Simultaneous Determination of Diosmin and Hesperidin in Pharmaceuticals by RPLC using Ionic Liquids as Mobile Phase Modifiers.

Diosmin and hesperidin are natural flavonoid glycosides found in various plant materials, mainly in citrus fruits in different concentrations. Diosmin for pharmaceutical use is obtained mainly semi-synthetically from hesperidin. Hesperidin often accompanies diosmin as a natural impurity in different pharmaceutical formulations; therefore, a simple, fast and precise method for the simultaneous assay of diosmin and hesperidin in pharmaceutical formulations has been developed to control their contents. Chromatographic resolution was performed using a column with C-18 packing and the following mobile phase: methanol/water (45:55, v/v) with 0.025% added didecyldimethylammonium lactate, which significantly affects retention, shortening analysis time and having a positive impact on the symmetry of resulting chromatographic peaks. The method shows linearity between 2.5 and 100 µg/mL, high repeatability (0.39 and 0.42% for diosmin and hesperidin, respectively) and accuracy of 96 to 102% for both the assayed compounds. Intraday and interday precision of the new method were less than RSD% 1, 2. The limit of detection of the assayed compounds is 2.5 and 1.2 µg/mL for diosmin and hesperidin, respectively. The method was tested on several pharmaceutical products available in Poland.

Determination of the Juglone Content of Juglans regia Leaves by GC/MS.

The constituents of walnut (Juglans regia L.) leaves are represented by tannins, phenolics, and naphthoquinones, the characteristic compound being juglone. The content of juglone in the methanolic extract of the leaves determined by the GC/MS method was 9.9 ± 0.2 mg/100 g; small amounts (1.3 ± 0.02 mg/100 g) were recorded in the infusion, whereas in the decoction it was not detected. As some studies indicate toxicity of juglone, only decoctions should be recommended for therapeutic use.

Coumarins in horse chestnut flowers: isolation and quantification by UPLC method.

The coumarins: scopoletin, esculetin and fraxetin were isolated from the flowers of horse chestnut (Aesculus hippocastanum L., Hippocastanaceae) and identified by spectrophotometric methods (UV, 1H, 13C NMR, ESI-MS). Their content, determined using the Ultra Performance Liquid Chromatography (UPLC), was 0.41, 0.13 and 0.05%, respectively.

Leonurus cardiaca L. (motherwort): a review of its phytochemistry and pharmacology.

Leonurus cardiaca is a perennial plant indigenous to central Europe and Scandinavia, but it is also found in the area spanning temperate Russia to central Asia. It has been introduced to North America and has become established locally in the wild. Motherwort (Leonuri cardiacae herba) consists of aerial parts of Leonurus cardiaca gathered during the flowering period, dried at 35 °C and, according to European Pharmacopoeia 7th edition, should contain a minimum of 0.2% flavonoids, expressed as hyperoside. Compounds belonging to the group of monoterpenes, diterpenes, triterpenes, nitrogen- containing compounds, phenylpropanoids, flavonoids and phenolic acids, as well as volatile oils, sterols and tannins, have been identified in motherwort. Traditionally, extracts of the herb have been used internally, mainly for nervous heart conditions and digestive disorders. However, they have also been used for bronchial asthma, climacteric symptoms and amenorrhoea, as well as externally in wounds and skin inflammations. Mild negative chronotropic, hypotonic and sedative effects can be attributed to the herb and preparations thereof. Pharmacological studies have confirmed its antibacterial, antioxidant, anti-inflammatory and analgesic activity, as well as its effects on the heart and the circulatory system. Sedative and hypotensive activity has been demonstrated in clinical trials.

Disseminated cutaneous Kaposi sarcoma in a patient receiving triptolide/tripdiolide for rheumatoid arthritis.

