Studies On Determination And Fingerprint Of Amino Acids In Mannatide Oral Solution

The paper is studied on resolving the lack of quality standard system on the amino acids in mannatide oral solution. An microwave hydrolysis method combining with the high performance liquid-pre column derivation was established for analyzing the amino acids in mannatide oral solution. The method was investgated by making the standard curves and methodological experiments. The fingerpringt of mannatide oral solution was established as well as the fingerprint of tcm. The results were shown below:1. The optimum conditions of microwave hydrolyzation were investigated. Obtained amino acids were derived by phenyl isothiocyanate, and were determined by high performance liquid chromatography. The optimum conditions of microwave hydrolyzation are:the volume ratio of HC1 and sample is 1:3, the temperature is 170℃, and the time is 10 min. The effect of microwave hydrolyzation was similar to the 24h traditional hydrolyzation. And 13 amino acids we got in mannatide oral solution. The method of amino acids determining was investigated. Chromatographic column:Shimadzu Shim-pack vp-ODS(150mm×4.6mm, 5μm); mobile phase A:pH6.0 acetate-acetonitrile(95:5), mobile phase B: acetonitrile-water(60:40), mobile phase C:pH5.0 acetate-acetonitrile(97:3); gradient elution; flow rate:1.0mL/min;detection wavelength:254nm. The calibration curves showed good linear relations, and correlation coefficient of 13 amino acids are in the range of 0.9922～0.9989. The average recovery of 13 amino acids are in the range of 98.4～100.9%, and RSD(n=6) are in the range of 1.1～2.8%. The samples are stable in 24h, and repeatability are well.2. The fingerprints of non-hydrolyzed sample and hydrolyzed sample were both established. There were 16 common peaks in chromatograms of 10 patches of non-hydrolyzed sample, 15 common peaks in chromatograms of 10 patches of hydrolyzed sample, and 13 peaks for amino acids. The similarity count of 10 patches of non-hydrolyzed sample was in the range of 0.953～0.998, and the hydrolyzed sample was in the range of 0.941～0.986. 3. The evaporative light scattering detector(ELSD) was applied to analyze the 18 amino acids standards and mannatide oral solution. Only 7 amino acids were absolutly separated. On this condition,6 amino acids in mannatide oral solution were determined, and the methodological experiments showed well.