Microsoft Word 吴映璇-二校 出1842、1843请不要动版

Transcription

1 6 5 Vol. 6 No Journal of Food Safety and Quality May., 2015 吴映璇 1,2, 林峰 1,2*, 欧阳少伦 1,2, 廖玮玲 3, 邵琳智 1,2, 邹游 1,2, 杜志峰 (1., ; 2., ; 3., ; 4., ) 4 摘要 : 目的,, 方法 - (HPLC-Q-TOF-MS), 1 17,,,, 结果 36, 2,, 结论 HPLC-Q-TOF-MS,, 2 关键词 : - ; ; Research on the biomarkers in partially hydrolyzed protein infant milk combined by high performance liquid chromatography-q-time of flight mass spectrometry WU Ying-Xuan 1,2, LIN Feng. 1,2*, OUYANG Shao-Lun 1,2, LIAO Wei-Ling 3, SHAO Lin-Zhi 1,2, ZOU You 1,2, DU Zhi-Feng 4 (1. Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou , China; 2. Key Laboratory of Animals and Plants and Food Import and Export of Technical Measures in Guangdong Province, Guangzhou , China; 3. South China Agricultural University, Guangzhou , China; 4. AB SCIEX Asia Pacific Application Support Center, Shanghai , China) ABSTRACT: Objective To compare partially hydrolyzed protein infant milk powder with ordinary infant milk powder, and explore potential biomarkers in fingerprint to effectively distinguish between two groups. Methods One partially hydrolyzed protein infant and 17 ordinary infant milk powders were analyzed by high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry method (HPLC-Q-TOF-MS), the fingerprints were compared with computer software in order to explore characteristic markers and verified the reliability of tracking samples by liquid chromatography-mass spectrometry techniques. Results Thirty-six kinds of component in samples might be potential biomarkers between 基金项目 : (2012IK150) Fund: Supported by the Research Program of Administration of Quality Supervision, Inspection and Quarantine (2012IK150) * 通讯作者 : linf@iqtc.cn *Corresponding author: LIN Feng, Professor, Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou , China. linf@iqtc.cn

2 5, : partially hydrolyzed protein infant milk powder and ordinary infant milk powder, 2 kinds of them were selected for verification. As the result, the tracking sample of partially hydrolyzed protein infant milk could be found and the ordinary infant formulas were not found. Conclusion This research on partially hydrolyzed protein infant milk combined with HPLC-Q-TOF-MS has better accuracy and repeatability, and that of 2 screening component are likely the potential biomarkers in partially hydrolyzed protein infant milk. KEY WORDS: high performance liquid chromatography-tandem-q-time of flight mass spectrometry; biomarker; partially hydrolyzed protein infant milk 1 引言,,, [1],,,,, [2-6],,,, [7-12],, -,, (MRM) [13-15], MRM,,,, 2 材料与方法 2.1 材料 3 19 ( ) 2.2 仪器与试剂 LC-20AD, AB 5600, (ESI), ( AB SCIEX ); LC-20AD, AB 5500QTRAP ( AB SCIEX );Milli-Q ( Millipore ); MS 3 basic ( IKA); 3-18K (Sigma );, ( ) : -d3, 99%, WITEGA Laboratorien Berlin-Adlershof GmbH : 50 ml 450 ml, 30 min, 2.3 色谱条件 : Atlantic T3 (150 mm 2.1 mm, 3 μm WATERS Co., Ltd.); ; : A: 5 mmol/l (V:V)+B: 5 mmol/l (V:V)=5:95 (V:V), : 0~1 min, 5% A; 1~18 min, 5%~100% A; 18~23 min, 100% A; 0.30 ml/min; : 40, : 5 μl 2.4 质谱条件 AB 5600 四极杆飞行时间质谱条件 : ESI(+) ; : m/z 100~1000; 379 kpa 207 kpa 448 kpa; 600, : 5.5 kv, (DP) : 80 V; AB 5500QTRAP 质谱条件 : (electrospray ionization, ESI+) ; : ; (CUR): 207 kpa; (GS1): 379 kpa; (GS2): 448 kpa; (CAD):

3 MEDIA; (IS): 5.5 kv; (TEM): 600 ; 2.5 实验方法 0.5 g( 0.01 g) 50 ml, 8 ml (65±5) 2 min, 8 ml, 30 s, r/min 10 min,, 2, 10 ml, 1 min, : (V:V=1:1) 25 ml,, r/min, 4 5 min, 950 μl 50 μl 1.5 ml,, r/min, 4 5 min, 0.2 μm 2.6 数据处理 17 4,, HPLC-Q-TOF-MS, MarkerView TM, -d3, ( ), P, P, P, P < 结果与讨论 3.1 部分水解婴幼儿奶粉与普通婴幼儿奶粉样本差异组分的筛选 MarkerView TM,, 100~ ,,, 1,, G 2 G, G t P,, P<0.05, 36, 部分水解婴幼儿奶粉与普通婴幼儿奶粉样本差异组分的验证 715.4(P=1.21e-92) 981.5(P=2.66e-88),, 715.4/ /360.2, MRM ( 3) MRM 18,,,, Fig Distribution of 18 milkpowder samples in first and second principle component analysis

