Geniposidic Acid Research Articles

Determination of Ten Components in Eucommia ulmoides Oliver by High-Performance Liquid Chromatography with Dual Detectors

Eucommia ulmoides Oliver is one of the oldest traditional Chinese medicines and food additives that has been widely used in China and other Asian countries. In this study, high-performance liquid chromatography with diode-array detection and evaporative light scattering detection was performed for the determination of chlorogenic acid, caffeic acid, pinoresinol-di-O-β-glucopyranoside, kaempferol, quercetin, isoquercitrin, aucubin, geniposide, genipin, and geniposidic acid in Eucommia ulmoides Oliver from different regions in China. The separation was performed by high-performance liquid chromatography with a C18 column using gradient elution with a mobile phase consisting of 0.1% formic acid–water and acetonitrile at a column temperature of 30°C. The diode array wavelength range was set from 210 to 400 nm and the optimized chromatographic wavelengths were 238 nm and 320 nm. All compounds were separated in 75 minutes and quantified with good linear calibration curves (determination coefficients R2 > 0.9965). The method showed good precision with overall inter-day coefficients of variation between 1.78% and 3.37%. The recoveries were 98.11%–103.85% for the analytes. This method was successfully applied to quantify ten components in Eucommia ulmoides Oliver samples from eight different regions in China and may provide quality assessment for this material.

Anti-atherosclerotic effect of geniposidic acid in a rabbit model and related cellular mechanisms

Context: Geniposidic acid, one of the main active ingredients in Gardenia jasminoides J. Ellis (Rubiaceae), may also possess important pharmacological activities for cardiovascular disorders similar to other derivatives, such as geniposide.Objective: To evaluate its anti-atherosclerosis (anti-AS) effect, the related pharmacological activities and possible cellular mechanisms were studied.Materials and methods: Thirty rabbits were randomly divided into normal control group, model control group, and geniposidic acid subgroups. In the AS model, its effects on the intima/media thickness ratio and aortic morphology were observed. In the study of primary cultured endothelial cells (ECs) and human umbilical artery smooth muscle cells (HUASMCs), its activities on both ECs and HUASMCs proliferation, HUASMCs' migration were also studied.Results: Compared with the model control group, the plaque area, intima/media thickness ratio, and intimal foam cells number in geniposidic acid (80, 160, and 240 mg/kg) subgroups were significantly improved (p < 0.05). By HE staining, the activities of geniposidic acid on relieving ECs shedding and improving aortic morphology disorders were also demonstrated. From the results of CCK-8 testing, only 100 μg/ml geniposidic acid performed significant inhibition on SMC proliferation. The relative IC50 of geniposidic acid on SMC inhibition was 87.73 μg/ml. Geniposide acid also showed promotion effect on ECs proliferation, and the related ED50 of geniposidic acid was 86.05 μg/ml. Besides, only 50 and 100 μg/ml geniposidic acid showed obvious inhibition on SMC migration from the upper chamber (p < 0.05).Discussion and conclusion: The effects of geniposidic acid on protecting vascular endothelium and reversing plaque formation in an atherosclerotic model were demonstrated.

UPLC-MS/MS determination of content of three iridoids of Xingnaojing oral preparation in rat brains and study on their brain pharmacokinetics

To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.

Effects of fructus psoraleae extract on the intestinal absorption kinetics of geniposide and geniposidic acid in rat.

Cortex Eucommia has been used as a kidney-tonifying herbal medicine with a long history of compatibility with Fructus Psoraleae. Geniposide (GP) and geniposidic acid (GPA) are the two main chemical components in Cortex Eucommia. In the present study, the effects of Fructus Psoraleae extract (FPE) on intestinal absorption kinetics of GP and GPA in rat were investigated. Twenty four male Sprague-Dawley rats were randomly assigned into four groups which were treated with GP, GPA, GP mixed with FPE and GPA mixed with FPE, respectively, by in situ intestinal perfusion for 3 h. The samples of intestinal perfusion solutions were collected every 30 min, and analyzed by ultra high performance liquid chromatography (UPLC). The curves of time and residual quantities of GP and GPA (lnx) in the intestinal perfusion solution and the cumulative absorption rate were obtained. The results showed that FPE exhibited different effects on the intestinal absorption of GP and GPA in rat: it increased the intestinal absorption of GP (p < 0.05), while demonstrated no significant effect on the absorption of GPA.

