Essential oil composition and variability of Hypericum perforatum from wild populations of northern Turkey

Context: Hypericum perforatum L. (Guttiferae) contains many bioactive secondary metabolites including hypericins, hyperforins, and essential oil.

Objective: The present study was conducted to determine the variation in composition of essential oil in H. perforatum accessions from Turkey.

Material and methods: At full flowering, aerial parts of 30 plants were collected from 10 sites of northern Turkey and assayed for essential oil components by GC-FID and GC-MS.

Results: The chemical analysis revealed that the main constituents of the all analyzed samples were hydrocarbon and oxygenated sesquiterpenes such as β-caryophyllene (4.08–5.93%), γ-muurolene (5.00–9.56%), β-selinene (5.08–19.63%), α-selinene (4.12–10.42%), d-cadinene (3.02–4.94%), spathulenol (2.34–5.14%), and caryophyllene oxide (6.01–12.18%). Monoterpenes, both hydrocarbon and oxygenated, were represented by scarce amounts of α- and β-pinene, myrcene, linalool, cis- and trans-linalool oxide, and α-terpineol. Principal component analysis was also carried out and, according to the results, the first nine principal components were found to represent 100% of the observed variation.

Discussion: The chemical variation among the populations is discussed as the possible result of different genetic and environmental factors.

Conclusions: The wild populations examined here are potentially important sources for breeding and improvement of the cultivated varieties.     

HPLC测定新疆贯叶金丝桃中金丝桃素的含量

目的　建立测定新疆贯叶金丝桃中金丝桃素含量的测定方法。方法　采用HPLC法 ,KromasilODS - 1色谱柱 (4 .6mm× 2 0 0mm ,5 μm) ,流动相为甲醇 -pH 6 .5磷酸盐缓冲液 (90∶10 ) ,流速 1ml·min- 1 ,柱温 4 0℃ ,检测波长 5 88nm。结果　金丝桃素和其他组分可达基线分离 ,线性范围为 0 .0 16 5～ 0 .0 82 5 μg(r=0 .9999) ,平均回收率为 10 1.1% ,RSD =3.7% (n =9)。 结论　本法操作简便快速 ,灵敏准确。新疆贯叶金丝桃药材中金丝桃素的含量略高于美国药典的要求。     

贯叶连翘提取物中金丝桃素的初步HPLC测定

采用ＨＰＬＣ荧光检测法检测围产贯叶连翘提取物中的线桃素。色谱柱为μ－Ｂｏｎｄａｐａｋ Ｃ１８,流动相为０．０５ｍｏｌ／Ｌ,磷酸盐缓冲液（ＰＨ７．０）－甲醇（３：７）和水－甲醇（３：７）,荧光检测激发波长Ｅｘ４７０ｎｍ,发射波簪Ｅｍ５９０ｎｍ。     

紫外分光光度法测定贯叶金丝桃胶囊中金丝桃素的含量

目的：建立制剂贯叶金丝桃胶囊中金丝桃素的含量测定方法。方法：采用紫外分光光度法，在590nm波长处检测。结果：金丝桃素在3.29—16.49mg.L^-1(r＝0．9999)范围内吸收值与其浓度呈良好的线性关系，方法平均回收率99．54％，RSD为0.44％(n＝6)。结论：该方法简便、快速、准确，可作为本制剂的质量控制方法。     

Role of hyperforin in the antidepressant-like activity of Hypericum perforatum extracts

Abstract Rationale. Hyperforin has been identified as an active constituent of Hypericum perforatum but its importance in the antidepressant effect of this plant's extracts is not really known. Objective. To evaluate the antidepressant-like activity of two extracts in relation to the content of hyperforin and its plasma and whole brain concentrations, compared with a stable salt of hyperforin (dicyclohexylammonium; DCHA). Methods. The effects of the extracts and hyperforin were evaluated in the rat forced swimming test. The specificity of the effects was demonstrated evaluating the rats' locomotor activity. Plasma and brain concentrations of hyperforin were determined by high performance liquid chromatography. Results. The 4.5% extract (but not the 0.5% extract) given as three IP injections in 24 h (3.12–6.25 mg/kg) reduced the total immobility of rats, yielding dose-related plasma concentrations of hyperforin. These concentrations were of a similar magnitude to those after hyperforin DCHA which also significantly reduced immobility when given on the basis of the hyperforin content of the 4.5% extract (0.14 and 0.28 mg/kg). However, hyperforin was undetectable in rat brain, possibly because of poor passage of the blood-brain barrier. Conclusion. These results support the view that hyperforin plays a key role in the antidepressant-like activity of Hypericum p. However, brain concentrations after effective doses are probably far from those active in vitro on the neurotransmitter mechanisms so far investigated. Electronic Publication     

