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Graphical abstract, Highlights • Furanocoumarins are the main CYP1A2 inhibitors in A. majus and A. archangelica. • The inhibitory potencies appear additive to CYP1A2 activity in vitro. • BMD determination for human furanocoumarin-consumption and CYP1A2 inhibition., Millions of individuals globally consume traditional herbal medicines (THMs), which contain abundant amounts of linear furanocoumarins. Linear furanocoumarins (i.e., 8-methoxypsoralen, 5-methoxypsoralen, and isopimpinellin) are inhibitors of cytochrome P450 (CYP) isoenzymes including 1A2, a major enzyme involved in drug metabolism and carcinogen bioactivation. Despite the high consumption of furanocoumarin-containing THMs, no studies have measured the furanocoumarin consumption level that triggers an inhibition to CYP1A2 activity in humans. The first objective was to verify if the potencies of the three furanocoumarins are additive towards the inhibition of CYP1A2 activity in vitro using concentration-addition and whole-mixture chemical-mixture-assessment models. A second objective was to determine the benchmark dose (BMD) with the mixtures of furanocoumarin oral doses, expressed as 8-MOP equivalents, and to assess the in vivo CYP1A2 activity, expressed as inhibition percentages. The in vitro results indicated that the three furanocoumarin inhibitory potencies were additive in the THM extracts, validating the use of the concentration-addition model in total furanocoumarin dose-equivalent calculations. Using the USEPA BMD software, the BMD was 18.9 μg 8-MOP equivalent/kg body weight. This information is crucial for furanocoumarin-related health-assessment studies and the regulation of THMs. Further studies should be performed for the remaining major metabolic enzymes to complete the safety profile of furanocoumarin-containing THMs and to provide accurate warning labelling.

A simple and efficient method based on ultrafiltration with liquid chromatography and mass spectrometry was used for the rapid screening and identification of ligands in the extracts of Stellera chamaejasme. The bound ligands, i.e. daphnoretin, isopimpinellin, chamaechromone, neochamaejasmin A, and chamaejasmine (purity of 96.8, 90.75, 91.41, 93.98, and 98.91%, respectively), were separated by semi-preparative high-performance liquid chromatography combined with high-speed counter-current chromatography. To the best of our knowledge, this is the first study to report the detection of potent lipoxidase and lactate dehydrogenase inhibitors in Stellera chamaejasme extracts. The results demonstrate that our method of ultrafiltration with liquid chromatography and mass spectrometry combined with mixed chromatography can be used to screen and confirm the bioactivity of all isolated compounds. This method also eliminates the need for separation of inactive compounds, thereby improving efficiency when studying bioactive substances. For some complex mixtures, neither semi-preparative high-performance liquid chromatography nor high-speed counter-current chromatography can purify all the target active compounds with high purity in a one-step separation. The combination of the two methods allow for efficient purification of target bioactive compounds with different polarities and physicochemical properties based on their complementary properties.

Wild parsnip (Pastinaca sativa) has been associated with livestock and human photosensitization. An investigation of a natural occurrence of photosensitization of grazing horses identified wild parsnip as a possible cause. HPLC-MS and MS/MS analysis of this plant identified five furanocoumarins i.e., xanthotoxin, bergapten, isopimpinellin, imperatorin and a putative methoxyimperatorin. Goats fed this wild parsnip were largely unaffected. Xanthotoxin was not detected in the serum of parsnip-fed goats or in the serum of goats dosed orally or intravenous with purified xanthotoxin. Cutaneous application produced severe photodermatitis in goats and a horse consistent with topical exposure as the likely route to produce wild parsnip-induced photosensitivity. Wild parsnip-induced superficial necrotizing dermatitis was consistent with photodermatitis with no evidence of other allergic or inflammatory components.

The parsnip webworm, Depressaria pastinacella, is restricted to two hostplant genera containing six structurally diverse furanocoumarins. Of these, imperatorin is detoxified by a specialized cytochrome P450, CYP6AB3. A previous whole-larva transcriptome analysis confirmed the presence of nine transcripts that belong to the CYP6AE subfamily. Here, by examining midgut-specific gene expression patterns we determined that CYP6AE89 transcripts were highly expressed and furanocoumarin-inducible. Computer docking and energy-minimization of a CYP6AE89 model with all six furanocoumarins showed that 5-methoxylated bergapten and 8-methoxylated xanthotoxin had the smallest distances from the heme to the proton-donor residue in the catalytic I-helix, and that the 5,8-dimethoxylated isopimpinellin and bergapten had the smallest energy-minimized distance from the heme oxygen to the furan ring double bond. To evaluate this prediction, we expressed the CYP6AE89 protein in an Escherichia coli system, and used it to detect high catalytic activity against the two mono-methoxylated linear furanocoumarins - bergapten and xanthotoxin - and weak activity against isopimpinellin. Thus, CYP6AE89, like CYP6AB3, is probably specialized for detoxifying only a subset of hostplant furanocoumarins. A maximum-likelihood tree built with six representative lepidopterans with manually annotated cytochrome P450s shows that CYP6AE89 may have evolved much faster than the other CYP6AE proteins, possibly indicative of host selection pressure.

Bioavailability of orally administered drugs can be influenced by many factors. Poor drug absorption across the intestinal membrane is one of the factors that contribute to low bioavailability of drugs. It has been suggested that the metabolism/active efflux in the small intestine is involved in the poor absorption of many drugs. Intestinal CYP3A4 and P-gp work coordinately to reduce the intracellular concentration of drugs. Recently, bioenhancers have been identified and extensively studied. The aim of this study was to evaluate natural furanocoumarins found in juices of common lime and kaffir lime as the potential enhancers for oral delivery by means of modulating CYP3A4 and/or P-gp activities. The role of isolated furanocoumarins on CYP3A4 was assessed by testosterone 6β-hydroxylation reaction, while the effect on P-gp was investigated using R123 and CAM uptake studies in Caco-2, as well as LLC-PK1 and LLC-GA5-Col300. In the present study, we demonstrated that isopimpinellin isolated from common lime is the best CYP3A4 inhibitor among 4 isolated furanocoumarins, implying that isopimpinellin would possibly act as a bioenhancer by inhibiting pre-systemic metabolism. 6’,7’-Dihydroxybergamottin found in kaffir lime is a dual inhibitor of CYP3A4 and P-gp, suggest that it could potentially be used as a bioenhancer by inhibiting both pre-systemic metabolism and efflux mechanism. However, in vivo study should be further conducted to confirm these effects in the body.

