CN105929033A - System and method for measuring trace lactic acid ion in lobaplatin by using column switching-ion chromatography method - Google Patents
System and method for measuring trace lactic acid ion in lobaplatin by using column switching-ion chromatography method Download PDFInfo
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- CN105929033A CN105929033A CN201610222810.0A CN201610222810A CN105929033A CN 105929033 A CN105929033 A CN 105929033A CN 201610222810 A CN201610222810 A CN 201610222810A CN 105929033 A CN105929033 A CN 105929033A
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- 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
Abstract
The invention relates to a system and method for measuring trace lactic acid ion in lobaplatin by using a column switching-ion chromatography method, and belongs to the technical field of medicine testing. The method uses a low-configure ICS-1500 ion chromatograph and an eluent generator cooperatively, and the only need of completing complete separation of the target lactic acid ions and the other ions is the first order change of the eluent concentration when the IonPac NG1 chromatographic column is used as a pretreatment column, and AS19 anion column used as an analytical column. Moreover, the eluent concentration in the method is low, which is energy-saving and environmentally friendly. The system and method established by the invention have less required sample, good universality, simple operation and method, and low requirement of device configuration, and are easy to automate.
Description
Technical field
The invention belongs to medicine detection technique field, be specifically related to trace breast in post switching-ion-chromatographic determination lobaplatin
The system and method for acid ion.
Technical background
Lobaplatin chemistry entitled 1,2-diaminomethyl-Tetramethylene .-lactic acid closes platinum, is that the diastereomeric of a kind of platinum (II) chromium complex is different
Structure mixture, is also a kind of novel antitumor drug, is mainly used in treating breast carcinoma, small cell lung cancer and chronic granulocyte white
Disorders of blood.In lobaplatin 1,2-diaminomethyl-Tetramethylene. is for stablize aglucon, and lactic acid is leaving group, its anti-tumor activity source DNA-medicine
Crosslinking in the chain of the formation of thing adduct, mainly GG and AG.Lobaplatin can affect the expression of C-myc gene, and the expression of C-myc
Generation, apoptosis and cell proliferation with tumor are relevant.Meanwhile, it can overcome some pre-clinical tumor models resistance to cisplatin.
Industrial, lobaplatin synthesis is main with 1,2-dicyano Tetramethylene. as raw material, anti-by reduction, condensation, cyclization 3 step
Should obtain.Owing to, in ring-closure reaction, lactic acid is combined generation lobaplatin as raw material with condensation product, therefore can be by detection
Lactic acid content determines the conversion ratio of ring-closure reaction.
The assay method of lactic acid mainly has titrimetry, polarimetry, UV enzymatic analysis, Enzyme Electrode and chromatography of ions at present
Method.Polarimetry minute is long, and means are loaded down with trivial details；UV enzyme process is only limitted to the L Plasma lactate that purity is high, and cost of determination is relatively
High；Although Enzyme Electrode specificity height, low cost, analysis soon, but are used for the mensuration of medical science blood lactase acid.These methods are to sample
Requirement relatively large, it is impossible to be applied to the occasion that sample size is few, and these methods be not suitable for trace analysis[7]。
For simple ion chromatography system, if by lobaplatin direct injected, it is possible to find there is the biggest matrix interference, formed one several
Minute Bao Feng, have a strong impact on the accuracy of measurement of lactic acid ion.For improving accuracy, using standard addition method, result obtains
A certain degree of improvement, but substrate impact can not be completely eliminated, simultaneously because standard addition method is for twice weighing of sample
Required precision is the highest, and anthropic factor impact strengthens, additionally, ion chromatographic column, suppressor are all polluted by substrate, and service life
It is greatly shortened.