Isolation and Purification of Geniposide, Crocin-1, and Geniposidic Acid from the Fruit of Gardenia jasminoides Ellis by High-Speed Counter-Current Chromatography

Geniposide, crocin-1, and geniposidic acid were simultaneously separated from the fruit of Gardenia jasminoides Ellis by one step of high-speed counter-current chromatography with the solvent system composed of ethyl acetate-n-butanol-water(1:4:5, v/v/v) within 130 min. The purities of the three compounds were 98.7%, 97.1%, and 90.4%, respectively, as determined by HPLC. Their structures were confirmed by MS, UV, 1H NMR, and13C NMR analysis.

LC/MS/MS determination and pharmacokinetic studies of six compounds in rat plasma following oral administration of the single and combined extracts of Eucommia ulmoides and Dipsacus asperoides

AimTo establish and apply a new LC/MS/MS method for the simultaneous, quantitative determination of six ingredients, aucubin (AU), geniposide (GP), geniposidic acid (GPA), pinoresinol diglucoside (PDG), secologanin (SLG), and loganin (LG) in single and combined extracts of Eucommia ulmoides and Dipsacus asperoides. MethodUsing the LC/MS/MS-ESI--MRM mode to detect the six compounds, chromatographic separation was achieved on an Agilent Eclipse plus C18 column, and the mobile phase consisted of solvent A (CH3CN) and solvent B (H2O containing 0.01% CH3COOH V/V). ResultsThis method was successfully applied to quantify the six compounds in rat plasma after oral administration, and showed good precision, accuracy, reproducibility, and linear regression (r2>0.99). ConclusionThe results showed that following the use of the two medicinal plants, for AU and GP, the values of tmax markedly increased, and the values of cmax markedly decreased. It was found that the compatibility of the medicinal plants might affect their pharmacokinetic properties of their constituents.

Development and validation of high liquid performance chromatography-tandem mass spectrometry method for simultaneous determination of geniposidic acid and aucubin in rat plasma for pharmacokinetic study after oral administration of Du-zhong tea extract.

A specific and sensitive high performance liquid chromatography coupled with tandem mass spectrometric (HPLC-MS/MS) method was developed and validated for the simultaneous determination of geniposidic acid and aucubin in rat plasma after oral administration of Du-zhong tea extract. The plasma samples were pretreated by protein precipitation with methanol and the chromatographic separation was performed on a Hypersil C18 column (4.6 mm×250 mm, 5 μm), using a gradient mobile phase system of water-methanol (0.05% formic acid). The detection was accomplished by multiple-reaction monitoring (MRM) scanning via electrospray ionization source operating in the negative ionization mode. The linear range was 1-1,000 ng/mL for geniposidic acid and 0.2-200 ng/mL for aucubin, respectively. The accuracy (relative error, R.E.%) were between -5.40 and 5.00%, while the intra-day and inter-day precisions were less than 7.95 and 7.87% for the two analytes, respectively. The method was fully validated for the sensitivity, selectivity, recovery, matrix effect and stability. Then this method was successfully applied to the pharmacokinetic study of geniposidic acid and aucubin after oral administration of Du-zhong tea extract to rats and the results indicated that this HPLC-MS/MS assay is a valuable method for the pharmacokinetic study of geniposidic acid and aucubin in rat plasma.

