CN101419201A - Method and special equipment for LC-MS analyzing melamine by on-line desalinisation - Google Patents
Method and special equipment for LC-MS analyzing melamine by on-line desalinisation Download PDFInfo
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- CN101419201A CN101419201A CNA2008102280923A CN200810228092A CN101419201A CN 101419201 A CN101419201 A CN 101419201A CN A2008102280923 A CNA2008102280923 A CN A2008102280923A CN 200810228092 A CN200810228092 A CN 200810228092A CN 101419201 A CN101419201 A CN 101419201A
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Abstract
The invention discloses a method for an online desalination LC-MS analysis on melamine and special equipment. The method comprises the following steps: separating a sample through a reversed phase ion pair chromatography and an ion exchange chromatography and concentrating the compositions to be tested through a trapping column; washing the trapping column to remove salt or an ion pair reagent in a mobile phase; and backwashing the trapping column with a salt-free mobile phase so that the compositions to be tested enter a mass spectrum to be analyzed. The equipment is provided with a multi-port valve; a first inlet of the multi-port valve is connected with a chromatographic column and is connected with a mobile phase input pump through a sampling valve; the first inlet is also connected with a desalting cleaning liquid input pump; a first outlet of the multi-port valve is connected with a waste liquid pool; a second inlet of the multi-port valve is connected with a salt-free mobile phase input pump; the second outlet of the multi-port valve is connected with a mass spectrum detector; and the trapping column is bridged between two valve holes of the multi-port valve. The equipment has the advantages of simple structure, convenient operation and low cost, thus the reversed phase ion pair chromatography and the ion exchange chromatography can be used together with a mass spectrometric analysis.
Description
Technical field:
The present invention relates to a kind of LC-MS analytical approach and device of melamine, especially a kind of simple in structure, easy to operate, with low cost, can realize the method and the specialized equipment of the online desalination LC-MS analyzing melamine of reversed-phase high-performance liquid chromatography and ion-exchange chromatography and mass spectrophotometry coupling.
Background technology:
Melamine (Melamine) is a kind of important azacyclo-Organic Chemicals, can be widely used in to produce plastics, floor tile, kitchen utensils, fire proofing firbre, commercial filter membrane, glue and fire retardant etc.But, U.S. food Drug Administration (FDA) had once found melamine on March 30th, 2007 in to the examining report of pet food first, find to contain melamine in 22 kinds of baby milk powders such as Sanlu, find all to exist in the liquid milk of multiple brands such as Erie and Mongolia Ox and the milk powder melamine in various degree in the detection subsequently again thereafter.Melamine itself is nontoxic, but facile hydrolysis produces the cyanuric acid that can cause kidney stone under acid condition, the food that contains excessive melamine is healthy with serious harm people or pet, is crucial for the accurate analysis that whether contains melamine in the food therefore.At present, the analytical approach to melamine in food or other materials mainly contains following two kinds, the problem of equal various degrees:
1. adopt reversed-phase high-performance liquid chromatography the melamine in the sample to be separated and be aided with the analytical approach that the UV, visible light detecting device detects with ion-exchange chromatography.This kind method is simple to operate, with low cost, be convenient to popularize, but can polluting with signal mass spectrometer, high salt that is adopted because of reversed-phase high-performance liquid chromatography and ion-exchange chromatography or the moving phase that contains ion-pairing agent disturbs, therefore can't obtain mass data with mass detector, carry out qualitative accurately, quantitative test, can only accuracy and reliability that melamine is analyzed have been reduced with the method for UV, visible light detecting device detection.
2. the special chromatographic column that adopts salt-free moving phase and be applicable to salt-free moving phase is separated the melamine in the sample and is detected with mass detector, i.e. the LC-MS analytical approach.This kind method can be obtained mass data with mass detector, carries out qualitative accurately, quantitative test, has overcome preceding method precision of analysis and the low problem of reliability.But because the singularity of its chromatographic column, at present can only be from external import, the cost height is not easy to promote the use of.
