The present work is to establish an HPLC characteristic chromatograms of Asarum heterotropoides var. mandshuricum(AH) and A. sieboldii(AS), combined with cluster analysis for the identification of the two species, and predict their potential anti-inflammatory related targets by network pharmacological method. Eighty-nine samples(12 batches of AS and 77 batches of AH) were analyzed, and 11 characteristic peaks were identified by reference substances, UV spectrum and LC-MS. Cluster analysis showed that AS and AH were divided into two groups, and the ratio of characteristic peak areas can be used to distinguish them. When the ratio of characteristic peak sarisan to kakuol was greater than 5, it was AS, and when the ratio was less than 2, it was AH. The network pharmacological analysis of 119 constituents of Asari Radix et Rhizoma suggested that the anti-inflammatory effect of Asari Radix et Rhizoma might be related to COX-2, COX-1, iNOS, MAPK14, NR3 C1, PPARG and TNF. Among them, COX-2 is a relatively key target, which interacted with the characteristic constituents, asarinin, sesamin, safrole, methyleugenol and sarisan. The characteristic constituents asarinin and sesamin also interacted with the iNOS and MAPK14. Safrole and sarisan can also interact with iNOS, COX-1 and LAT4 H. Methyleugenol also showed interaction with COX-1 and LAT4 H. Since asarinin and sesamin interacted with three targets, COX-2, iNOS and MAPK14, it implied that they were the main active constituents for the anti-inflammatory activity of Asari Radix et Rhizoma. The COX-2 inhibitory activities of asarinin and sesamin were further studied by molecular docking and bioassay. The HPLC method established was simple, feasible and reliable, with predicted anti-inflammatory targets and anti-inflammatory constituents, which could provide a reference for improving the quality evaluation system of Asari Radix et Rhizoma.
Anti-Inflammatory Agents/isolation & purification* ; Asarum/chemistry* ; Chromatography, High Pressure Liquid ; Molecular Docking Simulation ; Phytochemicals/isolation & purification* ; Rhizome/chemistry*

OBJECTIVETo explore the character of inorganic elements in Asari Radix et Rhizoma (Xixin).

METHODThe contents of 53 inorganic elements in Xixin samples from different localities and species were determined by ICP-AES and ICP-MS. The statistical data were made using SAS.

RESULTThe result demonstrated that Xixin has the high contents of Fe, Cr, Li. It has been observed that the content of Cu and Pb of the samples are much higher than the standard level. The results of hierarchical cluster analysis revealed two groups which correspond with the species of the samples. No correlations between the contents of the inorganic elements and the localities of the samples were found. Some characteristic elements were displayed in some specific areas. The difference of the contents of the 53 inorganic elements between root and rhizome of Xixin was reported for the first time. The primary form of inorganic elements in Xixin has been studied for the first time. The result demonstrated that the extraction rate between different elements varied, with the average extraction rate of (22.25 +/- 24.96)%.

CONCLUSIONThe inorganic elements analysis of Xixin can provide evidence of its identification, cultivation and application.

Asarum ; chemistry ; classification ; China ; Drugs, Chinese Herbal ; analysis ; Rhizome ; chemistry ; Trace Elements ; analysis

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

AIM: To develop a high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) and ultraviolet (UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang (DFT). METHOD: The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL·min(-1) and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV. RESULTS: All calibration curves showed good linear regression (r(2) ≥ 0.9990). The limits of detection and limits of quantification were 0.021-0.155 -g·mL(-1) and 0.076-0.520 -g·mL(-1), respectively. Overall precision RSD (intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%-101.45%, with RSD ranging from 1.54% to 3.01% for the analytes. CONCLUSION The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.
Aconitum ; chemistry ; Asarum ; chemistry ; Calibration ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Limit of Detection ; Quality Control ; Reproducibility of Results ; Rheum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spectrophotometry, Ultraviolet ; methods

