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1 5 7 Vol. 5 No Journal of Food Safety and Quality Jul., 2014 蔡明明 1, 陈启 1, 李爽 1, 任一平 (1., ; 2., ) 2* 摘要 : 目的方法 CarboPac PA20(3 mm 150 mm),, 结果, 0.2~20 mg/l, 0.999; ( 3) 10 mg/kg, ( 10) 30 mg/kg; 0.72%~1.53% 结论,,, 关键词 : ; ; Determination of lactose in human milk by ion chromatography CAI Ming-Ming 1, CHEN Qi 1, LI Shuang 1, REN Yi-Ping 2* (1. Beingmate Baby & Child Food Co., Ltd, Hangzhou , China; 2. Zhejiang Provincial Center for Disease Prevention and Control, Hangzhou , China) ABSTRACT: Objective To establish an analysis method by ion chromatography for the determination of lactose in human milk. Methods The modified gradient procedure was used to separate the lactose standard on a CarboPac PA20 (3 mm 150 mm) anion exchange column which was established automatic analysis method and used ion chromatography and external standard methods for quantitative analysis. Results Under optimal experimental condition, the lactose linear range of 0.2~20 mg/l, the correlation coefficient was greater than The method detection limit (S/N=3) was 10 mg/kg, the quantitative limit (S/N=10) was 30 mg/kg; lactose of human milk value was 0.72%~1.53%. Conclusion After method optimization, excluding impurities interference, lactose was determined more accurately. This method operation is simple, accurate and cheap, suitable for lactose in human milk analysis of the test. KEY WORDS: ion chromatography; human milk; lactose 1 引言,, [1],,,,,, [2], : [3], * 通讯作者 :,,, renyiping@263.net *Corresponding author: REN Yi-Ping, Senior Engineer, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou , China. renyiping@263.net

2 7, : 2055,, (GC) [4,5] (HPLC) [6-9] (TLC) [10,11] (CE) [12] (IC) [13,14],, [15],, [16],,,,, 2 实验部分 2.1 仪器试剂与材料 : (Dionex ICS 5000,, ); ( Millipore ); (FOSS, ) (SHA-B, ); (SK8210HP, ); (Allegra 64R, Beckman ); (BSA224S-CW, Sartorius ) : ( 99.5% ) Sigma-Aldrich ; Sigma-Aldrich : OnGuard RP Dionex, : OnGuard II RP (2.5 cc) 10 ml 15 ml 30 min 2.2 标准溶液与淋洗液的配制 10 mg( 0.01 mg) 50 ml,, 200 mg/l, 4,, 1; (250 mmol/l): 13.1 ml 50%, 1000 ml; (500 mmol/l): 20.5 g,, 2.6 ml 50%, 500 ml; 18.2 MΩ.cm 1 Fig. 1 Chromatogram of lactose standard 2.3 仪器工作条件 : : CarboPac PA20(3 mm 150 mm); PG20(3 mm 30 mm); :, Au, Ag/AgCl ; : 30 ; : 0.45 ml/min; : 10 µl; : A: ; B: 250 mmol/l ; C: 500 mmol/l ; : 人乳取样方法来源保存条件表 1 梯度洗脱条件 Table 1 Eluent gradient for HPAEC PAD analysis (min) A B C 0 80% 20% 0% % 20% 0% % 10% 80% % 10% 80% % 80% 0% % 80% 0% % 20% 0% % 20% 0% 5

3 2056 5,,,, 样品预处理 0.5 g( 0.1 mg) 100 ml, 80 ml,, 3 ml 50%, 100 ml, 30 ml 50 ml ;, 1 ml 50 ml, 50 ml 15 ml, OnGuard RP, 7 ml,, 0.22 μm, CarboPac PA20 CarboPac PA10, CarboPac PA20 CarboPac PA10 OH - CarboPac PA20,, CarboPac PA20 2 CarboPac PA20 CarboPac PA10, 2 CarboPac PA20,,, CarboPac PA 红外光谱人乳成分分析仪法 76, FOSS 3 结果与讨论 3.1 人乳样品提纯净化,,,,,,,,,, 20% 30% 50% 55%,,, 50%, 50%, OnGuard RP, OnGuard RP 3.2 色谱分析柱的选择, pka 11~13, ph,,, Fig. 2 2 Comparison of chromatograms with different columns,,, 3, 3 Fig. 3 3 Chromatograms of human milk before and after digestion 3.3 方法学验证线性范围和检出限 mg/l,,,,

4 7, : 2057,,, 0.2~20 mg/l, 0.999, 2,, 1:3, 1:10, 精密度和回收率,, 6,,, 3, 99.53%~102.40%, 0.72%~ 1.53%,,, 人乳成分分析仪与离子色谱法比较实际样品测定结果 76,,,,,, 4, 6.70 g/100 g, 7.02%, 7.50 g/100 g, 5.20 g/100 g; 6.61 g/100 g, 4.99%, 7.37 g/100 g, 5.40 g/100 g,,, 4 结论,, : CarboPac PA20,, 76 :,,,, Table 2 表 2 乳糖的线性范围标准曲线相关系数检出限和定量限 Linearity range, standard curve, correlation coefficient, limits of detection(lod) and limits of quantitation(loq) of lactose (mg/l) (mg/kg) (mg/kg) 0.2~20 y=0.9693x Table 3 表 3 乳糖的加标回收率和相对标准偏差 (n=6) Recoveries and relative standard deviation of lactose(n=6) 3 g/100 g 6 g/100 g 9 g/100 g Table 4 表 4 不同人乳检测方法结果比较 Comparison of results with different methods

2058 5 参考文献 [1]. [M]. :, 2001. Wang GC. Food Nutrition[M]. Beijing: China Agriculture Press, 2001. [2],. [J]., 2000, 12(4): 13 15. Chen WC, Feng JJ.

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g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min

g ml 10% ph 色谱条件 kinetex C μm 100 A 4. 6 mm 150 mm 25 5 μl A B 10 mmol /L ml /min 25 4 2013 8 监测技术 5 1 2 1 ( 1. 江苏省疾病预防控制中心, 江苏南京 210009; 2. 南京医科大学第二附属医院, 江苏南京 210029) 5 Phenomenex kinetex C 18 2. 6 μm 4. 6 mm 150 mm 10 mmol /L ph 2. 5 + 60 40 DAD 280 nm 5 20. 0 mg /L ~ 100 mg /L 0.