反相HPLC法测定余甘子果实中的游离氨基酸

应用反相HPLC法测定了余甘子果实中的游离氨基酸,色谱柱为爱尔兰产150mm×4．6mm，5μ RP18柱．流动相A为0．04mol／L KH2PO4（pH7．2±0．05）、B为水、C为乙腈，流速0．65mL／min，梯度洗脱，检测波长360nm，柱温40℃．结果显示，16种氨基酸标准品的线性范围为0．0156—0．2500μmol／mL，相关系数球=0．9949—0．9999．标准品在0—48h内经多次测定，保留时间和峰面积的相对标准偏差分别在0．03％-1．75％和0．54％-3．67％之间；回收率和回收率相对标准偏差分别在85．95％-102．18％和1．10％-6．54％之间．4种余甘子果实中检出有9种游离氨基酸，分别是天门冬氨酸、谷氨酸、丝氨酸、精氨酸、甘氨酸、苏氨酸、脯氨酸、丙氨酸和苯丙氨酸，含量为0．0042—0．5666mg／g干质量．

Determination of free amino acids in the emblica fruit by reversed-phase HPLC

Free amino acids in the emblica fruit were determined by a reversed-phase high performance liquid chromatography （RP-HPLC） method. Samples were injected into 5μ RP18 column （150 mm × 4. 6 mm） made in Ireland. Mobile phase A was 0. 04 mol/L KH2PO4 with pH 7. 2 ±0. 05, B was water and C was acetonitrile. Gradient elution was proceeded at the flow rate of 0. 65 mL/min. Operation conditions were set as follows： wavelength of 360 nm, column temperature of 40℃. The results showed that linear calibration of sixteen standard amino acid samples were within 0. 015 6 -0. 250 0 μmol/mL with correlation coefficient （ R^2） of 0. 994 9 - 0. 999 9. Relative standard deviation （RSD） of retained time and peak area were respectively within 0. 03% -1.75% and 0. 54% -3. 67% after repeated analyzed of standard samples were within 48 h. Recovery ratio was within 85. 95% - 102. 18% with RSD of 1.10% - 6. 54%. Nine free amino acids including Asp, Glu, Ser, Arg, Gly, Thr, Pro, Ala and Phe in emblica fruit were determined, and their contents were between 0. 1304 2 to 0. 566 6 mg/g dried mass.