Summary of the invention:
The present invention is in order to solve the above-mentioned technical matters of existing in prior technology, provide a kind of simple in structure, easy to operate, with low cost, can realize the method and the specialized equipment of the online desalination LC-MS analyzing melamine of reversed-phase high-performance liquid chromatography and ion-exchange chromatography and mass spectrophotometry coupling.
Technical solution of the present invention is: a kind of method of online desalination LC-MS analyzing melamine is characterized in that carrying out as follows:
A. sample is separated with ion-exchange chromatography and through trapping column enrichment component to be measured through reversed-phase high-performance liquid chromatography;
B. trapping column is washed to remove salinity or the ion-pairing agent in the moving phase;
C. towards trapping column component to be measured being entered in the mass spectrum with salt-free flowing opposite analyzes.
A kind of method specialized equipment of online desalination LC-MS analyzing melamine, it is characterized in that: be provided with two multiport valve V, be connected to chromatographic column 12 with first import of two multiport valve V, chromatographic column 12 is joined with moving phase front pump 10A by sampling valve 11, also be connected to desalination cleaning fluid front pump 10B with first import, export with first of two multiport valve V and be connected to waste liquid pool 15; Be connected to salt-free moving phase front pump 10C with second import of two multiport valve V, export with second of two multiport valve V and be connected to Mass Spectrometer Method instrument 16; Trapping column 14 is connected across between two valve openings of two multiport valve V and is communicated with first import of two multiport valve V, first outlet and second import, second outlet respectively.
Described chromatographic column 12, desalination cleaning fluid front pump 10B can join by first import of T-valve 13 and two multiport valve V.
Can be connected to mixer 17 between described moving phase front pump 10A and sampling valve 11, described desalination cleaning fluid front pump 10B joins by first import of mixer 17, sampling valve 11, chromatographic column 12 and two multiport valve V.
Described two multiport valve V can be two six-way valves, and first import, first outlet are respectively valve opening 6, valve opening 5; Second import, second outlet are respectively valve opening 3, valve opening 2; Trapping column 14 is connected across between the valve opening 1,4.
Described two multiport valve V are two ten-way valves, and first import, first outlet are respectively valve opening 1, valve opening 6; Second import, second outlet are respectively valve opening 8, valve opening 3; The first trapping column 14A is connected across between the valve opening 0,7, and the second trapping column 14B is connected across between the valve opening 2,5, and cross-over connection has normal open pipeline 18 between valve opening 4 and the valve opening 9.
Described trapping column 14 is silica matrix or polymer substrate reverse-phase chromatography filler.
The present invention uses reverse-phase chromatographic column as trapping column, utilize the different separating mechanisms of trapping column and reversed-phase high-performance liquid chromatography with the ion-exchange chromatography post, make component enrichment to be measured by trapping column, and the online purpose of component to be measured being washed and reached by salt-free moving phase replacement desalination or ion-pairing agent, solve reversed-phase high-performance liquid chromatography and ion-exchange chromatography and be difficult to problem with mass spectrometry, promptly have simple in structure, easy to operate, with low cost, the advantage of being convenient to promote the use of, also can obtain mass data simultaneously by mass detector, carry out qualitative accurately, quantitative test improves the main accuracy of sample quantitative test.
Description of drawings:
Fig. 1 is the embodiment of the invention 1 a device syndeton synoptic diagram.
Fig. 2 is the embodiment of the invention 2 device syndeton synoptic diagram.
Fig. 3 is the ultraviolet detection spectrogram of the embodiment of the invention 1.
Fig. 4 is the mass spectrogram of the embodiment of the invention 1.
