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  • Author or Editor: J.-Q. Zhu x
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Summary

A rapid, simple, and practical high-performance liquid chromatographic method (HPLC) was developed and validated for the simultaneous determination of norephedrine (NME), norpseudoephedrine (NMP), ephedrine (E), pseudoephedrine (PE), and methylephedrine (ME) in traditional Chinese medicines (TCM) which contained Ephedrae Herba (Ephedra). This analysis could be accomplished within 12.5 min with an Alltima Phenyl Column by isocratic elution using a mixture of KH2PO4 (20 mM)-acetonitrile (96:4, v/v) as the mobile phase at a flow-rate of 0.6 mL min−1 and a wavelength of 210 nm. This method was successfully applied to quantify ephedra alkaloids in both Ma-xing-gan-shi decoction and Ephedra decoction. The concentration of total ephedra alkaloids (4.62 mg mL−1) in Ma-xing-gan-shi decoction was much lower than that (7.10 mg mL−1) in Ephedra decoction. Furthermore, the concentration of NME, NMP, E, PE, and ME was significantly lower in Ma-xing-gan-shi decoction than that in Ephedra decoction, respectively. The method was easily acceptable and would be popular with most analytical laboratories.

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The study investigated the effects of environmental factors (salinity, pH, ions and activation media) on sperm motility (activation rate, duration of quick movement, and lifespan) and fertilization rate of Phascolosoma esculenta. The results showed that spermatozoa in the coelom and nephridium are able to move quickly. The optimal salinity was 14.64 to 43.35 and the optimal pH was 6.46 to 9.53 for sperm activation and motility, whereas the ranges for fertilization were narrower (18.56 to 30.3 for salinity and 6.46 to 8.61 for pH). Of the ions studied, Na+ was indispensable for sperm motility and fertilization, and Ca2+ and Mg2+ were necessary for fertilization. P. esculenta sperm could not fertilize eggs and have short lifespans in 200 to 600 mmol/L NaCl and KCl solutions. Furthermore, they could not be activated or move in 200 to 600 mmol/L CaCl2, MgSO4, and sucrose solutions.

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Summary

A rapid and sensitive method for the identification and quantification of yohimbine in Pausinystalia yohimbe is described. The method used is liquid chromatography-quadrupole ion trap mass spectrometry (LC-QIT/MS). The yohimbine standard solution was directly infused into the ion trap mass spectrometers (IT/MS) for collecting the MSn spectra. The major fragment ions of yohimbine were confirmed by MSn at m/z 355, 224, 212, and 144, in the positive-ion mode. The possible main fragment ion cleavage pathway was studied. Yohimbine provided good signals corresponding to the protonated molecular ion [M + H]+. The method is reliable and reproducible, and the detection limit is 0.1 ng mL-1. The method was validated in the concentration range 0.1–50 μg mL−1; the intra- and interday precision ranged from 1.36% to 2.73% and the accuracy was 96.5–108.2%. The mean recovery of yohimbine was 97.1–101% with a relative standard deviation (RSD) <1.93%. The LC-IT/MS method was successfully applied to determine the yohimbine in P. yohimbe.

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Seven compounds, including two flavanones, dihydrokaempferol (1) and naringenin (2), and five terpenoids, boscartol A (3), 3,7-dioxo-tirucalla-8,24-dien-21-oic acid (4), 3α-acetoxyl-7-oxo-tirucalla-8,24-dien-21-oic acid (5), 11-keto-β-boswellic acid (6), and acetyl-11-keto-boswellic acid (7), have been purified by high-speed counter-current chromatography (HSCCC) from olibanum. For the separation, from 250 mg of the crude extract, 3.1 mg of 1 (95.2% purity), 2.7 mg of 2 (96.1% purity), 9.1 mg of 3 (96.7% purity), 4.5 mg of 4 (95.3% purity), 5.4 mg of 5 (96.3% purity), 48.1 mg of 6 (96.8% purity), and 45.5 mg of 7 (98.1% purity) were obtained by HSCCC with petroleum ether–ethyl acetate–methanol–water (1:0.8:1.1:0.6, v/v). The structures of these seven compounds were elucidated by a combination of electrospray ionization mass spectrometry (ESI–MS) and extensive nuclear magnetic resonance (NMR) spectroscopic.

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A rapid and sensitive method for the identification and quantification of phillyrin (POG) in Forsythia suspense is described. The phillyrin standard solution was directly infused into the ion trap mass spectrometers (IT-MS) for collecting the MSn spectra. The electrospray ionization (ESI) mass spectral fragmentation pathway of phillyrin was proposed, and the ESI-MSn fragmentation behavior of phillyrin was deduced in detail. The major product ion at m/z 355 belongs to furofuran, which was formed by loss the glucopyranoside (180 Da), and the characteristic fragment ions m/z 473, 395, 337, 309, and 249 were observed. The loss of 18 Da could arise from two different fragmentation pathways, and the observed ion was composed of a mixture of two different structural ions. Quantification of phillyrin was assigned in positive-ion mode at a product ion at m/z 557 → 355 by liquid chromatography-mass spectrometry (LC-MS). The LC-MS method was validated for linearity, sensitivity, accuracy, and precision and then used to determine the content of the phillyrin. Lastly, the LC-MS method was successfully applied to determine phillyrin in real sample F. suspense and three of its medicinal preparations in the positive mode at the first time.