Isolation and characterisation of (−)-genifuranal: The principal antimicrobial component in traditional smoking applications of Eremophila longifolia (Scrophulariaceae) by Australian aboriginal peoples

Ethnopharmacological significanceEremophila longifolia is considered by some Australian Aboriginal tribal groups to be among the most significant of the medicinal plants in contemporary and traditional use. Usage modalities traditionally involved lipophilic extraction into animal fats and most importantly, ceremonial or medicinal smoking applications, involving the fumigation of mothers and infants following childbirth or boys following circumcision. An attempt was made to replicate the smoking modalities used by Australian Aboriginal people in the laboratory to identify bioactive compounds. Materials and methodsTwo methods were used to produce smoke extracts; smoke was channelled through a condenser then bubbled into solvent, or bubbled directly into H2O then partitioned into chloroform followed by butanol. Extracts were used, firstly for antimicrobial screening using micro-titre plate broth dilution to produce minimum inhibitory concentrations (MIC), and secondly for chemical analysis. Structure elucidation of an abundant compound isolated from the smoke extract was performed using 2D-NMR and derivatisation. ResultsSignificant antimicrobial activity (<1.0mg/ml) was produced using the smoke extracts against the Gram-positive species Staphylococcus aureus, Bacillus subtilis and the yeast Candida albicans. A major component of the smoke with strong antimicrobial activity (0.13–0.5mg/ml) was isolated which we have named (−)-genifuranal. Structure elucidation using 2D-NMR and derivatisation demonstrated genifuranal to be 5,6-dihydro-4H-cyclopenta[c]furan-4-ylacetaldehyde. Genifuranal is not observed in the leaves before heating, but is produced in the smoking or heating process and is thought to derive from hydrolysis and rearrangement of geniposidic acid or a related glycoside. Only geographically specific specimens of Eremophila longifolia produced (−)-genifuranal, which strongly supports previous hypothesised geographical variation in traditional usage, reflective of phytochemical variation. ConclusionIt would appear that genifuranal is the medicinal principal involved in traditional use of Eremophila longifolia when smoking modalities are used. Topical treatments traditionally produced by lipophilic extraction into animal fats are not likely to have had genifuranal present, as the mechanism for its formation requires heat.

Geniposide and its iridoid analogs exhibit antinociception by acting at the spinal GLP-1 receptors

We recently discovered that the activation of the spinal glucagon-like peptide-1 receptors (GLP-1Rs) by the peptidic agonist exenatide produced antinociception in chronic pain. We suggested that the spinal GLP-1Rs are a potential target molecule for the management of chronic pain. This study evaluated the antinociceptive activities of geniposide, a presumed small molecule GLP-1R agonist. Geniposide produced concentration-dependent, complete protection against hydrogen peroxide-induced oxidative damage in PC12 and HEK293 cells expressing rat and human GLP-1Rs, but not in HEK293T cells that do not express GLP-1Rs. The orthosteric GLP-1R antagonist exendin(9-39) right-shifted the concentration–response curve of geniposide without changing the maximal protection, with identical pA2 values in both cell lines. Subcutaneous and oral geniposide dose-dependently blocked the formalin-induced tonic response but not the acute flinching response. Subcutaneous and oral geniposide had maximum inhibition of 72% and 68%, and ED50s of 13.1 and 52.7 mg/kg, respectively. Seven days of multidaily subcutaneous geniposide and exenatide injections did not induce antinociceptive tolerance. Intrathecal geniposide induced dose-dependent antinociception, which was completely prevented by spinal exendin(9-39), siRNA/GLP-1R and cyclic AMP/PKA pathway inhibitors. The geniposide iridoid analogs geniposidic acid, genipin methyl ether, 1,10-anhydrogenipin, loganin and catalpol effectively inhibited hydrogen peroxide-induced oxidative damage and formalin pain in an exendin(9-39)-reversible manner. Our results suggest that geniposide and its iridoid analogs produce antinociception during persistent pain by activating the spinal GLP-1Rs and that the iridoids represented by geniposide are orthosteric agonists of GLP-1Rs that function similarly in humans and rats and presumably act at the same binding site as exendin(9-39).

