CN104849358A - Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system - Google Patents
Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system Download PDFInfo
- Publication number
- CN104849358A CN104849358A CN201510008851.5A CN201510008851A CN104849358A CN 104849358 A CN104849358 A CN 104849358A CN 201510008851 A CN201510008851 A CN 201510008851A CN 104849358 A CN104849358 A CN 104849358A
- Authority
- CN
- China
- Prior art keywords
- optimization
- concentration
- acid
- carry out
- hydrochloric acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses an optimization method for the parameters of an analytical instrument used for analysis of the forms of elemental arsenic based on an HPLC-HG-AFS system. The method comprises the following steps: optimization of high performance liquid chromatographic separation conditions; optimization of hydride generation conditions; and optimization of the parameters of an atomic fluorescence detector. Under the above-mentioned optimized conditions, three forms of arsenide, i.e., 4-aminobenzenearsonic acid, 4-hydroxy-3-nitrophenylarsonic acid and 4-nitrophenylarsonic acid respectively with a concentration of 0.1 to 2 [mu]g/mL are separately used for mixed-standard calculation of standard curves of the HPLC-HG-AFS system. Compared with the prior art, the method provided by the invention has good repeatability and high accuracy under the optimized conditions.
Description
Technical field
The present invention relates to a kind of analysis and testing technology of element organic form, particularly relate to arsenic element morphological analysis method.
Background technology
Chemical form Epidemiological Analysis is carried out to the different compounds of various element, for the toxicity understanding element in environment, geology and biological sample, metabolism and to transport performance be very important, therefore, morphological analysis in recent years more and more becomes an active research field.Arsenic is the indispensable element of human body, and is the basic constitution element of compound in many environment, biological sample, but for the arsenide of different shape, its toxicity but differs greatly.Generally, the toxicity of inorganic arsenic chemicals is greater than organoarsenium compound.Its toxicity of arsenide arsenide of different valence state is also different, and such as, it is reported, arsenious acid is soluble in water, and its toxicity will much larger than general arsenide.
In general, the toxicity of arsenide meets following order (being up to minimum toxicity): arsine > inorganic arsenite > organic trivalent arsenide compound (arsenooxides) > inorganic arsenic > organic pentavalent arsenic compound > arsonium compound > element arsenic [89-91].Arsenic betaine and arsenocholine are considered to nontoxic.In general, the toxicity of arsenic, normally with concentration dependent, under the dosage of correlation standard, can not affect the health of animal or human body.But due to genetoxic and the carcinogenicity of arsenic, international cancer research institution (IARC) mechanism is classified as material risk element, it is reported, the whole world has nearly 60-100 ten thousand people being exposed in the risk environment of Excessive Arsenic for a long time.
Part organo-arsenic is also the adjuvant that veterinary drug often uses, such as, arsanilic acid (the p-atoxylate of formal name used at school, 4-Aminobenzenearsonic acid, ASA), Nitarsone (4-nitrophenylarsonic acid, NIT), roxarsone (4-hydroxy-3-nitrophenylarsonic acid, ROX) are as veterinary drug, coccidia Enterozoa can be prevented and treated, so widely used [92] by livestock culture industry.Arsanilic acid and roxarsone for preventing and treating poultry coccidial parasites and livestock hemorrhagic enteritis, Nitarsone as veterinary drug for preventing and treating the histomoniasis (plague) caused by primary structure trichomonad.As feed addictive, the use of these organic arsenic preparations can cause arsenic element accumulation in animal body, the excreta of annual domestic animal is used for agricultural fertilizer, especially Rice Cropping, can cause Arsenic in Soil too high levels, the arsenic element in plant absorption soil, will enter human foods chain, there is report to query the actual effect of these organic arsenic preparations, and point out that these organic arsenic preparations can cause accumulation in mankind's body.In addition, a lot of report is pointed out, uses if this type of organic arsenic preparation exceeds standard, and some factors can cause it to be decomposed into forbid the poisonous arsenide of use.What is worse, have and report that ROX finally can be decomposed into the electrodeless arsenic of water-soluble hypertoxicity.
In order to ensure animal food safety, avoiding the abuse of this type of organic arsenic preparation, being necessary strictly to detect arsenic morphology in animal tissue, and for the effective extraction of organo-arsenic compound in animal derived food and detection method.
Summary of the invention
In order to overcome above-mentioned prior art, the present invention proposes a kind of arsenic element morphological analysis instrument parameter optimization method, achieve the instrument parameter optimization of HPLC-HG-AFS method for organo-arsenic morphological analysis method, the final typical curve that have detected three kinds of organic arsenic preparations under optimal conditions, verifies the method.
Apply three kinds of form arsenides such as high performance liquid chromatography-hydride generation-atomic fluorescence spectrometer Ponazuril, Xiao's phenylarsonic acid and roxarsone and carry out being separated the method detected, and instrument terms and conditions is optimized.
