CN106841488A - A kind of Liquid Chromatography-Tandem Mass Spectrometry method of sulfur-containing amino acid in non-derivative method detection blood plasma - Google Patents

A kind of Liquid Chromatography-Tandem Mass Spectrometry method of sulfur-containing amino acid in non-derivative method detection blood plasma Download PDF

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CN106841488A
CN106841488A CN201710129265.5A CN201710129265A CN106841488A CN 106841488 A CN106841488 A CN 106841488A CN 201710129265 A CN201710129265 A CN 201710129265A CN 106841488 A CN106841488 A CN 106841488A
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homocysteine
methionine
cysteine
blood plasma
amino acid
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曹云峰
孙晓宇
刘丽杰
刘明莉
赵姝琦
孙宏治
洪沫
高鹏
房中则
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Liaoning Runsheng Kangtai Biomedical Technology Co ltd
First Affiliated Hospital of Jinzhou Medical University
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Liaoning Runsheng Kangtai Biomedical Technology Co ltd
First Affiliated Hospital of Jinzhou Medical University
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    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
    • G01N2030/8818—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
    • G01N2030/8822—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving blood

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Abstract

In a kind of non-derivative method detection blood plasma the Liquid Chromatography-Tandem Mass Spectrometry method of sulfur-containing amino acid its belong to biochemical analysis detection field.The method is used and for blood plasma to add Isotopic Internal Standard solution, is well mixed, and dithiothreitol (DTT) is then added again（DTT）Reduction, then processed through protein precipitation, obtain the supernatant of sulfur-containing amino acid.Sample preprocessing is carried out using non-derivative method, method is simple, it is easy to operate, enormously simplify sample pretreatment operation；Isotopic Internal Standard reduces interference of the matrix to detecting, it is ensured that homocysteine, the degree of accuracy of cysteine and methionine detection.The method is detected to content methyllanthionine in blood plasma using LC MS/MS methods, using many reaction detection MRM scan modes, and found firstd 3 Methionine and homocysteine（136>90 passages）There is cross jamming, using the elution requirement of chromatogram gradient by its baseline separation, it is ensured that the accuracy of detection.The method sensitivity and specificity is strong, result is accurate.

