Valepotriates and valerenic acids in commercial preparations of valerian available in Australia

Thirty-one commercial valerian preparations available in Australia, including teas, tablets, capsules and liquids, were analysed by HPLC for valepotriates, valerenic acid and valerenic acid derivatives. The concentration of valerenic acid and its derivatives ranged from < 0.01 to 6.32 mg/g of product. Powder capsules, on average, contained the highest concentration of valerenic acids (2.46 mg/g) and liquids the lowest concentration (0.47 mg/ml). The mean concentration of valerenic acid in the five products standardized against valerenic acid (3.56 mg/g) was significantly higher than in the 26 non-standardized products (0.89 mg/g). There was a significant relationship between valerenic acids content and added valerian root for the standardized products but not for the non-standardised products. Valepotriates were found at low levels (< 1.0 mg/g) in some teas but were not detected in any of the finished products.     

Modulation of GABAA receptors by valerian extracts is related to the content of valerenic acid

Valeriana Officinalis L . is a traditionally used sleep remedy, however, the mechanism of action and the substances responsible for its sedative and sleep-enhancing properties are not fully understood. As we previously identified valerenic acid as a subunit-specific allosteric modulator of GABAA receptors, we now investigated the relation between modulation of GABAA receptors by Valerian extracts of different polarity and the content of sesquiterpenic acids (valerenic acid, acetoxyvalerenic acid). All extracts were analysed by HPLC concerning the content of sesquiterpenic acids. GABAA receptors composed of alpha 1, beta 2 and gamma 2S subunits were expressed in Xenopus laevis oocytes and the modulation of chloride currents through GABAA receptors (IGABA) by Valerian extracts was investigated using the two-microelectrode voltage clamp technique. Apolar extracts induced a significant enhancement of IGABA, whereas polar extracts showed no effect. These results were confirmed by fractionating a highly active ethyl acetate extract: again fractions with high contents of valerenic acid exhibited strong receptor activation. In addition, removal of sesquiterpenic acids from the ethyl acetate extract led to a loss of I (GABA) enhancement. In conclusion, our data show that the extent of GABAA receptor modulation by Valerian extracts is related to the content of valerenic acid.     

Hepatic Metabolism and Biliary Excretion of Valerenic Acid in Isolated Perfused Rat Livers: Role of Mrp2 (Abcc2)

The study was designed to investigate the hepatic metabolism and transport system of valerenic acid, a main active constituent of valerian, in isolated perfused livers from Wistar and Mrp2-deficient TR^? rats. After administration of 20 µM valerenic acid, the formation of seven valerenic acid glucuronides (M1–M7), namely two glucuronides of valerenic acid (M6, M7), four glucuronides of hydroxylated valerenic acid (M1, M3, M4, M5), and one glucuronide of hydroxylated dehydro-valerenic acid (M2) in bile and perfusate was quantified by HPLC. The hepatic extraction ratio and clearance of valerenic acid were very high in Wistar and TR^? rats (E: 0.983 ± 0.006 vs. 0.981 ± 0.004; Cl: 35.4 ± 0.21 mL/min vs. 35.3 ± 0.14 mL/min). However, biliary excretion and efflux of conjugates differed greatly in TR^? rats. While cumulative biliary excretion of unconjugated valerenic acid and the glucuronides M1–M7 dropped dramatically to 1–9%, their efflux into perfusate increased 1.5- to 10-fold. This indicates that valerenic acid and its glucuronides are eliminated into bile by Mrp2. In summary, valerenic acid was metabolized to several conjugates, whereby the canalicular transporter Mrp2 mediated biliary excretion of the parent drug and its glucuronides. © 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3839–3849, 2009     

Transport of a GABAA receptor modulator and its derivatives from Valeriana officinalis L. s. l. across an in vitro cell culture model of the blood-brain barrier