Summary of the invention
The present invention is directed to the above-mentioned problems in the prior art, utilize Vavle switching system, application reversed phase chromatography to disappear online
Except the interference of complex matrices, sample after eliminating substrate is collected, then uses the chromatography of ions to carry out ON-LINE SEPARATION detection, build
Stood a kind of can the chromatograph Vavle switching system of trace lactic acid ion in direct analysis complexity lobaplatin sample online, and apply this system
Directly measure the method for trace lactic acid ion in lobaplatin.Sample requirement is little, can purify sample by direct-on-line, eliminate complex matrices
Interference, efficiently solves the matrix interference problem that lactic acid ion runs in medicine lobaplatin that measures, reduce simultaneously ion chromatographic column and
The pollution effect of suppressor, increases the service life.
The present invention is achieved through the following technical solutions:
The system of trace lactic acid ion in a kind of post switching-ion-chromatographic determination lobaplatin, described system includes two gradients
Pump, injector, ten-way valve, quantitative loop, pretreatment column, infusion pump, six-way valve, collection ring, guard column, analytical column, suppressor and
Electric conductivity detector；Described injector and ten-way valve connect, and two gradient pumps are connected with ten-way valve, and pretreatment column two ends connect respectively
Connecing ten-way valve and six-way valve, infusion pump is connected with six-way valve, after six-way valve connects guard column, is sequentially connected with analytical column, suppressor
And electric conductivity detector.
The system of trace lactic acid ion in above-mentioned post switching-ion-chromatographic determination lobaplatin, it is preferred that pretreatment column is
IonPac NG1 chromatographic column；Guard column is IonPac AG19；Analytical column is IonPac AS19.
Present invention also offers a kind of method of trace lactic acid ion in post switching-ion-chromatographic determination lobaplatin, described
Method include the collection of baseline, elimination and the collection of lactic acid ion of interference substrate, the chromatographic isolation analysis of lactic acid ion and
The regeneration of pretreatment column, specifically comprises the following steps that
1) collection of baseline: connected by each parts in ion chromatographic column switched system, constitutes post switching-chromatography of ions system
System, is then continually entered pretreatment column by two gradient pumps by deionized water, is continually entered by leacheate by infusion pump
To guard column and analytical column, treat that it entirely reaches poised state, start to record background signal；
2) disturbing the elimination of substrate and the collection of lactic acid ion: testing sample injects ten-way valve by injector, quantitative loop controls
Sample capacity, after sample loads, ten-way valve and six-way valve are simultaneously switching to opposite location, two gradient pump conveying deionizations
Sample in quantitative loop is brought in pretreatment column by water, and low pole or non polar impurities are retained on pretreatment column, lactic acid ion
Enter and collect ring；
3) the chromatographic isolation analysis of lactic acid ion: sample introduction is after 3 minutes, the complete eluting of lactic acid ion on pretreatment column, leads to ten
Valve switches back into initial position together with six-way valve, and infusion pump conveying leacheate, to collecting ring, will collect the target cationic in ring
It is delivered to guard column, analytical column, suppressor and electric conductivity detector successively；
4) regeneration of pretreatment column: by inhaling on two gradient pump conveying mass fraction 50% acetonitrile solution eluting pretreatment columns
Attached impurity is to waste liquid, then spends from water balance pretreatment column, for sample introduction next time；
5) step 1)～4 is repeated), measure lactic acid ion standard solution and lobaplatin sample, draw lactic acid ion standard curve, then
The content of lactic acid ion in lobaplatin sample is calculated by standard curve.
Step 2) in, described quantitative loop controls sample capacity preferably 100 μ L.
Step 2) in, described collection ring 9 testing sample sample size preferably 2 ml；
In step 3), described leacheate is KOH-aqueous solution.
In step 3), in described leacheate, KOH concentration is: 0～18 min, 4 mmol/L；18～30 min, KOH
Concentration is increased to 30 mmol/L by 4 mmol/L gradients；30～35 min, 4 mmol/L.
