6 1 Vol. 6 No. 1 2015 1 Journal of Food Safety and Quality Jan., 2015 高 鹭 1, 董伟峰 1, 彭心婷 2, 史立娟 1, 李妍 1, 庞艳华 1, 徐静 1, 曹际娟 1* (1., 116001; 2., 834700) 摘要 : 目的 14, 6 方法, (inductively coupled plasma mass spectrometry, ICP-MS);, (atomic fluorescence spectrometry, AFS) ;, -- (high performance liquid chromatography-ultraviolet photo-oxidation-hydride generation-atomic fluorescence spectrometry, HPLC-(UV)-HG-AFS) 6 结果 14 0.038~46.2 mg/kg; 0.006~19.3 mg/kg; HPLC-(UV)-HG-AFS, As(III) As(V) DMA, MMA, AsB AsC 结论, HPLC-(UV)-HG-AFS,,, 关键词 : ; ; -- Determination of arsenic speciation in seaweeds using high performance liquid chromatography-ultraviolet photo-oxidation-hydride generation-atomic fluorescence spectrometry GAO Lu 1, DONG Wei-Feng 1, PENG Xin-Ting 2, SHI Li-Juan 1, LI Yan 1, PANG Yan-Hua 1, XU Jing 1, CAO Ji-Juan 1* (1.Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China; 2. Tacheng Entry-Exit Inspection and Quarantine Bureau, Tacheng 834700, China) ABSTRACT: Objective The content of total arsenic and inorganic arsenic were determined in 14 seaweeds and 6 kinds of arsenic species were determined at the same time. Methods The content of total arsenic was determined by inductively coupled plasma mass spectrometry (ICP-MS) after microwave digestion. According to the pretreatment method of national standard method, the content of inorganic arsenic was determined by atomic fluorescence spectrometry (AFS). The results of 6 arsenic species were studied by high performance 基金项目 : (201310141)(20121146) Fund: Supported by Quality Inspection Research in the Public Interest (201310141) and Liaoning Province Research Program for Young Scientist (20121146) * 通讯作者 :,, E-mail: cjj0909@163.com *Corresponding author: CAO Ji-Juan, Professor, Technical Center of Liaoning Entry-Exit Inspection & Quarantine Bureau, No.60, Changjiang East Road, Zhongshan District, Dalian 116001, China. E-mail: cjj0909@163.com
146 6 liquid chromatography-ultraviolet photo-oxidation-hydride generation-atomic fluorescence spectrometry (HPLC-(UV)-HG-AFS) after acid extraction, which were compared with the national standard method. Results In 14 seaweeds, the content of total arsenic ranged from 0.038 to 46.2 mg /kg and the content of inorganic arsenic ranged from 0.006 to 19.3 mg/kg. After the instrument optimization and methods of exploration, only As(III), As(V) and DMA were found in seaweed samples. The content of MMA was less. AsB and AsC were not detected. Conclusion Through the detection method of HPLC-(UV)-HG-AFS, some seaweed samples with complicated morphology can effectively avoid the transformation between organic arsenic and inorganic arsenic. It can reduce interference and increase the accuracy of the test. More arsenic speciation will be performed in seaweed samples. KEY WORDS: seaweed; arsenic speciation; high performance liquid chromatography-ultraviolet photooxidation-hydride generation-atomic fluorescence spectrometry 1 引言, [1],, [2],,,, : As( ) As( )(MMA) (DMA)(TMAO) (AsB) (AsC), [3], (IARc), [4],,, DNA, [5], [6,7],,, GB/T 5009.11-2003 [8],,,,,,,,,,, - (high performance liquid chromatographyultraviolet photo-oxidation-hydride generation-atomic fluorescence spectrometry, HPLC-(UV)-HG-AFS) [9-11], -(high performance liquid chromatography-inductively coupled plasma mass spectrometry, HPLC-ICP-MS) [12-15], -(high performance liquid chromatography-hydride generation-atomic absorption spectroscopy, HPLC-HG-AAS) [16] - (capillary electrophoresisinductively coupled plasma mass spectrometry, CE- ICP-MS) [17] HPLC,,,,,,, 2 材料和方法 2.1 仪器及试剂 ICP-MS 7700( ); MLS 1200(
