More than 80 aristolochic acids (AAs) and aristololactams (ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry (LC/MS(n)) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS(1) of AAs, the characteristic pseudomolecular ions were [M + NH4](+), [M + H](+), and [M + H - H2O](+). However, only [M + H](+) was found in the MS(1) of ALs, which was simpler than that of AAs. Distinct MS(n)fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants.
Aristolochiaceae ; chemistry ; Aristolochic Acids ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Tandem Mass Spectrometry ; methods

The article summarized the general situation of the study on the renal toxicity caused by aristolochic acids (AAs) and Chinese herbs containing AAs. The renal lesion induced by AAs and Chinese herbs containing AAs locates mainly in renal tubules, and glomeruluses have no obvious histological change. The short term administration of large doses causes acute renal epithelia denaturalization and tubular necrosis, but the long-term administration may result in chronically progressive interstitial fibrosis of the kidney. Renal failure may occur following both acute and chronic renal lesion. The renal function should be strictly monitored while one is using the Chinese herbs containing AAs, and the dosage and duration for the treatment must be limited to prevent renal toxicity.
Animals ; Aristolochiaceae ; chemistry ; Aristolochic Acids ; adverse effects ; isolation & purification ; toxicity ; Drugs, Chinese Herbal ; adverse effects ; isolation & purification ; toxicity ; Fibrosis ; Humans ; Kidney ; pathology ; Kidney Tubules ; pathology ; Nephritis, Interstitial ; chemically induced ; pathology ; Plants, Medicinal ; chemistry ; Renal Insufficiency ; chemically induced

OBJECTIVETo develop a HPLC method to determine the contents of aristolochic A in aristolochia debilis and Asarun spp..

METHODMethanol-water-formic acid extracts were separated on an Alltech C18 column with methanol-water-acetic acid (68:32:1) as mobile phase. The flow rate was 1.0 mL x min(-1). UV detection wavelength was 390 nm. Column temperature was 35 degrees C.

RESULTAristolochic acid A was separated well. The relationship of injection amounts and peak areas was linear (r = 0.9999) the range of 0.12-1.89 microg x g(-1) and the recovery rate was 101.8% (n = 5). 11 samples of aristolochia debilis which bought from different areas in China were determined, and the contents of aristolochic acid A varied from 0.9 to 2 mg x g(-1). The difference of the contents in Asarum spp. was obvious. The highest is 0.35, and aristolochic acid A couldn't be detected in one sample.

Aristolochia ; chemistry ; Aristolochic Acids ; analysis ; Asarum ; chemistry ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Bioassay-guided fractionation of the roots of Asarum sieboldii led to the isolation of the six compounds methylkakuol (1), sesamin (2), asarinin (3), xanthoxylol (4), and (2E,4E,8Z,10E/Z)-N-(2-methylpropyl) dodeca-2,4,8,10-tetraenamide (5/6). Among the isolates, xanthoxylol (4) exhibited significant cytotoxicity against human breast cancer cells MCF-7 and MDA-MB-231 in vitro with IC₅₀ values of 9.15 and 13.95 µM, respectively.
Asarum ; Breast Neoplasms ; Breast ; Humans ; In Vitro Techniques

To study the infection rate of leaf spot disease, the drying rate of root and volatile oil content of Asarum heterotropoides var. mandshuricum at the unwrapping stage, blooming stage, the initial fruit stage, fructescence and wither stage during the growth period under the different sunlight intensity of 100% (I), 50% (II), 28% (III), 12% (IV). The volatile oil content was measured according to the method of Chinese Pharmacopoeia and the oil composition was determined by GC-MS. The unwrapping stage, blooming stage and the early fruit stage postponed about 2 days with decrease of the sunlight intensity. The infection rate of leaf was 88.46%, 70.00%, 0.23%, 0.07% under light intensity of I, II, III and IV, respectively, the drying rate was 25.14%, 28.27%, 30.23%, 31.57% under light intensity of I, II, III and IV, respectively, and the volatile oil content was 18.1, 17.6, 16.3, 15.3 mL x kg(-1) under light intensity of I, II, III and IV, respectively. The composition of the oil determined by GC-MS was different between the groups, but the content did not changed significantly with the decrease of the light intensity.
Asarum ; growth & development ; radiation effects ; Light ; Regression Analysis

To explore the genetic diversity of Asarum sieboldii this study developed SSR markers based on transcriptome sequencing results and five populations of A.sieboldii from different regions were used as samples for genetic diversity assessment using software such as GenALEx 6.5, NTSYS 2.1, and Structure 2.3.4. The results showed that 16 SSR markers with high polymorphism and good repeatability were selected from the A.sieboldii transcriptome. Primers designed based on the flanking sequences of these markers successfully amplified 56 polymorphic fragments from 150 individual samples of the five A.sieboldii populations. On average, each primer amplified 3.5 polymorphic fragments, ranging from 2 to 8. The mean values of expected heterozygosity(H_e), Shannon's diversity index(I), Nei's gene diversity index(H), and the polymorphic information content(PIC) were 0.172, 0.281, 0.429, and 0.382, respectively. The mean population differentiation coefficient(F_(ST)) was 0.588, consistent with the analysis of molecular variance(AMOVA) results, which indicated greater genetic variation among A.sieboldii populations(69%) than that within populations(31%). The percentage of polymorphic loci(PPL) ranged from highest to lowest as SNJ>LN>SY>SZ>TB. Principal coordinate analysis(PCoA) and UPGMA clustering analysis further revealed genetic clustering of A.sieboldii individuals based on their geographical distribution, consistent with the results of the structure clustering analysis. In summary, the SSR markers developed from the transcriptome effectively assessed the genetic differentiation and population structure of natural A.sieboldii populations, revealing a relatively low genetic diversity in A.sieboldii, with genetic variation primarily observed at the population level and a correlation between population differentiation and geographic distance.
Humans ; Genetic Variation ; Asarum ; Transcriptome/genetics* ; Microsatellite Repeats/genetics* ; Phylogeny

