HPLC determination of the Shanghai area in differe

e707004304

HPLC determination of the Shanghai area in different harvesting of Citrus aurantium synephrine content

ol ~ amine modifier; column temperature: 35 ℃; flow rate: 1.0ml/min; detection wavelength: 224nm; injection volume: 10tA, external standard calculation point method Synephrine content. In the proposed chromatographic conditions, citrus aurantium synephrine are well separated in, HPLC chromatogram shown in Figure 1. A chromatogram of a 4681012tItrim Figure 1HPLC (A: Synephrine) 2.2 Preparation of reference substance solution accurately weighed amount of synephrine, adding 0.1mol / L phosphoric acid preparation of 0.1mg/ml of standard solutions, shaken, over 0.45m filter membrane, is, too. 2.3 Preparation of the test solution to take citrus aurantium crude drugs, dried in the sun to dry like the black continuous drying at 60 ℃ after 8h, crushed all over 60 mesh, 60 ℃ dried overnight. Weigh appropriate amount (equivalent to synephrine 5mg), set 50m1 flask, add 30m10.1mol / L phosphoric acid, ultrasonic extraction 30min, placed at room temperature, 0.1mol / L phosphoric acid diluted to the mark, shake, too 0.45m membrane, that is, too. 2.4 The standard curve precision drawing the synephrine reference solution 1, the set of 10 flask, with 0.1rml / L phosphoric acid diluted to the mark, too 0.010mg/ml solution, followed by injection 4,6, 8,10,20, measured peak area to peak area Y is vertical axis, standard sample size for the horizontal X 3,772,004 in September sitting of Shandong University of Traditional Chinese Medicine Volume 28, No. 5 standard regression analysis, the regression equation : Y = 6874,20981 X +4.77754,[link widoczny dla zalogowanych], r = 0.9999. Synephrine content of 0.04 to 0.2 good linear relationship. 2.5 precision experiments autosampler precision drawing synephrine reference solution 10l, 6 injection were repeated to determine Xinfu Raymond area, RSD = 0.50%. 2.6 reproduce the experiment with the number of samples taken, accurately weighed 5 copies determination method by sample extraction, analysis, results of Xin Fu Raymond area RSD was 1.12%. 2.7 Stability test the prepared test solution into the sample analysis, after 1 injection every 2h, measured 3 times and the results Synephrine content was 0.64% RSD, indicating that the content of the test solution little change within 6h. 2.8 The average recovery obtained experimental precision that a number of herbs with known content of 6 copies, respectively 加入辛弗林 reference substance, the test solution according to preparation method of treatment, and appropriate dilution, calculating recoveries, the results The average recovery was 99.07%, RSD = 1.23%. 2.9 Determination of the samples in different locations,[link widoczny dla zalogowanych], different times by picking citrus aurantium samples and chromatographic conditions the system of law which synephrine levels were measured, the results shown in Table 1. Table 1 in Shanghai during the picking of citrus aurantium different biomass, particle size and concentration on the ~ Synephrine. . . . .1 Application of the maximum absorption wavelength of the selected diode array detector DAD UV absorption wavelength of synephrine, 224nm of maximum absorption, when testing the baseline stability before and after the impurity synephrine small, so the choice of 224nm for the synephrine detection wavelength. 3.1.2 choice of mobile phase for the nitrogen-containing synephrine alkaloids, so the mobile phase ion-pair reagent to be added. The first experiment used acetonitrile water a phosphate sodium dodecyl sulfate (32:68:0.1:0.1) as mobile phase, but in this condition, Xin Fu Raymond occur, resolution is poor (Figure 2); with sodium lauryl sulfate instead of sodium dodecyl sulfonate Sodium, in the same ratio, no peak synephrine, but peak is 378 wide, serious peak tailing (Fig. 3); joined 5mmol / L of diethylamine modifier, the synephrine chromatography before and after the peak of the peak and the baseline separation of sharp peak, retention time constant,[link widoczny dla zalogowanych], the separation of satisfaction (Figure 4). 3.2 aurantium at each stage according to the average diameter size of the harvest can roughly determine the date of citrus aurantium, easy to harvest. In addition , May to mid-July early, citrus aurantium synephrine levels in descending order. The longer the larger particle size citrus aurantium, synephrine lower levels, but each and the absolute amount of synephrine in Citrus aurantium increases. From our interval of two weeks picking every situation, more than doubled its volume. Therefore, we believe that the best picking citrus aurantium in Shanghai in June should be the end of the period to between early July. [
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