茄尼醇同步提取皂化-超高效液相色谱测定方法研究

为了建立高效、准确检测烟草总茄尼醇的方法,研究了超声辅助条件下,同步提取、皂化烟草茄尼醇的溶剂、温度、液料比、皂化剂用量、提取时间及超高效液相色谱测定的仪器条件。结果表明,烟草样品以正己烷为萃取剂,液料比为50:1,0.1 moL/L氢氧化钠的乙醇溶液为皂化剂,在超声频率45 kHz,恒温水浴40 ℃,提取时间30 min条件下,完成烟草茄尼醇的提取、皂化,且使茄尼醇的提取、皂化以及与皂化剂的分离在同一离心管中完成。以ACQUITY UPLC BEH C18为色谱柱,甲醇-乙腈(50:50)为流动相,流速为0.5 mL/min,柱温为35 ℃,在波长208 nm条件下,超高效液相色谱进行检测。方法的线性范围为0.67~84.1 mg/L,方法检出限为0.07 mg/L,定量限为0.012%,空白及样品加标回收率分别在97.9%~104.7%、93.4%~102.3%范围内,相对标准偏差为1.34%~2.43%。该方法简便、快速,且节约有机溶剂,精密度和准确度较高,可以实现烟草茄尼醇批量高效检测。

The Simultaneous Extraction and Saponification of Tobacco Solanesol using Ultra Performance Liquid Chromatography

In order to established the method of determining total solanesol in tobacco efficiently and accurately ,we studied the conditions of simultaneously extracting and saponifying solanesol from tobacco, including solvent, temperature, liquid ratio, saponification dosage, extraction time and ultrasound-assisted instruments conditions of ultra-high performance liquid chromatography. Hexane was used as solvent for extraction with liquid-material ratio of 50:1 and 0.1 moL/L sodium hydroxide in ethanol solution was used as saponification agent. The ultrasonic frequency used was 45 kHz, temperature of extraction and saponification was 40 ℃, and time of extraction was 30 min. Under this condition, extracting and saponifying solanesol from tobacco were finished simultaneously, and saponification agent separation was done in the same centrifuge tube. The separation of target compound was performed on an ACQUITY UPLC BEH C18 column using methanol-acetonitrile (50:50) as mobile phase by gradient elution. Flow rate was set at 0.5 mL/min, column temperature was 35 ℃ and injection volume was 5 μL. The quantitative wavelength of UV detector was set at 208 nm. The results indicated that the calibration curve was linear in the range o f 0.67-84.1 mg/L, the limit of detection (LOD, S/N=3) was 0.07 mg/L, and the limit of quantification (LOQ, S/N=10) was 0.012%. The standard addition recoveries of blanks and samples were 97.9%-104.7% and 93.4%-100.0%, respectively, with relative standard deviations (RSDs) of 1.34%-2.43%. The method was simple, rapid, precisive, accurate, and solvent saving, suitable for batch testing .