꽃치자나무 추출물의 HIV-1 효소 억제 활성과 QSAR에 의한 활성인자 예측

꽃치자나무 추출물의 HIV-1 reverse transcriptase, protease 및 alpha-glucosidase에 대한 억제활성실험을 실시하였다. ELOSA 방법으로 실험한 역전사효소 억제활성 실험에서는 꽃치자나무 잎 MeOH 추출물 <TEX>$100{\mu}g/ml$</TEX> 농도에서 13.4%의 약한 억제활성이 관찰되었고, 줄기의 메탄올추출물에서는 32.5%의 높은 HIV-1 protease 억제활성과 줄기의 메탄올 추출물에서는 26.1%의 alpha-glucosidase 억제활성이 관찰되었다. 그리고 HIV-1 복제 억제활성은 MT-4 세포에 대한 HIV-1 유도 세포변성억제를 광학현미경으로 관찰하여 살펴본 결과 잎과 줄기의 모든 추출물에서 HIV-1 바이러스 증식억제에 억제활성이 관찰되지 않았다. In silico 실험결과 주요성분인 crocetin이 81.8%의 높은 역전사 효소활성이 예측되었으며, genipin이 55.39%, geniposidic acid aglycone이 64.5% 역전사효소 활성이 예측되었다. 주로 aglycone의 활성이 높게 예측되었으며 배당체의 경우 활성이 현저하게 저하되는 것으로 나타났다. In this study, we conducted the anti-HIV-1 enzymes assay in vitro and its active components were predicted by QSAR in silico for the elucidation of action mechanism on anti-HIV of natural resources. The extracts of Gardenia jasminoides var. radicans Makino were tested for their inhibitory effects on the reverse transcriptase (RT), protease and <TEX>${\alpha}$</TEX>-glucosidase. In the enzyme inhibition assay, the methanol extracts of Gardenia jasminoides var. radicans Makino stem showed a strong activity of 32.5% on the enzyme activity to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease. Moreover the methanol extracts of stem exhibited alpha-glucosidase inhibitory activities of 26.1%. The methanol extracts (<TEX>$100{\mu}g/ml$</TEX>) of stem showed a weak activity of 13.4% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. However, all extracts of leaf and stem didn't exhibit the HIV-1-induced cytopathic effects with IC (inhibitory concentration) of <TEX>$100{\mu}g/ml$</TEX> in HIV-1-infected human T-cell line. From these results, it is suggested that Gardenia jasminoides var. radicans Makino extracts may possibly be involved in the inhibition of reverse transcriptase, protease and alpha-glucosidase but can't vitally concerned with the viral replication in vitro.

Quality control and producing areas differentiation of Gardeniae Fructus for eight bioactive constituents by HPLC–DAD–ESI/MS

Gardeniae Fructus (G.Fructus), the fruit of Gardenia jasminoides Ellis (Rubiaceae), is a commonly used traditional Chinese medicine (TCM) that has been used for the treatment of hepatitis, jaundice, hypersonic, diabetes and hematuria. Numerous researches have demonstrated that the major active constituents in G.Fructus were responsible for the majority of medical effects of this fruit and their quantification were important for the quality control of G.Fructus. However, in the current quality control standard, only geniposide was used as characteristic marker of G.Fructus, which could not reflect the overall quality of this fruit. In order to identify more chemical makers for improving the quality control standard and evaluate producing areas differentiation of G.Fructus, in the present study, a novel and sensitive high-performance liquid chromatography–diode array detector coupled to an electrospray tandem mass spectrometer (HPLC–DAD–ESI/MS) was developed for the simultaneous determination of 8 major constituents, including geniposidic acid (1), chlorogenic acid (2), genipin-1-β-gentiobioside (3), geniposide (4), genipin (5), rutin (6), crocin-1 (7), crocin-2 (8) in G.Fructus. Moreover, chemometric analysis techniques with principal component constituent analysis (PCA) and cluster analysis (CA) involved were introduced in statistical analysis of 8 investigated constituents in the 34 batches samples to discriminate the samples from different producing areas.The results indicated that the contents of the 8 major bioactive constituents in G.Fructus varied significantly among different producing areas. From results of the loading plot from PCA analysis, genipin-1-β-gentiobioside may have more influence in discriminating the sample from different producing areas, and which was found to be the most abundant bioactive component besides geniposide in all the 34 batches samples, suggesting that it should be added as chemical marker for further investigation on the pharmacological actions and the quality control of G.Fructus.