BACKGROUND: To date, Kaposi sarcoma has not been mentioned among the adverse effects of triptolide/tripdiolide, ethyl acetate extracts or polyglycosides of the Chinese herbal remedy Tripterygium wilfordii Hook F. CASE REPORT: A patient was diagnosed with rheumatoid arthritis at the age of 29 years. She underwent treatment with corticosteroids, methotrexate and gold sodium thiosulfate, and was chronically taking ketoprofen. At the age of 59 years she started to take a powder (≈2 g/day) from a Chinese physician for treatment of rheumatoid arthritis. This powder was supplied to her regularly for 10 years. At the age of 69 years, multiple soft, violaceous to dark-red patches, plaques, nodules and blisters of varying sizes appeared on a background of severely edematous skin on her legs, and later on her arms. Biopsy specimens of the leg lesions were diagnostic for human herpesvirus 8-associated Kaposi sarcoma. Triptolide (235 µg/1 g) and tripdiolide were found in the Chinese powder by the use of Liquid Chromatography Electrospray Ionization Mass Spectrometry. Administration of the powder was stopped and medication with paclitaxel was introduced. General condition of the patient improved and skin lesions diminished significantly. CONCLUSIONS: This case indicates a possible association between triptolide/tripdiolide chronic intake and development of human herpesvirus 8-associated Kaposi sarcoma. Triptolide/tripdiolide could contribute to development of Kaposi sarcoma by reactivation of latent human herpesvirus 8, permitted by immunosuppression induced by triptolide.

[Phytotherapy of respiratory tract diseases]. / Fitoterapia chorób dróg oddechowych.

Herbal medicines have been used in cough due to their antitussive and expectorant activity. Antitussives act either centrally on the cough center of the brain or peripherally on the cough receptors in the respiratory passages. The antitussive effect of many herbs results from the content of mucilage, which exerts protective and demulcent activity. The activity of expectorant herbs results primarily from their influence on the gastric mucose (saponins and ipec alkaloids). This proves reflex stimulation which leads to an increase in the secretion of bronchial glands. Volatile-oil type expectorant herbs exert a direct stimulatory effect on the bronchial glands by means of local irritation with antibacterial activity. In colds and flu, herbs containing volatile oil can be used; also, volatile oils are ingredients of syrups and liquids as well as external phytomedicines in the form of liniments, ointments, and inhalations. The paper shows the herbs and phytomedicines present on the Polish market used for the treatment of respiratory tract diseases.

Flavonoids from the flowers of Aesculus hippocastanum.

The flavonoids, kaempferol derivatives: 3-O-alpha-arabinofuranoside, 3-O-beta-glucopyranoside, 3-O-alpha-rhamnopyranoside, 3-O-alpha-rhamnopyranosyl (1 --> 6)-O-beta-glucopyranoside and quercetin derivatives: 3-O-alpha-arabinofuranoside, 3-O-beta-glucopyranoside, 3-O-alpha-rhamnopyranosyl (1 --> 6)-O-beta-glucopyranoside, were isolated from the flowers of Aesculus hippocastanum and identified. The structures of these compounds were confirmed by a chemical analysis and spectrophotometric methods (UV, 1H-, 13C-NMR, ESI-MS). The presence of free aglycones: kaempferol and quercetin was confirmed chromatographically by comparison with standards.

Comparative analysis of pharmaceuticals and dietary supplements containing extracts from the leaves of Ginkgo biloba L.

Abstract: Chromatography (TLC and HPLC) tests were performed of 11 preparations containing dry extract of Ginkgo biloba leaves: three pharmaceuticals (preparations 1, 3 and 5) and eight dietary supplements (preparations 2, 4, 6-11), and dry extract of Ginkgo biloba leaves (preparation 12) as a standard certified for compliance with Eur. Ph. 6.1. and FP VIII (1, 2). Preparations registered in Poland as pharmaceuticals contained the major active ingredients (flavonoids and terpene lactones) in amount declared by their producers (and consistent with pharmacopoeial requirements) and acceptable level of potentially toxic ginkgolic acids (below 5 ppm). The concentration of active compounds in dietary supplements was varied. Some of them satisfied applicable quality criteria (mainly preparation 8), however, the majority had reduced levels of therapeutic compounds (4, 6, 7, 11) and increased concentration of ginkgolic acids (4, 9, 10, 11).

Effect of Aquilegia vulgaris (L.) ethyl ether extract on liver antioxidant defense system in rats.

