Analysis of nephroloxic and carcinogenic aristolochic acids in Aristolochia plants by capillary electrophoresis with electrochemical detection at a carbon fiber microdisk electrode

Aristolochic acids (AAs) are the main bioactive ingredients in the most of Aristolochia plants, which are used to make dietary supplements, slimming pills and Traditional Chinese Medicines (TCMs). Excessive ingestion of AAs can lead to serious nephropathy. Therefore, quantitative analysis and quality control for the plants containing AAs is of great importance. In this paper, capillary electrophoresis (CE) with electrochemical detection (ED) at a 33 mu m carbon fiber microdisk electrode (CFE) has been applied to detect AA-I and AA-II in Aristolochia plants. Under the optimum conditions: detection potential at 1.20 V, 2.0 x 10(-2) mol L-1 phosphate buffer solution (PBS) (pH 10.0), injection time 25 s at a height of 17 cm and separation voltage at 12.5 kV, the AA-I and AA-II were baseline separated within 5 min. Low detection limits for AA-I and AA-II were 4.0 x 10(-8) mol L-1 and 1.0 x 10(-7) mol L-1, respectively. Wide linear ranges were from 4.0 x 10(-8) mol L-1 to 1.9 x 10(-5) mol L-1 and 1.0 X 10(-7) mol L-1 to 5.0 x 10(-5) mol L-1 for AA-I and AA-II, respectively. The proposed method has been successfully applied to analyze AAs contents in plant extracts. The results indicated that the contents of AAs in each part of Aristolochia debilis Sieb. Et Zucc.

Examination of Antioxidative System's Responses in the Different Phases of Drought Stress and During Recovery in Desert Plant Reaumuria soongorica (Pall.) Maxim

The aim of this study was to test the protective roles of superoxide dismutases (SODs), guaiacol peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) against oxidative damage and their activities in different phases of the dry down process in Reaumuria soongorica (Pall.) Maxim. leaves. Drought stress was imposed during 100 consecutive days and rewatering after 16, 72, and 100 days. The concentration of hydrogen peroxide (H2O2), malondialdehyde, and SODs activities were elevated significantly with progressing drought stress. POD and CAT activities increased markedly in the early phase of drought and decreased significantly with further drought stress continuation, and POD activity was unable to recover after rewatering. Ascorbate, reduced glutathione, APX, and GR activities declined in the initial stages of drought process, elevated significantly with further increasing water deficit progression and recovered after rewatering. These results indicate that: (1) iron SODs-removing superoxide anion is very effective during the whole drought stress; (2) CAT scavenges H2O2 in the early phase of drought and enzymes of ascorbate-glutathione cycle scavenge H2O2 in further increasing drought stress; and (3) POD does not contribute to protect against oxidative damage caused by H2O2 under drought stress.

No C_4 Plants Found at the Haibei Alpine Meadow Ecosystem Research Station in Qinghai, China: Evidence from Stable Carbon Isotope Studies

Using the measurement of stable carbon isotopes in leaves as a tool to investigate photosyn-thetic pathway of 102 plant species grown at an alpine meadow ecosystem, at the foot of the Qilian Mountain, Qinghai, China. The results indicate that the δ~(3)C values of plants have a narrow range from -28.24‰ to -24.84‰, which means that none of the species examined belongs to C_4 and crassulaceous acid metabolism (CAM) photosynthetic pathway and all of these species perform photosynthesis through the C_3 pathway. This is likely due to a long-term adaptation to environments at the alpine meadow ecosystem.

Determination of salidroside in medicinal plants belonging to the Rhodiola L. Genus originating from the Qinghai-Tibet Plateau

The Rhodiola L. genus (Crassulaceae) is one of the most important medicinal plant products used by Tibetans in Chinese phytotherapy. Fourteen species were examined for their content of salidroside. A considerable quantitative variation was observed using high-performance liquid chromatography and this depended on species and regional factors. It was found that all samples contained salidroside at concentrations ranging between 0.02 mg g(-1) (R. sinuate) and 15.95 mg g(-1) (R. sacra), respectively. The content of salidroside in R. sacra was significantly higher than in other popular medicinal plants of this genus. This finding indicated that there may be more Rhodiola species present in the Qinghai-Tibet Plateau which may be used as a potential source of salidroside.