Fabrication of pH sensitive amphiphilic hot-melt pressure sensitive adhesives for transdermal drug delivery system

Based on the blend of styrene-isoprene-styrene (SIS) thermoplastic elastomer and acrylic resin Eudragit® EPO, amphiphilic hot-melt pressure sensitive adhesives (HMPSAs) were fabricated. Compatibility and micromorphology of SIS/EPO blends (SEBs) were analyzed with differential scanning calorimetry (DSC), atomic force microscopy (AFM) and scanning electron microscopy (SEM). The results showed that when the mass ratio of SIS to EPO was 1:1~1:2, bicontinuous structure was formed. Following the addition of an appropriate amount of polyethylene glycol (PEG), mineral oil and C5 resin, the amphiphilic HMPSAs were prepared. Because of the compatibility between SIS and EPO, as well as the hydrogen bond interaction between EPO and PEG, amphiphilic HMPSAs showed good thermostability. The adhesive performance of HMPSAs was measured with 180° peeling strength and holding power. Geniposide and oleanolic acid were used as model drugs to investigate drug release behavior. When the mass ratio of PEG to SEB was 13:30~16:30, the HMPSAs could maintain good adhesion performance and achieve continual release of both hydrophilic and lipophilic drugs. In weakly acidic conditions, the HMPSAs exhibited good hygroscopicity and release profile, it was shown that pH sensitive amphiphilic HMPSAs were more suitable for transdermal drug delivery system (TDDS).

Anti-Inflammatory Activity of Iridoids and Verbascoside Isolated from Castilleja tenuiflora

Castilleja tenuiflora (Orobanchaceae) has been used in Mexican traditional medicine as a treatment for cough, dysentery, anxiety, nausea and vomiting as well as hepatic and gastrointestinal diseases. The ethanolic extract of the aerial parts of Castilleja tenuiflora was separated by silica gel column chromatography. The fractions were evaluated using the induced edema acetate 12-O-tetradecanoylphorbol (TPA) anti-inflammatory activity model. The most active fraction was subjected to medium-pressure liquid chromatography (MPLC) with UV detection at 206 and 240 nm. The following iridoids were isolated: geniposidic acid, aucubin, bartioside, 8-epi-loganin, mussaenoside, and the phenylpropanoid verbascoside. The most active iridoid was geniposidic acid, which was more active than the control (indomethacin), and the least active iridoid was mussaenoside. 8-epi-Loganin, and mussaenoside have not been previously reported to be anti-inflammatory compounds. The results of these investigations confirm the potential of Mexican plants for the production of bioactive compounds and validate the ethnomedical use of Castilleja tenuiflora-like anti-inflammatory plants.

Simultaneous ligand fishing and identification of human serum albumin binders from Eucommia ulmoides bark using surface plasmon resonance-high performance liquid chromatography–tandem mass spectrometry

Eucommia ulmoides bark has long been known as tonic herb, however, the systematic biochemical fingerprint analysis and corresponding structural identification are inadequate so far. Here we describe on-line surface plasmon resonance-high performance liquid chromatography-tandem mass spectrometry (SPR-HPLC-MS/MS) to fish out and identify human serum albumin (HSA) binders from E. ulmoides bark. A total of 22 HSA binders, including 4 iridoids, 11 lignans, 3 flavonoids and 4 phenolic acids, were identified or tentatively characterized based on their retention times, UV spectra and MS/MS spectrum with references or literature data, among which, geniposidic acid, geniposide and chlorogenic acid were the predominant HSA binders. In addition, the major characteristic fragmentation pathways of iridoids and lignans were investigated. The higher reusability and stability of the immobilized HSA allow the method cost-effective and high-throughput. Compared with conventional HPLC-MS/MS for chemical fingerprint analysis, SPR-HPLC-MS/MS is simple and easy to provide biochemical fingerprint, moreover, it is easy to identify low-content HSA binders because of the sample clean-up/concentration procedure.

