Gardeniae Fructus Research Articles

Profiling and characterisation by liquid chromatography/multi-stage mass spectrometry of the chlorogenic acids in Gardeniae Fructus

The chlorogenic acids of Gardeniae Fructus used traditionally as a Chinese herbal medicine (zhizi) have been investigated qualitatively by liquid chromatography/multi-stage mass spectrometry (LC/MS(4)). Twenty-nine chlorogenic acids were detected and twenty-five characterised to regioisomer level on the basis of their fragmentation, twenty-four for the first time from this source. Assignment to the level of individual regioisomers was possible for three caffeoylquinic acids, three dicaffeoylquinic acids, three sinapoylquinic acids, four caffeoyl-sinapoylquinic acids, two feruloyl-sinapoylquinic acids, one p-coumaroyl-sinapoylquinic acid, three (3-hydroxy, 3-methyl)glutaroylquinic acids, two (3-hydroxy, 3-methyl)glutaroyl-feruloylquinic acids, one (3-hydroxy, 3-methyl)glutaroyl-dicaffeoylquinic acid, and one (3-hydroxy, 3-methyl)glutaroyl-caffeoyl-feruloylquinic acid. Six (3-hydroxy, 3-methyl)glutaroyl-caffeoylquinic acids were detected and two were tentatively assigned as 3-caffeoyl-4-(3-hydroxy, 3-methyl)glutaroylquinic acid and 3-caffeoyl-5-(3-hydroxy, 3-methyl)glutaroylquinic acid. The (3-hydroxy, 3-methyl)glutaroyl residue modifies the mass spectral fragmentation behavior and elution sequence compared with the chlorogenic acids that contain only a cinnamic acid residue(s). Fourteen of these twenty-nine chlorogenic acids have not previously been reported from any source.

Inhibitory activity of the Chinese herbal formula Huang Lian Jie Du Tang and its single components on Leukotriene and Prostaglandin synthesis

The Chinese herbal formula Huang Lian Jie Du Tang, consisting of Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri, and Fructus Gardeniae, is used in China as a decoction to clear heat and to relieve toxicity. According to Western medicine it is used in the treatment of gastritis, hypertension, cerebrovascular diseases, liver diseases, against eczema, chronic inflammations of the intestine, as well as for psychiatric disorders like schizophrenia and depression. Previous investigations demonstrated antioxidant and anti-cancer activities of the mixture, as well as its abilities to inhibit the progression of arterios progression of arteriosclerosis and to lower plasma triglycerides [1,2,3,4,5,6]. clerosis and to lower plasma triglycerides [1,2,3,4,5,6]. The aim of our study was to compare the effects of the mixture and of the single herbs concerning their ability to inhibit leukotriene biosynthesis in human granulocytes and prostaglandin formation by COX-1 and COX-2. Decoctions of the mixture and of the single herbs were lyophilized and tested. Moreover, decoctions were fractionated using liquid-liquid chromatography with n-heptane, dichlormethane, ethylacetate and n-butanol. Leukotriene and prostaglandin biosynthesis inhibition assays were performed as previously described [7,8]. The investigation showed that Huang Lian Jie Du Tang and decoctions of the single herbs are inhibiting the formation of LTB4 in human granulocytes, as recently described [9], but have hardly any inhibitory effect on cyclooxygenases (Fig.1)

Chronological aging experiments in yeast with the TCM formulation HLJDT

Similarities of the morphological and physiological changes to mammalian cells make yeast a simple model system for cellular and perhaps organismic aging [1]. During the study of the chronological aging experiments differences between cells of Saccharomyces cerevisiae wildtype BY4741 treated with the TCM formulation Huang-Lian-Jie-Du-Tang (HLJDT) and its isolated pure main compounds (baicalein, baicalin, berberine, geniposide, jatrorrhizine, wogonin) in comparison to untreated yeast cells were investigated. HLJDT consists of Gardeniae fructus (zhi zi; 1.5 parts), Scutellariae radix (huáng qín; 1 part), Phellodendri cortex (huáng bai; 1 part) and Coptidis rhizoma (huáng lián; 1.5 parts). In preliminary investigations the most effective concentrations of the lyophilized decoction and its main ingredients were figured out. Different liquid media seem to influence the results. A significant life-span extension detected as colony forming units (CFU) could be observed with the decoction over a period of 15 and 18 days, respectively. The pure compounds showed only weak effect on the prolongation of the life-span of the yeast cells. These results might underline the use of the TCM formulation HLJDT to prevent age-related diseases.