Determination of free fatty acids in bryophyte plants and soil by HPLC with fluorescence detection and identification by online MS

A method for the determination of long and short chain free fatty acids (FFAs), using 1-[2-(ptoluenesulfonate)-ethyll-2-phenylimidazole-[4,5-f-9,10-phenanthrene (TSPP) as labeling reagent, has been developed. Identification of FFA derivatives was carried out by HPLC-MS with atmospheric pressure chemical ionization (APCI) in positive ion mode. Gradient elution on an Agilent Eclipse XDB-C-8 column gave good separation of the derivatives. Excellent linear responses were observed and good compositional data could be obtained from as little as 200 mg of bryophyte plants and soil samples. Facile TSPP derivatization coupled with HPLC-APCI-MS analysis allowed the development of a highly sensitive method for the quantitative analysis of trace level of FFAs from biological and natural environmental samples.

Determination of free fatty acids in soil and bryophyte plants by precolumn derivatization via HPLC with fluorescence detection and tandem mass spectrometry (HPLC-MS/MS)

A sensitive method for the determination of 30 kinds of free fatty acids (FFAs, C-1-C-30) with 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4,5-f] 9,10-phenan- threne (TSPP) as labeling reagent and using high performance liquid chromatography with fluorescence detection and identification by online postcolumn mass spectrometry with atmospheric pressure chemical ionization (APCI) source in positive-ion mode (HPLC/MS/APCI) has been developed. TSPP could easily and quickly label FFAs in the presence of K2CO3 catalyst at 90 degrees C for 30 min in N,N-dimethylformamide (DMF) solvent, and maximal labeling yields close to 100% were observed with a 5-fold excess of molar reagent. Derivatives were stable enough to be efficiently analyzed by high performance liquid chromatography. TSPP was introduced into fatty acid molecules and effectively augmented MS ionization of fatty acid derivatives and led to regular MS and MS/MS information. The collision induced cleavage of protonated molecular ions formed specific fragment ions at m/z [MH](+)(molecular ion), m/z [M'+CH2CH2](+)(M' was molecular mass of the corresponding FFA) and m/z 295.0 (the, mass of protonated molecular core structure of TSPP). Fatty acid derivatives were separated on a reversed-phase Eclipse XDB-C-8 column (4.6 x 150 mm, 5 mu m, Agilent) with a good baseline resolution in combination with a gradient elution. Linear ranges of 30 FFAs are 2.441 x 10(-3) to 20 mu mol/L, detection limits are 3.24 similar to 36.97 fmol (injection volume 10 mu L, at a signal-to-noise ratio of 3, S/N 3:1). The mean interday precision ranged from 93.4 to 106.2% with the largest mean coefficients of variation (R.S.D.) < 7,5%. The mean intraday precision for all standards was < 6.4% of the expected concentration. Excellent linear responses were observed with correlation coefficients of > 0.9991. Good compositional data could be obtained from the analysis of extracted fatty acids from as little as 200 mg of bryophyte plant samples.Therefore, the facile TSPP derivatization coupled with HPLC/MS/APCI analysis allowed the development of a highly sensitive method for the quantitation of trace levels of short and long chain fatty acids from biological and natural environmental samples.

Determination of long-chain fatty acids in bryophyte plants extracts by HPLC with fluorescence detection and identification with MS