1, : 147 ); AFS-930 ( ); LC-15 ( ); (Sigma-aldrich ); Mini-Q synthesis ( ) Millipore (, Dima ), () 10 μg/ml Agilent Agilent (); 1000 μg/ml As( ); As(III) As(V) MMA DMA AsB AsC (, 32.4±0.7 124.3±1.2 46.2±1.5 97.4±3.3 92.2±2.7 91.6±3.7 μg/g) 14 :,,, 2.2 HPLC-(UV)-HG-AFS 分析条件,, ( 1)( 2) 2.3 实验方法 2.3.1 总砷和无机砷样品前处理, 0.5 g, 5 ml 0.5 h, 2 ml,, 50 ml GB/T 5009.11-2003, 6 mol/l 60 18 h,, Table 1 表 1 色谱实验参数 HPLC experimental parameters Hamilton PRP-X100(250 mm 4.1 mm, i.d.,10 µm) 25 mm 2.3 mm, i.d., 12~20 µm (Hamilton, Reno, NV). 15 mmol/l (NH 4 ) 2 HPO 4, ph6.0(10%) 1 ml/min 100 µl 表 2 氢化物发生原子荧光实验参数 Table 2 HG-AFS experimental parameters 1.5% KBH 4, 0.5%KOH, 6 ml/min 7%HCl, 6.0 ml/min UV : 0.1% K 2 S 2 O 8, 0.5% KOH, 6 ml/min, 193.76 nm 100 ma 45 ma 300 mv AFS 200 8 mm 400 ml/min 600 ml/min
148 6 2.3.2 砷形态检测提取方法 2 g( 0.01 g) 50 ml, 16 ml, 70 1.0 h, 16 ml 1.0 h, 8000 r/min 10 min, 4 ml, 4 ml, 70 20 min 0.22 µm, RP 3 结果与讨论 3.1 海藻中总砷和总无机砷含量测定 14 3,,,, ;, 14 1 mg/kg,, 91.4%~99.2% GB/T 5009.11-2003, 14,,,, 0.5 mg/kg, 82.6%~89.4% 9.24%~24.1% 3.2 HPLC-(UV)-HG-AFS 测定海藻样品中的 6 种砷形态 1 2, 6 10 min, 1 6 4 4 0.682~2.188 µg/l, 5%, 0.9990 5 5, 14 As(III) As(V) DMA, MMA, AsB AsC 3, 4, 4 3.26% 3.3 提取过程中砷形态稳定性及方法回收率, ( 6), 6 84.37%~90.17%, 5% Table 3 表 3 海藻中总砷和无机砷含量 The content of total arsenic and inorganic arsenic in seaweeds (mg/kg) (%) (mg/kg) (%) (%) 2.80 94.5 0.313 86.2 11.2 0.038 99.2 0.006 82.7 15.7 19.2 94.6 3.19 83.9 16.6 3.98 93.1 0.824 85.1 20.7 34.4 91.4 8.312 88.2 24.1 5.67 93.7 0.524 89.4 9.24 1.21 93.5 0.128 85.7 10.6 0.052 97.5 0.008 82.6 15.4 23.9 94.2 5.63 86.4 23.6 46.2 98.7 19.3 86.2 13.2 7.83 96.8 0.927 87.2 11.8 6.17 94.3 0.896 84.3 14.5 3.43 96.1 0.412 87.2 12.0 4.32 97.4 0.534 88.3 12.4
1, : 149 Fig. 1 1 Typical chromatograms of 6 arsenic species standard solutions Table 4 表 4 砷标准溶液的检出限 相对标准偏差和相关系数 Detection limit, relative standard deviation and relative coefficient of 6 arsenic standard solutions (µg/l) %(n=7) R As(V) 0.682 2.8 0.9994 As(III) 1.968 3.8 0.9992 MMA 0.987 3.8 0.9994 DMA 1.333 3.7 0.9991 AsC 2.188 3.4 0.9997 AsB 1.791 3.9 0.9996
150 6 表 5 不同海藻样品中形态砷的含量 (n=3) Table 5 The content of arsenic species in different samples (n=3) (mg/kg) As(III) As(V) MMA DMA AsB AsC 4(%) 0.025 0.010 N.D. * 0.029 N.D. N.D. 2.29 N.D. N.D. N.D. N.D. N.D. N.D. / 0.243 0.144 0.046 0.265 N.D. N.D. 3.64 0.052 0.026 N.D. 0.056 N.D. N.D. 3.37 0.375 0.230 0.114 0.416 N.D. N.D. 3.30 0.064 0.022 N.D. 0.062 N.D. N.D. 2.61 0.012 0.007 N.D. 0.014 N.D. N.D. 2.73 N.D. N.D. N.D. N.D. N.D. N.D. / 0.347 0.121 0.037 0.366 N.D. N.D. 3.64 0.670 0.310 0.162 0.752 N.D. N.D. 4.10 0.105 0.023 N.D. 0.122 N.D. N.D. 3.19 0.075 0.036 N.D. 0.088 N.D. N.D. 3.23 0.038 0.025 N.D. 0.041 N.D. N.D. 3.03 0.047 0.016 N.D. 0.049 N.D. N.D. 2.59 : N.D Table 6 表 6 羊栖菜中 6 种形态砷的添加回收率与相对标准偏差 (n=6) The recoveries and CV% about 6 arsenic species in Sargasum fusiforme (n=6) (µg/l) (µg/l) (%) RSD(%) As(III) 60 71.67 88.56 3.4 As(V) 60 62.84 90.17 3.4 MMA 60 58.37 89.72 4.5 DMA 60 75.25 90.12 3.5 AsB 100 87.52 87.52 4.2 AsC 60 50.62 84.37 3.5 4 结论 14 6, 5%, 1.5 mg/kg 5,,,,, [18],,,,, 参考文献 [1] Jerome ON. Arsenic in the environment, human health and ecosystem effects[ M]. John Wiley & Sons Inc, 1994.
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