The present invention proposes a kind of arsenic element morphological analysis instrument parameter optimization method based on HPLC-HG-AFS system, the method comprises the following steps:
5% methanol aqueous solution (containing 0.1% (v/v) trifluoroacetic acid, 0.05mol L-1 potassium dihydrogen phosphate) is adopted as mobile phase, in mobile phase, to add trifluoroacetic acid (TFA) adjusted to ph, add KH
2pO
4make buffer salt, carry out the optimization of high performance liquid chromatography separation condition in fact;
Carry out the optimization of hydride occurrence condition, specifically comprise:
Select hydrochloric acid and KBH
4peristaltic pump rotating speed is 120r min
-1, carry out peristaltic pump flowing rate, be under the condition of 5% (v/v) at concentration of hydrochloric acid, reductive agent KBH
4concentration elect 20g L as
-1, carry out reductant concentration optimization, adopt containing 5g L at reductive agent
-1the 20g L of KOH
-1kBH
4under the condition of solution, the concentration of hydrochloric acid elects 5% as, carries out concentration of hydrochloric acid optimization;
Carry out atomic fluorescence detector parameter optimization, specifically comprise:
Selected 330v is normalization standard value, carries out photomultiplier high pressure; Adopt 100mA hollow cathode lamp current, carry out the optimization of hollow cathode lamp current; Selected 300mL min
-1for the optimal value of flow rate of carrier gas, carry out flow rate of carrier gas optimization; Select minimum shield gas flow amount, reach best shielding action, carry out shield gas flow speed and optimize
Under the condition of above-mentioned optimization, choose 0.1 ~ 2ug mL respectively
-1arsanilic acid, roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides mix the typical curve that mark calculates HPLC-HG-AFS system.
Compared with prior art, under optimal conditions, the method is reproducible in the present invention, and accuracy is high.
Accompanying drawing explanation
Fig. 1 is experimental system structural representation of the present invention;
Fig. 2 is obtained spectrogram by be respectively the different condition of 95:5,90:10,80:20 tri-kinds in TFA/ methyl alcohol volume ratio under;
A (), employing different proportion TFA/ methyl alcohol detect spectrogram as the arsenic morphology of mobile phase; Wherein detect sample and adopt the mixed mark of 1.0ug mL-1 arsenic; (1) ASA; (2) NIT; (3) ROX.;
The relative signal intensity of three kinds of arsenides in (b), different mobile phase:
Mobile phase A: 95:5 v/v TFA/methanol pH=2.43; Mobile phase B: 90:10 v/v TFA/methanol pH=2.52; Mobile phase C:80:20 v/v TFA/methanol pH=2.72, other parameters are in table 2;
Fig. 3 is that reductive agent flow velocity affects schematic diagram for experimental result;
Fig. 4 is KBH
4the atomic fluorescence testing result of concentration optimization experiment, horizontal ordinate is KBH
4concentration, ordinate is spectrum peak area; Adopt 5% (v/v) hydrochloric acid, duplicate detection three times;
Fig. 5 is the atomic fluorescence testing result of concentration of hydrochloric acid Optimal Experimental, and horizontal ordinate is concentration of hydrochloric acid, and ordinate is spectrum peak area.Adopt the 20%KBH4 solution containing 5%KOH, duplicate detection three times;
Fig. 6 is the signal to noise ratio (S/N ratio) schematic diagram of signal under different photomultiplier negative high voltage;
Fig. 7 is the signal to noise ratio (S/N ratio) schematic diagram of detection signal under different hollow cathode lamp-current condition;
Fig. 8 is that flow rate of carrier gas affects schematic diagram for testing result;
Fig. 9 is the impact of shielding gas for testing result;
Figure 10 concentration range 0.1 ~ 2ug mL
-1arsenic standard solution examination criteria spectrogram; Testing conditions is as table 2;
Figure 11 concentration range 0.1 ~ 2ug mL
-1arsenic standard solution examination criteria curve map; Testing conditions is in table 2.Standard equation and related coefficient are in table 3; %RSD, n=3.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in detail, but practical range of the present invention is not limited thereto.
One, experimental apparatus used in the present invention and reagent
1, experimental apparatus
Test instrument used to mainly contain: AFS9600 non-dispersive hydride generation-atomic fluorescence spectrometric instrument (HG-AFS) (Beijing Haiguang Instrument Co., Ltd.), HPLC P600 efficient liquid-phase chromatographic pump (Labtech company of the U.S.), be equipped with 7725i six path sampling valve (U.S. Rheodyne, Cotati company), Phenomenon C18 chromatographic column (Luna5u, 250 × 4.60mm).HPLC-UV-HG-AFS system construction drawing as shown in Figure 1-2.