Description

The Liquid Chromatography-Tandem Mass Spectrometry of sulfur-containing amino acid in a kind of non-derivative method detection blood plasma Method
Technical field
The invention belongs to biochemical analysis detection field, it is related to a kind of detection homocysteine in blood plasma（Hcy）, half Cystine（Cys）And methionine（Met）Liquid Chromatography-Tandem Mass Spectrometry method.
Background technology
Amino acid is the organic compound that a class contains amino and carboxyl, is to constitute constituting into substantially for body tissue cell Point, it is also the base unit for constituting protein.Document report, in human plasma amino acid change and numerous diseases generation all In the presence of close correlation.Hcy is a kind of amino acid being present in blood plasma, is a centre in Met and Cys metabolic processes Product, but itself it is not involved in the synthesis of protein.Under normal condition, Hcy concentration is 5 ~ 15 in blood plasmaμmol/L.Heredity or Obtain sexual factor so that Hcy concentration is higher than persistently normal value limit high, i.e. referred to as hyperhomocysteinemiainjury, relatively early investigation point Analysis result shows that hyperhomocysteinemiainjury is coronary heart disease, the independent harm intervened of palsy and Deep vain thrombosis Factor[1,2], but perspective study result thinks recently, hyperhomocysteinemiainjury is only artery sclerosis with existing As[3,4], because rising and the cerebrovascular disease incidence of disease of homocysteine in plasma are proportionate, therefore to the Guang ammonia of homotype half Prevention of the measure of acid to cardiovascular and cerebrovascular disease is significant.
Homocysteine is converted by two kinds of approach under normal circumstances：One is methionine circulation, and Hcy synthesizes in methionine With vitamin B12 as confactor in the presence of enzyme, 5-methyltetrahydrofolate generates Met as methyl donor；Second approach To turn sulphur approach, Hcy in the presence of cystathionine synthetase with vitamin B6 as confactor and serine is condensed into cystathionie, Further generate Cys[5].It can be seen that Met, Cys play very heavy in the circulation of homocysteine from metabolic pathway The effect wanted, the measure to Cys and Met can more comprehensively react the overall condition of amino acid metabolism.
Current detection method is generally derivatization method, and concentrates on the measure only to homocysteine, derivatization method step It is rapid cumbersome, waste time and energy, it is relatively costly.The high performance liquid chromatography tandem mass spectrum method that the present invention is used（LC-MS/MS）, by chromatogram Colleges and universities' resolution capability and mass spectrographic special, sensitive, multi-analyte immunoassay ability organically combine.LC-MS/MS technologies are with its strong spy Interfering between the opposite sex, exclusion material, realizes measure of the amino acid under the conditions of non-derivative.Isotopic Internal Standard of the present invention Use, eliminate interference of the matrix to determinand, it is quantitative accurate.MRM detection modes, findd 3 - methionine and homotype half Cystine（136>90 passages）There is cross jamming, using the elution requirement of chromatogram gradient by its baseline separation, it is ensured that detection Multiple determinands are determined exclusive by accuracy simultaneously, accurately, sensitive, efficiently.
[1] Walds, Law M, Morris JK. Homocysteine and cardiovascular disease: evidence on causality from a metanalysis. BMJ, 2002, 325(7374): 1202-1206
[2] Homocysteine studies collaboration. Homocysteine and risk of ischemic heart disease and stroke: a metanalysis. JAMA, 2002, 288(16): 2015-2022
[3] Loscalzo J. Homocysteine trials-clear out comes for complex reasons N Engl J Med, 2006, 354(15): 1629-1632
[4] Hankey GJ. Is plasma homocysteine a modifiable risk factor for stroke. Nat Clin Pract Neurol, 2006, 2(1): 26-33
[5] Welch GN, Loscalzo J. Homocysteine and atherothrombosis. N Engl J Med, 1998,338(15):1042-1050。
The content of the invention
The invention provides a kind of non-derivative method detection homocysteine, the liquid matter connection of cysteine and methionine With method, the method is first using dithiothreitol (DTT)（DTT）Mating type amino acid is reduced, reprecipitation albumen, remove blood plasma In protein, obtain the supernatant containing Hcy, Cys and Met；Using LC-MS/MS methods simultaneously to Hcy, the Cys in blood plasma and Met is quantified, the interior interference for being designated as Isotopic Internal Standard, excluding matrix employed in method.Wherein deuterated methionine pair There is cross jamming in the measure of homocysteine, deuterated methionine and homocysteine are realized chromatogram point by this method From, it is ensured that the accuracy of detection.The method specificity, result are accurate, detection time is short, make up most using evaporative light pair at present Hcy carries out quantitative deficiency, and can more comprehensively react the overall condition of homocysteine metabolism.
