Extraction And Purification Of Squalene And Product Research Of Soft Capsules Carrying Oxygen

Squalene is a highly straight unsaturated chain of three terpenoids,has many important biological activity.It is an excellent blood oxygen agent.So,The research has a very importent significance.In this paper,a new method of high performance liquid chromatography?HPLC?for the determination of squalene was established.The results showed that the method was effective and accurate.The content of squalene in raw material?soybean oil deodorized distillate?was 1.92%.Secondly,NaOH-ethanol was used for saponification of soybean deodorizer distillate and extracted with n-hexane to obtain unsaponifiable matter containing 5.58% squalene in a yield of 92.02%.The unsaponifiable matter was used as the raw material and through the method which combined silver ion complexation extraction with column chromatography.The squalene content can be enriched to 94.16%.Finally,based on the study of the oxygen carrying capacity of squalene in vitro and in vivo,squalene soft capsules with improved hypoxia tolerance was developed to provide scientific basis for improving people's life and seeking alternatives to rhodiola.Specific research conclusions are reached as follows:?1?We have established a HPLC method for analysis of squalene,specific chromatographic conditions: C18 column,mobile phase of methanol and water?90:10,v/v?as A phase and isopropyl alcohol as B phase?A: B = 80:20?,the velocity is 1.0 mL/min;sample size is10 ml.Under these conditions,the retention time of squalene was 18.528 min,and the peak time was 8 min ahead of the current method.The optimum process of saponification: reaction time 50 min,reaction temperature is 70 ?,alkali concentration 1 mol/L,the solid-liquid ratio was 1: 3,squalene purity was 5.58%,the extraction rate was 92.02%.?2?For the first time using silver ions complexation extraction combined with the method of column chromatography separation of squalene.First of all,according to the central composite design principle of the Box-Benhnken optimization of silver ions complexation extraction process conditions for methanol volume fraction of 60%?v/v?,the temperature of 0 ?,time of 8 h,AgNO₃ concentration is 1.5 mol/L,squalene feeding ratio of 4:1;the optimum stripping time was 60 min,extraction temperature was 65 ?.In this condition,the concentration of squalene was highest,up to 46.52 g/ml,the yield of 77.27%.After complexation extraction,Then with silica gel as stationary phase and n-hexane as the elution solvent,investigation on the elution rate and sample weight,etc.,and the effects of factors on the chromatography.To determine the best technology of column chromatography purified squalene: the sample amount and ratio of 1:20,silicone velocity for 15 ml/min.Take each fraction zhenggan available squalene extract,purity is 94.16%,the recovery was 96.13%.?3?In vitro oxygen carrying test device of squalene was designed and manufactured.The process of oxygen-oxygen release was discussed and through the hypoxia tolerance test in mice,determine its carry oxygen saturation dose of 60 mg/kg·bw.The formulation of the product?1000 grains?: The raw material: Squalene extract 700 g?purity 94.16%?;Ingredients: soybean oil 1400 g?beeswax 84 g?polyglycerol ester 7g.Tests show that this product can significantly improve hypoxia tolerance.