2, experiment reagent
It is pure that all reagent used in experiment are analysis.Test the reagent used: NaOH (Tianjin Feng Chuan chemical reagent company limited), KBH
4(Tianjin chemical reagent research institute), KH
2pO
4(chemical reagent company limited of Chinese Medicine group), hydrochloric acid (Duksan Chun Yao company of Korea S), methyl alcohol (Dikma Science and Technology Ltd. of the U.S.), trifluoroacetic acid (Tianjin Aifa Aesar Reagent Company), silica sand (Shanghai Tian Lian chemical reagent company limited), arsanilic acid (German Dr.Ehronstorfer GmbH laboratory, purity 99.0%), roxarsone (German Dr.Ehronstorfer GmbH laboratory, purity 99.0%), Nitarsone (Japanese Wako Wako Pure Chemical Industries, Ltd.), ultrapure water instrument (Millipore company limited of the U.S.).
The collocation method of Standard Reserving Solution: accurately take arsanilic acid standard items 50mg, be placed in 50mL volumetric flask, add deionized water and be settled to 50mL, ultrasonic water bath 30min, arsanilic acid standard items are fully dissolved, then crosses 0.45um membrane filtration, be namely configured to 1000ug mL
-1arsanilic acid Standard Reserving Solution, is placed in brown bottle, is put in 4 DEG C of preservations in refrigerator.The configuration of roxarsone and Xiao's arsenobenzene acid standard reserving solution also adopts same step.
Three kinds of form arsenide mixed mark solution matching while using, compound method: respectively get the above-mentioned arsanilic acid of 500uL, roxarsone and Xiao's arsenobenzene acid Standard Reserving Solution in 50mL volumetric flask, add deionized water and be settled to 50mL, be namely configured to 10ug mL
-1arsanilic acid, roxarsone and Xiao's benzene arsenic acid three kinds of arsenide mixed standard solutions.Get this arsenide mixed mark 200uL and be placed in 2mL color comparison tube, add deionized water and be settled to 2mL, be namely configured to 1ug mL
-1three kinds of mixed marks of arsenide.Adopt identical method, configuration 0.1ug mL
-1, 0.4ug mL
-1, 1ug mL
-1, 2ug mL
-1the mixed mark of three kinds of different shape arsenides of four kinds of variable concentrations.
Table 1 arsanilic acid, roxarsone and Xiao's benzene arsenic acid mix target compound method
Mobile phase matching while using, adopt 5% methanol aqueous solution (containing 0.1% (v/v) trifluoroacetic acid, 0.05mol L
-1potassium dihydrogen phosphate).Collocation method: accurately take 3.402gKH2PO4 in 500mL deionized water, sonic oscillation 30min hydrotropy, crosses 0.45um membrane filtration, then add 0.5mL trifluoroacetic acid, after mixing, get above-mentioned solution 475mL, mix with the pure methyl alcohol of 25mL, ultrasonic degas 30min, for subsequent use.
The collocation method of hydride generating system reaction dissolvent:
Current-carrying: adopt 5% hydrochloric acid.Standard but measures 25mL pure hydrochloric acid (electronic pure, extremely low concentration hydrogenation element), mixes rear stand-by with 475mL deionized water.
Reductive agent: adopt the 20g L-1KBH4 solution containing 5g L-1KOH.Compound method: accurately take 2.5gKOH (top grade is pure) and be dissolved in 500mL deionized water, gradation slowly adds 10gKBH until completely dissolved
4(top grade is pure), stand-by after dissolving completely.
Two, the optimization of experiment condition
1, the optimization of high performance liquid chromatography separation condition
Application ASA, NIT, ROX tri-kinds of arsenide standard solution optimize HPLC condition.Here C18 Reversed Phase High Performance has been selected.The present invention finds, just in adjustment mobile phase the ratio of first alcohol and water for improving peak shape and separating effect effect is little, so, have studied and add trifluoroacetic acid (TFA) adjusted to ph in mobile phase, add KH
2pO
4make the method for buffer salt.When the volumetric concentration of trifluoroacetic acid in mobile phase (v/v) is 0.1%, resolution and signal to noise ratio (S/N ratio) are all relatively good, and 50mM KH
2pO
4peak shape can be helped improve as buffer salt.As Fig. 2, compare and obtain spectrogram under TFA/ methyl alcohol volume ratio is respectively the different condition of 95:5,90:10,80:20 tri-kinds.
As seen from the figure, when methanol concentration is higher, owing to rinsing excessive velocities, it is very fast that each one-tenth separates peak, and retention time is shorter, but as shown in Fig. 2 (b), through measuring AFS signal spectrum peak area, go out peak mobile phase faster, its detection sensitivity is lower; And 5% (v/v) methanol aqueous solution is as mobile phase, separating effect is better, and its detection sensitivity is also higher, so, adopt 5% methanol aqueous solution (containing 0.1% (v/v) trifluoroacetic acid, 0.05mol L
-1potassium dihydrogen phosphate) as mobile phase.
2, hydride occurrence condition is optimized
The flow velocity of hydrochloric acid and reductive agent and concentration directly affect hydrogenation efficiency, by adopting univariate method, and each one of them variable of change, the optimal combination of research hydrochloric acid and reductive agent.
