葡萄酒中生物胺多组分的UVD/FLD串联HPLC方法研究

目的建立葡萄酒中多组分生物胺的柱前衍生反相高效液相色谱-荧光/紫外串联检测器的测定方法。方法以正丁醇/氯仿(1∶1)为萃取液提取葡萄酒中生物胺,丹磺酰氯为衍生剂,1,7-二氨基庚烷为定量内标,采用C18色谱柱分离,甲醇/水为流动相梯度洗脱,流速1.5ml/min,紫外检测波长254nm,荧光激发波长(Ex)350nm,发射波长(Em)520nm。结果色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、精胺和亚精胺在30分钟内得到良好分离。在给定的浓度范围内,各组分生物胺呈现良好线性相关(r>0.99)。在葡萄酒中添加生物胺混合标准溶液,平均回收率为79.2%~127.9%,相对标准偏差RSD小于10%。结论采用荧光/紫外检测器串联方法检测,满足了葡萄酒中多组分生物胺检测的需要。

Method of determination of biogenic amines in wines by high-performance liquid chromatography with fluorescence detector and ultraviolet detector

OBJECTIVE: A pre-column derivation reversed-phase high-performance liquid chromatography method was developed for the determination of biogenic amines (tryptamine, phenylethyamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) in wines with fluorescecse detector (FLD) and ultraviolet detector (UVD). METHODS: Liqiud-liqiud extraction (LLE) was used to clean up wine samples with chloroform/butanol prior to derivatization with dansyl chloride, biogenic amines were separated using gradient elution. Mobile phase A was methanol and mobile phase B was water with the flow rate 1.5 ml/min. The column used was a CAPCELL PAK C18 MG (4.6 mm I.D. x 150 mm, 5 microm) and gradient elution at constant column tempereture with 30 degrees C under fluorescence detector with Ex 350nm and Em 520nm, The method and ultraviolet detector with 254nm. 1,7-Diaminoheptane was used as the internal standard (IS). RESULTS: was linear for the amines studied at concentration ranging from 0.05 to 25 mg/L, except for spermidine and spermine, which ranging from 0.05 to 15 mg/L. The average recoveries ranged from 79.2% to 127.5% for all amines, The RSDs were less than 10%. With this method, 5 wine samples were determined, the results were favorable. CONCLUTION: method can be applied for determination of biogenic amines in wines.