4 第 5 期 吴映璇, 等: 应用高效液相色谱-四极杆飞行时间高分辨质谱联用技术研究部分水解蛋白婴幼儿奶粉中的标记物 1845 图 2 18 个奶粉样品的可视化统计分析 Fig. 2 Visual statistical analysis of 18 milk samples Table 1 表 1 有统计意义的组分 The statistically significant components 序号 No. 相对分子量 m/z 保留时间 Ret. Time P值 P-value E E E E E E E E E E E E E E E E E-81

5 No. m/z Ret. Time P P-value E E E E E E E E E E E E E E E Fig. 3 3 The extracted ion chromatogram, and primary and secondary spectra spectrum of Characteristic component

6 5, : 验证样品的检测, 2, MRM :,, 4 4 结论 -, 36, 2, MRM, ;,, 5 后续研究,,,, Fig. 4 4 Extracted ion chromatogram of trace samples

7 参考文献 [1],,,. [J]., 2011, 39(4): Cui L, Huang MN, Zhang L, et al. Functional domestic milk market situation and development trend analysis [J]. Jiangsu Agric Sci, 2011, 39(4): [2],,,. - [J]., 2009, 19(12): , OuYang SL, Lin F, Wu YX, et al. Determination of praziquantel residues in milk products by liquid chromatography-tandem mass spectrometry [J]. Chin J Health Lab Technol, 2009, 19(12): , [3],,,. - [J]., 2014, 42(4): Zhang TT, Zeng MF, Jin Y, et al. Determination of nucleosides and nucleotides in breast milk and infant formula by reversed phase liquid chromatography-tandem mass spectrometry [J]. Chin J Anal Chem, 2014, 42(4): [4] GB/T [S]. GB/T Determination of ivermectin, abamectin, doramectin and eprinomectin residues in milk and milk powder-lc-ms-ms method [S]. [5] GB/T [S]. GB/T Determination of streptomycin, dihyrostrptomycin and kanamycin residues in milk and milk powder-lc-ms-ms method [S]. [6] GB C [S]. GB Determination of vitamin C in foods for infants and young children, milk and milk products [S]. [7],,, [J]., 2014, 33(3): Wang ML, Zeng L, Yan HF, et al. Rapid screening and confirmation of illegally added phosphodiestrase-5 inhibitors and analogues in health foods by liquid chromatography-ion trap-time of flight tandem mass spectrometry [J]. J Instrum Anal, 2014, 33(3): [8],,,. - / 111 [J]., 2014, 33(5): Liu H, Zhang YL, Ding T, et al. Screening and quantification of 111 pesticide residues in wines by high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry [J]. J Instrum Anal, 2014, 33(5): [9],,, [J]., 2014, 33(4): Xie HB, Wang YG, Zhou MY, et al. Simultaneous screening and determination of five kinds of 26 veterinary drug residues in pork by liquid chromatography quadrupole time of flight mass spectrometry [J]. J Instrum Anal, 2014, 33(4): [10],,,. UFLC-IT-TOF-MS 12 [J]., 2013, 32(11): Zhang XB, Huang LY, Chen XZ, et al. Determination of 12 kinds of estrogenic and progesterone hormones in infant formula milk powder by ultra fast liquid chromatography-ion trap-time of flight mass spectrometry [J]. J Instrum Anal, 2013, 32(11): [11],,. SELDI-TOF-MS [J]., 2012, 22(13): Su NL, Mou WF, Wang B. SELDI-TOF-MS screening serum protein markers in hepatocel lularcarcinoma [J]. China J Mod Med, 2012, 22(13): [12],,,. UFLC/Q-TOF MS [J]., 2014, 33(3): Wang Q, Fu L, Zhang Y, et al. Rapid identification of phospholipids in rat plasma by UFLC/Q-TOF MS [J].J Instrum Anal, 2014, 33(3): [13],,,. - [J]., 2009, 28(5): Wu YX, Lin F, Lin HD, et al. Determination of azoxystrobin residues in legume using high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry [J]. J Instrum Anal, 2009, 28(5): [14],,. - [J]., 2013, 4(5): Wu YX, Lin F, Yao YX. Simultaneous determination of streptomycin and dihydrostreptomycin residues in fish using hydrophilic interaction liquid chromatography coupled with

8 5, : electrospray ionization tandem mass spectrometry [J]. J Food Saf Qual, 2013, 4(5): [15],,, [J]., 2008, 36(12): Ma Q, Wang C, Wang X, et al. Simultaneous determination of 22 sulfonamides in cosmetics by ultra performance liquid chromatography tandem mass spectrometry [J]. Chin J Anal Chem, 2008, 36(12): ( 责任编辑 : 李振飞 ) 作者简介 吴映璇, 高级工程师, 主要研究方向为食品安全分析 wuyx@iqtc.cn 林峰, 研究员, 主要研究方向为食品安全分析 linf@iqtc.cn 食品中农药残留 兽药残留检测技术 专题征稿,,,,,,, JMPR CCRVDF, ,, 投稿方式 : : jfoodsq@126.com 食品安全质量检测学报 编辑部