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1、北京市海淀区产品质量监督检验所北京市海淀区产品质量监督检验所国家食品质量安全监督检验中心国家食品质量安全监督检验中心20112011年年0606月月2323日日质谱分析方法质谱分析方法定性与定量定性与定量 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspecti
2、on Center China National Food Quality Safety Supervision and Inspection Center The birth of VenusLC and MS:A Match Made in HeavenJames Jorgenson，University of North Carolina59th，ASMS，June 5，Denver，USA China National Food Quality Safety Supervision and Inspection Center China National Food Quality Sa
3、fety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center The Arnolfini Wedding阿尔诺菲尼的婚礼 扬.凡.艾克 1434 China National Food Quality Safety Supervision and Inspection Center China Nation
4、al Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 主要内容主要内容一、质谱基础及相关术语一、质谱基础及相关术语二、定性（确证）方法二、定性（确证）方法三、定量方法三、定量方法 China National Food Quality Safety Supervi
5、sion and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 真空系统真空系统进样系统进样系统检测器检测器数据系统数据系统质量分析器质量分析器离子源离子源大气压大气压质谱仪的基本结构质谱仪的基本
6、结构GCLCICCEEI、CIESIAPCI、APPIMALDIDART-四级杆四级杆离子阱离子阱飞行时间飞行时间磁质谱磁质谱FT-ICRFT-ICR一、质谱基础及相关术语一、质谱基础及相关术语 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Ce
7、nter China National Food Quality Safety Supervision and Inspection Center 各种离子化方法的使用范围各种离子化方法的使用范围 选择的依据选择的依据挥发性挥发性 热稳定性热稳定性 极性极性 NeutralNeutral10101 110102 210103 310104 410105 5Molecular WeightMolecular WeightIonicIonicAnalyte PolarityESI or MALDIESI or MALDIAPCI/APPIAPCI/APPIGC/MSGC/MS China Nati
8、onal Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 质量范围质量范围质谱仪所能测定的离子质荷比的范围质谱仪所能测定的离子
9、质荷比的范围不同仪器不同仪器不同应用不同应用质谱仪主要性能指标质谱仪主要性能指标 扫描速度扫描速度 分辨率分辨率色谱峰的数据点数目和峰宽、扫描时间有关色谱峰的数据点数目和峰宽、扫描时间有关 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Cente
10、r China National Food Quality Safety Supervision and Inspection Center 质谱仪主要性能指标质谱仪主要性能指标 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center Chi
11、na National Food Quality Safety Supervision and Inspection Center 质谱仪主要性能指标质谱仪主要性能指标 磁质谱的分辨率定义最为严格：对两个相等强度磁质谱的分辨率定义最为严格：对两个相等强度的相邻峰，当两峰间的峰谷不大于其峰高的相邻峰，当两峰间的峰谷不大于其峰高10%10%时，则时，则认为两峰已经分开，其分辨率：认为两峰已经分开，其分辨率：磁质谱中，磁质谱中，R R不随不随m/zm/z变化，在测定小分子时有优势变化，在测定小分子时有优势eg.R=5000eg.R=5000，500500与与500.1 5000500.1 5000与
12、与50015001 分辨率分辨率 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 有机质谱仅要求
13、有机质谱仅要求5050峰谷峰谷刚刚分开就算分开（这时刚刚分开就算分开（这时称为有机质谱的单位质量称为有机质谱的单位质量分辨）分辨）简化为用单峰法表示，即简化为用单峰法表示，即测定一个峰半峰高处的全测定一个峰半峰高处的全峰宽峰宽Full width half Full width half MaximumMaximum（简写为（简写为FWHMFWHM）分分 辨辨 率率eg.FWHM=0.25m/z=100，R=400m/z=1000，R=4000 China National Food Quality Safety Supervision and Inspection Center China