Embodiment:
Embodiment 1:
As shown in Figure 1: a kind of method specialized equipment of online desalination LC-MS analyzing melamine, be provided with two six-way valve V (outsourcing), can be manual or electronic six-way valve V, six valve openings 1～6 are arranged, by manual or motor-operated control valve core, with realize communicating in twos in the valve opening 1～6, obstructed two positions, both I position, II position in twos, two-wire pipeline UNICOM during the I position, short-term pipeline connection during the II position.Be respectively first import, first outlet with valve opening 6, valve opening 5; Valve opening 3, valve opening 2 are respectively second import, second outlet, and trapping column 14 is connected across between the valve opening 1,4, and 14 of trapping column are communicated with valve opening 6, valve opening 5 and valve opening 3, the valve opening 2 of two six-way valve V respectively.Be connected to chromatographic column 12 with first import of two six-way valve V, chromatographic column 12 is reverse-phase paired ion chromatography or ion-exchange chromatography, can be silica gel bonded reverse-phase chromatography filler, polymkeric substance chromatograph packing material, ion-exchange chromatography filler or ion exchange resin etc., and trapping column 14 is reverse-phase chromatographic columns, can be silica matrix or polymer substrate reverse-phase chromatography filler, as SCX trapping column etc., can also be other inorganic oxide matrix chromatograph packing materials.Chromatographic column 12 is joined with moving phase front pump 10A by sampling valve 11, also is connected to desalination cleaning fluid front pump 10B with first import, exports with first of two six-way valve V and is connected to waste liquid pool 15; Be connected to salt-free moving phase front pump 10C with second import of two six-way valve V, export with second of two six-way valve V and be connected to Mass Spectrometer Method instrument 16; Chromatographic column 12, desalination cleaning fluid front pump 10B can join by first import (valve opening 6) of T-valve 13 and two six-way valve V, and pump 10A, 10B, 10C can use high pressure constant flow pump.Described Mass Spectrometer Method instrument 16 used ion guns can be electron impact ionization (EI), chemical ioni zation (CI), field ionization (FI), field desorptiion (FD), fast atom bombardment (FAB), secondary ion mass spectrum (SIMS), laser desorption (LD), electrofluid effect ionization (EH), electro-spray ionization (ESI), thermal spray ionization (TSI) etc.
Can realize the method for online desalination LC-MS analyzing melamine by embodiment 1 described equipment, promptly carry out as follows:
A. sample is separated with ion-exchange chromatography and through trapping column enrichment component to be measured through reversed-phase high-performance liquid chromatography;
B. trapping column is washed to remove salinity or the ion-pairing agent in the moving phase;
C. towards trapping column component to be measured being entered in the mass spectrum with salt-free flowing opposite analyzes.
Concrete operation method is as follows:
1. operating conditions:
Chromatographic column: Elite MSP 5 μ m, ID4.6mm * 250mm, moving phase: acetonitrile/damping fluid, 10/90 (containing the 10mM citric acid in the damping fluid, the 10mM sodium heptanesulfonate), flow: 1.0mL/min sample size: 10 μ L, column temperature: 40 ℃, detect wavelength: UV240nm.
Above-mentioned reversed-phase high-performance liquid chromatography and ion-exchange chromatography condition are taken from the Yi Lite website.
The mass spectrum condition:
Mass spectrum ionization mode: electron spray ionisation source (ESI); Dry gas temperature: 350 ℃; Atomization gas pressure: 35psi; Cracked voltage: 100V; Dry gas flow velocity: 12L/min; Capillary voltage: 3500V; Select ion detection (SIR), mass-to-charge ratio (m/z): 127,85,60 (take from: Lai Biqing, Zheng Xiaohang, Han Yintao, high performance liquid chromatography one quadrupole rod mass spectrometry is measured content of melamine in the feed, feed industry, 2008 the 29th the 4th phases of volume).
Mass?Range：40-450amu；
Scan?Mode：Full(Electron?Ionization)；
Filament?Delay：4.2min(splitless)；3.8min(split)；
Threshold：100；
MS?Quad：150℃；
MS Source:230 ℃ (taking from the condition that FDA GC-MS analyzes).
