The Study Of Functional Oral Solution With Compound Amino Acids And Compound Vitamins

The amino acid is the basic material which the human body vital activity must have,a protein can have more than 20 kinds of amino acids,of which 8 kinds of amino acids cannot synthesize in vivo,must supplement from food.The physiological effects of amino acids are:energy generation,promoting protein synthesis,promote collagen synthesis,promote growth hormone secretion and improve immune function, skin beauty and beauty,and so on.Vitamin is maintains kind of low molecular organic compound which the human body and the animal body normal physiology function must need,the human and the animal to the vitamin demand,although is not much,its function is very significant. Because the human and animal in vivo cannot synthesize the vitamin,must rely on food and the feed provides the enough vitamin.When the body's vitamin intake from the outside world can not meet the needs of their lives,it will cause metabolic functions of the disorder,leading to illness or even death.The main contents of this topic primary include the study of compound amino acids function oral solution and the study of compound vitamins function oral solution. In this paper,the work includes the following four components:The first part:Establish a HPLC analysis method for compound amino acids functional oral solutionUse Agilent 1100 HPLC instrument:VWD UV detector,G1354A pump(US Agilent company);chromatographic column:Diamonsil^TM C₁₈ column(250×4.6 mm, 5μm);precolumn C₁₈ column(10×4.6 mm,5μm);The mobile phase A was 0.1 moL·L^-1 sodium acetate,and the mobile phase B was acetonitrile;The flow rate was 1.0 mL·min^-1;The wavelengths for detection was 254 nm;The column temperature was 40℃and the injection volume was 20μL;The amino acids were precolumn derivatized with phenyl isothiocyanate(PITC) and were detected after separated under the condition of gradient elution.Experiment result showed that 16 amino acids were separated within 43 min; Within the concentration range,the correlation coefficients of 16 amino acids were 0.9996 - 0.9999;The intra-day RSD was not more than 1.87%,and the inter-day RSD was not more than 1.89%;The detectability of 16 amino acids were 0.61 - 2.50 ng,it shows well sensibility;The average recoveries were 95.97%- 103.99%;and the sample was stable and with well detection result.The method is efficient,sensitive and accurate,with well specificit,and convenient and useful.The second part:Preparation and stability investigation of compound amino acids function oral solutionPreparated functional oral solution included 16 amino acids,which contains 8 essential amino acids.This oral solution was sterilizated under the 120℃high-temperature for 20 min.Inspects 60℃high temperature influencing factor test and 45001x±5001x illumination glare influencing factor test,to influence the stable key inspection project of the amino acids oral solution;The acceleration test of the oral solution which according to the stable key inspection project was taken under 40℃high temperatures for 6 months,long-term test was simultaneously carried on.The results showed that the oral solution could be sterilizated under the 120℃high-temperature for 20 min,60℃influencing factor test indicated when the amino acids oral solution exposured to 60℃for 10 d,the stable key inspection project of the amino acids oral solution:the solution color,Smell,taste,pH value,clarification, the content of animo acids are essentially unchanged.45001x±5001x illumination influencing factor test indicated when the amino acids oral solution exposured to 45001x±5001x illumination for 10 d,the stable key inspection project of the amino acids oral solution:the solution color,Smell,taste,pH value,clarification,the content of animo acids are essentially unchanged.The high temperature acceleration test showed the oral solution was lay aside stably under 40℃conditions for 6 months. The long-term table showed the oral solution can lay aside 6 months under 25℃conditions(present to achieve 6 month data) stably.It means that the oral solution with well stability.The third part:Establish a HPLC analysis method for compound vitamins functional oral solution Use RP-HPLC method,and choose two chromatographic conditions to detect two kinds of fat-soluble vitamins(V_A and V_E,chromatographic conditions 2) and four water-soluble vitamins(V_B1,V_C,V_B6 and V_pp,chromatographic conditions 1), separately.Chromatographic conditions 1(detecting for V_B1,V_C,V_B6 and V_pp):Agilent 1100 HPLC instrument:VWD UV detector,G1354A pump(US Agilent company); chromatographic column:Diamonsil^TM C₁₈ column(250×4.6 mm,5μm,Diamond Technology Co.,Ltd);precolumn C₁₈ column(10×4.6 mm,5μm);The mobile phase was methanol - 0.05 moL·L^-1KH₂PO₄(include 0.3%triethylamine,pH6.0)(7:93); The flow rate was 1.0 mL·min^-1;The wavelengths for detection was 266 nm;The column temperature was 25℃and the injection volume was 20μL.Chromatographic conditions 2(detecting for V_A and V_E):Agilent 1100 HPLC instrument:VWD UV detector,G1354A pump(US Agilent company); chromatographic column:Diamonsil^TM C₁₈ column(150×4.6mm,5μm,Diamond Technology Co.,Ltd);precolumn C₁₈ column(10×4.6mm,5μm);The mobile phase was methanol-water(98:2 );The flow rate was 1.0 mL·min^-1;The column temperature was 25℃;The wavelengths for detection was 325 nm when 0 min,and 280 nm when 10 min;and the injection volume was 20μL.Experiment results showed that under chromatographic conditions 1,V_B1,V_C, V_B6 and V_pp were well separated within 15 min,under chromatographic conditions 2, V_A and V_E were well separated within 23 min;Within the concentration range,the correlation coefficients of 6 vitamins were 0.9997 - 0.9999;The intra-day RSD of 6 vitamins were not more than 1.55%,and the inter-day RSD was not more than 1.75%, indicated well precision;The detectability of 6 vitamins were 0.08 - 0.68 ng,it shows well sensibility;The average recoveries were 95.30%- 102.52%;and the sample was stable and with well detection result.The method is sensitive and accurate,with good linear correlation and recoveries,convenient and useful.The fourth part:Preparation and investigation of V_A and V_E nanoemulsionUse Tween-80 as surfactant,and use glycerin(Gly) as cosurfactant,the mixed surfactant was Tween-80:Gly(4:1);took mixed surfactant:mixed vitamins=7:1, (mixed vitamins oil is V_A:V_E=1:3,m/m ),prepared V_A and V_E nanoemulsion under 25℃and with magnetic stirring.The physical and chemical stability of V_A and V_E nanoemulsion were also investigated.The results showed that the nanoemulsion with small particle size, approximately 15 nm,and with narrow distribution was successfully prepared,Zeta potential of the nanoemulsion was -20.9±1.6 mV.The destruction of V_A in nanoemulsion is extremely serious when heated under 120℃,it indicated that compound vitamins oral solution cannot be sterilizated under the high-temperature.Influencing factor test under 60℃,40℃,30℃indicated that V_A was serious damaged when the temperature was above 30℃,but V_E was very stable under different temperature;when the nanoemulsion exposured to 25℃for 10 d,V_A content is 98.94%compare to 0 d,and V_E content is 101.65%,it indicated that V_A and V_E were stable under 25℃.45001x±5001x illumination influencing factor test showed that when the nanoemulsion was exposured to 45001x±5001x illumination for 10 d,approximately 10%of V_A content was damaged, but V_E content was not change.That means the nanoemulsion is physical stable,but is not chemical stable.Nanoemulsion preparation must be protected from light,and cannot stored under the temperature above 25℃.