液相色谱-质谱联用仪校准规范[1]
福建省地方计量技术规范 JJF(闽)1032-2010
液相色谱-质谱联用仪校准规范
Calibration Specification for Liquid Chromatography — Mass
Spectrometers
2010-03-01 发布 2010-03-10 实施
福 建 省 质 量 技 术 监 督 局 发布
JJF(闽)1032-2010
液相色谱-质谱联用
仪校准规范
Calibration Specification for Liquid
Chromatography — Mass spectrometers
JJF(闽)1032-2010
本规范经福建省质量技术监督局于2010年03月01日批准,并
自2010年03月10日起施行。
归 口 单 位:福建省质量技术监督局
起 草 单 位:福建省计量科学技术研究所
本规范由起草单位负责解释
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本规范主要起草人:
许 航(福建省计量科学技术研究所)
参加起草人:
杨乙强(泉州市产品质量检验所)
林 峰(福建省计量科学技术研究所)
陈小燕(福建省计量科学技术研究所)
蔡泽兵(漳州市计量所)
罗 峰(福建省计量科学技术研究所)
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目 录
1 范围 ··························································································································· (1) 2 引用文献 ···················································································································· (1) 3 术语和计量单位·········································································································· (1)
3.1大气压离子化(atmospheric pressure ionization, API)················································· (1)
3.2质荷比;m/z ············································································································· (1)
3.3质量范围(mass range)···························································································· (1)
3.4原子质量单位u(atomic mass unit) ·········································································· (1)
3.5 分辨力(resolution) ································································································ (1)
3.6 信噪比(signal-to-noise ratio, S/N)··········································································· (1) 4 概述 ··························································································································· (2) 5 计量性能要求 ············································································································· (2) 6 校准条件 ···················································································································· (3)
6.1实验室环境要求 ········································································································ (3)
6.2校准设备及标准物质 ································································································· (3) 7 校准项目和校准方法··································································································· (3)
7.1分辨力 ··················································································································· (3)
7.2质量准确性 ············································································································ (3)
7.3质量范围 ··············································································································· (3)
7.4信噪比 ··················································································································· (3)
7.4.1 ESI正离子 ············································································································· (4)
7.4.2 ESI负离子 ············································································································· (4)
7.4.3 APCI正离子··········································································································· (4)
7.5整机定量及定性重复性 ····························································································· (4) 8 校准结果的处理·········································································································· (4) 9 复校时间间隔 ············································································································· (5) I
JJF(闽)1032-2010
