|
Cordyceps [Cordyceps sinensis (Berk.) Sacc.] Is a traditional precious Chinese medicine has the role to protect the respiratory system , maintenance of renal function , anti-cancer , anti- oxidation . Its anamorph Hirsutella of China (Hirsutella sinensis Liu, Guo, Yu Zeng), has broad application prospects . Sterol Cordyceps main active ingredient , wherein the ergosterol is fungi characterized in sterols , has significant anticancer activity , the main subject of Chinese Hirsutella sterol extract the optimal conditions for the study . Total sterol content of phosphorus and sulfur iron law and vanillin - acetic acid colorimetric determination method studies , 2 ways color absorbance change by less than 1% after 1 hour , better stability ; precision (RSD) 1.9% and 3.6% , respectively ; recoveries were 126.3 ± 16.5% and 105.4 ± 3.6 % , higher accuracy of vanillin - acetic acid method ; Determination select vanillin - acetic acid method for Hirsutella total sterol content method. Single factor experiments examined saponification with alkali, ethanol concentration , extraction time , solid-liquid ratio , extraction times five factors of the ultrasonic assisted extraction of total sterol , found saponification with alkali, ethanol concentration , extraction times three factors adjacent level significant difference (P lt; 0.05), and extraction time , solid to liquid ratio ( 1/100 ) reference to extract the impact was not statistically significant sex ( P gt ; 0.05) ; the KOH / mycelium ratio were 0 , 0.5, 1 , ethanol concentration were 60% ??, 80% , 95% , extraction times were 1, 2, 3 of 3 factors and 3 levels orthogonal optimization of the ultrasonic assisted extraction conditions , was the best extraction conditions were as follows : KOH / mycelium ratio is 1 , and the ethanol concentration is 95% and extracted three times with a total sterol content of about 5.016 % ± 0.205% , and measured . Single factor tests show little effect of temperature on the heat reflux extraction effect ; to 3 factors ultrasonic 3 level orthogonal design to examine the thermal reflow method was optimum extraction conditions were : reflux temperature 50 ° C , KOH / mycelium body ratio of 0.5 , the ethanol concentration was 95% , extracted 2 times , each 1 hour . Validation studies have shown that the ultrasonic method is better than the thermal reflow method . By RP-HPLC China Hirsutella total ergosterol content Hypersil C18 reverse column (250 mm × 4.6 mm , 5 μm ) , pure methanol flow , sample volume 201μL, flow rate 1.0mL/min detection wavelength 282nm, measured total ergosterol content was 0.08% .
|