Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories

doi:10.3969/j.issn.1673-8640.2019.03.001

Abstract

Abstract:

Liquid chromatography-tandem mass spectrometry （LC-MS/MS） is an emerging technology which has developed rapidly in recent years. It has combined the separation properties of liquid chromatography and the high sensitivity and specificity of mass spectrometry,which is widely applied in various areas,such as chemistry,biology,pharmaceutical science,food,clinic and environmental science. Especially in the field of clinical laboratory and diagnostics,LC-MS/MS used as a complement to traditional diagnostic techniques can often provide more accurate and reliable testing results in accurate and rapid diagnosis of diseases. In this review,some internationally published LC-MS/MS method validation guidelines,related literature and practical experience were summarized,and some key processes of LC-MS/MS development were introduced. Using 25-hydroxyvitamin D₃[25（OH）D₃] as an example,the key elements of method validation were reviewed,in order to provide a reference for the establishment,verification and implementation of LC-MS/MS.

Key words: Liquid chromatography-tandem mass spectrometry, Method development, Method validation

CLC Number:

R446.1

Clinical Mass Spectrometry Committee, Chinese Medical Doctor Association of Laboratory Medicine. Consensus of method development and validation of liquid chromatography-tandem mass spectrometry in clinical laboratories[J]. Laboratory Medicine, 2019, 34(3): 189-196.

Figures/Tables 8

References 15

[1]	Food and Drug Administration. Bioanalytical method validation guidance for industry[Z]. 2018.
[2]	Clinical and Laboratory Standards Institute. Liquid chromatography-mass spectrometry methods[S]. C62-A,CLSI, 2014.
[3]	Clinical and Laboratory Standards Institute. Mass spectrometry in the clinical laboratory:general principles and guidance[S]. C50-A,CLSI,2007.
[4]	中华医学会检验医学分会,卫生计生委临床检验中心. 液相色谱—质谱临床应用建议[J]. 中华检验医学杂志,2017,40(10):770-779.
[5]	Clinical and Laboratory Standards Institute. Evaluation of precision of quantitative measurement procedures[S]. EP05-A3,CLSI,2014.
[6]	李水军,SIHE W.液相色谱-质谱联用技术临床应用[M]. 上海:上海科学技术出版社,2014:10.
[7]	VOGESER M,SEGER C.Pitfalls associated with the use of liquid chromatography-tandem mass spectrometry in the clinical laboratory[J]. Clin Chem,2010,56(8):1234-1244.
[8]	BOŽOVI A,KULASINGAM V. Quantitative mass spectrometry-based assay development and validation:from small molecules to proteins[J]. Clin Biochem,2013,46(6):444-455.
[9]	VAN DEN OUWELAND J M,KEMA I P. The role of liquid chromatography-tandem mass spectrometry in the clinical laboratory[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2012,883-884:18-32.
[10]	Scientific Working Group for Forensic Toxicology. Scientific Working Group for Forensic Toxicology(SWGTOX) standard practices for method validation in forensic toxicology[J]. J Anal Toxicol,2013,37(7):452-474.
[11]	BONFIGLIO R,KING R C,OLAH T V,et al.The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds[J]. Rapid Commun Mass Spectrom,1999,13(12):1175-1185.
[12]	TAYLOR P J.Matrix effects:the Achilles heel of quantitative high-performance liquid chromatography-electrospray-tandem mass spectrometry[J]. Clin Biochem,2005,38(4):328-334.
[13]	EI-KHOURY J M,BUNCH D R,REINEKS E,et al. A simple and fast liquid chromatography-tandem mass spectrometry method for measurement of underivatized L-arginine,symmetric dimethylarginine,and asymmetric dimethylarginine and establishment of the reference ranges[J]. Anal Bioanal Chem,2012,402(2):771-779.
[14]	International Organization for Standardization. Guide to the expression of uncertainty in measurement[S] .1995.
[15]	European Chemical Measurement Traceability and Quality Improvement of Network Organization,Chemical Analysis of International Traceability Cooperation Group. Quantifying uncertainty in analytical measurement[S]. 2000.

