活泼羰基化合物(reactive carbonyl species,RCS)是一类具有高反应活性的有害物质,白酒发酵过程及成品均发现存在这类物质,给白酒产品带来安全隐患。该文建立一种同时测定白酒中丙烯醛、丙酮醛、巴豆醛、5-羟甲基糠醛、甲醛和乙醛含量的高效液相色谱检测方法,并对不同香型白酒样品及加工过程中的酒体样品进行了综合分析。样品经稀释后以2,4-二硝基苯肼为衍生化试剂,在50 ℃条件下反应1.5 h,采用Beconsil C18色谱柱(250 mm × 4.6 mm,5 μm),以纯水为流动相A,乙腈为流动相B,梯度洗脱,流速为1.0 mL/min,柱温40 ℃,二极管阵列检测器,检测波长360、400 nm,在此色谱条件下,各组分都得到很好分离,线性范围较宽,相关系数均在0.999以上,检出限1~8 ng/mL,定量限3~25 ng/mL,加标回收率85.39%~105.78%,相对标准偏差均<4%。采用该方法测得6种主要香型白酒中酱香型和芝麻香型白酒中活泼羰基化合物含量较高;酱香型白酒8轮发酵,7次取酒过程中,RCS总体含量最高的为第3、4和5次;浓香型白酒原料出窖堆积过程中产生的RCS升至最高,液体发酵酒原液中双轮底和中层发酵酒样品含量分列前2。该液相分析方法操作简便,稳定性好,灵敏度高,适用于成品白酒及白酒加工过程中活泼羰基化合物的检测及监控。
Reactive carbonyl species (RCS) are toxic and highly reactive materials in liquor fermentation process which bring potential safety hazards to liquor products. In this study, a high performance liquid chromatography analysis method was established to simultaneously determine acrolein, methylglyoxal, crotonaldehyde, 5-hydroxymethylfurfural, formaldehyde and acetaldehyde in Chinese Baijiu. A comprehensive analysis of different flavor liquor samples and wine samples during processing was carried out. The diluted sample was derivatized with 2, 4-dinitrobenzene hydrazine at 50 ℃ for 1.5 h. A Beconsil C18 chromatographic column (250mm×4.6 mm, 5 μm) was used. The mobile phase was acetonitrile and water under gradient elution at a flow rate of 1.0 mL/min. The column temperature was set to 40 ℃, and the diode array detector was selected as dual-channel detection, with detection wavelength of 360 nm (ACR, MGO, CRO, FOR, ACE) and 400 nm (5-HMF), respectively. This method exhibited good linearity with correlation coefficients all above 0.999. The detection limits were in the range of 1-8 ng/mL and the quantitate on limit were in the range of 3-25 ng/mL. The spiked recoveries ranged from 85.39%-105.78%, with RSDs less than 4%. The level of RCS in Maotai-flavor Baijiu and sesame-flavor Baijiu was higher, which was speculated to be related to the high temperature fermentation. In the process of 8 rounds of fermentation and 7 times of liquor extraction, the RCS concentration in Maotai-flavor Baijiu was the highest in the 3rd, 4th and 5th rounds. The RCS of strong-flavor liquor rose to the highest in the process of “accumulation out of cellar”, and the RCS level in the sample of “the double bottom of liquid fermentation” and “the middle layer of liquid fermentation” were ranked first and second, respectively. Our method is stable and sensitive, which is easy to operate, and suitable for detecting RCS in liquor and semi-finished products in fermentation processing.
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