To study the chemical constituents of Asarum himalaicum, fifteen compounds were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, and HR-ESI-MS, these compounds were identified as 4-demethoxyaristolochic acid BII (1), aristolochic acid I (2), aristolochic acid Ia (3), 7-hydroxyaristolochic acid I (4), aristolochic acid IV (5), aristolic acid II (6), debilic acid (7), aristololactam I (8), 9-hydroxyaristololactam I (9), 7-methoxyaristololactam IV (10), (2S)-narigenin-5, 7-di-O-beta-D-pyranosylglucoside (11), 4-hydroxybenzoic acid (12), 3, 4-dihydroxybenzoic acid (13), 4-hydroxycinnamic acid (14), and beta-sitosterol (15). All of these compounds (1-15) were obtained from A. himalaicum for the first time. Among them, 1 was identified as a new compound, and compounds 3-6, 9, 12-14 were isolated from Asarum genus for the first time. Since the kidney toxicity of aristolochic acids and aristololactams has been reported, the result of this investigation suggests that it should be cautioned to use A. himalaicum as a medicine.
Aristolochic Acids ; chemistry ; isolation & purification ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Coumaric Acids ; chemistry ; isolation & purification ; Hydroxybenzoates ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Parabens ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Propionates ; Sitosterols ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo compare the anti-inflammatory and anti-nociceptive effects of the different water extracts which were prepared by regular decoction time or removing volatie oil and ethanol extracts which were prepared in different concentration ethanol of Xixin (the roots and rhizomes of Asarum heterotropoides var. mandshuricum), and then to investigate the anti-inflammatory and antinociceptive mechanisms.

METHODXylene-induced ICR mouse ear edema and hot plate test were utilized to evaluate the anti-inflammatory and anti-nociceptive effects of Xixin at different dose level (water extracts of regular decoction time or removing volatie oil at the dose of 0.8 g x kg(-1) and 1.6 g x kg(-1); 95% ethanol extracts at the dose of 0.91 g x kg(-1) and 1.82 g x kg(-1); 50% ethanol extracts at the dose of 0.76 g x kg(-1) and 1.52 g x kg(-1)). Xylene-induced adrenalectomized mouse ear edema model was used to study the anti-inflammatory mechanisms. To reveal the anti-nociceptive mechanisms, mice were pretreated with naloxone in the hot plate test.

RESULTThe water extracts significantly reduced the weight of ear edema as compared with control group. Inhibition ratios were 43.20% and 63.69% at the higher dose, respectively. The ethanol extracts also significantly reduced the weight of ear edema and the inhibition ratios were 61.86% and 52. 56% at the higher dose, respectively. Mice administered water extracts delayed the latency times in the hot plate test. The anti-nociceptive effects of water extracts peaked at 2.0 h after i.g. administration. The hot plate latency times were increased by 51.27%, 62.78%, 60.08% and 68.00% at peak time, respectively. Regular decoction time group showed more significant effects in both models. The anti-inflammatory effect of 95% ethanol extracts was similar to 50% ethanol extracts. The water extracts were not effective in reducing xylene-induced adrenalectomized mouse ear edema. The anti-nociceptive effect of water extracts was blocked by naloxone.

CONCLUSIONBoth the water extracts and ethanol extracts of Xixin showed considerable anti-inflammatory potency against xylene induced inflammation. The water extracts produced anti-nociception in thermal model. The water extracts prepared in regular decoction time showed better anti-inflammatory and anti-nociceptive effects. Both the 95% ethanol and 50% ethanol extracts showed similarly anti-inflammatory effects. The anti-inflammatory effect of water extracts related to adrenal gland. The anti-nociceptive effect of water extracts was involved in activating opioid receptor.