(1) peristaltic pump flowing rate
Hydrochloric acid and reductive agent are driven by peristaltic pump, and under the condition that pump line is determined, the rotating speed of peristaltic pump determines the flow velocity of various reagent.First the impact for experimental result of hydrochloric acid and reductive agent different in flow rate is have studied in experiment.
Hydrochloric acid, as the current-carrying of HPLC effluent and reaction reagent, first determines that the peristaltic pump rotating speed of hydrochloric acid is 120r min
-1, study the impact of different reductive agent flow velocity for experimental result.Change KBH
4peristaltic pump rotating speed by 70r min
-1gradient increases, and composes the testing result of peak area as shown in Figure 3 under different reductive agent flow velocity.
As seen from Figure 3, KBH is worked as
4when flow velocity is less, because reductive agent is not enough, cause tested hydrogenation element hydrogenation incomplete, so detection signal strength is not high, detection sensitivity is lower.Continuous increase reductive agent flow velocity, detects spectral strength and increases gradually, and spectrum peak area increases, and sensitivity increases.But along with the increase of reductive agent flow velocity, hydrogenation is further strong, when peristaltic pump rotating speed is more than 120r min
-1after, owing to reacting violent, occur foam in gas-liquid separator, detection signal noise increases, and signal to noise ratio (S/N ratio) declines.When peristaltic pump rotating speed is 120r min
-1time, peak value is the highest, and sensitivity is higher, now hydrogenation moderate strength, and signal noise is not high, so be 120r min at current-carrying peristaltic pump rotating speed
-1time, selected KBH
4peristaltic pump rotating speed is 120r min
-1.
For the determination of hydrochloric acid flow velocity, in experiment, determine that KBH4 peristaltic pump rotating speed is 120r min
-1, change hydrochloric acid peristaltic pump rotating speed, find when hydrochloric acid flow velocity is less, spectral strength is very low, and sensitivity is poor, mainly because hydrogenation will in acid condition, and now KBH
4flow velocity is comparatively large, causes concentration of hydrochloric acid in reaction block very little, so reaction not exclusively, measured object detected value is less.Along with the increase of concentration of hydrochloric acid, spectral strength increases, but meanwhile hydrogenation is further violent, peristaltic pump rotating speed 120r min
-1after, occur foam in gas-liquid separator, detection signal noise increases, and peak shape becomes very poor, and signal to noise ratio (S/N ratio) is very low.At hydrochloric acid peristaltic pump rotating speed 120r min
-1before and afterwards, spectrum peak area all reduces, so selected hydrochloric acid and KBH
4peristaltic pump rotating speed is 120r min
-1.
(2) reductant concentration optimization
Selected salts acid concentration 5%, changes KBH
4concentration, determine KBH by series of experiments
4optium concentration.
Detect sample and all adopt concentration 1ug mL
-1arsanilic acid, the mixed mark of roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides.We change KBH respectively
4concentration be 10g L
-1, 15g L
-1, 20g L
-1, 25g L
-1with 30g L
-1, utilize atomic fluorescence spectrometer to detect the result of spectrum peak area as shown in Figure 4, find in experiment, under concentration of hydrochloric acid is 5% condition, when just starting, along with KBH
4the increase of concentration, the sensitivity of each composition is all risen, and is 20g L to concentration
-1time reach maximum, subsequently along with KBH
4the increase of concentration, the detection sensitivity of three kinds of compositions starts to decline, and its reason may be along with KBH
4the increase of concentration, hydrogenation becomes more and more violent, and the hydrogen of generation gets more and more, the air-flow velocity sending into AFS detecting device after gas-liquid separator separates is excessive, and ratio shared by sample gas is more and more less, detection is affected, result in the decline of detection sensitivity.
As seen from Figure 4, under the condition of employing 5% (v/v) hydrochloric acid, work as KBH
4concentration be 20g L
-1time, the detection sensitivity of analyte is the highest, KBH
4concentration too high or too muting sensitivity all reduce.So, be under the condition of 5% (v/v) at concentration of hydrochloric acid, reductive agent KBH
4concentration elect 20g L as
-1.
(3) concentration of hydrochloric acid optimization
Reductive agent adopts containing 5g L
-1the 20g L of KOH
-1kBH
4solution, is changed the concentration of hydrochloric acid, is determined the optium concentration of hydrochloric acid by series of experiments.
Detect sample and all adopt concentration 1ug mL
-1arsanilic acid, the mixed mark of roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides.The concentration that we change hydrochloric acid is respectively 2%, 5%, 8% and 10%, utilizes atomic fluorescence spectrometer to detect the result of spectrum peak area as shown in Figure 5.
