| High Performance Liquid Chromatography determination technology with its characteristics such as highefficiency, rapidity, highsensitivity and so on, has become a perfect separation and analysis method. It coupled with various detectors has been widely applied in biological science, food science and environmental detection, etc.The paper takes health food of Ophiocordyceps Sinensis and fish as detected object, the main research content includes four parts:(1)Determination of polysaccharide peptides in Ophiocordyceps Sinensis by HPLC-PDAOBJECTIVE:To establish a direct determination method of polysaccharide peptides(PSP) in Ophiocordyceps sinensis by HPLC-PDA. METHODS: Using HPLC-PDA and water extraction and alcohol precipitation method, chromatographic column was Atlantis T3(4.6×250mm, 5μm),column temperature was 35℃, the mobile phase was the solution of CH3OH:H2O=5:95, the flow velocity was 1.0ml/min, the sample size was 10μL, and PDA scanning wavelength was between 190 nm and 460 nm. RESULTS:The average recovery rate was 96.4%, and the RSD was 0.78%, the content of PSP in sample is 16.8 mg/g. CONCLUSION:The proposed method is simple, rapid, and accurate, and has high recovery, it can be used to offer a reliable method for the determination of PSP in health products of Ophiocordyceps Sinensis.(2)Simultaneous determination of adenosine and cordycepin in Ophiocordyceps Sinensis by HPLC-PDAOBJECTIVE: To develop a method with HPLC and PDA to simultaneously determine adenosine and cordycepin in Ophiocordyceps Sinensis. METHODS: The adenosine and cordycepin in products were ultrasonically extracted with mobile phase and separated with UltimateTM AQ-C18 column(4.6×250mm, 5mm) and CH3OH(methanol): KH2PO4(potassium dihydrogen phosphate aqueous solution, 0.01 mol/L)=(10:90,V/V) mobile phase,, the column temperature was 30℃, the flow velocity was 1.0m L/min, the sample size was 20μL, and PDA scanning wavelength was 260 nm. RESULTS: The linearity of adenosine and cordycepin both were good in the range of 1.0~100.0μg/m L.The recovery of adenosine and cordycepin were 99.4%, 97.8%, and their relative standard deviation(RSD) were 0.48%, 3.85%, respectively.The detection limitwas 0.1μg /m L., the content of adenosine and cordycepin in sample are respectively. CONCLUSION: The method has simple mobile phase and high recovery, and can be applied for simultaneous, rapid and accurate detection of adenosine and cordycepin in Ophiocordyceps Sinensis and its health foods.(3) Simultaneous determination of adenosine, cordycepin and cordyceptic acid in Ophiocordyceps Sinensis by HPLC-ESI-MSOBJECTIVE: To develop a method with HPLC and electron spray ionization mass spectrometry(ESI-MS) to simultaneously determine adenosine, cordycepin and cordyceptic acid in Ophiocordyceps Sinensis. METHODS: Using HPLC-MS methord, Selective Ion Monitoring(SIM) and ESI.The adenosine, cordycepin and ordyceptic acid in products were ultrasonically extracted with pure water and separated with Thermo Bio Basic-18 PIONEER(150 mm×2.1 mm, 5 μm) and formic acid(0.05mol/L):methanol=(94:6,V/V) mobile phase, the column temperature was 30℃, the flow velocity was 0.2m L/min, the sample size was 5μL. RESULTS: The linearity of adenosine and cordycepin both were good in the range of 10~50μg/m L, while the linearity of cordyceptic acid was good in the range of 100~500μg/m L.The average recovery of adenosine, cordycepin and cordyceptic acid were 96.56%, 96.27%, and 97.45% respectively, their RSD were 2.75%, 2.60%, and 2.34% respectively.The detection limit were 0.16μg /m L,0.23μg /m L and 7.3μg /m L respectively, the content of adenosine, cordycepin and cordyceptic acid in sample are 0.74mg/g, 0.22mg/g and 5.1mg/g respectively. CONCLUSION: The method was simple but rapid, which can be applied for quality control of Sinensis heath food and fast identification of its adulterants.(4)Determination of methylmercury(Me Hg) in fish by HPLC-AFSOBJECTIVE: To establish a determination method of Me Hg residues in fish by HPLC-AFS. METHODS:Use the methrod of HPLC-AFS, chromatographic column was Solid Phase Extraction Column C18(150×4.6 mm,5μm), column temperature was 30 ℃, the mobile phase of solution was acetonitrile(5%, V/V)+ammonium acetate(0.462%, m/V)+cysteine(0.12%, m/V), the flow velocity was 1.0m L/min, the sample size was 10μL, and the conditions of atom fluorescence spectrometry were that total current was 30 m A, the negative high-voltage was 270 V, the flow rate of carrier gas was 600 m L/min, and the flow rate of shield gas was 1000 m L/min. RESULTS:The recoveries ranged from 94.3% to 101.4%, and the RSD ranged from 2.24% to 4.70%; when the limits of detection(LOD) for methylmercury was 0.003 mg/kg(S/N=3), and the limits of quantification(LOQ) was 0.006 mg/kg(S/N=10). CONCLUSION:The proposed method has good reproducibility and high recovery, which can be used for the determination of methylmercury in aquatic product(fish).The main contribution includes the following: ①proposing the method which can be used to determine polysaccharide peptides in Ophiocordyceps Sinensi, which is simple, rapid, accurate and has high recovery, similar method had not been reported so far; ②proposing the methods which can be used to simultaneously determine the content of two reactive components-adenosine and cordycepin, or three reactive components-adenosine, cordycepin and cordyceptic acid in Ophiocordyceps Sinensis, exploring and optimizing the experimental parameters, which achieves good experiment results; ③proposing the method which can be used to determine,with experimental tests of fish samples taken from local market, this method is simple, and convenient, and has good result. |
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