Angiogenesis is the formation of new blood vessels from an existing vasculature through a series of processes such as activation, proliferation, and directed migration of endothelial cells. Angiogenesis is instrumental in the metastatic spread of tumors. Isopimpinellin, a furanocoumarin group of phytochemicals, is an anticarcinogenic agent. However, no studies have proven its antiangiogenic effects. The current study thus aimed to screen the antiangiogenic effect of isopimpinellin. Human Umblical Vein Endothelial Cell (HUVEC) as an in vitro model and zebrafish embryos as an in vivo model was used in this study. The experimental results showed that isopimpinellin effectively inhibited HUVEC proliferation, invasion, migration, and tube formation, which are the key steps in angiogenesis by markedly suppressing the expression of pro-angiogenic genes VEGF, AKT, and HIF-1α. In addition, isopimpinellin exerts its anti-angiogenic effect through the regulation of miR-15b-5p and miR-542-3p. Furthermore, in zebrafish embryos, isopimpinellin inhibited the development of intersegmental vessels (ISVs) through the significant downregulation of all pro-angiogenic genes vegf, vegfr2, survivin, angpt-1, angpt-2, and tie-2. Collectively, these experimental findings offer novel insights into the antiangiogenic nature of isopimpinellin and open new avenues for therapeutic approaches.

Due to global warming, invasive species have spread across the world. We therefore studied the impact of short-term (1 day or 2 days) and longer (7 days) heat stress on photosynthesis and secondary metabolites in Heracleum sosnowskyi, one of the important invasive species in the European Union. H. sosnowskyi leaves exposed to short-term heat stress (35 °C/1 d) showed a decrease in chlorophyll and maximum potential quantum efficiency of photosystem II (Fv/Fm) compared to control, 35 °C/2 d, or 30 °C/7 d treatments. In turn, the high level of lipid peroxidation and increased H2O2 accumulation indicated that the 30 °C/7 d stress induced oxidative damage. The contents of xanthotoxin and bergapten were elevated in the 2 d and 7 d treatments, while isopimpinellin was detected only in the heat-stressed plants. Additionally, the levels of free proline and anthocyanins significantly increased in response to high temperature, with a substantially higher increase in the 7 d (30 °C) treatment. The results indicate that the accumulation of proline, anthocyanins, and furanocoumarins, but not of phenolic acids or flavonols, contributes to protection of H. sosnowskyi plants against heat stress. Further studies could focus on the suppression of these metabolites to suppress the spread of this invasive species.

Daily consumption of caffeinated beverages is considered safe but serious health consequences do happen in some individuals. The Apiaceous and Rutaceae families of plant (ARFP) products are popular foods and medicines in the world. We previously reported significant amounts of furanocoumarin bioactive such as 8-methoxypsoralen, 5-methoxypsoralen, and isopimpinellin in ARFP products. As both caffeine and furanocoumarin bioactive are metabolized by the same hepatic CYP1A1/2 isozyme in humans, caffeine/ARFP product interactions may occur after co-administration. The objectives of the present study were to study in vivo loss of caffeine metabolizing activity by comparing the pharmacokinetics of caffeine in volunteers before and after pre-treatment with an ARFP extract, study the correlation between the decrease in hepatic CYP1A2 activity and the content of furanocoumarin bioactive in ARFP extracts, characterize CYP1A2 inactivation using in vitro incubations containing 14C-caffeine, a furanocoumarin bioactive, and human liver microsomes (HLMs), and provide a mechanistic explanation for both in vivo and in vitro data using the irreversible inhibition mechanism. The study results showed pre-treatment of volunteers with four ARFP extracts increased the area-under-the-concentration-time-curve (AUC0-inf) ratio of caffeine in the plasma ranging from 1.3 to 4.3-fold compared to the untreated volunteers indicating significant caffeine metabolism inhibition. The increases in AUC0-inf ratio also were linearly related to the effect-based doses of the furanocoumarins in the ARFP extracts, a finding which indicated caffeine metabolism inhibition was related to the content of furanocoumarin bioactive in an ARFP product. In vitro incubation studies also showed individual furanocoumarin bioactive were potent inhibitors of caffeine-N-demethylation; the IC50 for 8-methoxypsoralen 5-methoxypsoralen, and isopimpinellin were 0.09, 0.13, and 0.29 µM, respectively. In addition, CYP1A2 inactivation by individual furanocoumarin bioactive was concentration- and time-dependent involving the irreversible inhibition mechanism. The proposed irreversible inhibition mechanism was investigated further using 14C-labeled 8-methoxypsoralen and HLMs. The formation of 14C-adducts due to 14C-8-MOP-derived radioactivity bound to HLMs confirmed the irreversible inhibition of CYP1A2 activity. Thus, furanocoumarin bioactive metabolism in humans would result in reactive metabolite(s) formation inactivating CYP1A2 isozyme and inhibiting caffeine metabolism. Once the CYP1A2 isozyme was deactivated, the enzymic activity could only be regained by isozyme re-synthesis which took a long time. As a result, a single oral dose of ARFP extract administered to the human volunteers 3.0 h before still was able to inhibit caffeine metabolism.