14、National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 分分 辨辨 率率例如：要区分以下离子，分辨率要达到例如：要区分以下离子，分辨率要达到 CO+CO+27.994927.9949 N2+N2+28.006128.0061 ArCl+ArCl+74.9
15、31274.9312 As+As+74.921674.9216 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection
16、 Center attoatto摩尔级的灵敏度；摩尔级的灵敏度；优于优于2 2 ppmppm的的MSMS质量准确度质量准确度优于优于5 5 ppmppm的的MS/MSMS/MS质量准确度质量准确度线性动态范围可以达到五个数量级线性动态范围可以达到五个数量级 ；质量分辨率至质量分辨率至20,00020,000高速数据采集能力（每秒获得高速数据采集能力（每秒获得1010张张MS/MSMS/MS谱图），更好地兼容超快速谱图），更好地兼容超快速液相色谱液相色谱宽质量范围：宽质量范围：m/zm/z范围从范围从252520,000 u20,000 u 举例：举例：China National Food Q
17、uality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 质谱峰类型质谱峰类型分子离子分子离子必须是化合物谱图中质量最高的离子必须是化合物谱图中质量
18、最高的离子必须是奇电子离子、符合氮规则必须是奇电子离子、符合氮规则必须能通过丢失合理的中性离子，产生谱图中高质量区的重要离子必须能通过丢失合理的中性离子，产生谱图中高质量区的重要离子准分子离子准分子离子碎片离子碎片离子同位素离子同位素离子由于天然同位素的存在，因此在质谱图上由于天然同位素的存在，因此在质谱图上出现出现M+1M+1，M+2M+2等峰，由这等峰，由这些同位素所形成的峰称之为同位素峰些同位素所形成的峰称之为同位素峰一、质谱基础及相关术语一、质谱基础及相关术语 术语术语 China National Food Quality Safety Supervision and Inspect
19、ion Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 1-Cl2-Cl China National Food Quality Safety Supervision and Inspection Center Chin
20、a National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 1-Br2-Br China National Food Quality Safety Supervision and Inspection Center China National Food
21、 Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 3-Br4-Br China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety
22、 Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center Beynon Beynon 表表 例如：例如：MM=150=150化合物化合物 MM+1 +1 MM+2+2 化合物化合物 MM+1 +1 MM+2+2C6H14NOCl 8.15 0.49C6H14NOCl 8.15 0.49 C7H11N4 9.25
23、 0.38 C7H11N4 9.25 0.38C6H14O4 6.86 1.0C6H14O4 6.86 1.0 C8H6 O3 8.36 0.95 C8H6 O3 8.36 0.95C7H2O4 7.75 1.06C7H2O4 7.75 1.06 C8H8NO2 9.23 0.78 C8H8NO2 9.23 0.78C7H4NO3 8.13 1.06C7H4NO3 8.13 1.06 C8H11N2O 9.61 0.61 C8H11N2O 9.61 0.61C7H6N2O2 8.50 0.72C7H6N2O2 8.50 0.72 C8H12N3 9.98 0.45 C8H12N3 9.98 0
24、.45C7H8N3O 8.88 0.55C7H8N3O 8.88 0.55 C9H10O2 9.96 0.84 C9H10O2 9.96 0.84 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food
25、 Quality Safety Supervision and Inspection Center 二、定性（确证）方法二、定性（确证）方法质谱法只有在与色谱分离在线或脱机联用时才能作为质谱法只有在与色谱分离在线或脱机联用时才能作为确证方法确证方法在分析仪器上的进样顺序是：空白溶剂、阴性控制样在分析仪器上的进样顺序是：空白溶剂、阴性控制样品、要确证的样品、阳性控制样品再进样，最后是阴品、要确证的样品、阳性控制样品再进样，最后是阴性控制样品性控制样品 1.1.基本原则基本原则 China National Food Quality Safety Supervision and Inspectio