In vitro & in vivo assessment of a herbal formula used topically for bone fracture treatment

Aim of the study A novel topical paste used for fracture healing (FH), consisting of the extracts of six herbs, Radix Dipsaci, Ramulus Sambucus Williamsii, Rhizoma Notoginseng, Flos Carthami, Rhizoma Rhei and Fructus Gardeniae, was developed according to the classical theory of traditional Chinese medicine. This study aimed to determine the effectiveness of this formula, and some of its important chemical components in the promotion of fracture healing. The transdermal transport of FH was also examined. Materials and methods The osteogenic, angiogenic and nitric oxide suppressing effects of FH and its important chemical marker components were assessed by using osteoblastosacroma UMR-106 cells, human umbilical vein endothelial cells (HUVEC) and murine macrophage RAW264.7 cells, respectively. The bone healing effects of the FH paste and its transdermal absorption were determined using a rabbit fracture model. The callus sizes, bone specific alkaline phosphatase levels and biomechanical properties of the healed bone were assessed. Results FH significantly increased the cell proliferation in UMR-106 and HUVEC cells and inhibited the nitric oxide production in murine macrophage in dose-dependent manner. Its important chemical components asperosaponin VI, ginsenoside Rg1 and emodin were shown to be acting positively in the respective in vitro studies. FH paste significantly improved the bone healing in the rabbit fracture model, as was indicated by the increases in callus size at weeks 2–5, and the elevations in bone specific alkaline phosphatase activities at weeks 5–6. The analysis using LC/MS/MS also showed the presence of important chemical marker components of the FH formula in the plasma after 8 weeks of topical treatment. Conclusion This study presents the first scientific evidence of the efficacy of a herbal paste in the promotion of fracture healing. There were evidences of transdermal transport of the chemical components, control the inflammation through nitric oxide inhibition, promotion of angiogenesis, and bone healing in the in vitro tests, as well as in the experimental animal.

Optimization of combination modes of medical materials in extractof Huanglian Jiedu decoction by semi-bionic extraction

To optimize the combination modes of medical materials in extract of Huanglian Jiedu decoction. Divided the medical materials of Huanglian Jiedu decotion into 15 groups and extracted by semi-bionic estraction (SBE), the contents of berberine, baicalin, geniposide, total alkaloid, total flavonoid, total integral calculus area of HPLC and extract of 1 000 were determined, standardized, weighted and evaluated, respectively. The maximum evaluated value Y for the extracting technique was AD + BC. The optimal combination mode of the perscription is that Rhizoma Coptidis and Fructus Gardeniae, Radix Scutellariae and Cortex Phellodendri Amurensis are decocted together respectively then the two decoctions are merged.

In-vivo and in-vitro studies on the effect of Huang-Lian-Jie-Du-Tang on nimodipine transport across rat blood-brain barrier

Huang-Lian-Jie-Du-Tang (HLJDT), an aqueous extract of Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri and Fructus Gardeniae (3:2:2:3) is an important multi-herb remedy in traditional Chinese medicine (TCM). The aim of this study was to evaluate the effect of HLJDT on nimodipine transport across rat blood-brain barrier (BBB). It was found that in-vivo the brain concentrations of nimodipine significantly increased when rats were pretreated with HLJDT. In-vitro, the serum of HLJDT-treated rats increased the accumulation of nimodipine in primary cultured rat brain microvessel endothelial cells (rBMECs) and decreased the expression of P-glycoprotein (P-gp) on rBMECs. Our previous study showed that the peak concentration of baicalin and berberine in rats after administration of HLJDT was 5 mug mL(-1) and 10 ng mL(-1), respectively. This study showed that 5 mug mL(-1) baicalin significantly increased the accumulation of nimodipine in rBMECs, while 10 ng mL(-1) berberine had no effect on the accumulation of nimodipine in rBMECs. Both the in-vivo and in-vitro experimental findings indicated that HLJDT pretreatment may alter the transport of nimodipine across rat BBB.