(1) Cisplatin (CDDP) is used in melanoma chemotherapy, but it has many side effects. Hence, the search for natural substances that can reduce the dose of CDDP, and CDDP-related toxicity, is highly desired. Coumarins have many biological properties, including anticancer and antiproliferative effects. (2) An in vitro 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay on two human melanoma cell lines (FM55P and FM55M2) examined the antitumor properties of CDDP and five naturally occurring coumarins (osthole, xanthotoxin, xanthotoxol, isopimpinellin, and imperatorin). The antiproliferative effects produced by combinations of CDDP with the coumarins were assessed using type I isobolographic analysis. (3) The most potent anticancer properties of coumarins were presented by osthole and xanthotoxol. These compounds were characterized by the lowest median inhibitory concentration (IC50) values relative to the FM55P and FM55M2 melanoma cells. Isobolographic analysis showed that for both melanoma cell lines, the combination of CDDP and osthole exerted synergistic and additive interactions, while the combination of CDDP and xanthotoxol exerted additive interactions. Combinations of CDDP with xanthotoxin, isopimpinellin, and imperatorin showed antagonistic and additive interactions in two melanoma cell lines. (4) The combination of CDDP and osthole was characterized by the most desirable synergistic interaction. Isobolographic analysis allows the selection of potential candidates for cancer drugs among natural substances.

Photochemotherapy is one of the most interesting current therapeutic approaches for the treatment of melanoma. Different classes of naturally occurring phytochemicals demonstrated interesting photoactive properties. The aim of this study was to evaluate the photocytotoxic potential of two Cachrys species from Southern Italy: C. sicula and C. libanotis (Apiaceae). The enriched-coumarin extracts were obtained from aerial parts through both traditional maceration and pressurized cyclic solid-liquid (PCSL) extraction using Naviglio extractor&reg, Qualitative and quantitative analyses of furanocoumarins were performed with GC-MS. The photocytotoxic effects were verified on C32 melanoma cells irradiated at a dose of 1.08 J/cm2. The apoptotic responses were also assessed. Moreover, phenolic content and the in vitro antioxidant potential were estimated. Xanthotoxin, bergapten, and isopimpinellin were identified. All the samples induced concentration-dependent photocytotoxic effects (IC50 ranging from 3.16 to 18.18&thinsp, &mu, g/mL). The C. libanotis sample obtained with Naviglio extractor&reg, was the most effective one (IC50 = 3.16&thinsp, &plusmn, 0.21 &mu, g/mL), followed by C. sicula sample obtained with the same technique (IC50 = 8.83 &plusmn, 0.20 &mu, g/mL). Both Cachrys samples obtained through PCSL induced up-regulation of apoptotic signals such as BAX (Bcl2-associated X protein) and PARP (poly ADP-ribose polymerase) cleavage. Moreover, these samples proved to be more photoactive, giving a greater upregulation of p21 protein in the presence of UVA radiation. Obtained results suggest that investigated species could be promising candidates for further investigations aimed to find new potential drugs for the photochemotherapy of skin cancer.

Melanoma is the most aggressive form of skin cancer. Photochemotherapy, combining the action of a light source and a chemical photosensitizer, is one of the most interesting current therapeutic approaches. Plants represent a rich source of photoactive compounds, and furanocoumarins are some of the most important naturally occurring phytoconstituents. The aim of this study was to evaluate the photocytotoxic potential of Cachrys libanotis L. (Apiaceae) from Southern Italy. This species belongs to a genus rich in furanocoumarins and widely distributed in Europe. The aerial parts were extracted through both traditional maceration and pressurized cyclic solid-liquid (PCSL) extraction using Naviglio extractor®. Qualitative and quantitative analyses were performed to detect the coumarins content using GC-MS, and the photocytotoxic effects of the extracts were assessed on UVA-irradiated C32 melanoma cells. The apoptotic responses were also evaluated. Furthermore, phenolic content and the in vitro antioxidant potential were also estimated. Xanthotoxin, bergapten and isopimpinellin were identified and quantified. Both extracts affected cell viability in a concentration-dependent manner after irradiation for 1 hour at a dose of 1.08 J/cm2. Sample obtained through PCSL extraction was the most effective, with an IC50 equal to 3.16 μg/mL, a very interesting value if compared with the positive control bergapten. This extract induced up-regulation of apoptotic signals such as BAX and PARP cleavage and, in the presence of UVA radiation, it caused a greater upregulation of p21 protein. Obtained results suggest that investigated species could be a good candidate for further studies aimed to find new drugs with photocytotoxic potential.

Ventilago denticulata is an herbal medicine for the treatment of wound infection, therefore this plant may rich in antibacterial agents. UHPLC-ESI-QTOF-MS/MS-Based molecular networking guided isolation and dereplication led to the identification of antibacterial and antifungal agents in V. denticulata. Nine antimicrobial agents in V. denticulata were isolated and characterized, they are divided into four groups including (I) flavonoid glycosides, rhamnazin 3-rhamninoside (7), catharticin or rhamnocitrin 3-rhamninoside (8), xanthorhamnin B or rhamnetin 3-rhamninoside (9), kaempferol 3-rhamninoside (10) and flavovilloside or quercetin 3-rhamninoside (11), (II) benzisochromanquinone, ventilatones B (12) and A (15), (III) a naphthopyrone ventilatone C (16) and (IV) a triterpene lupeol (13). Among the isolated compounds, ventilatone C (16) was a new compound. Moreover, kaempferol, chrysoeriol, isopimpinellin, rhamnetin, luteolin, emodin, rhamnocitrin, ventilagodenin A, rhamnazin and mukurozidiol, were tentatively identified as antimicrobial compounds in extracts of V. denticulata by a dereplication method. MS fragmentation of rhamnose-containing compounds gave an oxonium ion, C6H9O3+ at m/z 129, while that of galactose-containing glycosides provided the fragment ion at m/z 163 of C6H11O5+. These fragment ions may be used to confirm the presence of rhamnose or galactose in mass spectrometry-based analysis of natural glycosides or oligosaccharide attached to biomolecules, that is, glycoproteins.

