The Research On Extraction Of Natural Hormone From Antler Velvet With Supercritical CO₂

Antler velvet has been used in traditional Chinese medicines for over two thousand years. It has long been recognized as one of the most effective and powerful invigorants for strengthening the kidney, sexual-reinforcing, and pro-longing life. The products can have certain effects and be safe. Antler velvet contains various active ingredients including natural estrogen and progesterone which can efficiently guard against and treat the menopause syndrome. The extraction of natural hormone from antler velvet has important academic value and application value for developing products which can prevent and cure menopause syndrome.Based on the previous research results in our laboratory, supercritical CO₂ (SC-CO₂) extraction technology was firstly used and the process of extraction of natural hormone from antler velvet was systematically studied. Influences of concentration of ethanol, dosage of co-solvent, extraction pressure and temperature on the extraction yield and the contents of estradiol and progesterone were investigated. The experimental results present that the extraction yield can reach 1.56% when 85% ethanol as static co-solvent, the ratio of co-solvent (mL) and the quality of antler velvet (g) is 1. The contents of estradiol and progesterone in the extracts are 3.07ng/g and 776.18ng/g, respectively. The extraction yield can reach 3.88% when adding 85% ethanol by pump, the ratio of co-solvent (mL) and the quality of antler velvet (g) is 15. Under such conditions, the contents of estradiol and progesterone in the extracts are 3.86ng/g and 588.34ng/g. Traditional methods like the refluxing extraction technology, soxhlet method, oscillation method and the ultrasonic wave method were also investigated. Then SC-CO₂ and the refluxing extraction technology were also compared together and the results show that supercritical CO₂ extraction by adding co-solvent by pump has better effect.At the same time, the extracts from antler velvet with supercritical CO₂ were also systematically studied by various analytical methods as follows. The content of estradiol and progesterone was determined by radioimmunoassay (RIA). GC-MS was employed to analyze lipids of the extracts. O-phthaladehyde colorimetry method was established to determine the content of cholesterol in extracts. The phospholipids were analyzed by thin layer chromatography (TLC). The content of hypoxanthine and p-hydroxybenzaldehyde were identified by HPLC. The establishment of analytical methods can provide some essential application basis data for controlling the quality of products.In addition, antler velvet polysaccharide was then separated from the remainder of supercritical CO₂ extraction by hot water extraction, diluted alkali solution extraction and diluted alkali solution combined with enzymolysis extraction, respectively. The results show that diluted alkali solution combined with enzymolysis extraction is a good choice to make full use of the remainder of extraction for future research.