Effects of gardeniae fructus extract and geniposide on promoting ligament cell proliferation and collagen synthesis

Gardeniae Fructus is a traditional medicine used for the treatment of contusion such as ankle sprain. Geniposide is one of the main components of Gardeniae Fructus with diverse biological activities. In order to gain further insight into the therapeutic action of Gardeniae Fructus extract (GFE) and geniposide on ligament injuries, a new in vitro model was developed in the present study. Rat hind ankle ligament fibroblasts (RHALFs) derived from Sprague-Dawley rats were cultured, and the cell proliferation and collagen content were examined by MTT and a Sirius Red-based colorimetric assay after stimulating with each drug. The cell growth of RHALFs was promoted by culturing with 37.5-150 microg/mL of GFE and 25-200 microM of geniposide. The content of collagen in the RHALFs was significantly increased up to 131.4% and 124.2% of the control value by culturing with the GFE and geniposide, respectively. By contrast, both cell growth and collagen content were impaired by adding 25-200 microM of diclofenac, one of the common medications for ligament injuries. The findings suggest that GFE and geniposide may ameliorate the treatment of ligament injuries by proliferating ligament fibroblasts and promoting the synthesis of collagen. However, the use of diclofenac to treat acute ligament injuries should be reassessed although it possesses a potential effect on relieving symptoms.

Active ingredients of traditional Japanese (kampo) medicine, inchinkoto, in murine concanavalin A-induced hepatitis

Aim of the study The traditional Japanese (kampo) medicine inchinkoto (ICKT) is used in Eastern Asia as a choleretic and hepatoprotective agent. Previously, we reported that ICKT ameliorates murine concanavalin A (con A)-induced hepatitis via suppression of interferon (IFN)-γ and interleukin (IL)-12 production. In the present study, we investigated the active ingredients of ICKT. Materials and methods ICKT and extracts of its component herbs were fractionated, and their effects on liver injury and cytokine production in vivo (biochemical markers of liver injury and cytokine levels in serum) and in vitro (cytokine and nitrite production in the cultures of splenocytes and peritoneal macrophages). Results Decoctions of component herbs, Artemisiae Capillari Spica (Artemisia capillaris Thunberg: ‘Inchinko’ in Japanese), Gardeniae Fructus (Gardenia jasminoides Ellis: ‘Sanshishi’) and Rhei Rhizoma (Rheum palmatum Linné: ‘Daio’) were administered orally. Inchinko and Sanshishi decreased serum transaminases and IFN-γ concentrations. Examination of fractions of component herbs suggested that capillarisin, a component of Inchinko, has potent hepatoprotective activity in vivo. In in vitro studies, capillarisin and genipin, an intestinal metabolite of geniposide that is contained in Sanshishi, were examined. IFN-γ production was significantly suppressed by capillarisin and genipin in con A-stimulated splenocyte culture. Genipin also suppressed IL-1β, IL-6, and IL-12p70 synthesis. Capillarisin and genipin decreased nitrite release from IFN-γ-stimulated macrophages. Conclusions These results suggested that both Inchinko and Sanshishi may contribute to the protective effects of ICKT against con A hepatitis. Capillarisin was found to be potently hepatoprotective, and genipin may also contribute, especially via modulation of cytokine production.

The Effect of Hydrolyzed Gardeniae Fructus Extract Hydrogel on the Treatment of Ecchymoses in a Rat Model

Gardeniae fructus (GF) has been used in traditional medicine for the treatment of inflammatory disease. To evaluate the therapeutic effect of GF gel on the resolution of ecchymoses in rats. Fifty hind limbs (in 25 Sprague-Dawley rats) were evaluated. The ecchymoses were produced by dropping a 100-g weight from a height of 20 cm on the posterior side of the hind limbs 25 times. Then, 0.5 g of hydrolyzed GF extracted gel was applied to the right hind limbs and 0.5 g of gel without GF extract powder was applied to the left hind limbs. The area of the ecchymosis was measured, and histological analysis was performed. The area affected by the ecchymosis after 5 days was 15% in the control group and 2% in the GF gel group (p=.002). The mean duration for the ecchymosis was 5.8 days in the control group and 4.5 days in the GF gel group. The extravasated red blood cells and inflammation were less prominent in the GF gel group than in the control group. The results of this study showed that hydrolyzed gel of GF extract, containing genipin, was effective for the treatment of ecchymoses in a rat model.

