鹿角脱盘及其5种提取部位21种氨基酸含量对比分析

目的：建立RP-HPLC法同时检测鹿角脱盘及其提取部位中21种水解氨基酸含量，分析鹿角脱盘5种提取部位氨基酸含量和组成。方法：采用RP-HPLC法，异硫氰酸苯酯（PITC）作为衍生剂，乙腈-水（4：1）为流动相A，0.1 mol&#183;L^-1乙酸钠溶液（乙酸调pH 6.5）-乙腈（97：3）为流动相B，检测波长254 nm，流速1 mL&#183;min^-1，柱温36 ℃，以梯度洗脱的方法（0 min，100%B；11 min，98.5% B；21.7 min，92.4%B；23.9 min，89%B；27 min，86.6%B；39 min，70%B；42 min，30%B；45 min，0%B；52 min，0%B；55 min，100%B；70 min，100%B）检测鹿角脱盘及其5种提取部位中水解氨基酸含量。结果：21种氨基酸质量浓度在0.001 9~2.700 0 μmol&#183;mL^-1范围内线性关系良好，R²均>0.992 0，水解氨基酸的加样回收率在93.2%~102.8%，RSD在0.65%~2.6%之间；5种提取部位的氨基酸含量均显著高于鹿角脱盘粉（P<0.05），但水提取部位、碱提取部位和醇提取部位间氨基酸含量无显著性差异（P>0.05）；药用氨基酸的比例达60%以上；鹿角脱盘碱提取部位中必需氨基酸与总氨基酸的含量比值和必需氨基酸与非必需氨基酸的含量比值为各组最高。结论：本方法适合鹿角脱盘及其提取部位水解氨基酸含量的测定；5种提取部位较鹿角脱盘有更高的医学和营养学价值。

Objective:To establish an RP-HPLC method for simultaneous determination of 21 hydrolyzed amino acids in deer antler base and its extractions,and compare the composition and contents of amino acids in 5 kinds of extractions.Methods:The samples were derivated with PITC and analyzed by RP-HPLC.The mobile phase consisted of acetonitrile-water(4:1)(A)and 0.1 mol&#183;L^-1 sodium acetate buffer solution(pH 6.5)-acetonitrile(93:7)(B)with gradient elution(0 min,100%B;11 min,98.5%B;21.7 min,92.4%B;23.9 min,89%B;27 min,86.6%B;39 min,70%B;42 min,30%B;45 min,0%B;52 min,0%B;55 min,100%B;70 min,100%B)at a flow rate of 1 mL&#183;min^-1.The column temperature was 36 ℃ and the detection wavelength was 254 nm.Results:In the range of 0.001 9-2.700 0 μmol&#183;mL^-1,the 21 amino acid had good linearity(the R² were all over 0.992 0).The average recoveries of hydrolyzed amino acids were 93.2%-102.8%(RSD=0.65%-2.6%).The amino acids contents in extractions were significantly above those in powdered deer antler base(P<0.05).But,there was no significant difference among the water extraction,the alkali extraction and the alcohol extraction.The medical amino acids accounted for more than 60% of the total contents.The content ratio of essential amino acids to total amino acids and the content ratio of essential amino acids to non-essential amino acids in the alkali extraction were higher than those in other samples.Conclusion:The established method was suitable for determination of amino acids in deer antler base and its extractions.The five extractions had higher medicinal and nutritional value than the deer antler base.