Find in experiment, adopt containing 5g L at reductive agent
-1the 20g L of KOH
-1kBH
4under solution condition, along with concentration of hydrochloric acid increases by 2%, the sensitivity of each composition is all risen, when concentration is 5%, reach maximum, subsequently along with the increase of concentration of hydrochloric acid, experiment finds that reaction becomes more and more violent, and generate hydrogen and get more and more and create a lot of foam, reaction is continued until in gas-liquid separator.Detected by atomic fluorescence and find, more than 5%, the detection sensitivity of later three kinds of compositions starts to decline concentration of hydrochloric acid, its reason may be that hydrogenation is violent, the hydrogen produced gets more and more, finally cause salt excessive acid, the hydrogenation and analyte has no chance to occur completely, and make to enter AFS and be detected sample and incur loss.In figure, when concentration of hydrochloric acid reaches 10%, the sensitivity of roxarsone continues to reduce, but arsanilic acid and Xiao's benzene arsenic acid rise on the contrary to some extent, and its reason may be that now hydrogenation is more violent, the gas flow rate entering AFS detection is very large, causing the vibration of flame of atomic fluorescence detector Atom device severe especially, making spectrogram noise very large, when calculating spectrum peak area, cause very big error, so now the deviation of three experimental results is also larger.
In hydrogenation, reductive agent should be excessive, to ensure that hydrogenation element farthest hydrogenation occurs.In experimenting, we find out that, adopts containing 5g L at reductive agent
-1the 20g L of KOH
-1kBH
4under the condition of solution, select 5% hydrochloric acid time detection sensitivity maximum, concentration of hydrochloric acid is too high or too low all makes sensitivity reduce, so, reductive agent adopt contain 5g L
-1the 20g L of KOH
-1kBH
4under the condition of solution, the concentration of hydrochloric acid elects 5% as.
3, atomic fluorescence detector parameter optimization
In atomic fluorescence detector, Photomultiplier tube voltage, hollow cathode lamp current, carrier gas and shield gas flow amount all can have influence on final detection sensitivity, we pass through the series of experiment research optimal conditions of atomic fluorescence detector major parameter, detect sample and are concentration 1ug mL
-1arsanilic acid, the mixed mark of roxarsone and Xiao's benzene arsenic acid three kinds of arsenides.In the optimal value process of assessment parameters, the signal to noise ratio (S/N ratio) under adopting normalized method to evaluate various Parameter Conditions, what signal to noise ratio (S/N ratio) was higher is its optimal value.
(1) photomultiplier high pressure
Photomultiplier mainly amplifies the fluorescence signal detected, the size being added to the negative high voltage above it determines its enlargement factor.In experiment, select six Grad such as 290v, 300v, 310v, 320v, 330v and 340v respectively, selected 330v is normalization standard value, and under each parameter, the signal to noise ratio (S/N ratio) of detection signal as shown in Figure 6.
As seen from Figure 6, when photomultiplier negative high voltage is less, fluorescence signal is more weak, and spectral strength is lower, and partial spectrum peak is even submerged in noise signal, and now, signal to noise ratio (S/N ratio) is lower; Along with the increase of negative high voltage, the enlargement factor of fluorescence signal increases, and the intensity of noise signal also increases, but relative fluorescence signal, its degree increased is less, so signal to noise ratio (S/N ratio) promotes very fast, is that the spectral strength of the arsenide of various form increases from spectrogram.But, when negative high voltage constantly increases, although spectral strength increases always, but the intensity of code book floors, various noise signal also increases always, to 330v, although continue to increase negative high voltage, fluorescence signal intensity continues to increase, but signal to noise ratio (S/N ratio) but starts to decline, because the degree that now noise signal increases becomes increasing, so, we select 330v to be the optimal value of photomultiplier negative high voltage, because now signal to noise ratio (S/N ratio) is maximum, signal intensity is higher, and background values is also lower.
(2) hollow cathode lamp current is optimized
Hollow cathode lamp is used for fluorescence excitation signal, tested Elements Atom absorbs the energy of hollow cathode lamp radiation, transit to higher energy level, when getting back to ground state, just fluorescence signal is sent, so the size of hollow cathode lamp radiation intensity can have influence on the atomic quantity of radiation-absorbing energy, then has influence on the intensity of the fluorescence signal of radiation.Hollow cathode lamp adopts boundling pulsed to power, and the size of pulse current determines the height of emittance.
In experiment, constantly strengthen the lamp current of hollow cathode lamp, increase from 80mA gradient, using the signal to noise ratio (S/N ratio) of detection signal during 100mA as normalization standard value, gained testing result as shown in Figure 7.
As can be seen from the figure, when hollow cathode lamp current is less, the fluorescence signal excited is more weak, and spectral strength is lower, and partial spectrum peak is even submerged in noise signal, and now, signal to noise ratio (S/N ratio) is lower; Along with the increase gradually of hollow cathode lamp current, the intensity of detection signal also increases gradually, when increasing to 100mA, signal to noise ratio (S/N ratio) reaches maximum, then along with the increase of lamp current, signal to noise ratio (S/N ratio) starts to decline on the contrary, its reason may be that lamp current there occurs self-priming after 100mA, thus fluorescence signal is weakened on the contrary, and lamp current is larger, spontaneous imbibition phenomena is more serious, and it is more weak to compose peak-to-peak signal; In addition, excessive lamp current may affect the serviceable life of hollow cathode lamp, even causes damage.So, adopt 100mA hollow cathode lamp current.