Investigation of impact of storage conditions on Hypericum perforatum L. dried total extract

Hypericum perforatum L. (Hypericaceae) has been widely prescribed for mild to moderate depression following the release of promising results in clinical trials. However, it is known that its constituents may be affected by milieu. The stability complexities of the constituents of H. perforatum have gained interest in recent years. The aim of the present study was to examine the impact of storage conditions on H. perforatum total extract simultaneously under different storage conditions. Temperature, humidity, and light conditions were evaluated. Comparative analyses of methanol extracts were conducted using high performance liquid chromatographydiode array detection for chlorogenic acid, rutin, hyperoside, isoquercitrin, quercitrin, quercetin, amentoflavone, pseudohypericin, hyperforin, and hypericin. Analysis and extraction were performed using a validated method. The fluctuation of the constituents of the plant extract has been demonstrated. Among these components, chlorogenic acid was the most stable. Hyperforin, hypericin, and pseudohypericin were more stable than the flavonoids at −20°C, in the 6^th month. As estimated, decay was lowest at −20°C and highest at 40°C–75% relative humidity for the analyzed constituents. Except for hyperforin, light protection decreased the breakdown of components within 4 months. However, at the 6^th month, equivalent changes were seen for all constituents. Degradation of the constituents at −20°C indicates the importance of stability tests in analysis studies covering time and storage conditions.     

RP-HPLC测定贯叶连翘中金丝桃素、芦丁和槲皮素的含量

目的:采用高效液相法测定贯叶连翘中的金丝桃素、芦丁和槲皮素的含量方法.方法:色谱柱为Symmtry C18(4.6mm×250 mm,5μm),检测波长分别为588 nm,359 nm,流动相分别为甲醇-乙腈-1.0%磷酸二氢钠溶液(340:12:7)和甲醇-水(40:60).结果:线性范围分别在0.02704～0.1352μg和0.264～1.32μg,0.052～0.260μg内良好,回收率分别为98.39%(n=3)、98.56%(n=3)和98.85%(n=3).RSD分别为0.78%和1.01%和1.02%.结论:该法简便,精密度高,结果准确可靠.     

贯叶连翘提取物抗炎镇痛作用实验研究

目的：探讨贯叶连翘提取物的抗炎镇痛作用。方法：采用二甲苯致小鼠耳廓肿胀、大鼠棉球肉芽肿实验观察贯叶连翘提取物抗炎作用;通过小鼠甲醛致痛实验观察其镇痛作用。结果：贯叶连翘提取物能够明显抑制二甲苯致小鼠耳廓肿胀,抑制大鼠棉球肉芽肿的增重,减少小鼠在注射甲醛后0～5min,15～30min两个时相内舔足时间、减轻甲醛所致小鼠足跖肿胀。结论：贯叶连翘提取物具有明显的抗炎镇痛作用。     

RP—HPLC法测定贯叶连翘片剂中有效成分金丝桃素和金丝桃苷的含量

采用ＲＰ－ＨＰＬＣ法测定贯叶连翘片剂中有效成分金丝桃素和金丝桃苷的含量。方法分别使用ＯＤＳ和ＯＤＳ－２色谱柱,流动相分别是１．５６％磷酸二氢钠－甲醇－醋酸乙酯（１：４：１）及甲醇－０．５％磷酸,检测波长分别为５９０ｎｍ,３６０ｎｍ。结论本方法有良好的重性及精密度。     

浊点萃取法提取贯叶连翘总黄酮的研究

目的建立浊点萃取提取贯叶连翘中总黄酮的工艺。方法采用浊点萃取法进行总黄酮提取,并用紫外分光光度法测定其含量。用正交实验法确定表面活性剂质量浓度、液固比、离子强度、平衡时间等关键因素对提取率的影响,优化提取工艺。结果采用非离子表面活性剂聚氧乙烯脂肪醇醚Genapol-X 80,质量浓度为50.0g.L-1,液固比为100∶1,NaCl浓度为3.0mol.L-1,在65℃下,萃取40min,达到最高的提取率,相对于传统工艺,提取率大大提高。结论该方法稳定易行,操作简单,绿色环保,避免高温破坏,提取率高。     

Adverse effects profile of the herbal antidepressant St. John's wort (Hypericum perforatum L.)

