isobavachalcone and bavachin

A method for the simultaneous quantification of 13 bioactive compounds (psoralen, isopsoralen, isobavachin, bakuchalcone, neobabaisoflavone, bavachin, corylin, psoralidin, isobavachalcone, bavachinin, corylifol A, bavachalcone, and bakuchiol) by ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry has been developed and validated in rat plasma. Osthol was used as an internal standard and plasma samples were pretreated with one-step liquid-liquid extraction. These analytes were separated using a gradient mobile phase system of water and acetonitrile at a flow rate of 0.2 mL/min on a reverse-phase C18 column and analyzed in the selected multiple reactions monitoring mode. All calibration curves were linear (r > 0.9952) over the tested ranges. The intra- and interday accuracy and precisions of these analytes at three different concentration levels were within the acceptable limits of <15% at all concentrations. The mean recoveries of these analytes at three concentrations were more than 60.2% and the matrix effects were in the range of 85-115%. Stability studies proved that the analytes were stable under the tested conditions. The developed method was applied to evaluating the pharmacokinetic study of 13 bioactive compounds after oral administration of Psoraleae Fructus in rat of different genders. Some active compounds in Psoraleae Fructus had sex-related pharmacokinetics.

Topics: Animals; Benzofurans; Chalcones; Chromatography, High Pressure Liquid; Coumarins; Female; Ficusin; Flavones; Flavonoids; Furocoumarins; Male; Mass Spectrometry; Molecular Structure; Phenols; Psoralea; Rats; Rats, Sprague-Dawley

Enhancing the surface area of stationary phase is essential in chromatographic science. In this work, nanoscale NiAl-layered double hydroxides (NiAl-LDHs) with flower-like structure was used as a platform for supporting the stationary phase. Then strong hydrophobic p-naphtholbenzein molecule was immobilized onto the LDHs layer as sorbent for stir bar sorptive extraction (SBSE). The flower-like LDHs layer significantly increased the extraction efficiency through increasing the specific surface area and immobilized amounts of stationary phase. In addition, the LDHs can also provide anion exchange ability, which expanded the application of this stir bar for analysis of not only hydrophobic but also anionic analytes. For improving the workability, a poly(ether ether ketone) (PEEK) jacket stir bar with detachable dumbbell-shaped structure was employed. The PEEK jacket with high mechanical strength and dumbbell-shaped structure improved the durability of stir bar and the detectable design allowed elution to be realized with less solvent that enhanced the enrichment factor. The proposed stir bar showed good performance for the extraction of multiple analytes including flavonoids, non-steroid anti-inflammatory drugs and chlorophenoxy acids. By coupling with high performance liquid chromatography-ultraviolet detection (HPLC-UV), the SBSE-HPLC-UV method was applied for the extraction of three active components including bavachin, isobavachalcone and bavachinin in Psoralea corylifolia L. herb with low limit detection of 0.01-0.02 ng/mL.

Topics: Adsorption; Chalcones; Chromatography, High Pressure Liquid; Flavonoids; Hydroxides; Limit of Detection; Naphthols; Psoralea; Reproducibility of Results; Solid Phase Extraction; Spectrophotometry, Ultraviolet

As an edible traditional Chinese herb, Fructus psoraleae (FP) has been widely used in Asia for the treatment of vitiligo, bone fracture and osteoporosis. Several cases on markedly elevated bilirubin and acute liver injury following administration of FP and its related proprietary medicine have been reported, but the mechanism in FP-associated toxicity has not been well investigated yet. This study aimed to investigate the inhibitory effects of FP extract and its major constituents against human UDP-glucuronosyltransferase 1A1 (UGT1A1), the key enzyme responsible for metabolic elimination of bilirubin. To this end, N-(3-carboxy propyl)-4-hydroxy-1,8-naphthalimide (NCHN), a newly developed specific fluorescent probe for UGT1A1, was used to evaluate the inhibitory effects of FP extract or its fractions in human liver microsomes (HLM), while LC-UV fingerprint and UGT1A1 inhibition profile were combined to identity and characterize the naturally occurring inhibitors of UGT1A1 in FP. Our results demonstrated that both the extract of FP and five major components of FP displayed evident inhibitory effects on UGT1A1 in HLM. Among these five identified naturally occurring inhibitors, bavachin and corylifol A were found to be strong inhibitors of UGT1A1 with the inhibition kinetic parameters (Ki) values lower than 1 μM, while neobavaisoflavone, isobavachalcone, and bavachinin displayed moderate inhibitory effects against UGT1A1 in HLM, with the Ki values ranging from 1.61 to 9.86μM. These findings suggested that FP contains natural compounds with potent inhibitory effects against human UGT1A1, which may be one of the important reasons for triggering FP-associated toxicity, including elevated bilirubin levels and liver injury.

