ABSTRACT
@#The inhibition effects of the extracts of five Verbenaceae herbs, Clerodendrum fortunatum L. , Clerodendrum japonicum(Thunb. )Sweet, Clerodendrum philippinum Schauer var. simplex Moldenke, Callicarpa longissima(Hemsl. )Merr. and Pygmaeopremna herbacea, were investigated by cell hemolysis model in vitro on classical and alternative complement activation pathways. The water extracts of Pygmaeopremna herbacea, the water extract and the ethanol extract of Clerodendrum fortunatum L. , the water extract and the ethanol extract of Callicarpa longissima(Hemsl. )Merr. , the ethanol extracts of Clerodendrum japonicum(Thunb. )Sweet and Clerodendrum philippinum Schauer var. simplex Moldenke showed inhibition cell hemolysis effects on the classical pathway. Their CH50 values were 0. 092±0. 008, 0. 074±0. 008, 0. 088±0. 006, 0. 134±0. 017, 0. 123±0. 010, 0. 380±0. 080, and 0. 200±0. 015 g/L, respectively. The water extracts of Pygmaeopremna herbacea and Callicarpa longissima(Hemsl. )Merr. and the ethanol extract of Clerodendrum fortunatum L. showed inhibition cell hemolysis effects on alternative pathway. Their AP50 values were 0. 533±0. 033, 0. 758±0. 031, and 0. 362±0. 029 g/L, respectively. Five Verbenaceae herbs appear good anticomplementary effects in vitro. The ethanol extract of Clerodendrum fortunatum L. showed the best inhibitory activity.
ABSTRACT
An automated analytical method for simultaneous determination of vitamin A and E in livers, fortified infant formulae and eggs has been developed based on on-line solid phase extraction (SPE) coupled with a dual gradient high performance liquid chromatography system with column-switching. Firstly, food samples were centrifuged after saponified in mixture solution of anhydrous alcohol, potassium hydroxide and ascorbic acid at 80 ℃ for 30 min. Secondly, the saponified sample was loaded and washed on the first dimension extraction column using methanol-water (60∶40, V/V). Afterwards, the targeted analytes were trapped and enriched on the SPE column. Finally, the trapped analytes were transferred to the second dimension analysis column by valve-switching technique for the following separation and determination. Several key factors such as the type of SPE columns, elution buffer as well as pH of washing solution were optimized. The results showed that the calibration curves of vitamin A and E were linear in the range of 0 . 02-20 mg/L with correlation coefficient (R2) more than 0. 9998. In addition, the limits of detection (S/N=3) were found in the range of 3. 0-30. 0 μg/L. The spiked recoveries of the vitamin A and E from livers, eggs and fortified infant formulae ranged from 87 . 3% to 115 . 0% with the relative standard deviations ( RSDs ) of 1 . 8% -4 . 6%. The developed method is simple, sensitive and rapid to determine vitamins A and E in animal derived food.
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Objective To investigate the antioxidation activity and resistance of lipid peroxidation of panax notoginseng flower total saponins.Methods The panax notoginseng flower buds were extracted with ethanol. The hydroxyl free radical(?OH), 1,1-diphenyl-2-picrythydrazyl radical(DPPH)clearing rate and resistance of lipid peroxidation of rat liver induced by Fe2+-cysteine were determined by spectrophotometry. Results Half clearance of hydroxyl free radical and DPPH. by panax notoginseng flower total saponin was 0.035 mg/ml and 0.094 mg/ml, the maximum inhibition rate of lipid peroxidation of rat liver induced by Fe2+-cysteine was 89.31%, therefore moderate concentration of extracts had a strong inhibitory effect on lipid peroxidation. Conclusions Panax notoginseng flower total saponins have antioxidant activity and resistance of lipid peroxidation.
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A method for determination of residues of 26 β2-agonists in pork liver was developed using high performance liquid chromatography with tandem mass spectrometric ( HPLC-MS/MS ) . After enzymatic hydrolysis with β-Glucuronidase/Arylsulfatase for 12 hours, the pH of sample solution was adjusted to 1 using perchloric acid for protein precipitation. The precipitate was extracted with 0. 1mol/L perchloric acid aqueous. The extracts in the above two steps were combined and adjusted to pH 4 for the solid phase extraction ( MCX) . And then the 26 β2-agonists residues in the extracts were separated on a reversed phase HPLC column using a gradient elution program of 0. 1% formic acid aqueous solution (A) and 0. 1% formic acid in acetonitrile solution ( B) . Multiple reaction monitoring ( MRM) with positive polarity was selected to monitor qualitative and quantitative ion. Based on the optimized method, 26 β2-agonists could be analyzed in 15 min. The recoveries ranged from 64 . 0% to 112 . 7% for the 26 kinds ofβ2-agonists residues with three spiked levels of 5, 10 and 20 μg/kg. The relative standard deviations ( RSDs) were less than 15. 2%. The limits of detection (LOD) for the 26 kinds of β2-agonists were 0. 15-1. 35 μg/kg.
ABSTRACT
<p><b>OBJECTIVE</b>To establish the EST-SSR marker system for Cordyceps by using ESTs of C. bassiana and C. militaris.</p><p><b>METHOD</b>The ESTs of Cordyceps were downloaded from the public database of NCBI, and the redundant ESTs with low quality were removed. The EST-SSR primers were designed by Sequece Seiner 1. 2. And the primers were screened through PAGE-Electrophoresis.</p><p><b>RESULT</b>The 4 556 non-redundant ESTs which from C. bassiana with total length of 2 953 173 bp were selected. 718 EST-SSRs distributed in 616 ESTs were totally screened out, accounting for 15.8% of the non-redundant ESTs. It was discovered that the average distance of EST-SSSR was 1/4 096 bp in EST-SSRs distribution of C. bassiana. Trinucleotide repeats were the most abundant types with 419 repeated sequences. Regarding to C. militaris, totally 1 363 non-redundant ESTs were acquired, from which 1 117 EST-SSRs were screened, and rate of SSR sites in ESTs was 81.95%. The leading motif of SSR was nucleotide A. The 50 pairs of EST-SSR primers were designed according to the ESTs of C. bassiana, and preliminary test showed the 34 pairs of primers amplified clear fragments,accounting for 68% of all primers. Furthermore, the 39 of the 40 pairs of primers from the ESTs of C. militaris were found to be amplified as the clear fragments, accounting for 97.5%. The phylogenetic analysis revealed that different anamorph of Cordyceps spieces were divided into four branches.</p><p><b>CONCLUSION</b>The EST-SSR of Cordyceps had comparably higher utility value. The EST-SSR markers developed from ESTs of C. bassiana and C. militaris had well transferability in Cordyceps. And it was suggested that the EST-SSR markers should be an easy and effective way to assay molecular genetic structure of Cordyceps.</p>