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The aim of this study was to investigate the effects of maternal lead exposure on the learning and memory ability and expression of tau protein phosphorylation (P-tau) and beta amyloid protein (Aβ) in hippocampus of mice offspring. Pb exposure initiated from beginning of gestation to weaning. Pb acetate administered in drinking solutions was dissolved in distilled deionized water at the concentrations of 0.1%, 0.5% and 1% groups. On the 21 th of postnatal day, the learning and memory ability of the mouse pups was tested by Water Maze test and the Pb levels in blood and hippocampus of the offspring were also determined. The expression of P-tau and Aβ in hippocampus was measured by immunohistochemistry and Western blotting. The Pb levels in blood and hippocampus of all exposure groups were significantly higher than that of the control group ( P < 0.05). In Water Maze test, the performances of 0.5% and 1% groups were worse than that of the control group ( P < 0.05). The expression of P-tau and Aβ was increased in Pb exposed groups than that of the control group ( P < 0.05). Tau hyper-phosphorylation and Aβ increase in the hippocampus of pups may contribute to the impairment of learning and memory associated with maternal Pb exposure.

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Cereal Research Communications
Authors: N. Niu, Y.X. Bai, S. Liu, Q.D. Zhu, Y.L. Song, S.C. Ma, L.J. Ma, X.L. Wang, G.S. Zhang, and J.W. Wang

Studies of the pollen abortion mechanism in thermo-sensitive male sterile lines may provide a strong foundation for breeding hybrid wheat and establishing a theoretical basis for marker-assisted selection. To investigate the cause of pollen abortion in Bainong thermo – sensitive male sterile (BNS) lines, we analyzed the properties of pollen grains, changes in the tapetum and microspores in different anther developmental stages, and the distribution and deposition of nutrient substances in microspores. We found that tapetum degraded in the early uninucleate stage in sterile BNS (S-BNS), which was earlier than that of fertile BNS (F-BNS) tapetum. Large amounts of insoluble polysaccharides, lipids, and proteins were deposited until the trinucleate pollen stage in the nutritive cells in F-BNS. At the binucleate stage, the vacuoles disappeared and pollen inclusion increased gradually. At the trinucleate stage, these nutrients would help pollen grains mature and participate in fertilization normally. Therefore, early degradation of the tapetum, which inhibits normal microspore development, and the limited content of nutrient substances in pollen may be the main factors responsible for male sterility in BNS lines.

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An efficient and sensitive analytical method based on precolumn derivatization and gas chromatography—mass spectrometry—selected ion monitoring (GC—MS—SIM) was proposed and validated for analysis of two cembrenediols (CBDs) which are α-cembrenediol and β-cembrenediol in tobacco samples. CBDs in tobacco samples were extracted by sonication with 50 mL dichloromethane for 10 min before derivatized with 2:3 (v/v) bis(trimethylsilyl)trifluoroacetamide (BSTFA)—pyridine at 20 °C for 100 min. CBDs’ level in tobacco samples was analyzed by GC—MS—SIM and quantified by the internal standard method. The linear range for α-CBD and β-CBD was 13.6–554.6 μg mL−1 and 4.11–162.6 μg mL−1, and the correlation coefficients of both were 0.9998. The limit of detection (LOD) and limit of quantification (LOQ) of α-cembrenediol and β-cembrenediol were 0.40 μg g−1 and 1.34 μg g−1, and 0.27 μg g−1 and 0.90 μg g−1, respectively. Average recoveries of α-CBD and β-CBD were 94.4–99.9% and 91.9–98.2% while the relative standard deviations (RSDs, n = 5) were ranged from 2.67 to 5.6% and 2.04 to 4.22%, respectively. This proposed analytical method has been successfully applied to analyze CBDs in tobacco samples.

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In this study, a new substitution line, 12-5-1, with 42 chromosomes that was derived from BC3F2 descendants of the hybridization between Triticum aestivum cv. CN19 and Aegilops biuncialis was created and reported. The 12-5-1 was immune to both powdery mildew and stripe rust and has stable fertility. Multi-color fluorescence in situ hybridization indicated that 12-5-1 was a substitution line 1Mb(1B). The seed storage protein electrophoresis showed that 12-5-1 presented high molecular weight glutenin subunits (2 + 12) of CN19 and a new subunit designated as M which apparently originated from parent Ae. biuncialis, and absent 7 + 8 subunits. Additionally, the flour quality parameters showed that the protein content, Zeleny sedimentation value, wet gluten content, and grain hardness and mixing time of 12-5-1 were signifiantly higher than those of its parent CN19. Moreover, 5 pairs of the chromosome 1Mb-specifi polymerase chain reaction-based landmark unique gene markers, TNAC1021, TNAC1026, TNAC1041, TNAC1-02 and TNAC1-04, were also obtained. The new substitution line 1Mb(1B) 12-5-1 could be a valuable source for wheat improvement, especially for wheat end product quality and resistance to disease.

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