Objective: This paper provides a systematic review of adverse drug reactions (ADRs) associated with the use of extracts of the herb St. John's wort (Hypericum perforatum L.) for the treatment of mild to moderate depression. Methods: Searches of four computerized literature databases were performed for records of (ADRs). Manufacturers of hypericum products, the international drug monitoring centre of the World Health Organization (WHO) and the national drug safety monitoring bodies of Germany and the United Kingdom were also contacted for information. Results: Information on (ADRs) originates from case reports, clinical trials, post-marketing surveillance and drug monitoring studies. Collectively, the data suggest that hypericum is well tolerated, with an incidence of adverse reactions similar to that of placebo. The most common adverse effects are gastrointestinal symptoms, dizziness/confusion and tiredness/sedation. A potential serious adverse effect is photosensitivity, but this appears to occur extremely rarely. Conclusions: Hypericum has an encouraging safety profile. However, as most of the current data originate from short-term investigations, more long-term studies are desirable. Received: 2 February 1998 / Accepted in revised form: 8 June 1998     

The in vivo evaluation of anti-angiogenic effects of Hypericum essential oils using the chorioallantoic membrane assay

Context. Hypericum species including Hypericum confertum Choisy, H. hircinum L., H. hyssopifolium Chaix. subsp. elongatum (Ledeb.) Woron var. microcalycinum (Boiss. &; Heldr.) Boiss. and H. perforatum L. (Clusiaceae) are used as medicinal plants in Turkey.

Objective: The anti-angiogenic evaluation of Hypericum essential oils using the chick embryo chorioallantoic membrane (CAM) assay are performed with this study for the first time.

Materials and methods: The anti-angiogenic activity of Hypericum essential oils (0.5–5.0?mg/ml) was evaluated in vivo using the CAM assay, compared to standard anti-angiogenic substances at the same concentrations, in trice replicated independent assays. GC and GC-MS analyses were carried out simultaneously to identify the chemical compositions of the Hypericum essential oils.

Results: The CAM treated with H. perforatum essential oil showed anti-angiogenic effect (score 0.6?±?0.3) at 50?µg/pellet concentration, whereas other tested Hypericum essential oils showed no effect compared to the standards (e.g. suramin score 0.5?±?0.2). Furthermore, the tested oils showed neither membrane toxicity nor irritation at the tested concentrations. The major compound of the essential oil of H. confertum was identified as germacrene D (30.2%). The major compound of the essential oils of the H. hircinum. H. hyssopifolium subsp. elongatum var. microcalycinum and H. perforatum was identified as α-pinene (88.3, 57.8, 33.3%), respectively.

Discussion and conclusion. Hypericum species and in particular H. perforatum essential oil may have important effect toward wound healing and various inflammations. The data obtained in this experiment suggest further investigations on various cancers due to its anti-angiogenic effects observed.     

An in vivo study of Hypericum perforatum in a niosomal topical drug delivery system

The active compounds present in Hypericum perforatum L. (Hypericaceae) include hyperforin, hypericins and flavonoids, which are assumed to be responsible for the activity of the extract in the treatment of wounds and scars. The present study aimed to incorporate H. perforatum extract standardized to a known content of phloroglucinols, naphthodianthrones and polyphenolic compounds into an effective transdermal drug delivery system capable of entrapping both lipophilic and hydrophilic constituents in the form of niosomal gels for wound treatment. An 80% ethanol extract (HE) was prepared on a pilot scale using DIG-MAZ. An HPLC-DAD holistic profile was established for HE and was standardized to contain 3.4?±?4 rutin, 1.1?±?3 chlorogenic acid, 0.5?±?2 quercitrin, 2.8?±?2 hyperforin, and 0.51?±?3% w/w total hypericins. Niosomes were prepared using the modified reverse phase evaporation technique (REV). The wound healing effect of the gel was tested on 16 adult mongrel dogs. A significant decrease in the inflammatory cell count (18.4?±?5.3) was recorded in the niosomal gel 1.5% NaCMC-treated group at the 7th day post wounding. It induced a marked regression in the inflammatory phase and enhanced the early beginning of the proliferative phase of wound healing. After 21?days, it showed complete re-epithelization, formation of new matrix fibers and significant reduction in the wound size, compared to the control and the Panthenol® 2% cream treated groups. This is the first study of H. perforatum in a niosomal topical drug delivery system.     

