonin functions as a signal of changes in photoperiod in other species. In the current study, serotonin and melatonin were quantified during flower development. Higher concentrations of serotonin were found in flower buds at the tetrad stage of microspore development and higher melatonin concentrations were detected during uninucleate mircosporogenesis. Additionally, the regeneration potential of isolated anthers was highest in the same stage that had elevated melatonin concentrations. These data provide the first evidence of the presence of melatonin during flower development and raise many questions about the potential roles of serotonin and melatonin as regulatory molecules in the reproductive flexibility of higher plants.

Animals↗

Sulfonated xanthones from Hypericum sampsonii.

Xanthones, 1,3-dihydroxy-5-methoxyxanthone-4-sulfonate and 1,3-dihydroxy-5-O-beta-D-glycopyranosylxanthone-4-sulfonate, together with nine known compounds were obtained from H. sampsonii. This is the first report of sulfonated xanthonoids. Furthermore, compounds 1 and 2 exhibited significant cytotoxicity against the P388 cancer cell line.

Animals↗

Inhibitory effect of the antidepressant St. John's wort (hypericum perforatum) on rat bladder contractility in vitro.

OBJECTIVES: To evaluate the effect of St. John's wort (SJW), an effective and safe herbal antidepressant, on rat bladder contractility. Recent data have suggested a strong association between depression and urinary incontinence. METHODS: Strips were cut from the bladder body and placed in organ baths containing Krebs solution. Contractions were induced by electrical field stimulation (EFS) and, in some experiments, by exogenous alpha,beta (alpha,beta)-methylene adenosine triphosphate. RESULTS: St. John's wort was significantly more active in inhibiting the EFS-induced contractions than the alpha,beta-methylene adenosine triphosphate-induced contractions, suggesting both a presynaptic site of action and a direct inhibition of bladder smooth muscle. The inhibitory effect of SJW on EFS-induced contractions was unaffected by methysergide, haloperidol, phentolamine plus propranolol (antagonists that block the action of the neurotransmitters 5-hydroxytriptamine, dopamine, and noradrenaline on their own receptors), the L-type calcium channel antagonist verapamil, capsazepine (which blocks the vanilloid receptor), or cannabinoid CB1 receptor antagonist SR141716A. However, the opioid receptor antagonist naloxone significantly reduced the inhibitory effect of SJW on EFS-induced contractions. Among the chemical constituents of SJW tested, hyperforin and, to a lesser extent, the flavonoid kaempferol showed inhibitory effects. CONCLUSIONS: The results of our study demonstrated that SJW inhibits excitatory transmission of the rat urinary bladder and also directly inhibits smooth muscle contractility. The inhibitory effect on excitatory transmission could involve, at least in part, opioid receptors. SJW may be evaluated for its possible use in treating urinary incontinence in depressed patients.

Acetylcholine↗

Excipient compatibility study of Hypericum perforatum extract (St. John's wort) using similarity metrics to track phytochemical profile changes.

The formulation of botanical dietary supplements is challenging due to their complex activity-composition relationship, as well as physical and chemical stability issues. As excipient compatibility testing is a major component of sound formulation development, the objectives of this work were: (1) explore excipient compatibility storage paradigms; (2) determine interactions between phytochemicals of interest in Saint John's Wort (SJW) with several excipients; and (3) explore the application of similarity metrics to the data. Modifications to conventional isothermal stress testing paradigms included additional storage conditions of heat and moisture (5, 50 degrees C, 5 and 0% added water), as well as more rigorous controls. Binary blends of SJW and ten commonly used excipients were prepared and neat SJW was used as control. After 3 weeks, the percentage remaining of each phytochemical was determined by HPLC. Several similarity metrics were applied to the data. Common storage paradigms were suitable for excipient compatibility testing when controls of neat material are stored under similar conditions and the percentage of phytochemicals remaining in excipient:SJW blends and neat SJW are compared. Excipient incompatibilities were determined for SJW phytochemicals of interest. Similarity metrics applied to the phytochemical profiles conveniently summarized the data. This work allows logical decisions to be made regarding the formulation of SJW.

Drug Storage↗

Hyperforin a constituent of St John's wort (Hypericum perforatum L.) extract induces apoptosis by triggering activation of caspases and with hypericin synergistically exerts cytotoxicity towards human malignant cell lines.

Hyperforin (HP) is an abundant component of St John's wort with antibiotic and antidepressive activity. We report here the ability of HP and that of polyphenolic procyanidin B2 (PB-2) to inhibit the growth of leukemia K562 and U937 cells, brain glioblastoma cells LN229 and normal human astrocytes. HP inhibited the growth of cells in vitro with GI(50) values between 14.9 and 19.9 microM. The growth inhibitory effect of PB-2 was more pronounced in leukemia cell lines K562 and U937, the GI(50) concentrations being about 12.5 microM established after 48 h incubation differed significantly (P<0.05) from those of LN229 and normal human astrocytes (103.1 and 96.7 microM), respectively. Further, HP and hypericin (HY) (a naphthodianthrone from St John's wort) acted synergistically in their inhibitory effect on leukemic (K562, U937) cell growth. Cell death occurred after 24 h treatment with HP and PB-2 by apoptosis. A dose-dependent loss of membrane phospholipid asymmetry associated with apoptosis was induced in all cell lines as evidenced by the externalization of phosphatidylserine (PS) and morphological changes in cell size and granulosity by scatter characteristics. In leukemia U937 cells, HP increased the activity of caspase-9 and caspase-3 and in K562 cells caspase-8 and caspase-3. In addition, the broad spectrum caspase inhibitor z-VAD-fmk inhibited both the appearance of PS exposure and the activation of caspases, illustrating the functional relevance of caspase activation during HP-induced apoptosis. Cytocidal effects of HP and its cooperation with HY on tumor growth inhibition in a synergistic manner make the St John's wort an interesting option in cancer warranting further in vitro and in vivo investigation.