Studies on &lt;i&gt;In Vitro&lt;/i&gt; Release Performance of Hydrophilic Drugs and Lipophilic Drugs in Amphiphilic SIS-Based Hot-Melt Pressure Sensitive Adhesives

In order to fabricate a kind of amphiphilic hot-melt pressure sensitive adhesives (HMPSAs) suitable for transdermal drug delivery systems (TDDS) of natural medicines, SIS-based hot-melt pressure sensitive adhesives were modified by a melt-blending method, in which a kind of hydrophilic poly (ethyl acrylate-co-methyl methacrylate-co-trimethylammonioethyl methacrylate chloride) (RLPO) and polyethylene glycol 2000 (PEG2000) were utilized. Functional RLPO and its plasticizer PEG2000 worked as a hydrophilic skeleton of amphiphilic HMPSAs. SEM and FT-IR results indicated that RLPO and SIS were partially compatible with each other through n-π complex between the n electrons of the carbonyl group of RLPO and the π electrons of the benzene rings of SIS and their compound had a good thermal stability. The phase microscope images showed that PEG could improve the compatibility between RLPO phase and SIS phase. As the ratio of SIS/RLPO/PEG equaled to 1/2/1.6, their compounds obtained bi-continuous structures. Geniposide (logP&lt;0) and oleanic acid (logP=9.0) were chosen as representatives of hydrophilic drugs and lipophilic drugs, respectively. It was observed that both hydrophilic drugs and lipophilic drugs had a continuous release in the optimized amphiphilic HMPSAs. In addition, the release behavior of hydrophilic geniposide could be controlled by adjusting the ratio of RLPO to PEG.

Heat Shock Factor 1 Inducers from the Bark of Eucommia ulmoides as Cytoprotective Agents

The barks of Eucommia ulmoides (Eucommiae Cortex, Eucommiaceae) have been used as a traditional medicine in Korea, Japan, and China to treat hypertension, reinforce the muscles and bones, and recover the damaged liver and kidney functions. Among these traditional uses, to establish the recovery effects on the damaged organs on the basis of phytochemistry, the barks of E. ulmoides have been investigated to afford three known phenolic compounds, coniferaldehyde glucoside (1), bartsioside (2), and feretoside (3), which were found in the family Eucommiaceae for the first time. The compounds 1-3 were evaluated for their inducible activities on the heat shock factor 1 (HSF1), and heat shock proteins (HSPs) 27 and 70, along with four compounds, geniposide (4), geniposidic acid (5), pinoresinol diglucoside (6), and liriodendrin (7), which were previously reported from E. ulmoides. Compounds 1-7 increased expression of HSF1 by a factor of 1.214, 1.144, 1.153, 1.114, 1.159, 1.041, and 1.167 at 3 μM, respectively. Coniferaldehyde glucoside (1) showed the most effective increase of HSF1 and induced successive expressions of HSP27 and HSP70 in a dose-dependent manner without cellular cytotoxicity, suggesting a possible application as a HSP inducer to act as cytoprotective agent.

Identification and Quantification of Phytochemical Composition and Anti-inflammatory, Cellular Antioxidant, and Radical Scavenging Activities of 12 Plantago Species

Twenty-eight seed samples of 12 Plantago species were investigated for their chemical compositions and anti-inflammatory, cellular antioxidant, and radical scavenging properties. A new UPLC-UV procedure was developed and applied to quantify acteoside and geniposidic acid, the characteristic constituents of the genus Plantago. The amounts of acteoside and geniposidic acid ranged from 0.07 to 15.96 mg/g and from 0.05 to 10.04 mg/g in the tested samples, respectively. Furthermore, 26 compounds were tentatively identified by UPLC/Q-TOF-MS analysis. The Plantago samples significantly differed in their phytochemical compositions. The extracts of Plantago seeds also showed inhibitory effects on LPS-induced IL-1β, IL-6, and COX-2 mRNA expression in RAW 264.7 mouse macrophage cells. Additionally, significant variations were observed among different samples on cellular antioxidant activities in HepG2 cells, as well as DPPH and hydroxyl radical scavenging capacities. The results from this study may be used to promote the use of the genus Plantago in improving human health.