RP-HPLC法测定不同产地不同部位贯叶金丝桃中金丝桃苷的含量

目的：建立测定贯叶金丝桃中金丝桃苷含量的方法,并测定不同产地不同部位贯叶金丝桃中金丝桃苷的含量。方法：采用反相高效液相色谱法。色谱柱为Boston Breeze-C18（250mm×4.6mm,5μm）,流动相为甲醇-0.1%磷酸水溶液（40∶60,V/V）,检测波长为360nm,流速为1.0mL·min-1。结果：金丝桃苷进样浓度在0.14～2.80μg·mL-1范围内与峰面积积分值呈良好的线性关系（r=0.9994）,平均加样回收率为95.97%,RSD=1.16%（n=6）。四川产地贯叶金丝桃花、叶中的金丝桃苷含量最高。结论：本方法简便、准确、重复性好,可用于贯叶金丝桃的质量控制。     

正交试验优选贯叶金丝桃中金丝桃苷的乙醇提取工艺

目的:优选贯叶金丝桃中金丝桃苷的乙醇提取工艺。方法:以乙醇浓度、乙醇加入倍数、提取温度、提取次数为考察因素,以金丝桃苷提取率为指标,采用正交试验优选金丝桃苷乙醇提取最佳条件。结果:最佳提取工艺为20倍量的60%乙醇,90℃提取4次,每次2h。结论:该方法简便、准确、重复性好,可作为贯叶金丝桃中金丝桃苷的提取工艺。     

运用GC-MS结合PCA技术对川白芷与杭白芷挥发油成分的比较分析

目的:运用GC-MS结合主成分分析(PCA)技术对川白芷与杭白芷挥发油成分进行比较分析。方法:采用水蒸气蒸馏法提取川白芷和杭白芷挥发油,运用气相色谱-质谱联用技术,结合计算机检索对其挥发油进行分离鉴定,并结合PCA技术对结果进行验证。结果:共鉴定出川白芷和杭白芷中122个挥发油成分,其中有57个共有成分,两者所含的挥发油在组分和含量上存在一定的差异,PCA分析亦进一步验证了其异同。结论:川白芷与杭白芷挥发油含有大量共有成分,但在组分和含量上存在一定的差异,本研究为川白芷与杭白芷药材的鉴别及质量控制提供了重要依据。     

贯叶连翘对PC12细胞缺血性损伤的保护作用

目的:研究贯叶连翘对PC12细胞缺血性损伤的保护作用。方法:在离体培养的PC12细胞,用NaCN加缺糖造成拟缺血损伤模型,噻唑蓝(MTT)、LDH活力测定、细胞内肌酸激酶(CK)活力测定、流式细胞仪测定细胞凋亡率和线粒体跨膜电位。结果:在10-7~10-5mol.L-1范围内,贯叶连翘浓度依赖地增加MTT比色值,降低缺血性损伤所致培养介质内LDH的释放,增加细胞内CK活力。贯叶连翘还可剂量相关性地降低PC12细胞的凋亡百分率和稳定细胞线粒体跨膜电位。结论:贯叶连翘对PC12细胞缺血性损伤具有保护作用,其机制可能与增加细胞内CK活力和稳定细胞线粒体跨膜电位而抑制缺血性损伤诱导的PC12细胞的凋亡有关。     

贯叶连翘薄膜片中金丝桃素和金丝桃素类化合物的HPLC和UV测定

建立了RP-HPLC法和UV法分别测定贯叶连翘薄膜片中金丝桃素和总金丝桃素类的含量.RP-HPLC法采用ODS柱,流动相为5%甲醇(含0.1%磷酸,三乙胺调至pH 4.5)-甲醇-乙腈(15:40:45),检测波长588nm.UV法采用甲醇避光超声提取制备样品溶液,588 nm处测定.金丝桃素和总金丝桃素类的线性范围分别为0.04～0.24μg和2～12μg/ml.平均回收率分别为103.8%和103.3%.     

HPLC法测定不同产地贯叶连翘不同部位中金丝桃素的含量

目的:建立以高效液相色谱法测定贯叶连翘中金丝桃素含量的方法,并对不同产地贯叶连翘不同部位中金丝桃素的含量进行比较。方法:色谱柱为Boston pHlex ODSC18(150mm×4.6mm,5μm),流动相为甲醇-水(85:15,氨水调pH9.5),检测波长为588nm,流速为1.0mL·min-1。结果:金丝桃素检测浓度在0.14～3.60μg·mL-1范围内与峰面积积分值呈良好线性关系(r=0.9997);平均回收率为100.7%,RSD=1.98%(n=6)。不同产地贯叶连翘不同部位中金丝桃素的含量在0.005～0.562mg·g-1之间。结论:不同产地贯叶连翘药材不同部位中金丝桃素含量均不同。本方法简便、准确、重现性好,可用于贯叶连翘的质量控制。