6 8 Vol. 6 No. 8 2015 8 Journal of Food Safety and Quality Aug., 2015 L- * 叶少文 (, 519040) 摘要 : 目的, L-, L- 方法,,, Na, L- 结果 L- 0.3861~3.0648 mg/ml, r>0.999 80% 100% 120%, L- 97.2% 结论, L- 关键词 : ; ; L- Determination of free L-cysteine by automatic amino acid analyzer YE Shao-Wen * (By-Health Co., Ltd., Zhuhai 519040, China) ABSTRACT: Objective By comparison of the traditional detection method for sulfur-containing amino acids, formic acid oxidation method, this paper puts forward a more simple operation method with accurate calculation of free L-cysteine content for those health food adding L-cysteine. Methods The sample was dissolved with buffer ultrasonic, filtered by microporous membrane filter, and analyzed by automatic amino acid analyzer. Na type ion exchange chromatography column separation was used. The content was calculated by L-cysteine standard quantitative calculation. Results The concentration of L-cysteine in the range of 0.3861~3.0648 mg/ml had a good linearity with peak area, the correlation coefficient r was >0.999. At the level of adding 80%, 100%, and 120%, the average recovery of L-cysteine was 97.2%. Conclusion The method is simple, accurate, reproducible and applicable for the determination of L-cysteine in health food. KEY WORDS: sulfur-containing amino acid; automatic amino acid analyzer; L-cysteine 1 引言 -SH, -SH, L- [1] GB/T 15399-1994, GB/T 15399-1994 [2],,,, ( ), [3] L-,, L- [4], L-, GB/T 15399-1994 [5],,, L-, L- * 通讯作者 :,, E-mail: 442884571@qq.com *Corresponding author: YE Shao-Wen, Herbalists, By-Health Co., Ltd., Zhuhai 519040, China. E-mail: 442884571@qq.com
3130 6 [6] 2 材料与方法 2.1 试剂与仪器 L- (: 99.0%, :, : JS0925RC14) [7] ; [ (, )19.6 g+ (, )17.3 ml+(, )20 ml+(, )1.2 ml 1 L ]; 1[(, )19.6 g+ (, )13.5 ml+ (, )2 ml+(, )1.2 ml+ (, )20 ml 1 L ]; 2[ (, )19.6 g+ (, )9.3 ml+(, )1.2 ml 1 L ]; 3[(, )19.6 g+ (, )0.7 ml+ (, )58.4 g+(, )1.2 ml 1 L ]; 4[ (, )16 g 1 L ]; (biochrom, : 1065214) [8] ; L- (: biochrom 30+) : biochrom Na+ Sodium Column 2.2 分析测试条件 : 20μL : ( ) (min) (ml/h) 1 45 1 35 ON OFF Nin Rec commands 1 45 0 35 ON OFF Reset 1 45 1 35 ON OFF Load 1 45 1 35 ON OFF 1 45 0 35 ON OFF Reset 1 45 5 35 ON ON 2 47 5 35 ON ON 3 47 1 35 ON ON 3 98 5 35 ON ON 4 98 4 35 ON ON 1 98 3 35 ON ON 0 50 2 OFF OFF OFF 1 50 3 44 OFF OFF 1 45 2 35 ON OFF : Nin, Rec : ( ) (min) (ml/h) Nin Rec 3 98 5 35 ON ON 4 98 4 35 ON ON 1 98 3 35 ON ON 0 50 2 OFF OFF OFF 1 50 3 44 OFF OFF 1 45 2 35 ON OFF 2.3 标准品溶液的配制 L- 193.99 mg 25 ml,, 0.5 1.0 2.0 3.0 4.0 ml 10 ml, L- 0.3841 0.7682 1.5364 2.3046 3.0728 mg/ml [9] 2.4 供试品溶液的配制 30 mg 25 ml, (40 khz)10 min [10],, 0.45 μm, [11] 2.5 线性实验 L-, 0.45 μm,,, (X), (Y),, [12] 2.6 重复性实验 6, 2.4,, RSD(%) [13] 2.7 加标回收实验 15 mg 9, 25 ml, 3, 3, L- 6.0 7.5 9.0 mg, 2.4, 0.45 μm, [14] 3 结果与分析 3.1 标准曲线确证结果 L- 1,
8 : L- 3131 0.999, L- 0.3841~3.0728 mg/ml Table 1 表 1 标准曲线方程 The standard curve equation (r) L- Y=4919.5X 1275.98 0.999 3.2 检出限和定量限 LOD LOQ (S/N) LOD S/N=3, LOQ S/N=10 3.2.1 检出限 L- LOD=4.82 μg/ml, S/N=3, L- : 4.82 μg/ml 25 ml/1000/30 mg 100=0.004% 3.2.2 定量限 S/N=10 : LOQ LOD/3 10,, L- : 0.004/3 1 0.013% 3.3 重复性实验 L- 2, 1 2 3 4 5 12 h, 2, (RSD) 1.3%, 12 h, 3.4 回收率实验 L- 3 3, 3 L- : 97.2%(RSD) 1.3% 3.5 实际样品的测定 2.2, L-( 1), L- L-, 2,, 表 2 重复性的实验 (n=6) Table 2 The precision of the experiment (n= 6) h (%) (%) RSD(%) 1 50.53 2 50.43 L- 3 4 50.16 50.41 50.39 0.3 5 50.52 12 50.28 Table 3 表 3 加标回收率实验 (n=9) The standard addition recovery of experiment (n=9) (mg) (mg) (%) (%) RSD(%) 6.02 5.85 97.17 6.05 5.87 97.02 6.04 5.81 96.19 7.52 7.41 98.53 L- 7.48 7.26 97.05 7.56 7.37 97.59 9.13 8.91 97.59 9.06 8.84 97.57 9.14 8.84 96.71 97.2 0.7
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