The roots and rhizome of Valeriana officinalis L . s. l. are therapeutically used for their sedative and sleep-enhancing effects. Some of the active compounds found in commonly used extracts are the sesquiterpenic acids, especially valerenic acid, which was recently identified as a GABA (A) receptor modulator. To interact with this receptor in the brain, substances such as valerenic acid and its derivatives acetoxyvalerenic acid and hydroxyvalerenic acid have to cross the blood-brain barrier (BBB). The aim of our study was to obtain BBB permeability data of these compounds for the first time and to elucidate possible transport pathways across our BBB in vitro model. Transport of valerenic acid, acetoxyvalerenic acid and hydroxyvalerenic acid was compared with the permeability of the GABA (A) modulator diazepam, which is known to penetrate into the central nervous system transcellularly by passive diffusion. Experiments were carried out with an established Transwell in vitro model based on the human cell line ECV304. Results indicated clearly that all three acids permeated significantly slower than diazepam. The ranking was confirmed in group studies as well as in single-substance studies after normalization to diazepam. Valerenic acid (1.06 +/- 0.29 microm/min, factor 0.03 related to diazepam) was the slowest to permeate in the group study, followed by hydroxyvalerenic acid (2.72 +/- 0.63 microm/min, factor 0.07 related to diazepam) and acetoxyvalerenic acid (3.54 +/- 0.58 microm/min, factor 0.09 related to diazepam). To elucidate the contribution of the paracellular transport, studies were performed at different tightness status of the cell layers reflected by different transendothelial electrical resistance (TEER) values. Results showed an exponential correlation between transport and TEER for all three acids, whereas diazepam permeated TEER independently. In summary, it is hypothesized that the investigated compounds from Valeriana officinalis L. S. L. can probably only pass through the BBB by a still unknown transport system and not transcellularly by passive diffusion.     

Analysis of sesquiterpenes in Valeriana officinalis by capillary electrophoresis.

A capillary electrophoresis (CE) method permitting the determination of the main sesquiterpenes in Valeriana officinalis has been developed. A separation of valerenic acid and its hydroxy and acetoxy derivatives, three compounds characteristic for the species, was achieved using a 40 mM phosphate-borate buffer at pH 8.5, which contained 10% isopropanol as organic modifier. Applied temperature and voltage were 35 degrees C and 17.5 kV, respectively. This setup allowed a baseline separation of the three compounds within 8 min, with a detection limit of 5.8 micrograms/ml or less. Out of six market products analyzed, only one contained a detectable amount of the marker compounds, with 0.54% of hydroxyvalerenic acid and 0.13% valerenic acid, respectively. The quantitative results were comparable to those obtained by HPLC.     

基于JNK信号通路探讨缬草酸对癫痫孕鼠仔鼠海马神经元保护作用

目的:基于JNK通路探究缬草酸对癫痫仔鼠海马神经元保护作用。方法:18只大鼠建立癫痫模型并成功妊娠后,随机分为模型对照组及缬草酸低、高剂量组(各6只),另取6只大鼠成功妊娠后为正常对照组。其中缬草酸低、高剂量组灌胃60 mg·kg^-1·d^-1、180 mg·kg^-1·d^-1缬草酸溶液至孕20 d,模型对照组、正常对照组同时间灌胃等量0.9%氯化钠溶液。统计各组总仔鼠数及活仔鼠数;HE染色观察仔鼠海马组织病理学变化;TUNEL技术检测仔鼠海马CA1区神经元细胞凋亡情况;免疫印迹法检测仔鼠海马组织中JNK、p-JNK、p38 MAPK、p-p38 MAPK蛋白表达变化。结果:与模型对照组相比,缬草酸低、高剂量组总仔鼠数、活仔鼠数均升高,海马CA1区神经元细胞AI、海马组织中p-JNK/JNK及p-p38 MAPK/p38 MAPK表达水平均降低(P均＜0.05)。结论:缬草酸对癫痫仔鼠海马神经元具有保护作用,可能是通过抑制JNK通路实现的。     

Central nervous depressant activity of valerenic Acid in the mouse

Valerenic acid, isolated from VALERIANA OFFICINALIS L. s.l. affected the rotarod and traction performance of mice in a similar manner to that of pentobarbital which was used together with chlorpromazine and diazcpam as reference substance in both tests. A decrease of locomotility of mice was also noticed after administration of valerenic acid, in particular when this compound was administered to the same group of mice a week later than the vehicle. In addition valerenic acid was found to prolong the pentobarbital induced sleeping time. It can be concluded, that valerenic acid has aspecific central nervous depressent properties. This factor should be taken into account when considering the sedative action of Valerian preparations.     

