Strong polar organic matter molecular mass detection method in the coking waste water biochemical treatment discharged liquid
Technical field: the present invention relates to a kind of detection method of organic matter molecular mass, strong polar organic matter molecular mass detection method in particularly a kind of coking waste water biochemical treatment discharged liquid.The present invention adopts extracting n-butyl alcohol, the separation of liquid chromatography gradient elution, mass spectrum (ESI interface) to identify strong polarity colored organism molecular weight in the water outlet of coking chemical waste water after biological treatment.
Background technology: coking chemical waste water is a kind of industrial waste water that typically contains persistent organic pollutants, and its pollutant component complexity except that containing a large amount of organic compounds, also contains hazardous contaminants such as cyanogen, fluorine, ammonia nitrogen.The disposal route of coking chemical waste water mainly is divided into biochemical method and physico-chemical process.Wherein biochemical process can be removed multiple pollutant in single biological treatment system, and simple to operate, operating cost is also lower, and therefore, biochemical processing method is the main means of Treatment of Coking Effluent always.But coking chemical waste water is through behind the biological treatment, and indexs such as CODcr, colourity still do not reach emission standard, could discharge after also need adding multiple physico-chemical process, and expense is costliness relatively.Therefore, coking chemical waste water is analyzed through the composition of coloured organic contaminant in the water outlet after carrying out a biological disposal upon, understood the molecular weight size of organic contaminant, help the exploitation and the optimization of method for subsequent processing.
For organic analysis in the coking chemical waste water, " analysis of hardly degraded organic substance in the coking chemical waste water " (environmental engineering, 1991,9 (1): 31~33) propose: the mensuration of organic constituents can adopt the method for dichloromethane extraction and gas chromatography-mass spectrometry (GC-MS) in the coking chemical waste water." organic pollutant analysis in the coke-oven plant A/O water outlet " (Environmental Pollution and Control, 2001,23 (3): 140~142) also adopt dichloromethane extraction, organic principle has carried out qualitative analysis in the water outlet after gas chromatography and mass spectromentry coupling method (GC/MS) is carried out a biological disposal upon to the Baosteel chemical industry.This method adopts methylene chloride as extractant, is the organic compound method commonly used of separating water-insoluble and being slightly soluble in water, also is that one of method that adopts is recommended by EPA (EPA).But in the process of analyzing the water outlet of coking chemical waste water after biological anoxic Aerobic Process for Treatment, find methylene chloride to the extracting power of colored organism a little less than, this may be water-soluble stronger material with colored organism, relevant fully with low pole and the higher dichloromethane extraction of nonpolar organic matter percentage extraction are difficult to, therefore also need seek the extractant outside the methylene chloride.In addition, this method adopts gas chromatography-mass spectrometric hyphenated technique that organism is identified.Because gas chromatography (GC) can only be separated under the operating temperature and can gasify and Undec material, be mainly volatile organic matter (boiling point is lower than 200 ℃) and semi-volatile organic matter (200 ℃~300 ℃ of boiling points), these organic amounts are no more than 20%~30% of total organic carbon in the water.All the other 70%~80% mainly are higher boiling and strong polar organic matter, need to adopt high performance liquid chromatography (HPLC) to separate.In recent years along with analytical technology development, liquid chromatograph mass spectrography (HPLC-MS) technology begins more and more to be applied to the mensuration of thermal instability component and difficult volatile constituent.This makes in the water 70%~80% the higher boiling and the isolation identification of strong polar organic matter become possibility, also help us when extracting the stronger organism of polarity, can carry out isolation identification by the liquid chromatograph mass spectrography technology, understand the unmeasured organic molecular weight of GC/MS, provide foundation for improving follow-up advanced treating mode.
Summary of the invention: the objective of the invention is to overcome in the dichloromethane extraction water that exists in the analysis of coking chemical waste water organism the more weak and gas chromatography of strong polar organic matter ability and can only be separated in and can gasify under the operating temperature and the problem of Undec material, polar organic matter molecular mass detection method by force is provided in a kind of coking waste water biochemical treatment discharged liquid.Thinking of the present invention is to adopt the mode of extracting n-butyl alcohol, the separation of liquid chromatography gradient elution, mass spectrum (ESI interface) three combinations to identify strong polarity colored organism molecular weight in the water outlet of coking chemical waste water after biological anoxic Aerobic Process for Treatment.Identifying in the water outlet by the present invention the molecular weight of strong polarity colored organism can increase the understanding to strong polarity colored organism in the water outlet of coking chemical waste water after biological anoxic Aerobic Process for Treatment, is convenient to the optimization of subsequent treatment process.
