固相萃取-超高效液相色谱法测定黄酒中的有机酸  

Determination of organic acids in Chinese rice wine by solid phase extraction-ultra performance liquid chromatography

作　　者：王琳[1] 陈双[1] 徐岩 

机构地区：[1]江南大学,食品科学与技术国家重点实验室,工业生物技术教育部重点实验室,生物工程学院酿酒微生物与酶技术研究室,江苏无锡214122

出　　处：《食品与发酵工业》2016年第10期142-148,共7页Food and Fermentation Industries

基　　金：国家自然科学基金(31530055、21506074);国家高技术研究发展计划(863计划)(2013AA102108);江苏省自然科学基金(BK20140153)

摘　　要：确定了氨基酸/多肽和色素是黄酒有机酸色谱分析的主要干扰因素,筛选了对干扰因素有较强去除能力的固相萃取（SPE）小柱。建立了固相萃取-超高效液相色谱法（SPE-UPLC）快速测定黄酒中7种主要有机酸的方法。黄酒样品采用流动相稀释,经活化后的SCX-SPE小柱净化处理,并使用ACQUITY UPLC HSS T3柱对黄酒中的7种主要有机酸进行分离检测。流动相为20 mmol/L Na H2PO4（p H 2.7）,流速为0.25 m L/min,检测波长为210 nm。结果表明,该方法可在5 min内实现7种有机酸的完全分离,各有机酸在0.100-37 456.000 mg/L范围内线性相关性良好,回归方程的线性相关系数均在0.999 3以上,7种有机酸的加标回收率为94.96%-105.08%,相对标准偏差（RSD）为0.43%-1.79%（n=5）。该方法具有净化步骤简单,准确性高的特点。采用该检测方法对传统型黄酒（n=12）和清爽型黄酒（n=6）有机酸含量比较发现,传统型黄酒总酸含量显著（P〈0.05）高于清爽型黄酒,且这种差异是由于酒石酸、苹果酸和乳酸含量的显著差异（P〈0.05）造成的。The influence of major components in Chinese rice wine on the chromatographic analysis of organic acids was investigated. Amino acids,polypeptides and pigments were the major impurities which observably interfere with chromatographic separation of organic acids in Chinese rice wine. A method for the simultaneous determination of7 organic acids in Chinese rice wine was developed by solid-phase extraction and ultra performance liquid chromatography（ SPE-UPLC）. The sample was purified by the SCX-SPE cartridge after pretreatment,and then separated by ACQUITY UPLC HSS T3 column. 20 mmol / L Na H2PO4（ p H 2. 7） was used as the mobile phase at a flow rate of0. 25 m L / min. The detection was performed by a diode array detector at 210 nm. The results showed that 7 organic acids were completely separated and determined in 5 min. the linear correlation coefficients were above 0. 999 3 in the range of 0. 100-37 456. 000 mg / L. Under these conditions,the recoveries of 7 organic acids in rice wine were in the range of 94. 96%-105. 08% with the relative standard deviations（ RSDs,n = 5） of 0. 43%-1. 79%. Compared with traditional organic acid SPE＇s adsorption and elution operation,this method is feasible,accurate and applicable for the quantitative analysis of organic acids in Chinese rice wine. With this method,we analyzed the differences of the organic acids concentrations between Traditional and Qingshuang Chinese rice wine. The results showed that the total acids in Traditional Chinese rice wine was significantly higher（ P〈0. 05） than those in Qingshuang Chinese rice wine,while the difference was mainly caused by the remarkable difference（ P〈0. 05） in the concentrations of tartaric acid,malic acid and lactic acid.

关 键 词：超高效液相 固相萃取 有机酸 传统型黄酒 清爽型黄酒 

分 类 号：TS262.4[轻工技术与工程—发酵工程]