The roles of valerenic acid on BDNF expression in the SH-SY5Y cell

The roots of Valeriana officinalis L. (Valerianaceae) are used for treating sleep disorders and/or mild nerve tension. The effect of valerenic acid on brain-derived neurotrophic factor (BDNF) has not yet been studied, although it is known that gamma-amino butyric acid A (GABA_A) receptor is regulated by BDNF, which modulates the depressive-like behavior and neurogenesis. The purpose of this study is to determine the effect of V. officinalis root extract (VO), its main constituents valerenic acid (VA) and acetoxy valerenic acid (AVA) as well as valerenic acid-free (VAF), acetoxy valerenic acid-free (AVAF) extracts and increasing amounts of valerenic acid containing extracts on the BDNF expression in SH-SY5Y cell lines. The effect of methanolic extracts of VO, VA, AVA, VAF, AVAF, and the extracts whose amount of VA were increased gradually, were tested using a Human BDNF ELISA kit with 17β-estradiol as a positive control. The VO and VA extracts caused a significant (p?<?0.001) increase in the BDNF expression in SH-SY5Y cells compared to control. This effect completely disappeared when cells were treated with VAF extract. AVA alone did not show any significant change in the BDNF levels. The extracts with increasing amount of VA led to a concentration- dependent effect on the cells. In conclusion, our findings suggest that the antidepressant-like effect of the VO extract is also related to BDNF expression, and that this is mainly due to the presence of VA in the extract. Removing VA from VO extract leads to a loss of activity. Moreover, the concentration of VA plays a role for BDNF expressions in SH-SY5Y cells, which demonstrates the importance of quality control on the commercially available products.     

(-)-3 beta,4 beta-epoxyvalerenic acid from Valeriana officinalis

Chemical investigation of the root extract of Valeriana officinalis afforded a new bicyclic sesquiterpene acid, (-)-3 beta,4 beta-epoxyvalerenic acid together with valerenic acid and hexadecanoic acid. The structure of the new compound was elucidated by spectroscopic data and confirmed by partial synthesis of its methyl ester from valerenic acid. Methyl (-)-3 alpha,4 alpha-epoxyvalerenate was obtained as a minor product from the above reaction.     

GABA A receptors as in vivo substrate for the anxiolytic action of valerenic acid, a major constituent of valerian root extracts

Valerian extracts have been used for centuries to alleviate restlessness and anxiety albeit with unknown mechanism of action in vivo. We now describe a specific binding site on GABA_A receptors with nM affinity for valerenic acid and valerenol, common constituents of valerian. Both agents enhanced the response to GABA at multiple types of recombinant GABA_A receptors. A point mutation in the β2 or β3 subunit (N265M) of recombinant receptors strongly reduced the drug response. In vivo, valerenic acid and valerenol exerted anxiolytic activity with high potencies in the elevated plus maze and the light/dark choice test in wild type mice. In β3 (N265M) point-mutated mice the anxiolytic activity of valerenic acid was absent. Thus, neurons expressing β3 containing GABA_A receptors are a major cellular substrate for the anxiolytic action of valerian extracts.     

枳椇子脂肪油超临界CO_2流体萃取及GC-MS分析

目的:超临界CO2流体萃取枳椇子脂肪油并分析其组成成分。方法:采用超临界CO2流体萃取技术提取枳子脂肪油,用气相色谱-质谱联用技术进行分离和鉴定,以压力、温度、时间3因素确定其最佳提取条件。结果:枳椇子脂肪油的最佳提取条件为压力32MP、温度40℃、时间1h。枳子脂肪油含量为8.3%,其中不饱和脂肪酸约占37.6%。结论:超临界CO2流体萃取枳椇子脂肪油具有提取时间短、提取率高等优点,可为工业化生产提供理论依据。     

Stability control of valerian ground material and extracts: a new HPLC-method for the routine quantification of valerenic acids and lignans