A sensitive method for the determination of long-chain fatty acids (LCFAs) (>C20) using 1-[2-(p-toluenesulfonate)-ethyl]-2-phenylimidazole-[4.5-f]-9,10-phenanthrene (TSPP) as tagging reagent with fluorescence detection and identification with post-column APCI/MS has been developed. The LCFAs in bryophyte plant samples were obtained based on distillation extraction with 1: 1 (v/v) chloroform/methanol as extracting solvent. TSPP could easily and quickly label LCFAs at 90 degrees C in the presence of K2CO3 catalyst in DMF. Eleven free LCFAs from the extracts of bryophyte plants were sensitively determined. Maximal labeling yields close to 100% were observed with a five-fold excess of molar reagent. Separation of the derivatized fatty acids exhibited a good baseline resolution in combination with a gradient elution on a reversed-phase Eclipse XDB-C-8 column. Calculated detection limits from 1.0 pmol injection, at a signal-to-noise ratio of 3, were 26.19-76.67 fmol. Excellent linear responses were observed with coefficients of >0.9996. Good compositional data were obtained from the analysis of the extracted LCFAs containing as little as 0.2 g of bryophyte plant samples. Therefore, the facile TSPP derivatization coupled with HPLC/APCI/MS analysis allowed the development of a highly sensitive method for the quantitation of trace levels of LCFAs from biological and natural environmental samples. (c) 2006 Elsevier B.V. All rights reserved.

Independence of alpine meadow plants from soil organic matter for nitrogen supply: evidence from stable nitrogen isotopes

Stable nitrogen isotope signatures of major sources of mineral nitrogen ( mineralization of soil organic nitrogen, biological N-2 fixation by legumes, annual precipitation and plant litter decomposition) were measured to relatively define their individual contribution to grass assimilation at the Haibei Alpine Meadow Ecosystem, Qinghai, China. The results indicated that delta N-15 values (- 2.40 parts per thousand to 0.97 parts per thousand) of all grasses were much lower than those of soil organic matter (3.4 +/- 0.18 parts per thousand) and mineral nitrogen ( ammonium and nitrate together,7.8 +/- 0.57 parts per thousand). Based on the patterns of stable nitrogen isotopes, soil organic matter (3.4 +/- 0.18 parts per thousand), biological N-2 fixation (0 parts per thousand), and precipitation (- 6.34 +/- 0.24 parts per thousand) only contributed to a small fraction of nitrogen requirements of grasses, but plant litter decomposition (- 1.31 +/- 1.01 parts per thousand) accounted for 67%.

Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry

A pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS/MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEOC. BCEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. The derivatives showed an intense protonated molecular ion corresponding m/z (M + H)(+) under electrospray ionization (ESI) positive-ion mode with an exception being Tyr detected at negative mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of C-O bond of BCEOC molecule. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. Derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (BDC-OH) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (BCEOC-OH) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (BCEOC)(2). In addition, the detection responses for BCEOC derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (CEOC) in our laboratory. The ratios AC(BCEOC)/AC(CEOC) = 2.05-6.51 for fluorescence responses are observed (here, AC is relative fluorescence response). Separation of the derivatized peptides and amino acids had been optimized on Hypersil BDS C-18 column. Detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (Lys)-177.6 (His) fmol. The mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %CV < 7.5. The mean interday precision for all standards was < 10% of the expected concentration. Excellent linear responses were observed with coefficients of > 0.9999. Good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. Therefore, the facile BCEOC derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 Elsevier B.V. All rights reserved.

The effect of land management on carbon and nitrogen status in plants and soils of alpine meadows on the Tibetan plateau

Large-scale grassland rehabilitation has been carried out on the severely degraded lands of the Tibetan plateau. The grasslands created provide a useful model for evaluating the recovery of ecosystem properties. The purposes of this research were: (1) to examine the relative influence of various rehabilitation practices on carbon and nitrogen in plants and soils in early secondary succession; and (2) to evaluate the degree to which severely degraded grassland altered plant and soil properties relative to the non-disturbed native community. The results showed: (1) The aboveground tissue C and N content in the control were 105-97 g m(-2) and 3.356gm(-2), respectively. The aboveground tissue C content in the mixed seed treatment, the single seed treatment, the natural recovery treatment and the severely degraded treatment was 137 per cent, 98 per cent, 49 per cent and 38 per cent, respectively, of that in the control. The corresponding aboveground tissue N content was 109 per cent, 84 per cent, 60 per cent and 47 per cent, respectively, of that in the control. (2) Root C and N content in 0-20 cm depths of the control had an 2 2 average 1606 gm(-2) and 30-36 gm(-2) respectively. Root C and N content in the rehabilitation treatments were in the range of 26-36 per cent and 35-53 per cent, while those in the severely degraded treatment were only 17 per cent and 26 per cent of that in the control. (3) In the control the average soil C and N content at 0-20 cm was 11307 gm(-2) and 846 gm(-2), respectively. Soil C content in the uppermost 20 cm in the seeded treatments, the natural recovery treatment and the severely degraded treatment was 67 per cent, 73 per cent and 57 per cent, respectively, while soil N content in the uppermost 20cm was 72 per cent, 82 per cent and 79 per cent, respectively, of that in the control. The severely degraded land was a major C source. Restoring the severely degraded lands to perennial vegetation was an alternative approach to sequestering C in former degraded systems. N was a limiting factor in seeding grassland. It is necessary for sustainable utilization of seeding grassland to supply extra N fertilizer to the soil or to add legume species into the seed mix. Copyright (c) 2005 John Wiley & Sons, Ltd.