Background New agents for developing alternative or complementary medicine to treat the hepatitis C virus (HCV) are still needed due to high rates of HCV infection globally and the current limitations of available treatments. Treatment of HCV with a combination of direct acting antivirals have been shown to be approximately 90% effective but will be limited in the future due to the emergence of drug resistance and high cost. The leaves of Melicope latifolia have previously been reported to have anti-HCV activity and are a potential source of bioactive compounds for future novel drug development. This study aimed to evaluate the efficacy of the extract of M. latifolia fruit to treat HCV and to isolate its active compounds. Method M. latifolia fruit was extracted using methanol and purified using vacuum liquid chromatography (VLC) and Radial Chromatography. The anti-HCV activity was analyzed using cell culture lines Huh7it-1 and JFH1 (genotype 2a). Time-of-addition and immunoblotting studies were performed to identify the mode of action of the isolated active compounds. The structures of the active compounds were determined using nuclear magnetic resonance (NMR) spectra, UV, IR, and Mass Spectra. Results Six known compounds were isolated from M. latifolia fruit: O-methyloktadrenolon, alloevodionol, isopimpinellin, alloxanthoxyletin, methylevodionol, and N-methylflindersine. N-methylflidersine was the most active compound with IC50 value of 3.8 μg/ml while methylevodionol, isopimpinellin, and alloevodionol were less active. O-methyloktadrenolon and alloxanthoxyletin were moderately active with IC50 values of 10.9 and 21.72 μg/ml, respectively. N-methylflidersine decreased level of HCV NS3 protein expression in the cells. Conclusion The alkaloid compound, N-methylflindersine which was isolated from M. latifolia possesses anti-HCV activity through post-entry inhibition and suppressed NS3 protein expression.

Toddalia asiatica L., a significant medicinal plant in the family Rutaceae, has been applied to treat rheumatoid diseases for decades. Its pharmacological activities are mainly attributed to the existence of generous coumarins and alkaloids; however, the pharmacokinetics of Toddalia asiatica L. remain unclear. A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for the simultaneous determination of four coumarins, three alkaloids and one flavonoid (hesperidin, nitidine chloride, chelerythrine, toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine) in rat feces as well as four coumarins and one alkaloid (toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine) in rat plasma and urine. Chromatographic separation was accomplished on an Agilent ZORBAX SB-C18 column (2.1 × 150 mm, 5 μm) with acetonitrile (containing 0.1% formic acid) and 5 mmol/L aqueous ammonium formate for gradient elution. A correlation coefficient greater than 0.9925 reflected the excellent linearity of the analytical response. The lower limits of quantification were 30.0, 10.0, 10.0, 30.0, 5.0, 10.0, 2.5 and 2.5 ng/mL for hesperidin, nitidine chloride, chelerythrine, toddalolactone, isopimpinellin, pimpinellin, bergapten and dictamnine, respectively. The intra- and inter-day precision were less than 12.7%, and the accuracy was between -11.8% and 12.9%. In summary, this study is the first to study the pharmacokinetics and excretion of T. asiatica extract after oral administration, which may provide a scientific basis for its clinical applications.

Rhizoma corydalis and Radix Angelicae Dahurica (Yuanhu–Baizhi) herbal medicine pair has been used for thousands of years and has been reported to be potentially active in recent cancer therapy. But the exact active components or fractions remain unclear. In this study, a new comprehensive two-dimensional (2D) 3-aminopropyltriethoxysilane (APTES)-decorated MCF7-cell membrane chromatography (CMC)/capcell-C18 column/time-of-flight mass spectrometry system was established for screening potential active components and clarifying the active fraction of Yuanhu–Baizhi pair. APTES was modified on the surface of silica, which can provide an amino group to covalently link cell membrane fragments with the help of glutaraldehyde in order to improve the stability and column life span of the MCF7 CMC column. The comprehensive 2D MCF7-CMC system showed good separation and identification abilities. Our screen results showed that the retention components are mainly from the alkaloids in Yuanhu (12 compounds) and the coumarins (10 compounds) in Baizhi, revealing the active fractions of Yuanhu–Baizhi herbal medicine pair. Oxoglaucine, protopine, berberine, osthole, isopimpinellin and palmitic acid were selected as typical components to test the effects on cell proliferation and their IC50 were calculated as 38.17 μM, 29.45 μM, 45.42 μM, 132.7 μM, 156.8 μM and 90.5 μM respectively. Cell apoptosis assay showed that the drug efficacy was obtained mainly through inducing cell apoptosis. Furthermore, a synergistic assay results demonstrated that oxoglaucine (representative of alkaloids from Yuanhu) and isopimpinellin (representative of coumarins from Baizhi) showed significant synergistic efficacy with GFT, indicating that these components may act on other membrane receptors. The proposed 2D CMC system could also be equipped with other cells for further applications. Besides, the follow-up in-vitro experimental strategy using cell proliferation assay, cell apoptosis assay and synergistic assay proved to be a practical way to confirm the active fractions of herbal medicine. Keywords: Cell membrane chromatography, Comprehensive two-dimensional chromatography, Herbal medicine, Rhizoma corydalis and Radix Angelicae Dahurica, Anti-breast-cancer components