The technical solution adopted in the present invention is：The liquid phase color of Hcy, Cys and Met in a kind of non-derivative method detection blood plasma Tandem mass spectrum method is composed, detection method step is：
A. the preparation of plasma sample：Take in plasma sample to EP pipes, after adding inner mark solution, add DTT reduction, it is heavy to add Shallow lake agent, vortex, centrifugation carry out protein precipitation treatment, obtain containing homocysteine, the supernatant of cysteine and methionine Liquid.
B. above-mentioned supernatant is analyzed and is gathered using LC-MS/MS methods.
The concrete operations of protein precipitation are addition 600 in A in stepμL precipitating reagents, be vortexed 30 s, 14000 g centrifugations 3min。
The concrete operations of reduction reaction are addition 100 in step AμL DTT reducing agents, 30 DEG C of water after the 30 s mixings that are vortexed 15 min of bath reaction.
In step in A inner mark solution be deuterated homocysteine (d 4 - Hcy), deuterated cysteine（d 2 -Cys）With it is deuterated Methionine（d 3 -Met）Mixed solution.
The reducing agent is dithiothreitol (DTT)（DTT）.
The chromatographic column uses hydrophilic chromatographic post.
The sample is blood plasma.
It is described mixing inner mark solution be containingd 4 - homocysteine,d 2 - cysteine,d 3 The solution of-methionine.
It is described mixing inner mark solution concentration be：
d 4 - homocysteine： 10μmol/L
d 2 - cysteine： 100μmol/L
d 3 - methionine： 20μmol/L
The precipitating reagent is acetonitrile, water, the wherein mixed solution of watery hydrochloric acid/formic acid/acetic acid, acetonitrile：Water：Sour volume ratio is 100: 10~30:0.001~0.01。
Liquid phase chromatogram condition in B in step：Chromatographic column：Waters Xbridge 4.6 × 50mm, 3.5μM, column temperature：40 DEG C, sample introduction room temperature：4 DEG C, flow velocity：0.6 mL/min, sampling volume：1μL, mobile phase A:0.1% formic acid water, Mobile phase B: Acetonitrile, gradient elution；Mass Spectrometry Conditions：Ion gun is ESI+, Desolvention gas velocity：800 L/h；Desolventizing temperature：500℃；Hair Tubule voltage：3KV.
1) configuration of reducing agent：Under room temperature condition, accurate in 5 g reducing agents to add 100 mL purified waters, vortex makes its molten Solution.
2) standard curve（Quality-control product）Preparation method：Take 5μL standard working solutions（Quality-control product）In 1.5 mL centrifuge tubes, Sequentially add 45μL dilutions, 50μL internal standard solutions, 100μL reducing agents, after 30 s that are vortexed are mixed, 30 DEG C of water-baths 15 min.Add 600μL precipitating reagents, in the min of vortex oscillation 3, high speed centrifugation（14000g）3 min, take 100μL of supernatant liquid is carried out Tandem Mass Spectrometry Analysis.
3) preparation method of sample：Take 50μL sample solutions sequentially add 50 in 1.5mL centrifuge tubesμL internal standard solutions, 100 μL reducing agents, after 30 s that are vortexed are mixed, 30 DEG C of min of water-bath 15.Add 600μL precipitating reagents, vortex oscillation 3min In, high speed centrifugation（14000g）3min, takes 100μL of supernatant liquid carries out Tandem Mass Spectrometry Analysis.
Beneficial effects of the present invention：The method carries out sample preprocessing using non-derivative method, and method is simple, it is easy to grasp Make, enormously simplify sample pretreatment operation；Isotopic Internal Standard reduces interference of the matrix to detecting, it is ensured that the Guang ammonia of homotype half The degree of accuracy of acid, cysteine and methionine detection.The method uses LC-MS/MS methods to homocysteine in blood plasma, Cysteine and methionine are detected, using many reaction detection MRM scan modes, and found firstd 3 - methionine with Homocysteine（136>90 passages）There is cross jamming, using the elution requirement of liquid chromatogram gradient by its baseline separation, Ensure the accuracy of detection.The method sensitivity and specificity is strong, result is accurate.The method is scanned using MRM, high-specificity, Using Isotopic Internal Standard it is quantitatively accurate, to exclude matrix interference, detection time short.Make up only homocysteine is determined at present The deficiency of amount, can more comprehensively react the overall condition of homocysteine metabolism.
Brief description of the drawings
Fig. 1 is the MRM of Cys, Met hybrid standard product and each determinand inner mark solution containing Hcy described in the embodiment of the present invention Chromatogram.
Fig. 2 is the plasma sample and inner mark solution MRM chromatograms described in the embodiment of the present invention.
Specific embodiment
Below in conjunction with instantiation, the present invention is expanded on further.These application examples be merely to illustrate the present invention and without In limitation the scope of the present invention.
1. instrument and reagent
Liquid chromatograph-mass spectrometer：UHPLC-Xevo Waters
High speed freezing centrifuge：TECHCOMP model Cs T18RT
Constant temperature DL instrument：THERMO-SHAKER models AS20130565138
Ultrasonic cleaner：New sesame model SB-25-12D