For realizing the object of the invention, technical scheme of the present invention is: strong polar organic matter molecular mass detection method in a kind of coking waste water biochemical treatment discharged liquid, may further comprise the steps: at first in coking waste water biochemical treatment discharged liquid, take a sample, add the extractant normal butyl alcohol then, evaporation extracting n-butyl alcohol phase, obtain concentrate and add dissolve with methanol, adopt liquid chromatography to separate then, chromatographic column is selected little footpath C
18Post, moving phase is selected water/methanol solution, adopt the gradient elution mode to wash extract, to separate the eluent that obtains containing strong polar material by liquid chromatography and carry out mass spectrophotometry, liquid chromatography and mass spectrometer adopt ESI interface, select for use the negative ion electrospray electric mass spectrum mode of spraying to analyze the molecular weight that obtains strong polar material in the eluent.
The sample of coking waste water biochemical treatment discharged liquid needs the pH value is adjusted to 3~4; Flow rate of mobile phase 0.2ml/min during liquid-phase chromatographic analysis; The proportioning of methyl alcohol and concentrate all is dissolved in the methyl alcohol and is advisable to remove condensed matter behind the normal butyl alcohol.
The formulation of technical solution of the present invention is according to being: 1, selection of Extractant: the molecular weight that understand colored organism in the water outlet (hereinafter to be referred as biological water outlet) of coking chemical waste water after biological anoxic Aerobic Process for Treatment is formed, and at first need find a kind of solvent of effective extraction colored organism.We are judging basis to extract back water change color, use 1mol/L H
2SO
4, 1mol/L NaOH is respectively with biological water outlet furnishing acidity, neutrality, alkalescence, carry out liquid-liquid extraction with an amount of phenixin, carbon disulphide, methylene chloride, methenyl choloride, ether, sherwood oil, benzene, monochloro-benzene, nitrobenzene, normal butyl alcohol, normal hexane etc. with the immiscible organic solvent of water, normal butyl alcohol extracts colored organism when acid condition (pH3~4) effect is the most obvious, extraction back water colourity reduces half, and is better than methylene chloride effect.Explain from polarity index: normal butyl alcohol polarity index 4.0, methylene chloride polarity index 3.1, ether polarity index 2.8, benzene polarity index 2.7, phenixin polarity index 1.6, normal hexane polarity index 0.0, the polarity index of normal butyl alcohol are the highest in these solvents.According to the similar principle that mixes of solvent, it more can extract the stronger colored organism of polarity than methylene chloride.We finally determine the extractant of normal butyl alcohol as strong polarity colored organism in the biological water outlet of extraction.2, concentrating of sample:, need with sample introduction analysis after the sample concentration because organic concentration is relatively low in the biological water outlet.Because the normal butyl alcohol in the concentrate has absorption under the liquid chromatography UV-detector, the meeting interference measurement is used the dissolve with methanol concentrate instead after therefore normal butyl alcohol all need being removed.Extracting n-butyl alcohol is put into round-bottomed flask mutually, adopt the rotary evaporation method, (water-bath is heated, and temperature is controlled at 65 ℃ with the RE52-II rotary evaporator, absolute pressure 0.923~0.933Mpa) steams normal butyl alcohol fall, and the concentrate of staying in the bottle is to be measured with small amount of methanol dissolving back.3, sample separation: on the HP6890 gas chromatography with two kinds of the most frequently used pillars: HP-INNOWAX (polar column, 260 ℃ of the column temperature upper limits) and HP-5 (low-pole column, 325 ℃ of the column temperature upper limits) post respectively temperature programme analyze to upper temperature limit, all do not have the peak to occur.Illustrate that the organism that n-butyl alcohol extract and dichloromethane extraction go out is different, polarity is stronger or be difficult for vaporizing on gas chromatography, can't adopt gas chromatography analysis.Can only adopt liquid chromatography to separate, liquid-phase condition is: chromatographic column: can select little footpath C
18Post; Moving phase is selected water/methanol solution, flow velocity 0.2ml/min; The gradient elution mode; Because extract component complexity is only used constant composition moving phase wash-out, and all components wash-out is come out.And gradient elution is to make the solvent that contains two or more opposed polarity in the moving phase, in elution process continuously or be interrupted the composition that changes moving phase, to regulate its polarity, all components in the sample was flowed out in the shortest analysis time, and between peak and the peak certain separating arranged.We are by changing the composition of two kinds of solvents of first alcohol and water, the polarity that makes moving phase from by force → weak → change successively by force, the material that makes opposed polarity all wash-out comes out.Liquid chromatogram detector: photodiode array detector.4, Mass Spectrometer Method: the selection of interface: liquid chromatograph mass spectrography the most frequently used interface at present is electron spray ionisation (ESI) and Atmosphere Pressure Chemical Ionization (APCI) (APCI) interface.ESI is suitable for the analysis of middle polarity to the compound molecule of strong polarity, and APCI is suitable for the micromolecular of nonpolar or middle polarity, considers that n-butyl alcohol extract polarity is stronger, and we select ESI interface.The selection of positive and negative ion pattern: electron spray ionisation is divided into negative ion and two kinds of patterns of positive ion according to the ionization mode, and positive ion mode is applicable to basic sample solution, and negative ion mode is applicable to acid sample.We all attempt at two kinds of patterns, the result that positive ion mode obtains is not quite desirable, each peak has all produced abundant quasi-molecular ion peak under the negative ion mode, illustrate that the colored organism polarity that normal butyl alcohol extracts is stronger under acid medium, relatively be fit to the negative ion electrospray electric mass spectrum mode of spraying.