(3) flow rate of carrier gas optimization
The Main Function one of carrier gas purges the tested element hydrogenation gas produced in course of reaction, is sent in atomizer and realized after atomization detecting; Two is in atomizer, form argon-hydrogen flame with reacting together with the hydrogen that generates, makes tested element hydrogen compound formation atomic vapour.So carrier gas can have influence on the stability of atomization efficiency and argon-hydrogen flame, then have influence on the size of signal to noise ratio (S/N ratio).
In experiment, make flow velocity from 200mL min
-1continuous increase, composes the size of peak area under more different flow rate of carrier gas, i.e. the size of detection sensitivity, and acquired results as shown in Figure 8.
As can be seen from figure, we, compose peak area less, because when flow velocity is less time at the beginning, argon purge scarce capacity, it is excessively slow that speed is swept in measured object hydrogenation air-blowing, makes spectrum conditions of streaking more serious, there is very big error in the calculating of spectrum peak area, and spectral strength is lower, and signal to noise ratio (S/N ratio) is lower; And when argon flow amount is not enough, the hydrogen flowing quantity that reaction generates is unstable, make argon-hydrogen flame unstable, cause flame to shake, thus detection signal is shaken, noise increases, so signal to noise ratio (S/N ratio) is lower.Continuous increase carrier gas flux, after flow reaches 300mLmin-1, along with the increase of carrier gas flux, the speed that purges increases, although spectrum peak conditions of streaking is improved, but spectral strength but reduces more, this is owing to purging excessive velocities, makes tested element hydrogen compound fail complete atomization and is namely discharged, so spectrum peak area starts to reduce, flow rate of carrier gas is faster, and peak area is less, and this is due to carrier gas purge excessive velocities.As can be seen from Figure, when flow rate of carrier gas is 300mL min
-1time, spectrum peak area is maximum, and the speed that now purges is moderate, and argon-hydrogen flame is also more stable, and noise is less, so selected 300mLmin
-1for the optimal value of flow rate of carrier gas.
(4) shield gas flow speed is optimized
Shielding gas is mainly used for protecting atomization steam not oxidized in atomizer; when analyzed element hydrogen compound is oxidized; then not easily absorb the emittance of hollow cathode lamp; fluorescence signal is weakened; sensitivity decrease, and the too high shielding gas of flow velocity can not play more effect, so the flow of shielding device is selected mainly to consider from economic aspect; select minimum shield gas flow amount, reach best shielding action.
As shown in Figure 9, make shield gas flow speed from 700mL min
-1continuous increase, composes the size of peak area, i.e. the size of detection sensitivity under more different shield gas flow speed.
As can be seen from Figure, when shield gas flow speed is less, to detect the spectrum peak area obtained less, and noise signal is comparatively strong, illustrates that the oxygen in air and the gas after parts atomic there occurs oxidation reaction, excited fluorescence radiation deficiency.Continuous increase shield gas flow speed, spectrum peak area increases gradually, and noise signal strength reduces, when flow velocity reaches 1000mL min
-1time, spectrum peak area reaches maximum, and shield gas flow amount is at 900mL min
-1time and 1000mLmin
-1spectrum peak area under condition is more or less the same, and considers from economic aspect, selects 900mL min
-1for shield gas flow amount.
Finally, determine that the instrument parameter of HPLC-HG-AFS morphological analysis combined system is as shown in table 2.
Table 2 HPLC-HG-AFS systematic parameter condition
Under the condition of above-mentioned optimization, choose 0.1 ~ 2ug mL respectively
-1arsanilic acid, roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides mix the typical curve that mark calculates HPLC-HG-AFS system.Standard spectrogram and typical curve are respectively as shown in Figure 10 and Figure 11.As shown in table 3, the related coefficient of typical curve is all better than 0.9992.
The typical curve of table 3 ASA, NIT and ROX, the range of linearity and detectability
4.4.2HPLC-AFS the detectability of method
The computing method of the detectability of HPLC-HG-AFS morphological analysis system are shown in formula (3-1), detect 3 times of slopes divided by often kind of arsenide typical curve of the value of three times with zero standard solution.According to the measurement result of table 3, through calculating, the detectability of arsanilic acid, Xiao's benzene arsenic acid and roxarsone three kinds of form arsenides is respectively 0.24ngmL
-1, 0.74ng mL
-1with 0.41ng mL
-1.This detectability is better than the HPLC method adopting UV-detector.
Adopt concentration 1ug mL
-1arsanilic acid, roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides as measured object, continuous coverage 6 times under condition shown in table 2, calculate often kind of arsenide measured value, mean value and relative standard deviation, measurement result is as shown in table 4.