Formic acid（Chromatographic grade）, acetonitrile（Chromatographic grade）, water（Wahaha Pure Water）
2. the preparation of standard liquid
Hcy is taken respectively, Cys and Met standard items are appropriate, accurately weighed, are placed in EP pipes, and acetonitrile is dissolved, and is made containing Hcy, The hybrid standard product solution of Cys and Met, it is standby.Dilute above-mentioned hybrid standard product solution with 50% acetonitrile solution, be made as The series standard solution and QC solution of lower concentration, are shown in Table 2.
The series standard solution and QC solution of table 2 Hcy, Cys and Met
Hcy Cys Met
10 50 20
25 125 50
100 500 400
400 2000 800
1000 5000 2000
QC is low 20 100 40
In QC 200 1000 400
QC is high 800 4000 1600
3. chromatographic condition：
Chromatographic column：Waters Xbridge 4.6 × 50mm, 3.5μm；
Mobile phase：0.1% aqueous formic acid（A）, acetonitrile（B）；
Flow velocity：0.6 ml/min；
Column temperature：40℃；
Sample size：1μL
Gradient elution program：
Time（min） A（%） B（%）
0.0 10 90
2.5 50 50
2.6 10 90
3.0 10 90
4. Mass Spectrometry Conditions：
Ion gun：ESI+
Source temperature：150℃
Precipitation is vented one's spleen temperature：500 ℃；
Desolvention gas velocity：800 L/h；
Capillary voltage：3KV；
Determinand MRM sweep parameters are shown in Table 3.
Table 3 Hcy, Cys, Met and interior target MRM sweep parameters
Title Taper hole voltage（V） Impact energy（V）
Cys 122 76 30 10
Hcy 136 90 28 6
Met 150 104 30 8
140 94 30 10
124 80 28 10
153 107 30 8
5. pre-treating method：
5.1 standard working solutions/quality-control product pre-treating method：
Take 5μL standard working solutions（Quality-control product）In 1.5 mL centrifuge tubes, 45 are sequentially addedμL dilutions, 50μL internal standard solutions, 100 μL reducing agents, after 30 s that are vortexed are mixed, 30 DEG C of min of water-bath 15.Add 600μL precipitating reagents, the min of vortex oscillation 3 In, high speed centrifugation（14000g）3 min, draw 100μL of supernatant liquid enters chromatography.
5.2 plasma sample pre-treating methods：
Take 50μL sample solutions sequentially add 50 in 1.5 mL centrifuge tubesμL internal standard solutions, 100μL reducing agents, be vortexed 30 s After mixing, 30 DEG C of min of water-bath 15.Add 600μL precipitating reagents, in the min of vortex oscillation 3, high speed centrifugation（14000g）3 Min, draws 100μL of supernatant liquid enters chromatography.
6. linear equation
Sample is prepared according to standard working solution pre-treating method, LC-MS/MS analyses are carried out.Respectively with the dense of Hcy, Cys and Met It is abscissa to spend, with Hcy, Cys and Met with interior target peak area ratio as ordinate, with weighting（W=1/C2）Least square method is entered Row is returned and calculated, and least square method carries out regressing calculation, the linear regression equation tried to achieve as standard curve.Typical recurrence side Journey and the range of linearity are shown in Table 4
The retention time of table 4 Hcy, Cys and Met, linear equation and linearly dependent coefficient
Analyte Linear equation Retention time（min）
Hcy Y=0.177X ﹣ 0.046 0.9960 1.97
Cys Y=0.059X ﹣ 0.107 0.9994 2.41
Met Y=1.915X + 0.118 0.9970 1.82
7. precision test
Sample is prepared according to QC pre-treating methods, each concentration prepares 6 samples, METHOD FOR CONTINUOUS DETERMINATION 3 days, and and standard curve respectively Determined with batch, the concentration of QC samples is calculated with the standard curve on the same day, try to achieve the precision of method（RSD）And the degree of accuracy（RE）, The results are shown in Table 5.
The Precision test result of table 5
8. recovery test
Precision measures calf serum 45μL, adds 5μSeries standard solution of the L containing each determinand, prepares Hcy, Cys and Met The QC samples of basic, normal, high three concentration（Per the sample analysis of concentration five）, the peak area for measuring is designated as A1, while another precision is measured Calf serum 45μL, in addition to internal standard is not added with, by " treatment of plasma sample " item, similarly hereinafter method is tested, in the supernatant addition of acquisition The standard liquid each 5 of mark solution and respective concentrationµL, vortex mixing carries out LC-MS/MS analyses, obtains corresponding peak area （Five average values of measure）, A2 is designated as, with the peak area ratio A of two kinds of processing methods of each concentration1/A2× 100% calculating is carried Fetch yield.The results are shown in Table 6.
The recovery test result of table 6
9. the calculating of analysis result
The peak area of testing sample is read, according to the Hcy that step 4 is set up, Cys, Met standard curves are calculated sample Hcy, Cys, Met content are respectively 10μMol/L, 150μMol/L and 20μmol/L.Appearance time as shown in Figure 1 is respectively 1.81min, 2.14min, 1.67min.
Although above-mentioned be described with reference to accompanying drawing to specific embodiment of the invention, not to invention protection domain Limitation, on the basis of technical scheme, those skilled in the art make by need not paying creative work Various modifications or deform it is still within the scope of the present invention.