INTRODUCTION: The ethyl ether extract from Aquilegia vulgaris (L.) (Ranunculaceae) contains a lot of phenolic acids. Their hydroxyl groups are capable of donating hydrogen atoms at the initial stage of lipid peroxidation (LPO), which inactivates hydroxyperoxides formed from polyunsaturated fatty acids (PUFAs) and leads to breakdown of the propagation chain. MATERIAL AND METHODS: Rats pretreated with acetaminophen (APAP) (600 mg/kg b.w., p.o.) were given ethyl ether extract (100 mg/kg b.w., p.o.) obtained from A. vulgaris herb. The study parameters measured were microsomal lipid peroxidation, reduced glutathione, and the activity of hepatic antioxidant enzymes and some drug metabolizing enzymes. RESULTS: The treatment with ethyl ether extract of the herb produced a 87-95% decrease in uninduced and Fe2+/ascorbate-stimulated microsomal lipid peroxidation in the liver of rats receiving APAP. Hepatic glutathione level depleted by APAP increased significantly (by 18%) after the extract treatment. Antioxidant enzyme activity in the liver, inhibited by APAP, was found to increase after administration of the extract: catalase by about 36%, glutathione reductase by 27% and glutathione S-transferase by 29%. Glucose-6-phosphate dehydrogenase, which decreased after APAP administration, increased again by 26% after extract treatment. The extract tested did not affect the activity of DT-diaphorase. The cytochrome P450 content, depleted by APAP, increased as much as by 100% after the treatment. The activities of NADPH-cytochrome P450 reductase, aniline hydroxylase and aminopyrine N-demethylase were not affected. CONCLUSIONS: The protective effect of the Aquilegia vulgaris extract in APAP-induced liver injury was mediated by its antioxidant activity. The extract did not inhibit the formation of reactive intermediate metabolites of APAP.

Aquilegia vulgaris extract attenuates carbon tetrachloride-induced liver fibrosis in rats.

Six groups of male Wistar rats were treated as follows: in groups II, III and V liver damage was induced by CCl(4) (per os, 1590 mg/kg b.w.day) given 2 days a week for 6 weeks; group III was treated simultaneously with ethanol extract of Aquilegia vulgaris (100 mg/kg b.w.day) for 6 weeks; group V with silymarin, positive control, at a dose of 100 mg/kg b.w.day for 6 weeks; and groups IV and VI received only the extract or silymarin, respectively. Microsomal lipid peroxidation in the liver increased following CCl(4) treatment by 61-213% and was not changed significantly by the extract. The effect of silymarin was more pronounced, 19-52% decrease in the lipid peroxidation level. Hepatic glutathione was depleted by 22% in CCl(4)-treated rats. The extract tested did not change this parameter. The activity of antioxidant enzymes was significantly reduced after CCl(4) administration, by 42-63%. Co-administration of the extract or silymarin resulted in significant increase in these enzymes activity; however, the basal level was not reached. Hepatic hydroxyproline concentration was elevated over 5-fold in comparison with controls. Co-administration of the extract or silymarin decreased the level of hydroxyproline by 66% and 55%, respectively. Activity of serum hepatic enzymes was elevated in rats treated with CCl(4) by 47-8700%. Both the extract and silymarin reduced significantly these enzymes' activity. The extract caused a fall in bilirubin and cholesterol level in rats treated with CCl(4) by 42% and 17%, respectively. Histopathological examination revealed less-severe fibrosis in rats co-administered the extract or silymarin when compared to animals treated with CCl(4) alone.

Polyphenolic compounds from Abutilon grandiflorum leaves.

The flavonol glycosides: kaempferol 3-O-beta-(6"-p-coumaroyl)-glucopyranoside, kaempferol and quercetin 3-O-beta-glucopyranosides and 3-O-beta-rutinosides were isolated and identified from the leaves of Abutilon grandiflorum. Their structures were established by chemical and spectroscopic methods. The presence of phenolic acids: p-hydroxybenzoic, p-coumaric, syringic, and vanillic was confirmed by TLC.