Annexin A5↗

Supercritical fluid extraction and high-performance liquid chromatographic determination of phloroglucinols in St. John's Wort (Hypericum perforatum L.).

A small-scale supercritical fluid extraction (SFE) method was developed for the selective extraction of phloroglucinols from St. John's wort (SJW) leaf/flower mixtures using supercritical carbon dioxide (CO(2)). The extraction efficiency was investigated as influenced by pressure, temperature, time, and modifier. The optimized condition of SFE was carried out at 3.80 x 10(4) kpa (5500 psi) and 50 degrees C. Samples were held in static extraction for 10 min, followed by a dynamic extraction for 90 min at the flow rate of 1 mL/min. A simple and sensitive HPLC method was developed for the analysis of hyperforin and adhyperforin, the major phloroglucinols, in the SFE extract of SJW.

Chromatography, High Pressure Liquid↗

St John's wort (Hypericum perforatum): drug interactions and clinical outcomes.

AIMS: The aim of this work is to identify the medicines which interact with the herbal remedy St John's wort (SJW), and the mechanisms responsible. METHODS: A systematic review of all the available evidence, including worldwide published literature and spontaneous case reports provided by healthcare professionals and regulatory authorities within Europe has been undertaken. RESULTS: A number of clinically significant interactions have been identified with prescribed medicines including warfarin, phenprocoumon, cyclosporin, HIV protease inhibitors, theophylline, digoxin and oral contraceptives resulting in a decrease in concentration or effect of the medicines. These interactions are probably due to the induction of cytochrome P450 isoenzymes CYP3A4, CYP2C9, CYP1A2 and the transport protein P-glycoprotein by constituent(s) in SJW. The degree of induction is unpredictable due to factors such as the variable quality and quantity of constituent(s) in SJW preparations. In addition, possible pharmacodynamic interactions with selective serotonin re-uptake inhibitors and serotonin (5-HT(1d)) receptor-agonists such as triptans used to treat migraine were identified. These interactions are associated with an increased risk of adverse reactions. CONCLUSIONS: In Sweden and the UK the potential risks to patients were judged to be significant and therefore information about the interactions was provided to health care professionals and patients. The product information of the licensed medicines involved has been amended to reflect these newly identified interactions and SJW preparations have been voluntarily labelled with appropriate warnings.

Contraceptives, Oral↗

Effect of light on hypericins contents in fresh flowering top parts and in an extract of St. John's Wort (Hypericum perforatum).

St. John's Wort is a medicinal plant increasingly used for its antidepressive activity. Hypericins are considered as one of the compounds contributing to the activity of the extract. These naphthodianthrones exist in various forms in Hyperici herba. Protopseudohypericin and protohypericin (protopigments) are converted into pseudohypericin and hypericin (pigments) under the action of light. The aim of this work is to study the influence of light on the phototransformation of protopigments into pigments. Two experiments were carried out. The studies were performed on one hand, on plant material in order to know the proportion of these substances in various plant parts and the possibility of transforming the protopigments into pigments under the action of sunlight; on the other hand, in the extract to determine the optimal wavelength allowing this transformation. Three parts of the fresh plant (buds, flowers, leaves) were treated with sunlight on three levels of exposure. Liquid extracts were exposed to various types of light with wavelengths ranging between 480 and 660 nm by means of diodes. The flowering tops of St. John's Wort contain a share of approximately 30% hypericins in the form of protopseudohypericin and protohypericin: buds (48%), flowers (30%), leaves (17%). After an exposure of fresh buds to sunlight for 16 hours the share of protopigments was then 32%. In the extract, the transformation of the protopigments is total and requires less energy than in the plant material. The optimal wavelength for the transformation of the protopigments in the extract is around 515 nm (green), close to the optimum absorption level of protopigments.

Anthracenes↗

Molecular markers as a tool to verify sexual and apomictic off-spring of intraspecific crosses in Hypericum perforatum.

The aim of this project was to establish RAPD markers to determine the percentage of sexual off-spring of the facultative apomictic plant species H. perforatum. We did reciprocal crosses between four different accessions (A x B, B x A, C x D, D x C) by mechanical emasculation and hand pollination. Genomic DNA of the parents and the off-spring was isolated and PCR conditions were optimized in order to obtain reproducible bands with RAPD markers. Of the 260 screened RAPD primers 127 revealed polymorphism between the parental lines of A and B, whereas 53 revealed no amplification products. Each progeny was tested for the presence of paternal bands with three primers. We found no sexual off-spring among the 22 progenies of A x B, the nine progenies of B x A and the ten progenies of D x C. However, we detected six sexual off-spring among the 45 progenies of C x D. We have proved that RAPD markers can be used to distinguish between sexual and apomictic off-spring in H. perforatum and that sexual off-spring can be obtained from intraspecific crosses. The percentage of sexual progeny might depend on the genotype of the parental lines.

Base Sequence↗