2. operation steps:
When six-way valve (V) is in the I position, the two-wire pipeline connection, start moving phase front pump 10A, simultaneously with test sample by sampling valve 11 sample introductions, separate by ion-exchange (ion pair) chromatographic column 12, pass through T-valve 13, valve opening 6, valve opening 1, trapping column 14, valve opening 4, valve opening 5 more successively, component to be measured (containing melamine) enrichment on the column cap of trapping column 14, waste liquid enters in the waste liquid pool 15.Behind the component enrichment certain hour to be measured, stop moving phase front pump 10A, start desalination cleaning fluid front pump 10B, trapping column is cleaned to remove freshen with no salt solusion.Switch two six-way valve V to the II position, the short-term pipeline connection, start salt-free moving phase front pump 10C, make salt-free moving phase by valve opening 3,4 recoil trapping column 14, and the component to be measured of institute's enrichment is analyzed in Mass Spectrometer Method instrument 16 by valve opening 1,2 sample introductions with salt-free moving phase.
The present invention is simple in structure, processing ease, with low cost, detect accurately, the mass spectrogram that Fig. 4 promptly analyzes, detects through present embodiment 1 (spectrogram is taken from the condition that FDA GC-MS analyzes), Fig. 3 separates with the ion-exchange chromatography condition with the reversed-phase high-performance liquid chromatography of present embodiment 1, with the spectrogram of UV-detector detection.By contrast as can be seen, the accuracy of present embodiment 1 usefulness mass detector testing result and reliability are significantly better than the testing result with UV-detector.
Embodiment 2:
As shown in Figure 2: two ten-way valve V (outsourcing) are arranged, can be manual or electronic ten-way valve V, ten valve openings 0～9 are all arranged, by manual or motor-operated control valve core, with realize communicating in twos in the valve opening 0～9, obstructed two positions in twos, both I position, II position, two-wire pipeline UNICOM during the I position, short-term pipeline connection during the II position, cross-over connection has normal open pipeline 18 between valve opening 4 and the valve opening 9.Be respectively first import, first outlet with valve opening 1, valve opening 6; Valve opening 8, valve opening 3 are respectively second import, second outlet, and trapping column 14A is connected across between the valve opening 0,7, are communicated with valve opening 1, valve opening 6 and valve opening 8, the valve opening 3 of two ten-way valve V respectively; Simultaneously trapping column 14B is connected across between the valve opening 2,5, trapping column 14 also is communicated with valve opening 1, valve opening 6 and valve opening 8, the valve opening 3 of two ten-way valve V respectively.Be connected to chromatographic column 12 with first import of two ten-way valve V, chromatographic column 12 is reverse-phase paired ion chromatography or ion-exchange chromatography, can be silica gel bonded reverse-phase chromatography filler, polymkeric substance chromatograph packing material, ion-exchange chromatography filler or ion exchange resin etc.As trapping column 14A, 14B, can be silica matrix or polymer substrate reverse-phase chromatography filler with reverse-phase chromatographic column, as SCX trapping column etc., can also be other inorganic oxide matrix chromatograph packing materials.Chromatographic column 12 is joined with moving phase front pump 10A by sampling valve 11, mixer 17, and desalination cleaning fluid front pump 10B also joins with mixer 17.Be connected to waste liquid pool 15 with first outlet of two ten-way valve V; Be connected to salt-free moving phase front pump 10C with second import of two ten-way valve V, export with second of two multiport valve V and be connected to Mass Spectrometer Method instrument 16; Moving phase front pump 10A, desalination cleaning fluid front pump 10B join by first import (valve opening 1) of mixer 17, sampling valve 11, chromatographic column 12 and two ten-way valve V.Pump 10A, 10B, 10C can use high pressure constant flow pump.Described Mass Spectrometer Method instrument 16 used ion guns can be electron impact ionization (EI), chemical ioni zation (CI), field ionization (FI), field desorptiion (FD), fast atom bombardment (FAB), secondary ion mass spectrum (SIMS), laser desorption (LD), electrofluid effect ionization (EH), electro-spray ionization (ESI), thermal spray ionization (TSI) etc.