Claims (3)

1. a kind of non-derivative method detects the Liquid Chromatography-Tandem Mass Spectrometry method of sulfur-containing amino acid in blood plasma, and sulfur-containing amino acid is same Type cysteine, cysteine and methionine, it is characterised in that comprise the following steps：
（1）The preparation of plasma sample
In taking plasma sample to EP pipes, after adding Isotopic Internal Standard solution, add reducing agent reduction, add precipitating reagent, vortex, Centrifugation carries out protein precipitation treatment, obtains containing homocysteine, the supernatant of cysteine and methionine；The same position Plain inner mark solution be containingd 4 - homocysteine,d 2 - cysteine,d 3 The solution of-methionine, reducing agent is two sulphur threoses Alcohol, precipitating reagent is acetonitrile：Water：Sour volume ratio is 100:10~30:0.001 ~ 0.01 mixed solution, acid for watery hydrochloric acid, formic acid or Acetic acid；
（2）The preparation of standard curve
The concentration with homocysteine, cysteine and methionine is as abscissa respectively, with homocysteine, half Guang ammonia Acid and methionine and interior target peak area ratio are ordinate, and recurrence calculating, least square method are carried out with weighted least-squares method Regressing calculation is carried out, the linear regression equation tried to achieve as standard curve；The standard curve of homocysteine is Y=0.177X ﹣ 0.046, the standard curve of cysteine is Y=0.059X ﹣ 0.107, the standard curve of methionine for Y=1.915X+ 0.118；
（3）Above-mentioned supernatant is analyzed and gathered using LC-MS/MS
Take 50μL sample solutions sequentially add 50 in 1.5mL centrifuge tubesμL internal standard solutions, 100μL reducing agents, be vortexed 30 s After mixing, 30 DEG C of water-bath 15min；
Add 600μL precipitating reagents, in vortex oscillation 3min, with 14000 gHigh speed centrifugation 3min, draws 100μL of supernatant liquid enters Row Tandem Mass Spectrometry Analysis；
Liquid phase chromatogram condition is chromatographic column：Waters Xbridge 4.6 × 50mm, 3.5μm；Column temperature：40 DEG C, sample introduction room temperature Degree：4 DEG C, flow velocity：0.6 mL/min, sampling volume：1μL, mobile phase A:0.1% aqueous formic acid, Mobile phase B:Acetonitrile, ladder Degree wash-out, elution program is shown in Table 1；Mass Spectrometry Conditions are：Ion gun：ESI+；Desolvention gas velocity：800 L/h；Desolventizing temperature： 500℃；Capillary voltage：3KV；
The gradient elution program of table 1
Time（min） Mobile phase A（v%） Mobile phase B（v%） 0.0 10 90 2.5 50 50 2.6 10 90 3.0 10 90
Homocysteine, cysteine and methionine and interior target peak area ratio, bring homocysteine, half Guang into respectively The standard curve of propylhomoserin and methionine, obtains the concentration of homocysteine, cysteine and methionine.
2. a kind of non-derivative method according to claim 1 detects the Liquid Chromatography-Tandem Mass Spectrometry of sulfur-containing amino acid in blood plasma Method, it is characterised in that：The chromatographic column uses hydrophilic chromatographic post.
3. a kind of non-derivative method according to claim 1 detects the Liquid Chromatography-Tandem Mass Spectrometry of sulfur-containing amino acid in blood plasma Method, it is characterised in that：The Isotopic Internal Standard solution concentration be containingd 4 - homocysteine 10μMol/L, containsd 2 - half Guang ammonia Acid 100μMol/L, containsd 3 - methionine 20μmol/L。
CN201710129265.5A 2017-03-06 2017-03-06 A kind of Liquid Chromatography-Tandem Mass Spectrometry method of sulfur-containing amino acid in non-derivative method detection blood plasma Pending CN106841488A (en)

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107091894A (en) * 2017-06-14 2017-08-25 杭州博圣生物技术有限公司 The method of Liquid Chromatography-Tandem Mass Spectrometry detection methylmalonic acid, methyl citric acid and/or homocysteine
CN107907618A (en) * 2017-10-24 2018-04-13 重庆医科大学 The detection method of homocysteine and cysteine in rat plasma
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101738440A (en) * 2009-12-15 2010-06-16 大连依利特分析仪器有限公司 Method for detecting sulfur-containing amino acid
US20120318971A1 (en) * 2011-06-16 2012-12-20 Baylor Research Institute Analysis of total homocysteine and methylmalonic acid in plasma by lc-ms/ms from a plasma separator device (psd)
CN103163226A (en) * 2011-12-14 2013-06-19 刘丽宏 A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof
CN104215721A (en) * 2014-10-09 2014-12-17 中国科学院上海有机化学研究所 Application of phthalic acid in amino acid analysis
CN106093226A (en) * 2016-06-01 2016-11-09 辽宁润生康泰生物医药科技有限公司 A kind of method detecting common amino acid in immune t-cell and B cell

