反式茴脑生物合成茴香酸体系紫外分光光度法的测定方法研究

Determination method of the system of trans-anethole biosynthesis anisic acid by UV spectrophotometry

根据反式茴脑和茴香酸及其混合物的紫外光谱特性,利用等吸收点建立了一种监测生物合成过程的紫外定量分析方法。混合物在250.0 nm处的吸光度与其总摩尔浓度的标准曲线A250.0=16.931Ct+0.012 8,线性范围为0.01～0.06 mmol/L;当总摩尔浓度为0.04 mmol/L时,混合物在268.5 nm处的吸光度与反式茴脑的摩尔分数的标准曲线为A268.5=0.342 7X+0.311 2。据某样品稀释前后的A250.0、A'268.5、A'250.0和A''268.5及光谱模拟得到该样品浓度为0.04 mmol/L的A268.5=(0.675 6-A'250.0)(A'268.5-A''268.5)/(A250.0-A'250.0)+A''268.5。反式茴脑和茴香酸的平均回收率分别为98.40%和96.47%,相对标准偏差分别为0.68%和0.80%,与高效液相色谱法比较的t值均小于结果的临界值1.724 7(α=0.05),表明该方法与高效液相色谱法无显著性差异,准确可靠,可为紫外分光光度法监测生物合成过程提供参考。

A quantitative analysis method by UV spectrophotometry monitoring biosynthesis was established with isoabsorptive point, being based on the UV spectral characteristics of trans-anethole and anisic acid and its mixture. Standard equation of absorbance at 250 nm and total molar concentration of mixture was A250.0 = 16.931 C, ＋ 0. 012 8, and linear range at 0. 01 - 0. 06 mmol/L; while the total molar concentration was 0. 04 mmol/L,standard equation of absorbance at 268.5 nm of mixture and molar fraction of trans-anethole was A268. 5 = 0. 342 7X ＋ 0.311 2. According to spectral simulation and A250. 0,A＇268.5. 5 ,A＇250. 0 and A ＂268. 5 before and after one sample dilution, A268. 5 = （ 0. 675 6 - A＇250. 0 ） （ A＇248.5 - A ＂268.5 ） / （ A250.0 - A＇250.0） ＋A ＇268.5 while the sample total molar concentration at 0.04 mmol/L. The average recovery of trans-anethole and anisic acid was 98.40% and 96.47% respectively,and the relative standard deviation of them was 0.68% ,0.80%. Comparing with HPLC,the t value test was less than the critical value of 1. 724 7 （ a = 0.05 ）. The results showed that the method was accurate and reliable, and had no significant difference with HPLC, which provided reference for monitoring biosynthesis process by UV spectrophotometry.