Can realize the method for online desalination LC-MS analyzing melamine by embodiment 2 described equipment, promptly carry out as follows:
A. sample is separated with ion-exchange chromatography and through trapping column enrichment component to be measured through reversed-phase high-performance liquid chromatography;
B. trapping column is washed to remove salinity or the ion-pairing agent in the moving phase;
C. towards trapping column component to be measured being entered in the mass spectrum with salt-free flowing opposite analyzes.
Concrete operation method is as follows:
1. operating conditions:
With embodiment 1.
2. operation steps:
When (1) two ten-way valves (V) being in the I position, the two-wire pipeline connection, this moment can be with moving phase front pump 10A, desalination cleaning fluid front pump 10B simultaneously as the moving phase front pump.Start pump (10A, 10B), flow communication is crossed mixer 17 and is mixed, simultaneously with test sample by sampling valve 11 sample introductions, separate by ion-exchange (ion pair) chromatographic column 12, again successively by valve opening 1, valve opening 2, trapping column 14B, valve opening 5, valve opening 6, component to be measured (containing melamine) enrichment on the column cap of trapping column 14B, waste liquid enters in the waste liquid pool 15.Behind the component enrichment certain hour to be measured, change the gradient condition of double pump (10A, 10B), import no salt solusion trapping column (14B) is washed, to slough the mass spectrometric salinity of easy interference.
(2) switch two ten-way valve V to the II position, the short-term pipeline connection, start salt-free moving phase front pump 10C, make salt-free moving phase by valve opening 8,9, normal open pipeline 18,4,5 recoil trapping column 14B, and the component to be measured of institute's enrichment is analyzed in Mass Spectrometer Method instrument 16 by valve opening 2,3 sample introductions with salt-free moving phase.
(3) when two ten-way valve V are in the II position, can change double pump 10A, 10B gradient condition again, and simultaneously as the moving phase front pump, with test sample by sampling valve 11 sample introductions, separate by ion-exchange (ion pair) chromatographic column 12, pass through valve opening 1, valve opening 0, trapping column 14A, valve opening 7, valve opening 6 more successively, component to be measured (containing melamine) enrichment on the column cap of trapping column 14A, waste liquid enters in the waste liquid pool 15.Behind the component enrichment certain hour to be measured, change the gradient condition of double pump (10A, 10B), import no salt solusion trapping column (14A) is washed, to slough the mass spectrometric salinity of easy interference.
(4) switch two ten-way valve V once more to the I position, the two-wire pipeline connection, start salt-free moving phase front pump 10C, make salt-free moving phase by valve opening 8,7 recoil trapping column 14A, and the component to be measured of institute's enrichment is analyzed in Mass Spectrometer Method instrument 16 by valve opening 0,9, normal open pipeline 18,4,3 sample introductions with salt-free moving phase.
Meanwhile, can make component to be measured enrichment on the column cap of trapping column 14B according to above-mentioned (1) step ...
Switch repeatedly, make trapping column 14B, 14A alternate operation, can obviously improve the efficient of sample analysis, and can detect two kinds of different samples simultaneously.
The present invention remove can be used for melamine LC-MS analytically, can also realize the desalting processing on liquid chromatography and other Instrument crosslinkings, perhaps be used for the desalting processing of two dimension or multidimensional liquid chromatography.
Claims (7)
1. the method for an online desalination LC-MS analyzing melamine is characterized in that carrying out as follows:
A. sample is separated with ion-exchange chromatography and through trapping column enrichment component to be measured through reversed-phase high-performance liquid chromatography;
B. trapping column is washed to remove salinity or the ion-pairing agent in the moving phase;
C. towards trapping column component to be measured being entered in the mass spectrum with salt-free flowing opposite analyzes.