Jiangella gansuensis gen. nov., sp nov., a novel actinomycete from a desert soil in north-west China

A novel actinomycete strain, designated YIM 002(T), was isolated from a desert soil sample in Gansu Province, north-west China. This actinomycete isolate formed well-differentiated aerial and substrate mycelia. In the early stages of growth, the substrate mycelia fragmented into short or elongated rods. Chemotaxonomically, it contained LL-2,6-diaminopimelic acid in the cell wall. The cell-wall sugars contained ribose and glucose. Phospholipids present were phosphatidylinositol mannosides, phosphatidylinositol and diphosphatidylglycerol. MK-9(H-4) was the predominant menaquinone. The major fatty acids were anteiso C-15:0 (35.92%), anteiso C-17:0 (15.84%), iso C-15:0 (10.40%), iso C-16:0 (7.07%) and C(17:10)w8c (9.37%). The G+C content of the DNA was 70 mol%. Phylogenetic analysis and signature nucleotide data based on 16S rRNA gene sequences showed that strain YIM 002(T) is distinct from all recognized genera of the family Nocardioidaceae in the suborder Propionibacterineae. On the basis of the phenotypic and genotypic characteristics, it is proposed that isolate YIM 002(T) be classified as a novel species in a new genus, Jiangella gansuensis gen. nov., sp. nov. The type strain is YIM 002(T) (= DSM 44835(T) = CCTCC AA 204001(T) = KCTC 19044(T)).