Topics: Bilirubin; Chalcones; Chemical and Drug Induced Liver Injury; Dose-Response Relationship, Drug; Flavones; Flavonoids; Fruit; Glucuronosyltransferase; Humans; Isoflavones; Liver; Microsomes, Liver; Plant Extracts; Psoralea

Bavachin and isobavachalcone are the isomeric compounds in the fructus Psoraleae. The ion trap mass spectrometric fragmentation pathways of the bavachin and isobavachalcone in negative ion mode were elucidated for the identification. A novel method for determination of isomeric bavachin and isobavachalcone has been developed by capillary electrophoresis coupled with mass spectrometric detection. The effects of several factors such as concentration, pH of ammonium acetate buffer, separation voltage, composition and flow rate of the sheath liquid were investigated. Under optimal conditions, the linear concentration range for bavachin and isobavachalcone were 0.8-100 μg/mL with the correlation coefficient of 0.996 and 0.995, respectively. Relative standard deviations of migration time and peak areas were lower than 5%. The limits of detection (signal/noise = 3) were 60 ng/mL. The proposed method can be successfully applied to the determination of bavachin and isobavachalcone in the fructus Psoraleae and six fructus Psoraleae-containing preparations.

Topics: Chalcones; Electrophoresis, Capillary; Flavonoids; Isomerism; Mass Spectrometry; Plant Extracts

A simple, sensitive and selective method of high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed for simultaneous determination of bavachin and isobavachalcone in Fructus Psoraleae. At optimized conditions, bavachin and isobavachalcone could be well separated within 15 min at a detection potential of +0.80 V with 0.03 mol/L acetate buffer solution (pH 5.17)/acetonitrile (2:3, v/v) as the mobile phase. The relationships between peak areas and concentrations were linear from 8.26 × 10(-7) to 1.21 × 10(-4) mol/L for bavachin, and from 1.01 × 10(-8) to 1.61 × 10(-4) mol/L for isobavachalcone, respectively. The method offered excellent linearity with regression coefficient R(2) >0.995. The method presented detection limits (S/N = 3) of 8.81 × 10(-9) mol/L for bavachin and 1.17 × 10(-10) mol/L for isobavachalcone. It indicates that the sensitivity of electrochemical detection is ten times higher than that of diode array detection (DAD). The mean recoveries around 98% with a relative standard deviation less than 3.1% for the two analytes have been obtained. The proposed separation and detection procedures were successfully applied to the simultaneous determination of bavachin and isobavachalcone in traditional Chinese medicine.

Topics: Chalcones; Chromatography, High Pressure Liquid; Electrochemical Techniques; Fabaceae; Flavonoids; Limit of Detection; Plant Extracts

An EtOH extract of fruits of Piper longum was found to exhibit a potent inhibitory effect against alpha-melanocyte-stimulating hormone (alpha-MSH)-induced melanin production in B16 mouse melanoma cells. Bioassay-directed fractionation led to the isolation of prenylated phenolic compounds bakuchiol, bavachin, and isobavachalcone. These compounds and the crude extract of the fruits of P. longum may have suppressive effects against pigmentation by melanin in the skin.