乙醇基质			血清基质（提取后）
样本名称	组分名称	峰面积	样本名称	组分名称	峰面积
Sample1-1	D3-IS	93 240	Sample3-1	D3-IS	259 600
Sample1-1	D3-IS	97 270	Sample3-1	D3-IS	272 200
Sample1-1	D3-IS	97 910	Sample3-1	D3-IS	270 100
Sample1-2	D3-IS	111 300	Sample3-2	D3-IS	260 900
Sample1-2	D3-IS	111 000	Sample3-2	D3-IS	261 900
Sample1-2	D3-IS	112 500	Sample3-2	D3-IS	262 300
Sample1-3	D3-IS	97 200	Sample3-3	D3-IS	256 800
Sample1-3	D3-IS	99 710	Sample3-3	D3-IS	273 500
Sample1-3	D3-IS	96 880	Sample3-3	D3-IS	261 100
Sample1-4	D3-IS	105 500	Sample3-4	D3-IS	274 100
Sample1-4	D3-IS	111 500	Sample3-4	D3-IS	272 400
Sample1-4	D3-IS	111 200	Sample3-4	D3-IS	278 100
Sample1-5	D3-IS	106 800	Sample3-5	D3-IS	276 200
Sample1-5	D3-IS	106 700	Sample3-5	D3-IS	278 600
Sample1-5	D3-IS	108 900	Sample3-5	D3-IS	272 000
Sample1-6	D3-IS	114 100	Sample3-6	D3-IS	270 100
Sample1-6	D3-IS	112 900	Sample3-6	D3-IS	263 000
Sample1-6	D3-IS	113 300	Sample3-6	D3-IS	268 800
均值		105 995	均值		268 428
CV（%）		6.61	CV（%）		2.55

乙醇基质			血清基质（提取后）
样本名称	组分名称	峰面积	样本名称	组分名称	峰面积
Sample1-1	D3-IS	93 240	Sample3-1	D3-IS	259 600
Sample1-1	D3-IS	97 270	Sample3-1	D3-IS	272 200
Sample1-1	D3-IS	97 910	Sample3-1	D3-IS	270 100
Sample1-2	D3-IS	111 300	Sample3-2	D3-IS	260 900
Sample1-2	D3-IS	111 000	Sample3-2	D3-IS	261 900
Sample1-2	D3-IS	112 500	Sample3-2	D3-IS	262 300
Sample1-3	D3-IS	97 200	Sample3-3	D3-IS	256 800
Sample1-3	D3-IS	99 710	Sample3-3	D3-IS	273 500
Sample1-3	D3-IS	96 880	Sample3-3	D3-IS	261 100
Sample1-4	D3-IS	105 500	Sample3-4	D3-IS	274 100
Sample1-4	D3-IS	111 500	Sample3-4	D3-IS	272 400
Sample1-4	D3-IS	111 200	Sample3-4	D3-IS	278 100
Sample1-5	D3-IS	106 800	Sample3-5	D3-IS	276 200
Sample1-5	D3-IS	106 700	Sample3-5	D3-IS	278 600
Sample1-5	D3-IS	108 900	Sample3-5	D3-IS	272 000
Sample1-6	D3-IS	114 100	Sample3-6	D3-IS	270 100
Sample1-6	D3-IS	112 900	Sample3-6	D3-IS	263 000
Sample1-6	D3-IS	113 300	Sample3-6	D3-IS	268 800
均值		105 995	均值		268 428
CV（%）		6.61	CV（%）		2.55

样本	峰面积比值				CV（%）	偏差（%）
样本	第1次测定	第2次测定	第3次测定	均值	CV（%）	偏差（%）
乙醇基质	1.52	1.53	1.49	1.51	1.38	1.96
血清基质	0.53	0.52	0.53	0.53	1.10
1:1混合物	1.01	1.01	0.97	1.00	2.32

样本	峰面积比值				CV（%）	偏差（%）
样本	第1次测定	第2次测定	第3次测定	均值	CV（%）	偏差（%）
乙醇基质	1.52	1.53	1.49	1.51	1.38	1.96
血清基质	0.53	0.52	0.53	0.53	1.10
1:1混合物	1.01	1.01	0.97	1.00	2.32

样本	25（OH）D₃	L-L	H-L
L1	1.11
L2	1.13	1.13
L3	1.09	1.09
H1	52.80
H2	53.45
L4	1.16		1.16
H3	53.60
H4	52.56
L5	1.10		1.10
L6	1.14	1.14
L7	1.09	1.09
L8	1.05	1.05
H5	52.28
H6	53.48
L9	1.09		1.09
H7	52.14
H8	53.09
L10	1.15		1.15
H9	52.79
H10	53.02
L11	1.12		1.12
均值		1.10	1.12
携带污染率（%）		-1.82