Table 4 1ug mL-1 tri-kinds of form arsenides mix mark 6 measurement results
Visible, adopt HPLC-HG-AFS method to detect these three kinds of different shape arsenide same samples 6 times, the relative standard deviation of its detected value all controls within 5%, illustrates that this method is reliable, and repeatability better.
Although invention has been described for composition graphs above; but the present invention is not limited to above-mentioned embodiment; above-mentioned embodiment is only schematic; instead of it is restrictive; those of ordinary skill in the art is under enlightenment of the present invention; when not departing from present inventive concept, can also make a lot of distortion, these all belong within protection of the present invention.
Claims (1)
1., based on an arsenic element morphological analysis instrument parameter optimization method for HPLC-HG-AFS system, it is characterized in that, the method comprises the following steps:
Adopt containing 0.1%v/v trifluoroacetic acid, 0.05mol L
-15% methanol aqueous solution of potassium dihydrogen phosphate, as mobile phase, adds trifluoroacetic acid adjusted to ph, adds KH in mobile phase
2pO
4make buffer salt, carry out the optimization of high performance liquid chromatography separation condition in fact;
Carry out the optimization of hydride occurrence condition, specifically comprise:
Select hydrochloric acid and KBH
4peristaltic pump rotating speed is 120r min
-1, carry out peristaltic pump flowing rate, be that under the condition of 5%v/v, the concentration of reductive agent KBH4 elects 20g L as at concentration of hydrochloric acid
-1, carry out reductant concentration optimization, adopt containing 5g L at reductive agent
-1the 20g L of KOH
-1kBH
4under the condition of solution, the concentration of hydrochloric acid elects 5% as, carries out concentration of hydrochloric acid optimization;
Carry out atomic fluorescence detector parameter optimization, specifically comprise:
Selected 330v is normalization standard value, carries out photomultiplier high pressure; Adopt 100mA hollow cathode lamp current, carry out the optimization of hollow cathode lamp current; Selected 300mL min
-1for the optimal value of flow rate of carrier gas, carry out flow rate of carrier gas optimization; Select minimum shield gas flow amount, reach best shielding action, carry out shield gas flow speed and optimize;
Under the condition of above-mentioned optimization, choose 0.1 ~ 2ug mL respectively
-1arsanilic acid, roxarsone and Xiao's benzene arsenic acid three kinds of form arsenides mix the typical curve that mark calculates HPLC-HG-AFS system.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510008851.5A CN104849358A (en) | 2015-01-08 | 2015-01-08 | Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510008851.5A CN104849358A (en) | 2015-01-08 | 2015-01-08 | Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104849358A true CN104849358A (en) | 2015-08-19 |
Family
ID=53849158
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510008851.5A Pending CN104849358A (en) | 2015-01-08 | 2015-01-08 | Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104849358A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548429A (en) * | 2015-12-04 | 2016-05-04 | 通威股份有限公司 | Method for simultaneously detecting organic arsenic and inorganic arsenic in aquatic feed |
| CN106323932A (en) * | 2016-10-26 | 2017-01-11 | 武汉大学 | Method for rapidly detecting arsanilic acid and roxarsone in water sample |
| CN108489955A (en) * | 2018-05-18 | 2018-09-04 | 江苏省农业科学院 | A kind of non-chromatographic detection method detecting four kinds of arsenic morphologies in rice flour based on HG-AFS |
| CN109557232A (en) * | 2019-01-25 | 2019-04-02 | 黑龙江省兽药饲料监察所 | The detection method of arsanilic acid, nitre arsenobenzene acid and roxarsone in a kind of egg |
| CN109580863A (en) * | 2019-01-25 | 2019-04-05 | 黑龙江省兽药饲料监察所 | The detection method of arsanilic acid, nitre arsenobenzene acid and roxarsone in a kind of feed |
| CN110687241A (en) * | 2019-11-06 | 2020-01-14 | 天津海关动植物与食品检测中心 | Method for detecting roxarsone and 8 metabolites thereof in animal body |
-
2015
- 2015-01-08 CN CN201510008851.5A patent/CN104849358A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548429A (en) * | 2015-12-04 | 2016-05-04 | 通威股份有限公司 | Method for simultaneously detecting organic arsenic and inorganic arsenic in aquatic feed |
| CN106323932A (en) * | 2016-10-26 | 2017-01-11 | 武汉大学 | Method for rapidly detecting arsanilic acid and roxarsone in water sample |
| CN106323932B (en) * | 2016-10-26 | 2018-10-26 | 武汉大学 | A kind of method of arsanilic acid and roxarsone in quick detection water sample |
| CN108489955A (en) * | 2018-05-18 | 2018-09-04 | 江苏省农业科学院 | A kind of non-chromatographic detection method detecting four kinds of arsenic morphologies in rice flour based on HG-AFS |
| CN108489955B (en) * | 2018-05-18 | 2020-11-06 | 江苏省农业科学院 | Non-chromatographic detection method for detecting four arsenic forms in rice flour based on HG-AFS |