A new HPLC-method for the separation of medium polar and nonpolar compounds in preparations of Valeriana officinalis was established for stability control. Powdered valerian root and a commercial ethanolic valerian extract were investigated for apparent differences in stability behaviour. Storage conditions were chosen according to the ICH-guidelines. Changes in composition of valerenic acids and lignans were observed depending on storage conditions and packaging materials. Hydroxyvalerenic acid, pinoresinol and hydroxypinoresinol were identified as degradation products in Valerian root, especially during accelerated testing. Ethanolic extracts appeared not to be as sensitive for chemical degradation under climatic influences compared to the crude plant material, and showed no increase in the amounts of lignan-aglyka. In comparison, extracts showed high sensitivity on changes of physical properties like loss on drying and viscosity.     

Telemetry as a tool to measure sedative effects of a valerian root extract and its single constituents in mice

Valeriana officinalis L. is a popular herbal treatment for mild sleep disorders. Clinical and non-clinical studies found contradictory results for valerian extracts and single constituents regarding the influence on sleep parameters. It was the aim of this study to investigate the sedative effects of a valerian root extract. Therefore, locomotor activity and core body temperature were recorded in male mice using radiotelemetry. A 70 % ethanolic extract prepared from the roots of V. officinalis (s. l.) and some of its single constituents, valerenic acid, linarin, and apigenin, were tested for effects on locomotion and body temperature over 180 minutes after oral administration. The extract was tested in a dose range of 250-1000 mg/kg, and only a dose of 1000 mg/kg valerian extract showed a mild short-term sedative effect with reduced locomotor activity between 66-78 min minutes after administration. Paradoxically, an increased activity was observed after 150 minutes after gavage. A dose of 1 mg/kg valerenic acid produced an intermittent stimulation of activity. However, a mild short-term sedative effect was found for linarin at 12 mg/kg and apigenin at 1.5 mg/kg. Considering the cumulative locomotor activity over the observation period of 180 min, it is concluded that neither the extract nor one of the compounds had considerable sedative effects. More precisely, the observed short-term changes in activity pattern indicate that valerian extract as well as the flavonoids linarin and apigenin are rather effective to reduce sleep latency than to act as a sleep-maintaining agent.     

亚麻种子油提取工艺及脂肪酸组成的GC-MS分析

目的：建立亚麻种子油的最佳提取工艺及测定其组成成分的最佳脂肪酸甲酯化方法。方法：分别采用不同烘干温度、提取时间、提取溶剂等对亚麻子进行种子油提取,比较其得油率,并对不同脂肪酸甲酯化方法得到的脂肪酸进行GC-MS分析。结果：种子在40℃恒温箱中烘干8 h,以石油醚-乙酸乙酯（6∶1）为溶剂,索式提取8 h为最佳提取方案。脂肪酸甲酯化以酸碱法效果最好。结论：本研究建立的亚麻种子油提取及脂肪酸甲酯化方法简单科学,适用于亚麻种子油的研究。     

Pharmacological Screening of Valerenal and some other Components of Essential Oil of Valeriana officinalis.

In order to clarify the pharmacological properties of some constituents of the essential oil of VALERIANA OFFICINALIS L. s. l. four pure compounds (valerenal, valerenic acid, valeranone and isoeugenyl-isovalerate), a hydrocarbon fraction, an oxygen fraction and the essential oil itself were submitted to a screening test. This test is an elaboration of the screening test of C AMPBELL and R ICHTER [11] to which several symptoms mentioned by I RWIN [12] and some other features have been added. From the results of this so-called Syndrome test valerenal and valerenic acid, which show some structural correspondence with the valepotriates, are pharmacologically active at the lowest dose levels. Together with valeranone, both compounds are likely to play a significant role in the depressive action of the essential oil whereas the activity of isoeugenyl-isovalerate is of relatively minor importance.     

枇杷果渣果胶最佳提取条件研究

目的 研究从枇杷果渣中提取果胶的最佳条件.方法 以不同的酸进行酸解,再进行正交试验以优化提取条件.结果 亚硫酸（H2SO3）是最适宜的酸解剂.结论 从枇杷果渣中提取果胶的最佳条件为：pH1.5、提取温度95℃、提取时间2.0h、固液比1∶30,在该条件下,果胶得率为15.29％,经脱色、脱蛋白、透析、盐沉淀、脱盐、洗涤干燥后,果胶得率为9.50％.     