The invention has the beneficial effects as follows: by liquid chromatography-electron spray-mass spectrometric hyphenated technique (HPLC-ESI-MS), the molecular weight of having determined the strong polarity colored organism that extracting n-butyl alcohol comes out is basically below 500.Understand coking chemical waste water through these organic molecular weight sizes in the biological treatment back water outlet, we are considered better advanced treating mode, during as membrane technology, membrane material, membrane aperture size, parent/hydrophobic selection have certain significance for reference.
Description of drawings:
Fig. 1 separates spectrogram for the PDA of n-butyl alcohol extract of the present invention
Fig. 2 is ESI (-) mass spectrogram of 1.38 minutes materials for appearance time of the present invention
Fig. 3 is ESI (-) mass spectrogram of 3.83 minutes materials for appearance time of the present invention
Fig. 4 is ESI (-) mass spectrogram of 18.07 minutes materials for appearance time of the present invention
Fig. 5 is ESI (-) mass spectrogram of 21.78 minutes materials for appearance time of the present invention
Fig. 6 is ESI (-) mass spectrogram of 32.97 minutes materials for appearance time of the present invention
Fig. 7 is ESI (-) mass spectrogram of 36.90 minutes materials for appearance time of the present invention
Embodiment:
Further understand essence of the present invention by following examples:
Get 1.6 liters of the coking chemical waste water of coke-oven plant after biological anoxic Aerobic Process for Treatment, CODcr is 300mg/l.Use 1mol/L H
2SO
4Water outlet is transferred to after PH is 3~4, heavily steam normal butyl alcohol with 400ml and divide aqueous phase extracted three times, organic phase after the merging places-clean round-bottomed flask in, remove normal butyl alcohol through the rotary evaporation method and (adopt the model of rotary evaporator: RE52-II, the water-bath heating, temperature is controlled at 65 ℃, absolute pressure 0.923~0.933Mpa).Stay the bottle in a concentrate fully dissolve with 10ml methyl alcohol after, be transferred in the centrifuge tube from round-bottomed flask, be used for the analysis.
Taking the n-butyl alcohol extract of dissolve with methanol analyzes to the HP1100 liquid chromatograph.Adopt WatersSymmetry C
18(1.0 * 150mm, 3.5um) post, flow velocity 0.2ml/min, gradient elution method is separated extract, and elution program is: described number percent is weight percentage.
0~10 minute: water 100% → 77%, methyl alcohol 0% → 23%,
10~20 minutes: water 77% → 60%, methyl alcohol 23% → 40%,
20~40 minutes: water 60% → 20%, methyl alcohol 40% → 80%,
40~45 minutes: water 20% → 0%, methyl alcohol 80% → 100%,
45~48 minutes: water 0%, methyl alcohol 100% remains unchanged,
48~50 minutes: water 0% → 100%, methyl alcohol 100% → 0%,
50~55 minutes: water 100%, methyl alcohol 0 remained unchanged.
Under this gradient elution mode, in 180~600nm ultraviolet light and wavelength of visible light scope, detect tens at peak with photodiode array detector (PDAD), the appearance time that obtains the stronger material of uv absorption was respectively 1.38 minutes, 3.83 minutes, 18.07 minutes, 21.78 minutes, 32.97 minutes, 36.90 minutes, with reference to Fig. 1.
Taking the n-butyl alcohol extract of dissolve with methanol analyzes to HP1100/FINNIGAN MAT95 liquid chromatograph-mass spectrometer.According to above-mentioned gradient elution program, make extract on the HP1100 liquid chromatography, obtain separating, under negative ion mode, carry out mass spectrum by electron spray ionisation (ESI) interface respectively according to the different elution order in each peak and identify.Because n-butyl alcohol extract polarity is stronger, and is acid sample, therefore be fit to very much ESI (-)-this mass spectrum mode of MS, each peak has all produced abundant quasi-molecular ion peak.The molecular weight of several main matter sees Table 1:
Table 1: the molecular weight of main matter in the n-butyl alcohol extract
Appearance time |
Molecular weight |
1.38 minute |
148 |
3.83 minute |
154 |
18.07 minute |
250 |
21.78 minute |
295 |
32.97 minute |
212 |
36.90 minute |
466 |
Annotate: under ESI (-) mode, the quasi-molecular ion peak of material+1 is the molecular weight of material.With reference to Fig. 2,3,4,5,6,7.