2. the method specialized equipment of an online according to claim 1 desalination LC-MS analyzing melamine, it is characterized in that: be provided with two multiport valve V, be connected to chromatographic column (12) with first import of two multiport valve V, chromatographic column (12) is joined by sampling valve (11) and moving phase front pump (10A), also be connected to desalination cleaning fluid front pump (10B) with first import, export with first of two multiport valve V and be connected to waste liquid pool (15); Be connected to salt-free moving phase front pump (10C) with second import of two multiport valve V, export with second of two multiport valve V and be connected to Mass Spectrometer Method instrument (16); Trapping column (14) is connected across between two valve openings of two multiport valve V and is communicated with first import, first outlet and second import, second outlet with two multiport valve V respectively.
3. the method specialized equipment of online desalination LC-MS analyzing melamine according to claim 2 is characterized in that: described chromatographic column (12), desalination cleaning fluid front pump (10B) join by T-valve (13) and first import of two multiport valve V.
4. the method specialized equipment of online desalination LC-MS analyzing melamine according to claim 2, it is characterized in that: be connected to mixer (17) between described moving phase front pump (10A) and sampling valve (11), described desalination cleaning fluid front pump (10B) joins with first import of two multiport valve V by mixer (17), sampling valve (11), chromatographic column (12).
5. the method specialized equipment of online desalination LC-MS analyzing melamine according to claim 3 is characterized in that: described two multiport valve V are two six-way valves, and first import, first outlet are respectively valve opening (6), valve opening (5); Second import, second outlet are respectively valve opening (3), valve opening (2); Trapping column (14) is connected across between valve opening (1), (4).
6. the method specialized equipment of online desalination LC-MS analyzing melamine according to claim 4 is characterized in that: described two multiport valve V are two ten-way valves, and first import, first outlet are respectively valve opening (1), valve opening (6); Second import, second outlet are respectively valve opening (8), valve opening (3); First trapping column (14A) is connected across between valve opening (0), (7), and second trapping column (14B) is connected across between valve opening (2), (5), and cross-over connection has normal open pipeline (18) between valve opening (4) and the valve opening (9).
7. according to the method specialized equipment of claim 2 or 3 or 4 or 5 described online desalination LC-MS analyzing melamines, it is characterized in that: described trapping column (14) is silica matrix or polymer substrate reverse-phase chromatography filler.
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| CN103706146A (en) * | 2013-12-10 | 2014-04-09 | 核工业北京地质研究院 | Column extraction system and extraction method for organic materials in solid particles |
| CN103706146B (en) * | 2013-12-10 | 2016-05-18 | 核工业北京地质研究院 | Organic column extracting system and extracting process in a kind of solid particulate matter |
| CN104991028A (en) * | 2015-07-31 | 2015-10-21 | 北京市药品检验所 | Method for reducing content of nonvolatile buffer salts in LC-MS test substance |
| CN112400109A (en) * | 2018-07-11 | 2021-02-23 | 沃特世科技公司 | Chromatography system and method for trap-elute mixed mode chromatography |
| CN109060992A (en) * | 2018-09-03 | 2018-12-21 | 大连技嘉科技有限公司 | The glycolated hemoglobin analysis of detection accuracy can be improved |
| CN109060992B (en) * | 2018-09-03 | 2024-02-27 | 苏州博睿嘉晟医疗科技有限公司 | Glycosylated hemoglobin analyzer capable of improving detection accuracy |
| WO2019228157A1 (en) * | 2019-01-14 | 2019-12-05 | 中国科学院南海海洋研究所 | Lc/ms and ms automatic switching high-throughput continuous sample feeding apparatus |
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Application publication date: 20090429 |