26、n Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 保留时间保留时间实验室条件下分析物的最小可接受保留时间，实验室条件下分析物的最小可接受保留时间，2t 2t0 0气相色谱（气相色谱（GCGC）或液相色谱（）或液相色谱（
27、LCLC）时，半峰宽应在原来峰）时，半峰宽应在原来峰宽的宽的90-110%90-110%以内，保留时间的变化应在以内，保留时间的变化应在5%5%以内以内二、定性（确证）方法二、定性（确证）方法 2.2.色谱分离色谱分离 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and I
28、nspection Center China National Food Quality Safety Supervision and Inspection Center 保留时间保留时间 测试部分中分析物的保留时间（或相对保留时间）测试部分中分析物的保留时间（或相对保留时间），应在一个特定的保留时间窗口范围内与校正标准的，应在一个特定的保留时间窗口范围内与校正标准的保留时间相符。保留时间窗口与该色谱系统的分辨能保留时间相符。保留时间窗口与该色谱系统的分辨能力相当。分析物和内标的保留时间之比，应与校正溶力相当。分析物和内标的保留时间之比，应与校正溶液的相对保留时间一致，液的相对保留时间一致，G
29、C GC 偏差为偏差为0.5%0.5%，LC LC 偏差为偏差为2.5%2.5%二、定性（确证）方法二、定性（确证）方法 2.2.色谱分离色谱分离 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food
30、 Quality Safety Supervision and Inspection Center GBT 16631-2008 GBT 16631-2008 高效液相色谱法通则：同样条件下重复试验高效液相色谱法通则：同样条件下重复试验的保留数据的分散性，的保留数据的分散性，RSD3%RSD3%JJG705-2002 JJG705-2002 液相色谱仪检定规程：定性测量重复性（液相色谱仪检定规程：定性测量重复性（6 6次测量）次测量）RSD1.5%RSD1.5%JJFJJF（闽）（闽）103210322010 2010 液相色谱液相色谱-质谱联用仪校准规范：定性重质谱联用仪校准规范：定性重复性
31、复性 RSD2%RSD2%JY/T 0211996 JY/T 0211996 分析型气相色谱方法通则：整机重复性分析型气相色谱方法通则：整机重复性 保留保留时间时间RSD5RSD5二、定性（确证）方法二、定性（确证）方法 关于保留时间的讨论关于保留时间的讨论 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safe
32、ty Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center GB/T 22260-2008 GB/T 22260-2008 饲料中甲基睾丸酮的测定饲料中甲基睾丸酮的测定 高效液高效液相色谱串联质谱法相色谱串联质谱法 试样保留时间与标准品的相对偏差不大于试样保留时间与标准品的相对偏差不大于15%15%GB/T 22147-2008 GB/T 22147-2008 饲料中沙丁胺醇、莱克多巴胺和盐饲料中沙丁胺醇、莱克多巴胺和盐酸克仑特罗的测定酸克仑特
33、罗的测定 液相色谱质谱联用法液相色谱质谱联用法 样品与标准品保留时间的相对偏差不大于样品与标准品保留时间的相对偏差不大于0.5%0.5%二、定性（确证）方法二、定性（确证）方法 关于保留时间的讨论关于保留时间的讨论 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Ins
34、pection Center China National Food Quality Safety Supervision and Inspection Center 全扫描全扫描 scanscan选择离子监测选择离子监测 SIMSIMMS-MS-MSMSn n技术，如多反应监测技术，如多反应监测 MRMMRM二、定性（确证）方法二、定性（确证）方法 3.3.质谱检测质谱检测 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Superv
35、ision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 全扫描全扫描 scanscan 用记录全扫描质谱图进行质谱测定时，在标准品参用记录全扫描质谱图进行质谱测定时，在标准品参考图谱中所有相对丰度考图谱中所有相对丰度10%10%（分子离子、分子离子的特（分子离子、分子离子的特征加成物、特征碎片离子、同位素离子等）的特征离子
36、征加成物、特征碎片离子、同位素离子等）的特征离子都必须检测都必须检测选择离子监测选择离子监测 SIMSIM 用碎片离子色谱图进行质谱测定时，分子离子最好用碎片离子色谱图进行质谱测定时，分子离子最好是一个选择的检测离子。选择的检测离子并不一定要源是一个选择的检测离子。选择的检测离子并不一定要源于分子的同一部分。每个检测离子的信噪比应于分子的同一部分。每个检测离子的信噪比应 3 31 1二、定性（确证）方法二、定性（确证）方法 3.3.质谱检测质谱检测 China National Food Quality Safety Supervision and Inspection Center Chin