Problematics for the validation of analyses of TCM herbal drugs and herbal drug preparations: the case Fructus Gardeniae (Zhizi) and its preparata

Fructus Gardeniae (Zhizi) is the dried ripe fruit of Gardenia jasminoides Ellis (Fam. Rubiaceae). According to Traditional Chinese Medicine (TCM) it reduces pathogenic fire, eases the mind, eliminates damp-heat, promotes diuresis and removes heat-toxicity from blood [1]. These preparations are widely used for treatment of many diseases, such as hepatitis, inner fever, hypertension, and diabetes [2]. Zhizi contains a large amount of iridoid glycosides to which can be related the activity of the herbal drug. However, other classes of constituents such as crocins and caffeoyl quinic acids are also present and can contribute in part to the activity [3]. Different preparata represented by the herbal drug dried after steaming are also present on the market and during the heat-treatment some constituents can modify their structures. The aim of this work was the validation of an HPLC/DAD/ESI-MS method to be used for the complete characterization of Fructus Gardeniae (Zhizi) and its preparata. The proposed method was validated and because its simplicity, sensitivity, accuracy and reproducibility, can be conveniently used for the analysis of the characteristic iridoids, crocins and quinic acid derivatives. The iridoids identified were scandoside methylester, gardenoside, genipin gentiobioside, geniposide, acetylgeniposide. The crocins were: crocetin, crocin-1, 2 and 3. The quinic acid derivatives identified were 3,4-dicaffeoyl-5-(3-hydroxy-3-methylglutaroyl) quinic acid and 3-caffeoyl-4-sinapoylquinic acid.

In-Jin-Ho-Tang as a potential anti-cancer drug by induction of apoptosis in HepG2 cells

Hepatocellular carcinoma is the world's most common primary malignant tumor of the liver. In-Jin-ho-Tang (IJHT) has been used as a traditional Chinese herbal medicine since ancient times, and today it is widely used as a medication for jaundice associated with inflammation of the liver. In-Jin-Ho-Tang is a drug preparation consisting of three herbs: Artemisiae Capillaris Herba (Artemisia capillaries <TEX>$T_{HUNS}$</TEX>, Injinho in Korean), Gardeniae Fructus (Gardenia jasminodes <TEX>$E_{LLIS}$</TEX>, Chija in Korean) and Rhei radix et rhizoma (Rheum palmatum L., Daehwang in Korean). This study investigated whether or not methanol extract of IJHT could induce HepG2 cancer cell death. Cytotoxic activity of IJHT on HepG2 cells was measured using an XTT assay, with an <TEX>$IC_{50}$</TEX> value of <TEX>$700{\mu}g/ml$</TEX> at 24 h Apoptosis induction by IJHT in HepG2 cells was verified by the cleavage of poly ADP-ribose polymerase, and a decrease in procaspase-3, -8, -9. Treatment of IJHT resulted in the release of cytochrome c into cytosol, loss of mitochondrial membrane potential (<TEX>${\Delta}{\Psi}_m$</TEX>), decrease in anti-apoptotic Bcl-2, and an increase in pro-apoptotic Bax expression. Thus, IJHT induced apoptosis in HepG2 cells via activation of caspase and mitochondria pathway. These results indicate that IJHT has potential as an anti-cancer agent.