In step 5), the compound method of described standard solution: weigh sodium lactate standard substance, it is configured to 1000 mg/L's
Lactic acid ion standard reserving solution, preserves, by standard reserving solution stepwise dilution to concentration 0.01～50 mg/L during use at 4 DEG C
Lactic acid ion standard solution.
In said determination method, the overall flow rate of two gradient pumps is 0.5 mL/min, and infusion pump flow velocity is 1.0 mL/min.
Described assay method, the inventive method detection sensitivity is 0.0005 mg L-1；Detection is limited to 0.0015mg L-1。
The system and method for trace lactic acid ion, its beneficial effect in post switching-ion-chromatographic determination lobaplatin of the present invention
As follows:
1) needed for, sample size is few.The online treatment technology that the present invention sets up realizes the elimination of interference impurity, sample introduction by chromatographic column
Amount only needs 100 μ L can complete the detection of the analysis to target compound, and general interference impurity elimination process is that off-line eliminates,
Required sample size is big, and the method that the present invention is set up is particularly well-suited to trace or the detection of valuable sample.
2) versatility is good.The method that the present invention is set up can be applied to a series of similar dry by the replacing of chromatographic column
Disturb problem, by online treatment technology, eliminate interfering material, extend the range of application of chromatography of ions.
3) simple to operate, it is easy to automatization.
4) method is simple, and instrument configuration requires low.During ion chromatography analysis organic acid, main employing AS11 chromatographic column,
Eluent concentration uses multistage change, and instrument configuration requires height.
5) present invention uses the low ICS-1500 ion chromatograph joined, this instrument and leacheate generator with the use of time,
Eluent concentration is only capable of doing single order change, and range of application is restricted.Selecting IonPac NG1 chromatographic column is pretreatment column, AS19
During anion analysis post, eluent concentration only needs to do single order change can complete dividing completely of target lactic acid ion and other ion
From.Meanwhile, this method eluent concentration ratio is relatively low, energy-conserving and environment-protective.
Accompanying drawing explanation
Fig. 1 is the system flow schematic diagram of trace lactic acid ion in embodiment 1 post switching-ion-chromatographic determination lobaplatin；
In figure: 1 and 2, gradient pump, 3, injector, 4, ten-way valve, 5, quantitative loop, 6, pretreatment column, 7, infusion pump, 8, six
Logical valve, 9, collect ring, 10, protect, 11, analytical column, 12, suppressor.13, electric conductivity detector；
Fig. 2 is Chromatographic Comparison's spectrogram of embodiment 2 deionized water, lactate standard product and actual sample lobaplatin, and wherein peak 1 is lactic acid
Ion.
Detailed description of the invention
The following stated is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Embodiment 1
The system (as shown in Figure 1) of trace lactic acid ion in a kind of post switching-ion-chromatographic determination lobaplatin, including two gradients
Pump 1,2, injector 3, ten-way valve 4, quantitative loop 5, pretreatment column 6, infusion pump 7, six-way valve 8, collect ring 9, guard column 10, analyze
Post 11, suppressor 12 and electric conductivity detector 13.Described injector 3 and ten-way valve 4 connect, two gradient pumps 1,2 and ten-way valve 4
Connecting, pretreatment column 6 two ends connect ten-way valve 4 and six-way valve 8 respectively, and infusion pump 7 is connected with six-way valve 8, and six-way valve 8 connects to be protected
After guard post 10, it is sequentially connected with analytical column 11, suppressor 12 and electric conductivity detector 13；
Wherein, pretreatment column 6 is IonPac NG1 chromatographic column；Guard column 10 is IonPac AG19；Analytical column 11 is IonPac
AS19；Ion chromatograph is ICS-1500 ion chromatograph.
Embodiment 2
Measure trace lactic acid ion in lobaplatin, specifically comprise the following steps that
1) solution preparation:
Standard solution: weigh sodium lactate standard substance, is configured to the lactic acid ion standard reserving solution of 1000 mg/L, protects at 4 DEG C
Depositing, by the lactic acid ion standard solution of standard reserving solution stepwise dilution to concentration 0.01～50 mg/L during use, concentration is successively
For；0.01,0.05,0.25,1,5,25,50 mg/L.