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

The compatible use of Veratrum nigrum with asarum, is one of the eighteen incompatible pairs. To research the toxic regularity of the compatible use of V. nigrum and asarum, this experiment adopted the uniform design combining with acute toxicity test, with the number of died animals as the observation indicator. The results showed that the toxicity came from the common effect of V. nigrum and asarum, and V. nigrum after the compatible use, and V. nigrum made a greater contribution to the toxicity caused by the compatible use. But the toxicity did not absolutely enhance with the increase in use of V. nigrum.
Animals ; Asarum ; toxicity ; Drug Incompatibility ; Female ; Lethal Dose 50 ; Male ; Mice ; Veratrum ; toxicity

PURPOSE: The purpose of this study was to evaluate bone response to anodized titanium implants coated with the extract of black cohosh, Asarum Sieboldii, and pharbitis semen. MATERIALS AND METHODS: Forty anodized titanium implants were prepared as follows: group 1 was for control; group 2 were implants soaked in a solution containing triterpenoids extracted from black cohosh for 24 hours; group 3 were implants soaked in a solution containing extracts of black cohosh and Asarum Sieboldii for 24 hours; group 4 were implants soaked in a solution containing extracts of pharbitis semen for 24 hours. The implants from these groups were randomly and surgically implanted into the tibiae of ten rabbits. After 1, 2, and 4 weeks of healing, the nondecalcified ground sections were subjected to histological observation, and the percentage of bone-to-implant contact (BIC%) was calculated. RESULTS: All groups exhibited good bone healing with the bone tissue in direct contact with the surface of the implant. Group 2 (52.44 +/- 10.98, 25.54 +/- 5.56) showed a significantly greater BIC% compared to that of group 3 (45.34 +/- 5.00, 22.24 +/- 2.20) with respect to the four consecutive threads and total length, respectively. The BIC% of group 1 (25.22 +/- 6.00) was significantly greater than that of group 3 (22.24 +/- 2.20) only for total length. CONCLUSION: This study did not show any remarkable effects of the extract of black coshosh and the other natural products on osseointegration of anodized titanium implants as coating agents. Further studies about the application method of the natural products on to the surface of implants are required.
Animals* ; Asarum ; Biological Products ; Bone and Bones ; Cimicifuga* ; Methods ; Osseointegration ; Rabbits ; Semen ; Tibia ; Titanium*

Chlorophyll content,leaf mass to per area,net photosynthetic rate and bioactive ingredients of Asarum heterotropoides var. mandshuricum,a skiophyte grown in four levels of solar irradiance were measured and analyzed in order to investigate the response of photosynthetic capability to light irradiance and other environmental factors. It suggested that the leaf mass to per area of plant was greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in the decrease of leaf mass to per area at every phenological stage. At expanding leaf stage,the rate of Chla and Chlb was 3. 11 when A. heterotropoides var. mandshuricum grew in full light irradiance which is similar to the rate of heliophytes,however,the rate of Chla and Chlb was below to 3. 0 when they grew in shading environment. The content of Chla,Chlb and Chl( a+b) was the greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in its decreasing remarkably( P<0. 05). The rate of Chla and Chlb decreased but the content of Chla,Chlb and Chl( a+b) increased gradually with continued shading. The maximum value of photosynthetically active radiation appeared at 10: 00-12: 00 am in a day. The maximum value of net photosynthetic rate appeared at 8: 30-9: 00 am and the minimum value appeared at 14: 00-14: 30 pm at each phenological stage if plants grew in full sunlight. However,when plants grew in shading,the maximum value of net photosynthetic rate appeared at about 10: 30 am and the minimum value appeared at 12: 20-12: 50 pm at each phenological stage. At expanding leaf stage and flowering stage,the average of net photosynthetic rate of leaves in full sunlight was remarkably higher than those in shading and it decreased greatly with decreasing of irradiance gradually( P < 0. 05). However,at fruiting stage,the average of net photosynthetic rate of leaves in full sunlight was lower than those in 50% and 28% full sunlight but higher than those in 12% full sunlight. All photosynthetic diurnal variation parameters of plants measured in four kinds of different irradiance at three stages were used in correlation analysis. The results suggested that no significant correlation was observed between net photosynthetic rate and photosynthetically active radiation,and significant negative correlation was observed between net photosynthetic rate and environmental temperature as well as vapor pressure deficit expect for 12% full sunlight. Positive correlation was observed between net photosynthestic rate and relative humidity expect for 12% full sunlight. Significant positive correlation was observed between net photosynthetic rate and stomatal conductance in the four light treatments. Only,in 12% full sunlight,the net photosynthetic rate was significantly related to photosynthetically active radiation rather than related to environmental temperature,vapor pressure deficit and relative humidity. In each light treatment,a significant positive correlation was observed between environmental temperature and vapor pressure deficit,relative humidity as well as stomatal conductance. Volatile oil content was 1. 46%,2. 16%,1. 56%,1. 30% respectively. ethanol extracts was 23. 44%,22. 45%,22. 18%,21. 12% respectively. Asarinin content was 0. 281%,0. 291%,0. 279% and 0. 252% respectively. The characteristic components of Asarum volatile oil of plant in different light treatments did not change significantly among different groups.
Asarum ; physiology ; radiation effects ; Chlorophyll ; analysis ; Photosynthesis ; Plant Leaves ; radiation effects ; Sunlight