Protective effect of Aquilegia vulgaris (L.) on carbon tetrachloride-induced oxidative stress in rats.

The ethyl ether extract of A. vulgaris inhibited in vitro microsomal lipid peroxidation (IC50 58.8 microg/ml) and showed moderate ability to scavenge superoxide radicals and to chelate iron ions. The extract (100 mg/kg body weight, po) decreased uninduced and enzymatic microsomal lipid peroxidation in the liver of male rats pretreated with CCl4 (1 ml/kg body weight) by 27 and 40%, respectively. Activity of antioxidant and related enzymes (catalase and glucose-6-phosphate dehydrogenase) inhibited by CCl4 was significantly restored after administration of the extract. The extract itself significantly enhanced superoxide dismutase activity. There was no effect of the extract on hepatic glutathione level and cytochrome P450 content, both were decreased by CCl4. Neither CCl4 nor the tested extract affected activities of NADPH-cytochrome P450 reductase and two monooxygenases, aniline hydroxylase and aminopyrine n-demethylase. It can be concluded that the protective effect of the A. vulgaris extract in CCl4-induced liver injury is mediated by inhibition of microsomal lipid peroxidation and restoring activity of some antioxidant and related enzymes.

Sylviside, a diterpene glucoside derivative from Gnaphalium sylvaticum.

A new diterpene glucoside (1), named sylviside, was isolated from the aerial parts of Gnaphalium sylvaticum. Its structure was elucidated as 2beta,15alpha,20alpha-trihydroxy-19,20-dicarboxy-ent-kaur-16-ene 2beta-O-(2'-angelate)-beta-D-glucopyranoside, on the basis of spectroscopic analysis ((1)H NMR, (13)C NMR, HMQC, HMBC, NOESY), and was confirmed by X-ray crystallographic analysis. Sylviside (1) displayed weak cytotoxicity against HeLa WT (human epitheloid cervical carcinoma) cells and was also evaluated for its effects on reversing multidrug resistance in HeLa cells overexpressing MDR1.

Protective effect of Aquilegia vulgaris L. on aflatoxin B(1)-induced hepatic damage in rats.

The aim of the study was to investigate the effect of ethanol and ethyl acetate extract obtained from Aquilegia vulgaris L. on microsomal lipid peroxidation, reduced glutathione level and antioxidant enzymes activity in the liver of rats intoxicated with aflatoxin B(1) (AFB(1)). Animals were pretreated with 12 daily p.o. doses of the extracts tested (100mg/kg body weight). Then AFB(1) was administered intraperitoneally at a single dose of 1.5mg/kg b.w. to evoke the liver damage. α-Tocopherol was used as a positive control. Reduced glutathione (GSH) was depleted in aflatoxin-treated rats by 80% in comparison with that in the controls. The extracts restored the GSH concentration up to the basal level. Microsomal lipid peroxidation stimulated by Fe(2+)/ascorbate (assessed by measuring TBARS) was enhanced in AFB(1)-treated rats by 28% as compared to that in the control group. The extracts caused a decrease in TBARS level by 40% and 27%. Only two antioxidant enzymes were affected by AFB(1) administration. The activity of catalase was reduced by 24% and the activity of glutathione-S-transferase (GST) was increased by 33%. The pretreatment with ethyl acetate and ethanol extract reduced the GST activity by 76% and 30%, respectively. No significant changes in the activity of other antioxidant enzymes were observed in rats treated with the extracts and AFB(1). It can be concluded that multiple pretreatment with the extracts obtained from A. vulgaris attenuated aflatoxin B(1)-induced hepatic damage as evidenced by inhibition of lipid peroxidation and preventing reduced glutathione depletion.

Phenolic acids in the flowers of Althaea rosea var. nigra.

Distribution of phenolic acids in the flowers of Althaea rosea var. nigra has been studied by 2D-TLC and HPLC methods. The phenolic acids occurring in these fractions have been identified as ferulic, vanillic, syringic, p-coumaric, p-hydroxybenzoic, p-hydroxyphenylacetic and caffeic acids. By means of the HPLC methods the contents of major phenolic acids were estimated. From among the phenolic acids analyzed the syringic, p-hydroxybenzoic and p-coumaric acids are dominant. Total content of phenolic acids was determined by the Arnov's method.