We previously showed rats fed with apiaceous vegetables, but not with their putative chemopreventive phytochemicals, reduced colonic DNA adducts formed by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), a dietary procarcinogen. We report here the effects of feeding apiaceous and cruciferous vegetables versus their purified predominant phytochemicals, either alone or combined, on prostate and pancreatic PhIP-DNA adduct formation. In experiment I, male Wistar rats received three supplemented diets: CRU (cruciferous vegetables), API (apiaceous vegetables), and CRU+API (both types of vegetables). In experiment II, rats received three diets supplemented with phytochemicals matched to their levels in the vegetables from experiment I: P + I (phenethyl isothiocyanate and indole-3-carbinol), FC (furanocoumarins; 5-methoxypsoralen, 8-methoxypsoralen, and isopimpinellin), and COMBO (P + I and FC combined). After 6 days of feeding, PhIP was injected (10 mg/kg body weight) and animals were killed on day 7. PhIP-DNA adducts were analyzed by LC-MS/MS. In prostate, PhIP-DNA adducts were reduced by API (33%, P

Toddalia asiatica (L.) Lam. (Rutaceae) has shown a broad spectrum of biological properties, such as anti-inflammatory, antioxidant, antimicrobial, anti-HIV, and anticancer properties. The present study is concerned with the separation of the main components with broad partition coefficients (KD values) from T. asiatica, using linear gradient high-speed counter-current chromatography (LGCCC) combined with an off-line two-dimensional (2D) mode. Similar to the binary gradient HPLC, the LGCCC mode is operated by the adjustment of the proportion between the mobile phase of 5:5:1:9 (v/v) (pump A) and 5:5:4.5:5.5 (v/v) (pump B) in an n-hexane/ethyl acetate/methanol/water solvent system. The off-line 2D-CCC mode was used in this study for the secondary separation of two similar KD value compounds with n-hexane/ethyl acetate/methanol/water (5:5:4:6, v/v). Notably, six coumarins, namely, tomentin (1), toddalolactone (2), 5,7,8-trimethoxycoumarin (3), mexoticin (4), isopimpinellin (5), and toddanone (6), were efficiently separated. The structures of the pure compounds were elucidated by spectral techniques and compared with the literature.

Mutual adulterations of lemon and lime juices may be detected using coumarins and psoralens as markers. Poor manufacturing practices or legal but mechanically intense processing of lemons were recently suspected to lead to false accusations of deliberate adulterations with lime juices due to potentially unspecific markers. Therefore, we studied coumarin and psoralen profiles in carefully dissected flavedo, albedo, and endocarp of lime and lemon as well as in juices produced under variable mechanical stresses at laboratory and pilot plant scale. Although the marker herniarin was detectable in juices from lime and harshly extracted lemons at low levels, isopimpinellin, bergapten and the herein proposed, tentatively assigned 5-geranyloxy-8-methoxypsoralen represented unambiguously lime-specific markers. Coumarin and psoralen data also allowed differentiating juices produced at differing degrees of mechanical stress. The latter was also possible using quantitative 1H-NMR spectroscopy, which yielded best results when combined with HPLC data on coumarins and psoralens. In the future, the reported approach may be used for establishing a robust database prior to being used in industrial practice.

The concentration of furocoumarins, known to induce mutagenesis, carcinogenesis and photodermatitis under UV radiation, is limited by the EU cosmetic regulation in consumer products such as cosmetics, sun protection and bronzing formulations. The bergapten concentration is also limited by the International Fragrance Association. Analytical methods have been proposed for their quantification using high-performance liquid chromatography (HPLC) coupled with UV detection or mass spectrometry with limits of detection (LOD) and quantification (LOQ) in accordance with the regulation. These methods are time and solvent consuming for routine quality control analyses. In this context, simple and fast HPLC methods in one- (1-D) and two- (2-D) dimensions were developed for the simultaneous analysis of the 15 most common furocoumarins reported in Citrus essential oils: psoralen, bergapten, xanthotoxin, isopimpinellin, oxypeucedanin, oxypeucedanin hydrate, byakangelicol, byakangelicin, heraclenin, 8-geranyloxypsoralen, bergamottin, imperatorin, isoimperatorin, phellopterin, epoxybergamottin along with five other furocoumarins known to be present in Angelica, bergamot or Heracleum species: angelicin, bergaptol, isobergapten, psora-4 and 5-(4-phenoxybutoxy)psoralen. The 1-D method was validated for the quantification of bergapten using a Camag TLC Scanner as a detector. The LOD was determined at 2 mg/kg. The method was linear from 2 to 100 mg/kg and the method range was defined using accuracy profiles at with an 80% tolerance interval, with a 30% error, to be 5–100 mg/kg in acetonitrile and 13–100 mg/kg in matrix.

Allergic inflammation is induced by allergens and leads to various allergic diseases, including rhinitis, asthma and conjunctivitis. Histamine is important in the pathogenesis of an immunoglobulin E-dependent allergic reaction and results in the secretion of cytokines associated with inflammation. Angelica dahurica (A. dahurica) is a medicinal plant widely used in China for the treatment of symptoms related to allergic inflammation. The present study investigated the chemical constituents from A. dahurica and evaluated their reductive effect on allergic inflammation. As a result, 15 compounds including 13 coumarins have been identified as isoimperatorin (1), imperatorin (2), oxypeucedanin (3), oxypeucedanin hydrate (4), bergapten (5), byakangelicin (6), phellopterin (7), byakangelicol (8), isopimpinellin (9), xanthotoxol (10), xanthotoxin (11), pimpinellin (12), scopoletin (13), β-sitosterol (14) and daucosterol (15). Compounds 1-13 were able to reduce the release of histamine, with compounds 4-6 exhibiting the most potent activity. Furthermore, compounds 1-12 were able to inhibit the secretion of tumor necrosis factor-α, interleukin (IL)-1β and IL-4, with compounds 5 and 7 exhibiting the strongest inhibitory effects. These compounds implemented the inhibitory effects on the expression of inflammatory cytokine genes through the inhibition of nuclear factor-κB activation. Virtual screening by a docking program indicated that compound 3 is a potent histamine H1 receptor antagonist. Additionally, the calculated physicochemical properties of these compounds support most furanocoumarins to be delivered to binding sites and permeate the cell membrane. The present findings contribute to understanding how A. dahurica attenuates allergic inflammation.