Analgesics ; administration & dosage ; chemistry ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; chemistry ; Asarum ; chemistry ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Ethanol ; chemistry ; Female ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Plant Roots ; chemistry ; Random Allocation ; Rhizome ; chemistry ; Water ; chemistry

To develop an UPLC-UV method for the determination of aristolochic acid A in the aerial and underground parts of Asarum, an AcquityUPLC HSST3 column (50 mm x 2.1 mm ID, 1.8 microm) was used with the mobile phase of acetonitrile-0.1% FA and eluted in gradient mode. The flow rate was 0.5 mL min(-1), the detection wavelength was 251 nm, and the column temperature was 40 degrees C. The calibration curve was linear in the range of 18 - 1450 ng of aristolochic acid A with correlation coefficient of 0. 9999 and the minimum detective concentration was 18.25 ng mL (-1). The method developed is accurate and reproducible, and can be applied in the determination of aristolochic acid A in Asarum.
Aristolochic Acids ; analysis ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Plant Components, Aerial ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrophotometry, Ultraviolet ; methods

OBJECTIVETo test the effect of asarinin, the extract of Herba Asari, on the acute heart transplantation rejection and the expression of adhesion molecule.

METHODAsarinin was extracted from herba asari. 64 SD rats undergone heart transplantation were divided into four groups: group A (control group), group B (Cyclosporine A treated), group C (Asarinin treated), and group D (1/2 CsA and 1/2 Asarinin). Some rats were used to examine survival time (n = 8) and the others were used to observe the pathological injury and the expression level of interrellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-I (VCAM-1) by using immunohistochemistry.

RESULTAsarinin could prolong the survival time of allografts, which was similar to CsA group (P > 0.05). Asarinin could relieve the damage of cardiomyocytes of the transplanted. Asarinin could also decrease the level of ICAM-1 and VCAM-1 in the allografts.

CONCLUSIONAsarinin may play important roles in suppressing the immune rejection, prolong the allografts survival time and protect the donor organ, which was similar to CsA. The expression level of ICAM-1 and VCAM-1 is increased in suppressing the course of acute rejection and asarinin can inhibit their expression level. Asarinin can decrease the dosage of CsA.

Animals ; Asarum ; chemistry ; Dioxoles ; isolation & purification ; pharmacology ; Graft Rejection ; metabolism ; Graft Survival ; drug effects ; Heart Transplantation ; Intercellular Adhesion Molecule-1 ; metabolism ; Lignans ; isolation & purification ; pharmacology ; Male ; Myocardium ; metabolism ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo study the influence of Xinqin tablets on guinea-pig nasal hypersensitivity.

METHOD2,4-Toluene Diisocyanate (TDI) was selected as antigen and used in nose to establish guinea-pig allergic rhinitis. The effects of Xinqin tablets on symptoms of nasal hypersensitivity in guinea-pigs, histamine content of nasal mucosa and activity of nitric oxide synthase (NOS) were examined.

RESULTXinqin tablets could significantly relieve the pathological symptoms of nasal hypersensitivity in guinea-pig, reduce histamine content of nasal mucosa and inhibit the activity of nitric oxide synthase.

CONCLUSIONXinqin tablets have significant effect on nasal hypersensitivity, and prevent the occurrence of allergic rhinitis.

Animals ; Asarum ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Guinea Pigs ; Histamine ; metabolism ; Male ; Nasal Mucosa ; drug effects ; metabolism ; pathology ; Nitric Oxide Synthase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; chemically induced ; metabolism ; pathology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

OBJECTIVETo establish a method for GC fingerprint determination of the chemical constituents in Herba Asari.

METHODGC and GC-MS were used to optimize the fingerprint determination method, and identify the main peaks in the GC fingerprint.

RESULTA preferable method for GC fingerprint determination of the chemical constituents in Herba Asari was established.

CONCLUSIONA general acquaintance of the chemical constituents in Herba Asari can be obtained by using the preferable GC fingerprint determination method, which is useful for quality evaluation of the crude drug of Herba Asari.

Anisoles ; analysis ; Asarum ; chemistry ; classification ; Gas Chromatography-Mass Spectrometry ; methods ; Monoterpenes ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Safrole ; analysis