中国北方典型生态区表土孢粉组合特征及其与现代植被和气候的关系

Surface pollen assemblages and their relationhips with the modern vegetation and climate provide a foundation for investigating palaeo-environment conditions by fossil pollen analysis. A promising trend of palynology is to link pollen data more closely with ecology. In this study, I summarized the characteristics of surface pollen assemblages and their quantitative relation with the vegetation and climate of the typical ecological regions in northern China, based on surface pollen analysis of 205 sites and investigating of modern vegetation and climate. The primary conclusions are as follows:The differences in surface pollen assemblages for different vegetation regions are obvious. In the forest communities, the arboreal pollen percentages are more than 30%, herbs less than 50% and shrubs less than 10%; total pollen concentrations are more than 106 grains/g. In the steppe communities, arboreal pollen percentages are generally less than 5%; herb pollen percentages are more than 90%, and Artemisia and Chenopodiaceae are dominant in the pollen assemblages; total pollen concentrations range from 103 to 106 grains/g. In the desert communities, arboreal pollen percentages are less than 5%. Although Chenopodiaceae and Artemisia still dominate the pollen assemblages, Ephedra, Tamaricaceae and Nitraria are also significant important in the pollen assemblages; total pollen concentrations are mostly less than 104grains/g. In the sub-alpine or high and cold meadow communities, arboreal pollen percentages are less than 30%. and Cyperaceae is one of the most significant-taxa in the pollen assemblages. In the shrub communities, the pollen assemblages are consistent with the zonal vegetation; shrub pollen percentages are mostly less than 20%, except for Artemisia and Hippophae rhamnoides communities.There are obvious trends for the pollen percentage ratios of Artemisia to Chenopodiaceae (A/C), Pinus to Artemisia (P/A) and arbor to non-arbor (AP/NAP) in the different ecological regions. In the temperate deciduous broad-leaved forest region, the P/A ratios are generally higher than 0.1, the A/C ratios higher than 2 and the AP/NAP ratios higher than 0.3. In the temperate steppe regions, the P/A ratios are generally less than 0.1, the A/C ratios higher than 1 and the AP/NAP ratios less than 0.1. In the temperate desert regions, the P/A ratios are generally less than 0.1, the A/C ratios less than 1, and the AP/NAP ratios less than 0.1.The study on the representation and indication of pollen to vegetation shows that Pinus, Artemisia, Betula, Chenopodiaceae, Ephedra, Selaginella sinensis etc. are over-representative in the pollen assemblages and can only indicate the regional vegetation. Some pollen types, such as Quercus, Carpinus, Picea, Abies, Elaeagus, Larix, Salix, Pterocelis, Juglans, Ulmus, Gleditsia, Cotinus, Oleaceae, Spiraea, Corylus, Ostryopsis, Vites, Tetraena, Caragana, Tamaricaceae, Zygophyllum, Nitraria, Cyperaceae, Sanguisorba etc. are under-representative in the pollen assemblages, and can indicate the plant communities well. Populus, Rosaceae, Saxifranaceae, Gramineae, Leguminosae, Compositae, Caprifoliaceae etc. can not be used as significant indicators to the plants.The study on the relation of pollen percentages with plant covers shows that Pinus pollen percentages are more than 30% where pine trees exist in the surrounding region. The Picea+Abies pollen percentages are higher than 20% where the Picea+Abies trees are dominant in the communities, but less than 5% where the parent plants are sparse or absent. Larix pollen percentages vary from 5% to 20% where the Larix trees are dominant in the communities, but less than 5% where the parent plants are sparse or absent. Betula pollen percentages are higher than 40% where the Betula trees are dominant in the communities" but less than 5% where the parent plants are sparse or absent. Quercus pollen percentages are higher than 10% where the Quercus trees are dominant in the communities, but less than 1% where the parent plants sparse or absent. Carpinus pollen percentages vary from 5% to 15% where the Carpinus trees are dominant in the communities, but less than 1% where the parent plants are sparse or absent. Populus pollen percentages are about 0-5% at pure Populus communities, but cannot be recorded easily where the Populus plants mixed with other trees in the communities. Juglans pollen accounts for 25% to 35% in the forest of Juglans mandshurica, but less than 1% where the parent plants are sparse or absent. Pterocelis pollen percentages are less than 15% where the Pterocelis trees are dominant in the communities, but cannot be recorded easily where the parent plants are sparse or absent. Ulmus pollen percentages are more than 8% at Ulmus communities, but less than 1% where the Ulmus plants mixed with other trees in the communities. Vitex pollen percentages increase along with increasing of parent plant covers, but the maximum values are less than 10 %. Caragana pollen percentages are less than 20 % where the Caragana plant are dominant in the communities, and cannot be recorded easily where the parent plants are sparse or absent. Spiraea pollen percentages are less than 16 % where the Spiraea plant are dominant in the communities, and cannot be recorded easily where the parent plants are sparse or absent.The study on the relation of surface pollen assemblages with the modern climate shows that, in the axis 1 of DCA, surface samples scores have significant correlation with the average annual precipitations, and the highest determination coefficient (R2) is 0.8 for the fitting result of the third degree polynomial functions. In the axis 2 of DCA, the samples scores have significant correlation with the average annual temperatures, average July temperatures and average January temperatures, and the determination coefficient falls in 0.13-0.29 for the fitting result of the third degree polynomial functions with the highest determination coefficient for the average July temperature.The sensitivity of the different pollen taxa to climate change shows that some pollen taxa such as Pinus, Quercus, Carpinus, Juglans, Spiraea, Oleaceae, Gramineae, Tamariaceae and Ephedra are only sensitive to the change in precipitation.