Sample solution: precision weighs lobaplatin sample 25 mg and is placed in 5 mL measuring bottles, with deionized water dissolving and be settled to carve
Degree.
2) collection of baseline: connected by each parts in ion chromatographic column switched system, constitutes post switching-chromatography of ions
System, during original state, ten-way valve 4 is in Inject position, and six-way valve 8 is in Load position；Then by gradient pump 1 and 2 will go from
Sub-water continually enters pretreatment column 6 according to overall flow rate 0.5 mL/min, by infusion pump 7 by KOH leacheate with flow velocity 1.0
ML/min continually enters to guard column 10 and analytical column 11, treats that it entirely reaches poised state, starts to record background signal；
3) elimination of substrate and the collection of lactic acid ion are disturbed: testing sample injects ten-way valve 4 by injector 3, and quantitative loop 5 is controlled
Sample capacity processed, after sample loads, ten-way valve 4 is switched to Load position (0-1.0 min), and six-way valve 8 is switched to Inject
Position (0-3.0 min), gradient pump 1 and 2 conveying deionized water the sample in quantitative loop 5 is brought in pretreatment column 6, low pole or
Non polar impurities is retained on pretreatment column 6, and lactic acid ion enters collects ring 9；
4) the chromatographic isolation analysis of lactic acid ion: after sample introduction 3min, the complete eluting of lactic acid ion on pretreatment column 6, leads to ten
Valve 4 switches back into initial position together with six-way valve 8, and infusion pump 7 carries leacheate (KOH, 0-18 min 4 mmol/L, 18-30
Min 4-30 mmol/L, 30-35 min 4 mmol/L) to collecting ring 9, the target cationic collected in ring 9 is carried successively
To guard column 10, analytical column 11, suppressor 12 and electric conductivity detector 13；
5) regeneration of pretreatment column: after sample introduction 3-8min, carries mass fraction 50% acetonitrile solution eluting by gradient pump 1 and 2
On pretreatment column 6, the impurity of absorption is to waste liquid, then with deionized water balance pretreatment column 6, for sample introduction next time；
6) step 2 is repeated)～4), measure lactic acid ion standard solution and lobaplatin sample, draw lactic acid ion standard curve, then
The content of lactic acid ion in lobaplatin sample is calculated by standard curve.(as shown in Figure 2)
Embodiment 2 applies trace lactic acid ion method performance parameter in post switching-ion-chromatographic determination lobaplatin to be shown in Table 1, detection
Interpretation of result is shown in Table 2.
Table 1
Table 2
Comparative example 1: trace lactic acid ion in ion-chromatographic determination lobaplatin, parameter is shown in Table 3.
Table 3
From embodiment and comparative example interpretation of result, this method and trace breast in prior art intermediate ion chromatography determination lobaplatin
Acid ion method is compared, and detection limit is low, and the range of linearity is wider.
Claims (10)
1. the system of trace lactic acid ion in a post switching-ion-chromatographic determination lobaplatin, it is characterised in that: described system
Including two gradient pumps, injector, ten-way valve, quantitative loop, pretreatment column, infusion pump, six-way valve, collection ring, guard column, analysis
Post, suppressor and electric conductivity detector；Described injector and ten-way valve connect, and two gradient pumps are connected with ten-way valve, pretreatment
Post two ends connect ten-way valve and six-way valve respectively, and infusion pump is connected with six-way valve, and six-way valve connects after guard column, are sequentially connected with point
Analysis post, suppressor and electric conductivity detector.
System the most according to claim 1, it is characterised in that: pretreatment column is IonPac NG1 chromatographic column；Guard column is
IonPac AG19；Analytical column is IonPac AS19.