| CN109557232A (en) * | 2019-01-25 | 2019-04-02 | 黑龙江省兽药饲料监察所 | The detection method of arsanilic acid, nitre arsenobenzene acid and roxarsone in a kind of egg |
| CN109580863A (en) * | 2019-01-25 | 2019-04-05 | 黑龙江省兽药饲料监察所 | The detection method of arsanilic acid, nitre arsenobenzene acid and roxarsone in a kind of feed |
| CN109557232B (en) * | 2019-01-25 | 2020-10-16 | 黑龙江省兽药饲料监察所 | Method for detecting arsinic acid, nitrendic acid and roxarsone in eggs |
| CN110687241A (en) * | 2019-11-06 | 2020-01-14 | 天津海关动植物与食品检测中心 | Method for detecting roxarsone and 8 metabolites thereof in animal body |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104849358A (en) | Optimization method for parameters of analytical instrument used for analysis of forms of elemental arsenic based on HPLC-HG-AFS system | |
| Dimitrova et al. | Flow injection on-line preconcentration of palladium by ion-pair adsorption in a knotted reactor coupled with electrothermal atomic absorption spectrometry | |
| Xu et al. | Metal–organic frameworks of zeolitic imidazolate framework-7 and zeolitic imidazolate framework-60 for fast mercury and methylmercury speciation analysis | |
| CN103196880B (en) | Method for determining content of arsenic in iron ore by using hydride generation-atomic fluorescence spectroscopy | |
| Jiménez et al. | Performance of different preconcentration columns used in sequential injection analysis and inductively coupled plasma-mass spectrometry for multielemental determination in seawater | |
| CN100498287C (en) | Nano argentum spectrophotometry for detecting hydroxy free radical | |
| Sun et al. | Determination of boron in serum, plasma and urine by inductively coupled plasma mass spectrometry (ICP-MS). Use of mannitol-ammonia as diluent and for eliminating memory effect | |
| Carvalho et al. | Multi-element determination of copper, iron, nickel, manganese, lead and zinc in environmental water samples by ICP OES after solid phase extraction with a C18 cartridge loaded with 1-(2-pyridylazo)-2-naphthol | |
| Segura et al. | Evaluation of atomic fluorescence and atomic absorption spectrometric techniques for the determination of arsenic in wine and beer by direct hydride generation sample introduction | |
| CN110132709A (en) | Arsenic, mercury content method in micro-wave digestion-Atomic fluorophotometry measurement agricultural product | |
| CN106248609B (en) | A kind of method that ultraviolet specrophotometer measures hexafluorophosphoric acid lithium content in lithium-ion battery electrolytes | |
| CN103543133A (en) | Method for determining content of bismuth in iron ores by hydride generation-atomic fluorescence spectrometry method | |
| Su | Direct and simultaneous determination of molybdenum and vanadium in sea-water using a multielement electrothermal atomic absorption spectrometer | |
| Liao et al. | Cloud point extraction combined with flow injection vapor generation inductively coupled plasma mass spectrometry for preconcentration and determination of ultra trace Cd, Sb and Hg in water samples | |
| Feng et al. | Flow-injection spectrophotometric determination of mercury (II) in water by the catalytic decomposition of ferrocyanide | |
| Peng et al. | Determination of trace amounts of beryllium (II) in drinking water and of beryllium vapor in air by graphite-furnace atomic absorption spectrophotometry using acetylacetone as a chelating agent | |
| dos Santos Depoi et al. | Methodology for Hg determination in honey using cloud point extraction and cold vapour-inductively coupled plasma optical emission spectrometry | |
| Asan et al. | Flow injection spectrofluorimetric determination of iron (III) in water using salicylic acid | |
| Alacakoc et al. | Determination of trace nickel in spinach samples using the combination of vortex-assisted deep eutectic solvent-based liquid phase microextraction and slotted quartz tube-flame atomic absorption spectrometry | |
| Santelli et al. | Determination of cobalt at low levels in silicate rocks by atomic absorption spectrometry using a continuous on-line precipitation-dissolution procedure based on 1-nitroso-2-naphthol | |
| Chooto et al. | Determination of inorganic arsenic species by hydride generation–inductively coupled plasma optical emission spectrometry | |
| CN111189956A (en) | By means of H2O2Method for detecting content of nitrite in sodium chloride sample by combining oxidation method with ion chromatography | |
| CN103543134A (en) | Method for determining lead content in iron ore through hydride generation-atomic fluorescence spectrometry | |
| Li et al. | Direct determination of trace lead in seawater by inductively coupled plasma mass spectrometry after photochemical vapor generation | |
| Wang et al. | Determination of beryllium and selenium in human urine and of selenium in human serum by graphite-furnace atomic absorption spectrophotometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150819 |
|
| WD01 | Invention patent application deemed withdrawn after publication |