Flavonoids from Abutilon theophrasti flowers.

Nine flavonoid compounds: kaempferol 3-0-beta-(6"-p-coumaroyl)-glucopyranoside, myricetin 3-O-beta-glucopyranoside, quercetin 3-0-beta-glucopyranoside, quercetin 3-0-alpha-rhamnopyranosyl (1-->6)-beta-glucopyranoside, kaempferol 3-0-beta-glucopyranoside, kaempferol 3-0-alpha-rhamnopyranosyl (1-->6)-beta-glucopyranoside, quercetin 7-0-beta-glucoside, quercetin 7-0-beta-diglucoside, kaempferol 7-0-beta-diglucoside were isolated and identified from the flowers of Abutilon theophrasti.

Antioxidant activity of isocytisoside and extracts of Aquilegia vulgaris.

Two extracts (ethyl acetate and ethanol) and isocytisoside obtained from Aquilegia vulgaris were tested for their antioxidant and free radical scavenging activity in vitro. Inhibition both non-enzymatic (IC50: 150-219 microg/ml) and enzymatic (IC50: 23-60 microg/ml) microsomal lipid peroxidation was observed, the extracts being more active than isocytisoside. The substances tested appeared to be weak hydroxyl radical scavengers, showed very low TEAC values and moderate iron chelation ability. However, all preparations at the concentration 25 microg/ml inhibited superoxide anion formation at the range 47-68%. Despite of the lack of a potent free radical scavenging ability the substances tested demonstrated significant antioxidant activity. Relationship between this parameter and the content of phenolic groups was noticed.

Antimicrobial activity of Viola tricolor herb.

The antimicrobial activity of infusion, decoction, ethanol extract and fractions obtained by successive extraction of Viola tricolor herb with dichloromethane, ethyl acetate and methanol was evaluated. The infusion, decoction and ethanol extract were found to be most effective against the tested microorganisms.

Protective effect of Aquilegia vulgaris (L.) on APAP-induced oxidative stress in rats.

Rats pretreated with acetaminophen (N-acetyl-p-aminophenol, APAP) (600 mg/kg b.w., p.o.) were administered with ethanol and ethyl acetate extracts as well as with isocytisoside (100 mg/kg b.w., p.o.) obtained from Aquilegia vulgaris (L.) (Ranunculaceae) herb. The substances tested decreased enzymatic, non-enzymatic and uninduced microsomal lipid peroxidation (LPO) in the liver of rats treated with APAP by 18-48%. Activity of the antioxidant enzymes in the liver inhibited by APAP was increased in the majority of groups after administration of the substances tested: catalase (CAT) by 55%, glutathione peroxidase (GPx) by 50%, glutathione reductase (GR) by 35% and glutathione S-transferase (GST) by 60%. Hepatic glutathione level depleted by APAP was only slightly increased by the substances tested. The cytochrome P450 contents, and the activities of NADPH-cytochrome P450 reductase and two monooxygenases were not affected by the extracts and isocytisoside. It can be concluded that the protective ability of the substances tested in APAP-induced liver injury is mediated by amelioration of microsomal lipid peroxidation and restoring antioxidant enzymes activity. Inhibition of enzymes responsible for metabolic activation of APAP is not involved in this process.

[Preventive role of curcumin in lung cancer]. / Prewencyjne znaczenie kurkuminy w nowotworach pluc.

Carcinogens from cigarette smoke form the link between nicotine addiction and lung cancer, which is the leading cause of cancer-related mortality in the world. One of the most frequently studied chemopreventive agents is a curcumin, a natural compound extracted from turmeric that inhibits cell proliferation and induces apoptosis in human leukaemia, prostate cancer, and non-small cell lung cancer. Curcumin (diferuoylmethane) is a major yellow pigment in turmeric (Curcuma longa) and is widely used as a spice. Curcumin exhibits a variety of pharmacological effects, and has been reported to have anti-inflammatory and anti-tumor activities.