Topics: Animals; Cell Line, Tumor; Cell Survival; Chalcones; Flavonoids; Melanins; Melanoma, Experimental; Mice; Phenols; Piper; Plant Extracts

Acyl-coenzyme A: cholesterol acyltransferase (ACAT) catalyzes cholesterol esterification and plays important roles in intestinal absorption of cholesterol, hepatic production of lipoproteins and accumulation of cholesteryl ester within macrophages and smooth muscle cells. Ethanol extract of Psoralea corylifolia showed a significant inhibition of ACAT enzyme. Via bioactivity-guided fractionation of the ethanol extract of Psoralea corylifolia, two prenylated flavonoids were isolated. Their structures were determined as bavachin (1) and isobavachalcone (2) by spectroscopic analysis ((1)H-, (13)C-NMR, 2DNMR, and ESI-MS). The IC(50) values were 86.0 (1) and 48.0 (2) microM in the ACAT assay system using rat liver microsome. Compound 2 also decreased cholesteryl ester formations in HepG2 cells. In addition, this compound showed a noncompetitive type of inhibition of ACAT.

Topics: Animals; Cell Line, Tumor; Chalcones; Cholesterol Esters; Dose-Response Relationship, Drug; Enzyme Inhibitors; Flavonoids; Humans; Indicators and Reagents; Kinetics; Liver; Magnetic Resonance Spectroscopy; Microsomes, Liver; Psoralea; Rats; Seeds; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Ultraviolet; Sterol O-Acyltransferase

Fructus Psoraleae, a widely used traditional Chinese medicine, is well known as a health supplement ingredient. In our study, an improved and comprehensive HPLC fingerprint of Fructus Psoraleae was established. Two important new benzofuran glycosides, psoralenoside and isopsoralenoside, were identified as characteristic constituents for the first time. HPLC separation was performed on an RP-C8 column. The mobile phase was acetonitrile and 0.1% acetic acid solution with linear gradient change of acetonitrile from 10 to 82% in 40 min. The flow rate was 1.0 mL/min, and the detection wavelength was set at 310 nm. The HPLC chromatograms of twenty-six samples from different regions of China showed a similar pattern. Twelve peaks were selected as characteristic peaks and further identified as psoralenoside, isopsoralenoside, psoralen, isopsoralen, bavachromene, corylifolin, corylin, psoralidin, isobavachalcone, bavachinin, corylifol A, and bakuchiol, respectively. Nine of them were simultaneously quantitatively analyzed for the first time. A more comprehensive analytical method was established for the fingerprint of Fructus Psoraleae. It is very useful for authentication and quality assessment of the crude drug.

Topics: Benzofurans; Chalcones; Chromatography, High Pressure Liquid; Coumarins; Drugs, Chinese Herbal; Ficusin; Flavonoids; Fruit; Furocoumarins; Glycosides; Magnetic Resonance Spectroscopy; Mass Spectrometry; Molecular Structure; Phenols; Psoralea; Solvents

A meroterpene and four flavonoids were isolated from the seeds of Psoralea corylifolia as antioxidative components. Their structures were elucidated by spectral data and identified as bakuchiol (1), bavachinin (2), bavachin (3), isobavachin (4) and isobavachalcone (5). In particular, meroterpene 1 and flavonoids 4 and 5 showed broad antioxidative activities in rat liver microsomes and mitochondria. They inhibited NADPH-, ascorbate-, t-BuOOH- and CCl(4)-induced lipid peroxidation in microsomes. They also prevented NADH-dependent and ascorbate-induced mitochondrial lipid peroxidation. Bakuchiol (1) was the most potent antioxidant in microsomes and the inhibition of oxygen consumption induced by lipid peroxidation was time-dependent. Furthermore, bakuchiol (1) protected human red blood cells against oxidative haemolysis. These phenolic compounds in P. corylifolia were shown to be effective in protecting biological membranes against various oxidative stresses.

Topics: Animals; Antioxidants; Chalcone; Chalcones; Erythrocytes; Flavonoids; Hemolysis; Humans; Lipid Peroxidation; Male; Microsomes, Liver; Mitochondria; Molecular Structure; Oxygen Consumption; Phenols; Plant Extracts; Psoralea; Rats; Rats, Wistar; Seeds