The first occurrences and dissemination of resistant microorganisms led to the inefficacy of many antibiotics, available in the market nowadays, therefore, the search for new substances with antimicrobial activity from natural sources has gained a great importance. The purpose of this work is to evaluate the antibacterial activity and modulation of drug resistance in Staphylococcus aureus by coumarins such as bergapten, xantotoxin, isopimpinellin and imperatorin obtained from two Rutaceae species (Metrodorea mollis and Pilocarpus spicatus). The antimicrobial activity was assessed based on the minimum inhibitory concentration (MIC), using the microdilution method. The MIC was >256 g/mL for all coumarins tested. Regarding the modulation of drug resistance assay, the isopimpinellin reducted the MIC of erytromicin by 4 times, whereas imperatorin exhibited the best result, reducing the MIC of tetracycline (2 times), erytomicin (4 times) and norfloxacin (4 times). By reducing the MIC of ethidium bromide, the imperatorin is consider in fact, as a putative efflux pump inhibitor of bacteria.

Seeds are classified as stimulatory, carminative, anti-inflammatory and extremely useful for vitiligo and leucoderma. Powdered seeds mixed with honey, that has been heated to remove scum, 10 g daily with warm water for fifteen days are claimed to cure vitiligo. It is also used as an emmenagogue to regulate menstruation, as a diuretic, and for the treatment of leprosy, kidney stones, and urinary tract infections. Since ancient times, a tea made from the fruit of Ammi visnaga (khella, as it is known in the Arab world) has been used as a remedy for renal colic due to kidney stones. In Israel and Morocco, the plant is used for the treatment of diabetes. Natural coumarins (furocoumarins), isopimpinellin, xanthotoxin (8-methoxypsoralen, 8-MOP), imperatorin, bergapten, umbelliprenin, maurin, alloimperatorin, ammirin, ammajin and marmesin, have been isolated from the fruits of A. majus. Khellin, visnagin, and khellol glycoside have been isolated from fruits of A. visnaga. 8-MOP is a purified extract of the root, that has been used in a crude form for centuries in the Middle East in the treatment of various skin diseases, and was first utilized in the treatment of vitiligo in 1948. Ethanol seed extract decreased TC, TGs and LDL-C, and increased HDL-C of rats, and also produced significant anti-inflammatory, analgesic, and antipyretic effects. Both a single oral dose and repeated administration of aqueous extract of A. visnaga significantly lowers blood glucose in normal and diabetic rats. Spontaneous passage of multiple ureteral stones in a patient, possibly as a result of using a khellin preparation was reported from Turkey.

An annual herb, a native of Egypt, but cultivated in Iran, India, Mediterranean region, and Europe. Ibn al-Baitar, quoting Dioscorides, said that the fruits (seeds) are diuretic, carminative, analgesic, anti-inflammatory, diaphoretic, galactagogue, and aphrodisiac. According to Razi, it is useful in ascites and relieves flatulence and borborygmus, and Avicenna called Anisun deobstruent for kidney, bladder, liver and uterine obstructions, useful in chronic fevers and swelling of face and feet. Ibn Jazlah described Anisun as fennel seeds, with a sweet taste, causes constipation, and is used to prevent obstruction of liver, and as antidote against poisons. Ibn Buţlān used it for cold stomach, against flatulence, and considered it diuretic, to relieve constipation, to increase postpartum milk production, and to improve eyesight. In Unani medicine, the fruits are included in diets of patients suffering from paralysis, facial palsy and epilepsy. Anise water and oil are topically applied to head in headache and to abdomen in flatulence and intestinal colic. Anise was not known to Hindu physicians and was introduced in India by Muslim physicians from Persia. In Iranian traditional medicine, the fruits are used as carminative, aromatic, disinfectant, antimicrobial, galactagogue, antioxidant, anticonvulsant, analgesic and muscle relaxant, and anise oil for treatment of convulsions. In Cayenne and French Guiana, whole plant is used as carminative and vermifuge, while the flowers and seeds decoctions are used for gas pain by Guyana Patamona people. The plant is used for respiratory disorders in Upper Egypt. In addition to anethol-glycol, creosol, anethol, acetaldehyde, isoamylamine, umbelliferone, bergaptene, isopimpinellin, isobergaptene and sphondin, anise seeds (fruit) contain protein, fat, carbohydrate, fiber, and minerals. The flavonoids reported include, quercetin-3-glucoronide, rutin, luteolin 7-glucoside, isoorientin, isovitexin, apigenin 7-glucoside, and a luteolin glycoside. Aqueous and ethanol extracts significantly increased milk production in rats by 68 and 81%, respectively. The weight gain by pups during the study period was correlated with the milk production. Aqueous suspension of powdered seeds (fruit) exhibited significant cytoprotective and antiulcer activities against gastric lesions. In a double-blind RCT of patients with functional dyspepsia, symptoms and quality of life were highly significantly improved after 12-weeks of treatment with anise powder.

Deverra tortuosa (Desf.) DC (Syn. Pituranthos tortusus (Desf.) Benth. & Hook.f.) is a species belonging to the Apiaceae family that is common in the Northern Region of Saudi Arabia. The plant is well known in traditional medicine along the Arabian ecoregion. In the framework of the present study, the crude extract of n-hexane and ethyl acetate of the seeds were fractionated to purify major bioactive secondary metabolites. Five compounds were identified for the first time from the seeds of D. tortuosa: Marmin 1, Pituranthoside 2, Isoimperatorin 3, Bergapten 4 and Isopimpinellin 5. Their structures were elucidated using 1D and 2D NMR, (ESI)-MS and IR spectroscopic analyses. The cytotoxic, α-glucosidase and antibacterial activities of the pure phytochemicals were also evaluated., Deverra tortuosa; Marmin; Furanocoumarins; cytotoxic activity; α-glucosidase inhibition; Antibacterial activities.