Temperature effect of rheological parameters and pharmaceutical availability of diclofenak sodium salt from hydrogel products made on carbopol base for topical use

The aim of the carried out investigations was to state temperature effect of conventional carried out results of pharmaceutical availability valuation. Rheological parameters were tested in temperature of 32 and 37 degrees C in market hydrogel products for topical use made on carbopol base with diclofenac sodium salt. The kinetics of diclofenac sodium salt release with Hanson extraction cells through a dialysis membrane SpectraPor was tested in vitro. Viscosity parameters were determined with cone-plate digital rheometer.     

Experimental design as a tool to evaluate chlorogenic and caffeic acids extracted from Cecropia glaziovii Sneth

The effects of different parameters, including ethanol concentration, time of drug:solvent contact, temperature and the presence of a preservative, on chlorogenic acid (CGA) and caffeic acid (CFA) yields in Cecropia glaziovii Sneth extracts were investigated using an experimental design. In order to quantify the phenolic acids in these extracts a high-performance liquid chromatography–diode array detection (HPLC–DAD) method was developed and validated. Extracts with 80% ethanol presented a higher CGA content, but low amounts of CFA. Extracts with 20% ethanol showed a higher CFA concentration, but a sharp reduction in CGA extraction yield. The presence of a preservative, under the same extraction conditions, resulted in a slight difference or no difference in the CGA and CFA extraction yields. When the temperature was controlled at refrigerator or room temperature, a slight alteration in the concentrations of the phenolics studied was observed. The present approach can be applied in order to determine the optimum conditions for the preparation of C. glaziovii Sneth extracts based on CGA or CFA extraction yield as a chemical marker.     

Effect of valerian, valerian/hops extracts, and valerenic acid on glucuronidation in vitro

Valerian preparations alone or in combination with hops are popular over-the-counter products used for sleep disturbances or anxiety. Therefore, it is important to characterize the effect of these products on the activity of human drug-metabolizing enzymes. The inhibitory effects of valerian and valerian/hops extracts as well as valerenic acid (a major constituent of valerian) on glucuronidation were evaluated in human liver microsomes and with expressed uridine 5'-diphosphate (UDP)-glucuronosyltransferases (UGT). Methanolic extracts of two herbal preparations caused significant reductions in the rate of formation of acetaminophen, oestradiol, morphine, and testosterone glucuronides. Oestradiol glucuronidation at the 3-hydroxy position was inhibited by nearly 87% in microsomal incubations. In addition, marked reductions in UGT1A1 and UGT2B7 activities were observed in the presence of the herbal extracts using oestradiol and morphine as probe substrates, respectively. Valerenic acid also demonstrated significant inhibitory effects on the glucuronidation of acetaminophen, oestradiol, and morphine with both microsomes and expressed UGTs. The relatively low IC50 values obtained for valerenic acid in microsomal incubations may indicate that this essential oil contributes to the effects observed with herbal extracts in inhibiting glucuronidation in vitro. Overall, these findings suggest that valerian-containing products may interfere with the glucuronidation of endo- and xenobiotics.     

高效液相色谱法测定木瓜水溶性有机酸含量的样品处理方法研究

目的 优化木瓜水溶性有机酸含量测定的样品处理方法。方法 采用经验证的高效液相色谱法,以苹果酸、莽草酸、绿原酸、原儿茶酸、奎尼酸、咖啡酸含量为评判指标,用单因素试验法考察提取方式(回流和超声提取),乙醇浓度和料液比对有机酸提取量的影响;提取后经净化、富集;用流动相中的水相溶解、进样分析。结果 样品最佳处理方法为50%乙醇、料液比为1∶20、超声提取60 min;在此条件下,木瓜有机酸提取量较高,非酸性成分干扰较少。色谱图基线平直,6种水溶性有机酸的色谱峰对称性和分离度良好。结论 该方法能有效提取样品中水溶性有机酸,降低HPLC分析中的基质效应,可为木瓜有机酸含量测定提供稳定可靠的供试品溶液制备方法。