Chunghyuldan attenuates brain microglial inflammatory response

Microglial cells are the prime effectors in immune and inflammatory responses of the central nervous system (CNS). During pathological conditions, the activation of these cells helps restore CNS homeostasis. However, chronic microglial activation endangers neuronal survival through the release of various proinflammatory molecules and neurotoxins. Thus, negative regulators of microglial activation have been considered as potential therapeutic candidates to target stroke and neurodegenerative diseases. Chunghyuldan, a combinatorial drug consisting of Scutellariae Radix, Coptidis Rhizoma, Phellodendri Cortex, Gardeniae Fructus, and Rhei Rhizoma, has an inhibitory effect on stroke recurrence in patients with small-vessel disease. It has also been reported to confer antihypertensive, antihyperlipidemic, and antiinflammatory effects. The aim of this study was to examine whether Chunghyuldan suppresses microglial activation. Chunghyuldan was effective at inhibiting LPS-induced nitric oxide (NO) release from rat brain microglia. Real-time reverse transcriptase PCR analysis revealed that pretreatment of rat brain microglia with Chunghyuldan attenuated the LPS-induced expression of mRNAs encoding inducible NO synthase, tumor necrosis factor (TNF)-alpha, interleukin-1beta, and cyclooxygenase-2. In rat brain microglia, Chunghyuldan reduced the LPS-stimulated production of TNF-alpha and prostaglandin E2. In addition, Chunghyuldan significantly decreased LPS-induced phosphorylation of the ERK1/2 and p38 signaling proteins. These results suggest that Chunghyuldan provide neuroprotection by reducing the release of various proinflammatory molecules from activated microglia.

Safety of an injection with a mixture of extracts from Herba Artemisiae annuae, Fructus Gardeniae and Flos Lonicerae

This study aimed to evaluate the safety for an injection with a mixture of extracts from Herba Artemisiae annuae, Fructus Gardeniae and Flos Lonicerae and to determine the risk factors that may affect its adverse drug reactions. A drug-oriented prospective observational study was performed. Physicians filled in clinical observation forms with detailed information of the patients including general information, drug information, therapeutic effects and adverse drug events. The adverse drug reaction factors were analyzed by both mono-factor and multiple-factor logistic regression methods. From April to July 2007, we collected 12,427 observation forms from 46 hospitals in Jiangsu Province of China. Among the 11,707 observation forms we analyzed, 8,074 patients were children younger than 14 years old (69%). Among 51 reported adverse drug events, 45 cases were adverse drug reactions. The total adverse drug reaction incidence of the injection was 0.38%. While most adverse drug reactions were previously known (e.g., rash, pruritus, vomiting and diarrhea), we observed three new ADR symptoms: shiver, phlebitis and anhelation. All the adverse drug reactions were controlled very well through the follow-up therapy, and none of them was life threatening. The mono-factor analysis showed that adverse drug reactions of the injection were significantly correlated with total medication dose (P = 0.0049) and combination medication (P = 0.0143), especially with antimicrobial drugs (P = 0.0079) and macrolides (P = 0.0017). The multiple factor analysis confirmed these results: medication dosage and combination medication had a crucial impact on adverse drug reactions of the injection; the risk was increased by 24.8% (the estimated value of relative risk was 1.248, 95% confidence interval: 1.054-1.479) and 89% (1.890, 1.001-3.566), respectively. The total adverse drug reaction incidence of the injection was 0.38% and lower than we expected. Moreover, we observed three new adverse drug reactions, none of which was severe.

Iridoid glycosides from Gardeniae Fructus for treatment of ankle sprain

The iridoid glycosides, genipin 1- O-β- d-isomaltoside ( 1) and genipin 1,10-di- O-β- d-glucopyranoside ( 2), together with six known iridoid glycosides, genipin 1- O-β- d-gentiobioside ( 3), geniposide ( 4), scandoside methyl ester ( 5), deacetylasperulosidic acid methyl ester ( 6), 6- O-methyldeacetylasperulosidic acid methyl ester ( 7), and gardenoside ( 8) were isolated from an EtOH extract of Gardeniae Fructus. The structures and relative stereochemistries of the metabolites were elucidated on the basis of 1D- and 2D-NMR spectroscopic techniques, high-resolution mass spectrometry, and chemical evidence. Geniposide ( 4), one of the main compounds of Gardeniae Fructus, was tested for treatment of ankle sprain using an ankle sprain model in rats. From the second to fifth day, the geniposide ( 4) (100 mg/ml) treated group exhibited significant differences ( p < 0.01) with ∼21–34% reduction in swelling ratio compared with those of the vehicle treated control group. This indicated the potential effect of geniposide ( 4) for the treatment of disorders such as ankle sprain.