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101738440A (en) * 2009-12-15 2010-06-16 大连依利特分析仪器有限公司 Method for detecting sulfur-containing amino acid
US20120318971A1 (en) * 2011-06-16 2012-12-20 Baylor Research Institute Analysis of total homocysteine and methylmalonic acid in plasma by lc-ms/ms from a plasma separator device (psd)
CN103163226A (en) * 2011-12-14 2013-06-19 刘丽宏 A simultaneous quantitative detection method of 30 amino acids and a preparation method thereof
CN104215721A (en) * 2014-10-09 2014-12-17 中国科学院上海有机化学研究所 Application of phthalic acid in amino acid analysis
CN106093226A (en) * 2016-06-01 2016-11-09 辽宁润生康泰生物医药科技有限公司 A kind of method detecting common amino acid in immune t-cell and B cell

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
潘晓华 等: "高同型半胱氨酸血症与缺血性脑血管病相关性", 《中国热带医学》 *
赵世晶 等: "高效液相色谱法同时测定血浆中S-腺苷甲硫氨酸和S-腺苷同型半胱氨酸", 《卫生研究》 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107091894A (en) * 2017-06-14 2017-08-25 杭州博圣生物技术有限公司 The method of Liquid Chromatography-Tandem Mass Spectrometry detection methylmalonic acid, methyl citric acid and/or homocysteine
CN107907618A (en) * 2017-10-24 2018-04-13 重庆医科大学 The detection method of homocysteine and cysteine in rat plasma
CN108362795A (en) * 2018-02-08 2018-08-03 杭州佰辰医学检验所有限公司 Content of homocysteine rapid detection method in dried blood spot
CN110018266A (en) * 2019-02-15 2019-07-16 广州市妇女儿童医疗中心 A kind of method of 48 kinds of amino acid of fast quantitative analysis
CN110018266B (en) * 2019-02-15 2022-03-04 广州市妇女儿童医疗中心 Method for rapidly and quantitatively analyzing 48 amino acids
CN110274973A (en) * 2019-07-16 2019-09-24 大连润生康泰医学检验实验室有限公司 The detection method and its enrichment material of sulfur-containing amino acid in a kind of serum
CN110274973B (en) * 2019-07-16 2021-03-26 大连润生康泰医学检验实验室有限公司 Detection method of sulfur-containing amino acid in serum and enrichment material thereof
CN110441457A (en) * 2019-08-02 2019-11-12 深圳市绿航星际太空科技研究院 A kind of method of homocysteine in detection urine
CN112964808A (en) * 2019-12-13 2021-06-15 中国科学院大连化学物理研究所 Biological body fluid total homocysteine detection kit and detection method
CN111983057A (en) * 2020-07-29 2020-11-24 上海润达榕嘉生物科技有限公司 Method and kit for detecting homocysteine in human plasma
CN112730723A (en) * 2020-12-30 2021-04-30 申友基因组研究院(南京)有限公司 Method for detecting 22 free amino acids in plasma by ultra-high performance liquid chromatography-tandem mass spectrometry
CN112730723B (en) * 2020-12-30 2022-11-04 申友基因组研究院(南京)有限公司 Method for detecting 22 free amino acids in plasma by ultra-high performance liquid chromatography-tandem mass spectrometry
CN112834677A (en) * 2020-12-31 2021-05-25 质谱生物科技有限公司 Method for simultaneously detecting homocysteine and metabolic related substances thereof
CN115825214A (en) * 2022-11-30 2023-03-21 厦门大学 MALDI-TOF-MS-based method for in-vitro screening of hMAT2A protein inhibitor and application thereof
CN117129605A (en) * 2023-10-25 2023-11-28 济南和合医学检验有限公司 Method for detecting 11 antihypertensive drugs and 3 metabolites by liquid chromatography-tandem mass spectrometry
CN117129605B (en) * 2023-10-25 2024-02-02 济南和合医学检验有限公司 Method for detecting 11 antihypertensive drugs and 3 metabolites by liquid chromatography-tandem mass spectrometry

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