附录 A 校准证书内页格式(供参考)·········································································· (6) 附录B 校准记录格式 ································································································· (7) 液相色谱-质谱联用仪校准记录····················································································· (7) 液相色谱-质谱联用仪校准记录(续页)······································································· (8) 附录C 不确定度评定 ································································································· (9) 附录D 碘化铯钠(NaCSI)离子质量(m/z) ····························································· (10) II
JJF(闽)1032-2010
液相色谱-质谱联用仪校准规范
1 范围
本规范适用于离子阱(QIT)、单级四极杆(Q)和三重四级杆(Q-Q-Q)液相色谱—大气压离子化质谱联用仪(Liquid Chromatography — Atmospheric Pressure Ionization Mass Spectrometers,简称LC-APIMS)的校准。其他类型液相色谱-质谱联用仪的校准检测可参照此规范进行。
2 引用文献
GB/T 6041-2002 《质谱分析方法通则》
JJG(教委)003-1996《有机质谱仪检定规程》
JJS K0136-2004 《高效液相色谱法-质谱分析法一般规则》
使用本规范时,应注意使用上述引用文献的现行有效版本。
3 术语和计量单位
3.1 大气压离子化(atmospheric pressure ionization, API)
指在大气压条件下的质谱离子化技术的总称,主要包括电喷雾离子化(electrospray ionization ESI)和大气压化学离子化(atmospheric chemical ionization, APCI)。
3.2 质荷比;m/z
离子的质量数和核电荷数之比。
3.3 质量范围(mass range)
质谱仪能检测的最低和最高质荷比。
3.4 原子质量单位u(atomic mass unit)
以12C质量的1/12为一个质量单位。
3.5 分辨力(resolution)
分辨两个相邻质谱峰的能力(R)。以某离子峰高50%处的宽度(简称半峰宽)表示,记为W1/2,单位u。
3.6 信噪比(signal-to-noise ratio, S/N)
在质量色谱图中,待测样品信号强度与基线噪声的比值,记为S/N。
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4 概述
液相色谱-大气压离子化质谱联用仪(Liquid chromatography-atmospheric pressure ionization mass spectrometry, LC-APIMS),主要由高效液相色谱仪,离子源,质量分析器,检测器,数据采集及分析系统组成(如图1.)。样品经色谱柱分离,由离子源离子化产生带有一定电荷、质量数的离子,离子在不同场中的运动行为不同,各种类型的质量分析器利用该原理把电离子按质荷比(m/z)分开,得到依质量顺序排列的质谱图。通过对质谱图的分析处理,进行定性、定量分析。
图1. LC-APIMS组成框图
5 计量性能要求
LC-APIMS各项技术指标见表1
表1 LC-APIMS主要技术指标
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6 校准条件
6.1 实验室环境要求
6.1.1 仪器室内不得有强烈的机械振动和电磁干扰,不得存放与实验无关的易燃、易爆和强腐蚀性气体或试剂。
6.1.2 实验室温度:20℃~30℃;相对湿度:≤70%。
6.2 校准设备及标准物质
6.2.1 利血平-70%乙腈水溶液:10.0 pg/μL;
6.2.2 氯霉素-70%乙腈水溶液:10.0 pg/μL;
6.2.3 碘化铯钠-异丙醇水溶液:2 μg/μL碘化钠,50 ng/μL碘化铯溶于50%异丙醇水溶液;
6.2.4 微量注射器:10 μL,分度为0.1 uL。
7 校准项目和校准方法
仪器稳定后,执行自动调谐,直到自动调谐通过后,开始以下校准。
7.1 分辨力
按照调谐的条件,注入调谐液,考察附录D中3~5个主要离子峰的分辨力,以半峰宽的最大值为仪器的分辨力W1/2。
7.2 质量准确性
仪器调谐过后,按照调谐的条件,注入碘化铯钠调谐液,记录附录D中的主要离子的实测质荷比,根据公式(1)计算质量准确性。
∆M=Mi测量-Mi理论 (1)
式中Mi测量——第i个离子三次测量平均值,u;
Mi理论——第i个离子理论值,u。
7.3 质量范围
将碘化铯钠调谐溶液由质谱进样口注入,并调入该类型的质量校准文件进行调谐和校准。使用标准荷质比范围(m/z为50-2200), 选择SCAN模式,以碘化铯钠溶液为调谐液进行全范围扫描,考察分辨力和质量准确性均满足技术指标要求的质荷比范围。
7.4 信噪比
以自动或手动调谐时确定的最佳值作为校准参数。采用70:30乙腈水溶液为 3
JJF(闽)1032-2010
流动相,使用C18或相当者为色谱分离柱。
7.4.1 ESI正离子
注入10 pg的利血平溶液,选择SIM模式(或MRM模式),提取m/z=609特征离子(或m/z609特征离子的m/z195子离子),根据公式(2)计算信噪比。
S/N=HH噪声 (2)
式中:H609——提取离子(m/z=609)的峰高;
H噪声——基线噪声。
7.4.2 ESI负离子
注入100 pg的氯霉素溶液,选择SIM模式(或MRM模式),提取m/z 321特
征离子(或m/z 321特征离子的m/z 152子离子),根据公式(2)计算信噪比。
7.4.3 APCI正离子
注入10 pg的利血平溶液,选择SIM模式(或MRM模式),提取m/z=609特征离子(或m/z 609特征离子的m/z 195子离子),根据公式(2)计算信噪比。
7.5 整机定量及定性重复性
以C18或相当者为色谱分离柱,采用70:30乙腈水溶液为流动相,参照7.4的
条件进行测量,分别记录质量色谱图中相应的峰面积和保留时间,重复测量6次,根据公式(3),以峰面积的相对标准偏差计算定量重复性,以保留时间的相对标准偏差计算定性重复性:
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∑(x
RSD=i=1i-)2
6-1⨯1
⨯100%(3)
式中:RSD——相对标准偏差,%;
xi——第i次测量峰面积或保留时间;
——峰面积或保留时间的算术平均值。
8 校准结果的处理
根据校准结果,发校准证书,所有校准项目及其结果均应在证书中反映。校准结果的表达按照JJF 1071-2000技术规范的要求,包含标题、实验室名称和地址、送校单位的名称和地址、校准日期、校准所用测量标准的溯源性及有效性说明、校准环境等方面内容。
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9 复校时间间隔
液相色谱-质谱联用仪的复校时间间隔由客户自定,建议不超过2年,更换重要部件、维修或对仪器性能有怀疑时,应随时校准。
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附录 A
校准证书内页格式(供参考)
本次校准所使用的主要计量标准器具
名称 型号规格 不确定度/准确度 证书编号
校准地点及环境条件 地点:
温度: 相对湿度: 其它: 本次校准所依据的技术规范(代号、名称):
校准数据/结果:
电离源: 质量分析器类型: 1. 质量范围 2. 质量准确性 3. 分辨力 4. 信噪比 5. 定量重复性 6. 定性重复性
附录B
校准记录格式
液相色谱-质谱联用仪校准记录
2页 第 1 页) (共
液相色谱-质谱联用仪校准记录(续页)
(共2 页 第 2 页)
附录C
不确定度评定
液相色谱-质谱联用仪校准的各项指标中,主要对信噪比进行不确定度评价,不确定度主要来自:
1)n次测量相对标准偏差,A类,记为:u1; 2)所采用标准物质的不确定度,B类,记为:u2。 因此,得到合成标准不确定度uc:
uc=u21+u22
将合成标准不确定度乘以包含因子k (k=2)得到扩展不确定度U扩展: U扩展=kuc
附录D
碘化铯钠(NaCSI)离子质量(m/z)