Effects of the Constituents of Gardenia Fructus on Prostaglandin and NO Production

The fruits of Gardenia jasminoides Ellis have been previously reported to possess anti-inflammatory activity. In this study, the constituents including geniposide, geniposidic acid, genipin and crocin were evaluated for their effects on prostaglandin and NO production in an attempt to establish anti-inflammatory cellular mechanisms. Among the constituents tested, only genipin significantly inhibited cyclooxygenase-2-mediated <TEX>$PGE_2$</TEX> and inducible nitric oxide synthase-mediated NO production from lipopolysaccharide-treated RAW 264.7 cells at 10-100 <TEX>${\mu}$</TEX>M. Genipin also inhibited nuclear transcription factor-<TEX>${\kappa}B$</TEX> activation. Moreover, genipin showed in vivo antiinflammatory activity on <TEX>${\lambda}$</TEX>-carrageenan-induced paw edema in mice (10.4-29.9% inhibition at 20-100 mg/kg, i.p.). All of these results suggest that genipin may contribute to anti-inflammatory activity of the fruits of G. jasminoides and an inhibitory action on prostaglandin and NO production is, at least, the part of anti-inflammatory mechanism of genipin.

마이크로웨이브로 추출한 용매별 치자(Gardeniae Fructus) 추출물의 생리활성

Dept. of Food Science and Technology, Kyungpook National University, Daegu 702-701, KoreaAbstractGardeniae Fructus (GF) is a fruit of Gardenia jasminoides Ellis (Rubiaceae) which has been used in traditional medicine. It contains not only geniposide but also resveratrol, a kind of stilbene as a natural antioxidant. In this study, we investigated physiological activities of GF extracts by measuring electron donating ability (EDA), nitrite scavenging ability (NSA), super oxide dismutase (SOD)-like activity and inhibitory effect of tyrosinase activity (ITA). As a solvent, it was extracted with water, 50 and 100% ethanol using microwave. At the concentration of 6 mg/mL, EDA was the highest at 50% ethanol extract but the lowest at 100% ethanol ±extract, 96.43±0.25% and 77.06 0.22% respectively. NSA showed the strongest activity at 50% ethanol ±extract like EDA (6 mg/mL: 75.26 0.28%). ITAs were below 20% in all samples and their activities reduced according to increasing concentration. Likewise, SOD-like activities decreased with increasing concentration of extracts in all samples. In conclusion, GF extracts showed low SOD-like activities and ITAs but EDAs and NSAs were fairly high; especially EDA of 50% ethanol extract (6 mg/mL) was high as those of 0.1 and 1% L-ascorbic acid. Therefore the results suggest that GF extracts may be useful as potential sources of natural antioxidants.Key words: Gardeniae Fructus, physiological activity, microwave-assisted extraction

Stir-baked Fructus gardeniae (L.) extracts inhibit matrix metalloproteinases and alter cell morphology

Matrix metalloproteinases (MMPs) play vital roles in many pathological conditions, including cancer, cardiovascular disease, arthritis and inflammation. Modulating MMP activity may therefore be a useful therapeutic approach in treating these diseases. Qing-Kai-Ling is a popular Chinese anti-inflammatory formulation used to treat symptoms such as rheumatoid arthritis, acute hypertensive cerebral hemorrhage, hepatitis and upper respiratory tract infection. In this paper, we report that one of the components of Qing-Kai-Ling, Fructus gardeniae, strongly inhibits MMP activity. The IC 50 values for the primary herbal extract and water extract against MMP-16 were 32 and 27 μg/ml, respectively. In addition, we show that the herbal extracts influence HT1080 human fibrosarcoma cell growth and morphology. These data may provide molecular mechanisms for the therapeutic effects of Qing-Kai-Ling and herbal medicinal Fructus gardeniae.