37、a National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 相对丰度（相对丰度（%基峰）基峰）EI-GC-MSEI-GC-MS（相对）（相对）CI-GC-MSCI-GC-MS、GC-MSnGC-MSn、LC-LC-MSMS、LC-MSnLC-MSn（相对
38、）（相对）50%50%10%10%20%20%20%to 50%20%to 50%15%15%25%25%10%to 20%10%to 20%20%20%30%30%10%10%50%50%50%50%相对离子丰度最大容许偏差相对离子丰度最大容许偏差二、定性（确证）方法二、定性（确证）方法 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National F
39、ood Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 二、定性（确证）方法二、定性（确证）方法2002/657/EC2002/657/EC规定：规定：要确证指令要确证指令 96/23/EC 96/23/EC 附录附录I-A I-A 组所列的物质，最少需要组所列的物质，最少需要4 4 个识别个识别点点，包括具有促合成作用和其他未允许的化合物，如二苯乙烯类，包括具有促合成作用和其他未允许的化合物，如二苯乙烯类化
40、合物及其衍生物、盐、酯等、抗甲状腺剂、甾醇、化合物及其衍生物、盐、酯等、抗甲状腺剂、甾醇、二羟基苯甲二羟基苯甲酸内酯如玉米赤霉醇、酸内酯如玉米赤霉醇、-受体激动剂等受体激动剂等要确证指令要确证指令 96/23/EC96/23/EC附录附录I-B I-B 组所列的物质，最少需要组所列的物质，最少需要3 3 个识别点个识别点，包括兽药和污染物，兽药如抗菌药、驱虫药、抗球虫药、氨基甲包括兽药和污染物，兽药如抗菌药、驱虫药、抗球虫药、氨基甲酸酯类、拟除虫菊酯类、镇静药、非甾体抗炎药等；其他环境污酸酯类、拟除虫菊酯类、镇静药、非甾体抗炎药等；其他环境污染物如有机氯、有机磷、真菌毒素、染料等染物如有机氯、
41、有机磷、真菌毒素、染料等 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center MSMS技术技术每种离
42、子的识别点数每种离子的识别点数低分辨质谱（低分辨质谱（LRLR）1.01.0LR-MSLR-MSn n母离子母离子1.01.0LR-MSLR-MSn n子离子子离子1.51.5高分辨质谱（高分辨质谱（HRHR）2.02.0HR-MSHR-MSn n母离子母离子2.02.0HR-MSHR-MSn n子离子子离子2.52.5质量碎片类型和识别点的关系质量碎片类型和识别点的关系 每个离子计算一次每个离子计算一次 EI-GC-MSEI-GC-MS、CI-GC-MSCI-GC-MS作为不同的技术计算作为不同的技术计算 用不同反应得到的衍生物，可增加识别点数用不同反应得到的衍生物，可增加识别点数 包括二级
43、离子和多级离子包括二级离子和多级离子 可最多结合三种不同的技术确定最小识别点数可最多结合三种不同的技术确定最小识别点数二、定性（确证）方法二、定性（确证）方法 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China Nationa
44、l Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Su
45、pervision and Inspection Center m/z+202m/z+202122122m/z-179m/z-17979.979.9ESI-ESI-ESI+ESI+甜蜜素甜蜜素SN/T 1948-2007 SN/T 1948-2007 进出口食品中环己基氨进出口食品中环己基氨基磺酸钠的检测方法基磺酸钠的检测方法 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Cent
46、er China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 应研究样品基质和检测器应研究样品基质和检测器性能引起的图谱改变性能引起的图谱改变 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision an
47、d Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 化合物保留时间（min）定性离子对定量离子对Fragmentor（V）CE（V）麻黄碱6.9166/148166/1488010166/13320伪麻黄碱8.0166/148166/1488010166/13320 China National Food Quality Safety
48、 Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 外标法外标法 内标法内标法 标准加入法标准加入法 基质效应基质效应三、定量方法三、定量方法 China Nation
49、al Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 三、定量方法三、定量方法 1.1.外标法外标法单点校正法单点校正法外标标
50、准曲线法外标标准曲线法 China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center China National Food Quality Safety Supervision and Inspection Center 三、定量方法三、定量方法