As a part of ongoing project on biological screening of about a hundred plant extracts from Iran, the significant inhibitory effect of n-hexane extract of roots of Heracleum persicum on α-glucosidase and its 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity were revealed. The present study was designed to evaluate the antidiabetic and antioxidant activities of various extracts of roots of Heracleum persicum and isolation of ten furanocoumarin and a rare furanocoumarin-dimer type compounds through “bioassay guided fractionation” method. The structures were elucidated by 1D and 2D NMR, EI-MS, and HRESI-MS data as psoralen (1), bergapten (2), xanthotoxin (3), isopimpinellin (4), angelicin (5), isobergapten (6), sphondin (7), pimpinellin (8), heratomin (9), 5-methoxyheratomin (10), moellendorffiline (11), and fraxetin (12). Moellendorffiline (11) with the IC50 value of 17.9 nM showed significant inhibitory activity against α-glucosidase, more potent than acarbose (IC50 = 23.5 nM) and also exhibited high antioxidant activity (IC50 = 0.2 μM), compared with butylated hydroxytoluene (IC50 = 0.1 μM).

Coumarins are a group of phytochemicals that may be beneficial or harmful to health depending on their type and dosage and the matrix that contains them. Some of these compounds have been proven to display pro-oxidant and clastogenic activities. Therefore, in the current work, we have studied the coumarins that are present in food sources extracted from the Phenol-Explorer database in order to predict their clastogenic activity and identify the structure-activity relationships and genotoxic structural alerts using alternative methods in the field of computational toxicology. It was necessary to compile information on the type and amount of coumarins in different food sources through the analysis of databases of food composition available online. A virtual screening using a clastogenic model and different software, such as MODESLAB, ChemDraw and STATISTIC, was performed. As a result, a table of food composition was prepared and qualitative information from this data was extracted. The virtual screening showed that the esterified substituents inactivate molecules, while the methoxyl and hydroxyl substituents contribute to their activity and constitute, together with the basic structures of the studied subclasses, clastogenic structural alerts. Chemical subclasses of simple coumarins and furocoumarins were classified as active (xanthotoxin, isopimpinellin, esculin, scopoletin, scopolin and bergapten). In silico genotoxicity was mainly predicted for coumarins found in beer, sherry, dried parsley, fresh parsley and raw celery stalks. The results obtained can be interesting for the future design of functional foods and dietary supplements. These studies constitute a reference for the genotoxic chemoinformatic analysis of bioactive compounds present in databases of food composition.

The combination of Glehniae radix (Beishashen, GR) and Ophiopogonis radix (Maidong, OR) has long been widely used as a traditional Chinese medicine (TCM) herb pair for the treatment of respiratory system and digestive system diseases. The pharmacokinetic interaction mechanism of these two herbs is still unclear. The aim of the study is to elucidate the potential herb–herb pharmacokinetic interaction between Glehniae radix and Ophiopogonis radix in rats. Three groups of Sprague-Dawley rats (n = 6) were given GR (20 g kg−1), OR (20 g kg−1) and GR–OR (20 + 20 g kg−1), respectively. To study the pharmacokinetic characteristics, nine components were determined in plasma with a newly developed superimposed multiple product ion (SMPI) liquid chromatography high resolution tandem mass spectrometry (LC-HR-MS/MS) method. The SMPI LC-HR-MS/MS method displayed high specificity and sensitivity. Co-administration of GR with OR could significantly increase the area under the concentration–time curve (AUC) of the nine analytes compared with the administration of a single herb (GR or OR). There were no obvious changes for the MRT0−∞ of the nine analytes except that the value for psoralen increased. The t1/2 values of the nine analytes did not display obvious differences between the GR–OR group and single herb (GR or OR) groups. The Cl/F of the nine analytes significantly decreased in the GR–OR group compared with that in the single herb (GR or OR) groups except for that of isopimpinellin. The present study showed that the GR–OR herb pair had an influence on the pharmacokinetic behaviors of the nine compounds, which could increase the bioavailabilities of the nine analytes, indicating that the co-administration of GR with OR can cause significant herb–herb pharmacokinetic interaction in rats. In addition, this study provides pharmacokinetic experimental evidence for the compatibility (Peiwu) study of the GR–OR herb pair.

In the present study, the chemical composition of the essential oil from aerial parts of Ononis reclinata L., a species not previously investigated, collected in Sicily was evaluated by GC and Gas chromatography-Mass spectrometry. The main components of O. reclinata were menthone (43.6%), isopimpinellin (38.4%) and pulegone (11.2%). The comparison with other studied oils of genus Ononis showed a peculiar and characteristic profile.

Ferulago carduchorum (Apiaceae) is an endemic plant of Iran. From the hexane and ethyl acetate extracts of F. carduchorum seven coumarins, one flavonoid and one steroid were isolated using column chromatography with silica gel and Sephadex LH20 as the stationary phases. Antimicrobial activity of the isolated compounds was examined by a broth microdilution method. Acetylcholinesterase (AChE) inhibitory activity of isolated coumarins was also investigated. The isolated compounds were identified as suberosin, suberenol, bergapten, xanthotoxin, isopimpinellin, prantschimgin, β-sitosterol and hesperetin by comparison of their NMR and MS spectral data with those reported in the literature. Evaluation of the minimum inhibitory concentration (MIC) for each compound showed that hesperetin (flavonoid) was the most potent antimicrobial agent against a gram-positive bacterium (Staphylococcus aureus) and among the coumarins, bergapten had the best activity against S. aureus and Candida albicans. All coumarins inhibited AchE enzyme, in which xanthotoxin showed the most inhibitory among them (IC50 = 39.64 µM). Our results indicate that isolated coumarins are effective against the tested bacterial strains and have AchE inhibitory activity suggesting their potential for commercial applications.