Quality control of medicinal herbs Fructus gardeniae, Common Andrographis Herb and their preparations for their active constituents by high-performance liquid chromatography–photodiode array detection–electrospray mass spectrometry

Dehydroandrographolide, andrographolide and geniposide are the main active constituents of many herbal medicines, e.g., Fructus gardeniae, Common Andrographis Herb. They are used as the markers to control the quality of such herbal medicines and their herbal preparations. In this paper, a simple and sensitive high-performance liquid chromatographic (HPLC) method coupled with photodiode array detection (DAD) and electrospray mass spectrometry (ESI/MS) were developed to determine the three compounds simultaneously in extracts of medicinal herbs and herbal preparations produced by different companies. The extracts were separated on a C 18 reversed phase HPLC column, with a gradient solvent system, the time for the separation of the three target analytes was 10 min. The abundance ions were recorded using selected ion monitoring (SIM) mode with m/ z 297.3, 297.3 and 411.1 for dehydroandrographolide, andrographolide and geniposide, respectively. The limit of detection for dehydroandrographolide, andrographolide and geniposide were 20, 30 and 150 ng mL −1, respectively. The proposed method was successfully applied to the determination of the contents of the compounds in related to medicinal herbs and preparations.

Determination and Pharmacokinetics of 6,7-Dimethoxycoumarin in Rat Plasma after Intragastric Administration of Different Decoctions of Yinchenhao Tang

A simple and sensitive reversed-phase high-performance liquid chromatography method with UV detection is developed and validated for the determination of 6,7-dimethoxycoumarin in rat plasma and comparative analysis of its pharmacokinetics after intragastric administration of 6,7-dimethoxycoumarin and three different decoctions of yinchenhao tang. The plasma samples are deproteinated with acetonitrile. The components are separated on a Kromasil C18 column (250 x 4.6 mm, 5 microm,) with methanol-1% acetic acid solution-tetrahydrofuran (30:63:7, v/v/v) as the mobile phase, and the UV detector is set at 340 nm. Coumarin is used as an internal standard. The linear calibration curve is obtained in the concentration range of 25-2500 ng/mL. The lower limit of quantitation of the method is 25 ng/mL. The intra- and inter-day precision are less than 12%, and the accuracy determined with relative error ranges from -2.9% to 1.7%. The data obtained from rat plasma are analyzed with Topfit 2.0 Pharmacokinetic Software. With pharmacokinetic analysis, the main parameters after intragastric administration of 6,7-dimethoxycoumarin, Herba Artemisiae Scopariae decoction, Artemisiae Scopariae decoction plus Radix et Rhizoma Rhei and Fructus Gardeniae decoction, yinchenhao tang are as follows: T(1/2) is 0.29, 1.30, 1.07, and 1.75 h, AUC(-->t) is 919.1, 1215.0, 2035.3, and 2537.9 ng-h/mL, AU(0-->) is 928.5, 1325.9, 2094.4, and 2612.6 ng x h/ mL, respectively.

Effects of QKL on the level of TNF-α, IL-1β, ICAM-1 after intracerebral hemorrhage

Aims: Inflammatory cytokines release and action are central in the pathogenesis of the inflammatory response that occurs after intracerebral hemorrhage (ICH). Among the inflammatory cytokines, TNF-α, IL-1β, intercellular adhesion molecule (ICAM)-1 is the key factors which lead to secondary brain damages e.g. hydrocephalus and neuron death. QKL is an injection of natural products (consisting of radix scutellariae, fructus gardeniae, cholic acid, radix notoginseng, concha margaritifera). The aim here was to detect the function of anti-inflammation of QKL after the ICH. Method: We established a rat model with collagenase-induced intracerebral hemorrhage. The QKL was injected from the tail vein 3mL every day. The ELISA and RIA were used to detect the level of TNF-α, IL-1β, ICAM-1 in the cerebral homogenate in different periods after ICH. Result: The level of the TNF-α, IL-1β, ICAM-1 is markedly decreased with treating in the period of 48 hours after occurrence of the ICH. But in the period of 24 hours and 72 hours it shows no visible effect. Therefore, QKL has a good effect to inhibit the inflammatory cytokines TNF-α, IL-1β, ICAM-1 in the period of 48 hours after the ICH and consequently protects the neurons from secondary damages.