3. a method for trace lactic acid ion in post switching-ion-chromatographic determination lobaplatin, described method includes baseline
Gather, disturb elimination and the collection of lactic acid ion, the chromatographic isolation analysis of lactic acid ion and the regeneration of pretreatment column of substrate, tool
Body step is as follows:
1) collection of baseline: connected by each parts in ion chromatographic column switched system, constitutes post switching-chromatography of ions system
System, is then continually entered pretreatment column by two gradient pumps by deionized water, is continually entered by leacheate by infusion pump
To guard column and analytical column, treat that it entirely reaches poised state, start to record background signal；
2) disturbing the elimination of substrate and the collection of lactic acid ion: testing sample injects ten-way valve by injector, quantitative loop controls
Sample capacity, after sample loads, ten-way valve and six-way valve are simultaneously switching to opposite location, two gradient pump conveying deionizations
Sample in quantitative loop is brought in pretreatment column by water, and low pole or non polar impurities are retained on pretreatment column, lactic acid ion
Enter and collect ring；
3) the chromatographic isolation analysis of lactic acid ion: sample introduction is after 3 minutes, the complete eluting of lactic acid ion on pretreatment column, leads to ten
Valve switches back into initial position together with six-way valve, and infusion pump conveying leacheate, to collecting ring, will collect the target cationic in ring
It is delivered to guard column, analytical column, suppressor and electric conductivity detector successively；
4) regeneration of pretreatment column: by inhaling on two gradient pump conveying mass fraction 50% acetonitrile solution eluting pretreatment columns
Attached impurity is to waste liquid, then spends from water balance pretreatment column, for sample introduction next time；
5) step 1)～4 is repeated), measure lactic acid ion standard solution and lobaplatin sample, draw lactic acid ion standard curve, then
The content of lactic acid ion in lobaplatin sample is calculated by standard curve.
Method the most according to claim 3, it is characterised in that: step 2) in, described quantitative loop controls sample capacity and is
100μL。
Method the most according to claim 3, it is characterised in that: step 2) in, described collection ring testing sample sample size
For 2ml.
Method the most according to claim 3, it is characterised in that: in step 3), described leacheate is KOH-aqueous solution.
Method the most according to claim 3, it is characterised in that: in step 3), in described leacheate, KOH concentration is: 0～
18 min, 4 mmol/L；18～30 min, 4～30 mmol/L；30～35 min, 4 mmol/L.
Method the most according to claim 3, it is characterised in that: in step 5), the compound method of described standard solution: claim
Extracting lactic acid sodium standard substance, are configured to the lactic acid ion standard reserving solution of 1000 mg/L, preserve at 4 DEG C, standard are stored up during use
Lactic acid ion standard solution for liquid stepwise dilution to concentration 0.01～50 mg/L.
Method the most according to claim 3, it is characterised in that: the overall flow rate of two gradient pumps is 0.5 mL/min, transfusion
Flow rate pump is 1.0 mL/min.
Method the most according to claim 3, it is characterised in that: detection sensitivity is 0.0005 mg L-1；Detection is limited to
0.0015mg L-1。
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| CN111721841A (en) * | 2019-03-19 | 2020-09-29 | 海南长安国际制药有限公司 | Detection of lobaplatin-related substances |
| CN111721840A (en) * | 2019-03-19 | 2020-09-29 | 海南长安国际制药有限公司 | Detection of related substances in lobaplatin |
| CN112946120A (en) * | 2021-02-01 | 2021-06-11 | 西安奕斯伟硅片技术有限公司 | Ion concentration detection equipment and method |
| CN115436555A (en) * | 2022-10-21 | 2022-12-06 | 广州谱临晟科技有限公司 | System and method for separating perchlorate or nitrite in food by using multidimensional liquid chromatography |
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| CN115436555A (en) * | 2022-10-21 | 2022-12-06 | 广州谱临晟科技有限公司 | System and method for separating perchlorate or nitrite in food by using multidimensional liquid chromatography |
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