The results of the study of extracts from the entire aerial part of the Sosnovsky Hogweed (Heracleum sosnowskyi, collected in the flowering phase, before and after the electric-discharge cavitation treatment using gas chromatography and chromatography-mass spectrometry) are presented. both aromatic and non-aromatic structure not characteristic of the native juice H. sosnowskyi. In the native juice of the plant identified from furanocumrins: metaxalene, isopimpinellin and angelicin. When electrically discharge cavitating the sap of a plant, these furanocoumarins are completely destructed. 3 benzyl alcohol. Among the organic acids in the juice of H. sosnowskyi after the electric discharge cavitation effect, 2-hydroxy-propionic and succinic acids are the absolute dominant. Depending on the phase of plant development, in which plant materials were harvested and the processing conditions of H. sosnowskyi juice, the component composition of organic substances in the final product can vary greatly, which is confirmed by the detection of robust acid in the second batch of H. sosnowskyi juice. Given the high content of succinic acid in the juice of H. sosnowskyi, subjected to cavitation effects, the authors consider it expedient to investigate in the future the growth-promoting properties of the obtained product when growing agricultural crops., Приведены результаты исследования экстрактов из всей надземной части борщевика Сосновского (Heracleum sosnowskyi), собранной в фазу цветения, до и после электроразрядной кавитационной обработки методами газовой хроматографии и хромато-масс спектрометрии. Установлено, что при кавитационном воздействии достигается селективная деструкция кумаринов и фуранокумаринов с образованием ряда соединений как ароматического, так и неароматического строения не характерных для нативного сока H. sosnowskyi. В нативном соке растения идентифицированы из фуранокумаринов: метаксален, изопимпинеллин и ангелицин. При электроразрядной кавитационной обработке сока растения указанные фуранокумарины полностью деструктируют. Продуктами деструкции фуранокумаринов являются: 2-фенил-этанол, 3-фенил-пропанол, 2-гидрокси-5-метил-ацетофенон, бутандиол-2,3 бензиловый спирт. Среди органических кислот в соке H. sosnowskyi после электроразрядного кавитационного воздействия, абсолютным доминатом являются 2-гидрокси-пропионовая и янтарная кислоты. В зависимости от фазы развития растения, в которую собрано было растительное сырье и условий обработки сока H. sosnowskyi, компонентный состав органических веществ в конечном продукте может сильно варьировать, что подтверждается обнаружением робустовой кислоты во второй партии сока H. sosnowskyi. Учитывая высокое содержание янтарной кислоты в соке H. sosnowskyi, подвергнутого кавитационному воздействию, авторы считают целесообразным исследовать в будущем ростостимулирующие свойства полученного продукта при выращивании сельскохозяйственных культур.

A sensitive and specific ultra-performance liquid chromatographic-tandem mass (UPLC-MS/MS) spectrometric method was established to investigate tissue distribution of fourteen coumarins of Angelica Dahurica cv. Yubaizhi roots (ADYR) in rat tissues, including isoimperatorin (1), imperatorin (2), isooxypeucedanin (3), byakangelicin (4), oxypeucedanin hydrate (5), bergapten (6), 2"R-neobyakangelicol (7), phellopterin (8), xanthotoxin (9), isopimpinellin (10), oxypeucedanin ethanolate (11), isobyakangelicol (12), columbianetin (13), (-)-marmesin (14). Detection was performed on a triple quadrupole mass spectrometer in multiple-reaction-mode (MRM). The method established in this assay was successfully applied to tissue distribution study of the selected 14 coumarins after oral administration of the extract of ADYR in rat tissues, including heart, liver, spleen, lung, kidney, stomach, small intestine, muscle, testis, and brain. Tissue distribution characteristics of the fourteen coumarins were clearly elucidated, and the results of this study indicated that the fourteen coumarins were distributed to rat tissues rapidly and could be detected in all of the selected tissues after oral administration. Concentrations of the coumarins were obviously higher in kidney, liver and stomach tissues, and lower in testis, brain and muscle tissues. As an important part of ADMET/Act. study on ADYR, the tissue distribution of multiple coumarins of ADYR in rats provides a significant basis for better evaluation of the metabolism and disposition process in vivo of the herb medicine. The information provided in this research is very useful for further understanding of the metabolic mechanism of ADYR in vivo.

Although the wild parsnip (Pastinaca sativa L. s.l.) fruits are known to contain linear and angular furanocoumarins, the individual components of the seeds have not been fully identified and quantitated, and, in the case of immature seeds, reported. In view of this, the main furanocoumarin compounds were extracted using pyridine, and were isolated using semi-preparative high-performance liquid chromatography. The structural elucidation of isolated compounds was done based on detailed spectral analysis conducted by liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI/MS), 1H and 13C NMR and, where possible, by gas chromatography–mass spectrometry (GC–MS). The quantitative analysis of furanocoumarin compounds in the wild parsnip was conducted by analytical ultra–performance liquid chromatography (UPLC–DAD), calculated against the standard curves of isolated compounds. The total yields of furanocoumarin compounds from the seeds after extraction with pyridine were 107.2–222.8 mg g−1 (fresh weight) and 50.2–66.4 mg g−1 (soluble dry matter). Thirteen furanocoumarins were identified. The main compounds (percentage in FW) in immature seeds were bergapten (40.8), pimpinellin (10.5), methoxsalen (5.7), isopimpinellin (4.3), imperatorin (3.2), and phellopterin (7.2). Seven constituents previously not described in P. sativa seeds and its products were identified, namely, byakangelicol (14.4), heraclenin (8.5), isobergapten (2.5), byakangelicin (1.3), heraclenol (0.5), psoralen (0.3), and isobyakangelicin (0.8). The latter is a new compound of the Apiaceae family. Extraction of immature seeds using pyridine gave a much higher yield and a greater variety of furanocoumarins. This indicates that the wild parsnip, along with other Apiaceae family plants, may be an important source of bioactive compounds.