超高效液相色谱-串联四极杆质谱法分析婴儿乳粉中的唾液酸  

Analysis of sialic acid in infant formulas using ultra performance liquid chromatography-triple quadrupole mass spectrometry

作　　者：解鸿蕾[1,2] 李春[1] 刘宁[1,2] 

机构地区：[1]东北农业大学食品学院,乳品科学教育部重点实验室,黑龙江哈尔滨150030 [2]国家乳业工程技术研究中心,黑龙江乳品工业技术开发中心,黑龙江哈尔滨150086

出　　处：《色谱》2013年第8期781-785,共5页Chinese Journal of Chromatography

基　　金：“十二五”国家科技支撑课题(2011BAD09B0304)

摘　　要：建立了超高效液相色谱-串联质谱(UPLC-MS/MS)测定婴儿配方乳粉中唾液酸含量的分析方法。利用酸水解方法释放出婴儿配方乳中的唾液酸,经HLB反相色谱固相萃取柱净化,采用BEH HILIC色谱柱分离,以0.1%甲酸水溶液和100%乙腈溶液作为流动相进行梯度洗脱,流速为0.25 mL/min,进样体积5μL,柱温30℃,电喷雾质谱检测,正离子多反应监测模式进行定性和定量分析。结果表明:唾液酸在0.05～5.0 mg/L范围内与唾液酸峰面积的线性关系良好(R2=0.998 9);以0.1、0.5、2.5和5.0 mg/L4个添加水平进行添加回收试验,唾液酸的平均回收率为84.3%～98.9%,相对标准偏差为4.9%～8.2%;唾液酸的检出限为0.01 mg/L。该方法简单、快速、重复性好、灵敏度高,可广泛用于奶粉、牛奶及母乳中唾液酸含量的分析测定。The method for analysing sialic acid in infant formulas by ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） has been established. Sialic acid in milk was released via acid hydrolysis, and purified by an HLB solid phase extraction car- tridge. The UPLC separation was performed on an ACQUITY UPLCTM BEH HILIC column （50 mm × 2. 1 mm, 1.7 μm） utilizing a gradient elution program of acetonitrile and water （ contai- ning 0. 1% formic acid） as the mobile phases at a flow rate of 0.25 mL/min. Injection volume and column temperature were set at 5 μL and 30 ℃, respectively. The identification and quanti- fication were achieved by using electrospray ionisation （ESI）-MS/MS in positive ion mode and multiple reaction monitoring （MRM） mode. The linear range was from 0.05 to 5.0 mg/L for sialic acid and the correlation coefficient （ R2 ） was greater than 0.99. The average recoveries spiked at the four concentration levels of 0. 1, 0.5, 2.5 and 5.0 mg/L ranged between 84.3% and 98.9% with the relative standard deviations from 4.9% to 8.2%. The limit of detection was 0.01 mg/L. Therefore, this method has the characteristics of simple operation, high reproduc- ibility and sensitivity. It can be widely applied to determine the total contents of sialic acid in infant formula, cow milk and human milk.

关 键 词：超高效液相色谱-串联质谱 唾液酸 婴儿配方乳 

分 类 号：O658[理学—分析化学]