| Hydroxysafflor yellow A?HYA?is an active ingredient extracted from safflower,which is used for cardiovascular disease in clinical,like angina,but also can lower blood pressure,and exhibits anti-tumor,anti-inflammatory,anti-oxidant,anti-aging and other pharmacological effects.However,the clinical oral application of HYA is limited for the poor permeability and low bioavailability.In this paper,HYA was prepared into hydroxysafflor yellow A nanoemulsion?HYANE?:oil-in-water HYANE?HYANE-O?and water-in-oil HYANE?HYANE-W?.We established a method for the determination of HYA in vivo and in vitro.And the quality of HYANE was evaluated by the physicochemical properties and pharmacokinetic experiments.The first section,determination of the content of HYA in HYANE.Objective:To establish the method determination of the content of HYA in HYANE.Method:HPLC method:the column was an Elite C18 column?250 mm×4.6 mm,5?m?;the detection wavelength was 403 nm;and the mobile phase was acetonitrile?A?-20 mmol/L phosphate buffer?pH 3.5,B?,gradient elution:03 min,10%A20%A;310 min,20%A;10.0120 min,10%balance.Results:The linear regression equation of HYANE was A=26852C-582,r=0.9998,and there was a good linear relationship in the concentration range of 1100?g·mL-1.The recovery rate were 95.00%,95.76%,99.28%,and RSD were 2.46%,0.92%,0.98%,respectively;the RSD of inter-day and within-day precision were 2.69%,0.90%,0.07%and1.25%,0.80%,1.65%,respectively.Conclusion:The method had high accuracy and good sensitivity,which could be used for the determination of HYA in HYANE.The second section,preparation and physicochemical properties of HYANE.Objective:To prepare HYANE-O and HYANE-W,and to evaluate their physicochemical properties.Methods:To prepare HYANE-O and HYANE-W,and to evaluate their physicochemical properties,such as appearance,transmission electron microscope,particle size and Zeta potential.Results:The appearance of HYANE-O and HYANE-W were all clear and transparent.Transmission electron microscope showed that HYANE-O was round,non-sticky and evenly distributed.The particle size of HYANE-O was 23.28±0.49 nm,and the PDI was 0.42±0.01,and the Zeta potential was-2.62±0.12 mV.There was no data about the particle size of HYANE-W,and the Zeta potential was about 0 mV.Conclusion:HYANE-O and HYANE-W were prepared.The third section,determination of the content of HYA in plasma samples.Objective:To establish the determination method of the content of HYA in plasma samples.Method:HPLC method:the column was an Elite C18 column?250 mm×4.6 mm,5?m?;the detection wavelength was 403 nm;and the mobile phase was acetonitrile?A?-20 mmol/L phosphate buffer?pH3.5,B?,gradient elution:03 min,10%A20%A;310 min,20%A;10.0120 min,10%balance.Rutin?RT?was used as an internal standard.Results:The linear regression equation of HYANE was Y=2.1393X+0.0631,r=0.9999,and there was a good linear relationship in the concentration range of 0.0520?g·mL-1.The extraction recovery rate were 98.68%,95.72%,97.10%,and RSD were 1.84%,0.51%,1.24%,respectively;the method recovery rate were 90.59%,97.29%,101.40%,and RSD were 2.49%,3.24%,1.00%,respectively;the RSD of inter-day and within-day precision were 1.26%,1.09%,0.68%,and 1.68%,1.96%,1.29%,respectively.Conclusion:The method was sensitive and reliable,which could be used for the determination of the content of HYA in plasma samples.The fourth section,pharmacokinetics evaluation of HYANE.Objective:To evaluate the pharmacokinetics of HYANE-W and HYANE-O in vivo.Methods:The pharmacokinetic parameters and bioavailability of HYA,HYANE-W and HYANE-O were investigated.Results:The peak concentration of HYANE-W was about 13.34 times of HYA,the peak time was about 2.27 times of HYA,and the bioavailability of HYANE-W was about 4.15 times of HYA;The peak concentration of HYANE-O was lower than that of HYA,the peak time of was higher than that of HYA,and the bioavailability of HYANE-O was lower than that of HYA.Conclusion:The preparation of HYA into HYANE-W can improve the bioavailability of HYA. |
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