Determination and pharmacokinetics of geniposidic acid in rat plasma after oral administration of Gardeniajasminoides fruit crude extract and Zhi‐zi‐chi decoction

A simple and efficient liquid chromatography-mass spectrometry method was developed and validated for the determination of geniposidic acid in rat plasma. After the addition of internal standard salidroside and acidification (0.1% formic acid, pH = 3.2), plasma samples were carried out by protein precipitation with acetonitrile and separated on a Kromasil C18 column (150 × 4.6 mm, 5 µm) within a run time of 9.0 min. Analysis was performed in selected ion monitoring mode with a positive electrospray ionization interface. No endogenous interference was observed at retention times of the analytes because of the high specificity of selected ion monitoring mode. The linear range was 0.02-4.0 µg/mL and the lower limit of quantification was 0.02 µg/mL. The mean extraction recoveries of geniposidic acid and internal standard from rat plasma were all >88.0% and the matrix effects were within acceptance criteria (90-110%). The validated method was successfully applied to the pharmacokinetic study of geniposidic acid in rat plasma after oral administration of G. jasminoides fruit crude extract and Zhi-zi-chi decoction, respectively.

Preparative isolation and purification of seven main antioxidants from Eucommia ulmoides Oliv. (Du-zhong) leaves using HSCCC guided by DPPH-HPLC experiment

Seven antioxidants were purified from Eucommia ulmoides Oliv. leaves using HSCCC guided by DPPH-HPLC experiment. HSCCC was successfully used to separate target antioxidants by three runs with different solvent systems after D101 column chromatography fractionation. Ethyl acetate–n-butanol–water (1:2:3, v/v/v) was selected as the optimum solvent system to purify geniposidic acid. Ethyl acetate–ethanol–water (4:1:5, v/v/v) was used to isolate caffeic acid, chlorogenic acid and ferulic acid. While three flavonoids, quercetin-3-O-sambubioside, rutin and isoquercitrin were purified by petroleum ether–ethyl acetate–methanol–water (1:5:1:5, v/v/v/v). The structures were identified by MS and NMR. Antioxidant activities were assessed, and compounds 2–7 showed strong antioxidant activities. This is the first report about separation of antioxidants from E. ulmoides leaves by HSCCC. The results indicated that the combinative methods using DPPH-HPLC and HSCCC could be widely applied for screening and isolation of antioxidants from complex extracts.

A sensitive liquid chromatographic–mass spectrometric method for simultaneous quantification of six iridoid glycosides from Zhi-zi-chi Decoction in rat plasma and its application to a pharmacokinetic study

A sensitive liquid chromatographic–mass spectrometric (LC–MS) method was developed and validated for simultaneous determination of geniposide, geniposidic acid, scandoside methyl ester, gardenoside, deacetyl asperulosidic acid methyl ester and genipin-1-β-gentiobioside after oral administration of Zhi-zi-chi Decoction in rat plasma. The six iridoid glycosides were extracted from plasma samples by protein precipitation, and then separated on an Apollo C18 column (250mm×4.6mm, 5μm) through the application of a gradient elution. The analytes were monitored in positive electrospray ionization by selected ion monitoring mode (SIM). The lower limits of quantitation (LLOQ) of the six analytes were all lower than 6ng/mL. The accuracy (relative error, RE%) was between −7.0% and 9.9%, while the intra- and inter-day precisions (relative standard deviation, RSD%) were less than 6.3% and 9.8% for the six analytes, respectively. The developed method was successfully applied to a comparative pharmacokinetic study of the six iridoids in rat plasma after oral administration of Zhi-zi-chi Decoction and Gardenia jasminoides extract.