CN113219087B - Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis - Google Patents

Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis Download PDF

Info

Publication number
CN113219087B
CN113219087B CN202110358321.9A CN202110358321A CN113219087B CN 113219087 B CN113219087 B CN 113219087B CN 202110358321 A CN202110358321 A CN 202110358321A CN 113219087 B CN113219087 B CN 113219087B
Authority
CN
China
Prior art keywords
chromatographic column
column
silver nitrate
mobile phase
chromatographic
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.)
Active
Application number
CN202110358321.9A
Other languages
Chinese (zh)
Other versions
CN113219087A (en
Inventor
李丽敏
季申
毛秀红
胡青
曹帅
周如洁
彭伟楠
程益清
付泽晓
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SHANGHAI INSTITUTE FOR FOOD AND DRUG CONTROL
Original Assignee
SHANGHAI INSTITUTE FOR FOOD AND DRUG CONTROL
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by SHANGHAI INSTITUTE FOR FOOD AND DRUG CONTROL filed Critical SHANGHAI INSTITUTE FOR FOOD AND DRUG CONTROL
Priority to CN202110358321.9A priority Critical patent/CN113219087B/en
Publication of CN113219087A publication Critical patent/CN113219087A/en
Application granted granted Critical
Publication of CN113219087B publication Critical patent/CN113219087B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8631—Peaks
    • G01N30/8634—Peak quality criteria
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P10/00—Technologies related to metal processing
    • Y02P10/20—Recycling

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Quality & Reliability (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)

Abstract

The invention discloses a method for recovering and treating reduced column efficiency of inorganic mercury chromatographic peak in mercury element form and valence state analysis, which aims at Hg 2+ The chromatographic peak column effect is reduced and the peak shape is deteriorated, a specially-made regeneration treatment reagent is provided, silver nitrate with a certain concentration is used as a water phase and is mixed with methanol in a certain proportion, and the problems of Hg in the existing mercury morphological analysis are solved by sequentially adopting the steps of pre-cleaning, regeneration treatment, replacement cleaning and the like 2+ The problems of poor and widened chromatographic peak shape, reduced chromatographic column efficiency and the like are not solved, and the pertinence is strong.

Description

Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis
Technical Field
The invention relates to the field of food and drug detection and analysis, in particular to a method for analyzing the form and valence state of mercury element by using octadecylsilane chemically bonded silica asBivalent mercury (inorganic mercury, hg) in the case of mercury form analysis by liquid chromatography column of filler 2+ ) A recovery processing method after the column efficiency of chromatographic peak is reduced.
Background
At present, the bivalent mercury (Hg) in the mercury element in a sample is analyzed by adopting a high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) technology 2+ Inorganic mercury), methylmercury (CH) 3 Hg), ethylmercuric acid (CH) 2 CH 3 Hg) and other element forms and valence states, and is widely applied in the fields of food and medicine.
In the method, a chromatographic column with octadecylsilane chemically bonded silica as a filler is generally adopted, and cysteine or mercaptoethanol and Hg are added into a mobile phase 2+ 、CH 3 Hg、CH 2 CH 3 Hg forms a complex for isolation. Hg is a mercury vapor 2+ Has stronger binding capacity to the residual silicon hydroxyl (-OH) of the octadecylsilane chemically bonded silica chromatographic column, and Hg is analyzed after a period of time due to the influence of a sample matrix 2+ The absorption and elution behaviors of chromatographic peaks are influenced, the chromatographic peaks are widened and trailing, the column efficiency is reduced and the like, and Hg is influenced 2+ Separated from the chromatographic peak of methyl mercury, which leads to inaccurate quantification and even Hg 2+ The chromatographic peak disappeared and analysis could not be performed.
To solve the problem, in the < 2322 of general rules of mercury, arsenic form and valence state analysis of the four parts of the design & ltChinese pharmacopoeia 2020 edition, note (3), it is mentioned that "when mercury form and valence state analysis is performed, hg is subjected to the incomplete end-capping Si hydroxyl group remaining in the chromatographic column 2+ Leading to a faster column efficiency loss. It is recommended to use columns with higher coverage of the end-caps and, if necessary, to continue the analysis after flushing the column with a high proportion of organic phase at certain sample injection intervals, using valve switching techniques ". In the existing liquid chromatographic analysis technology, when the column efficiency of a chromatographic column is reduced, leading to the situations of chromatographic peak broadening, tailing and the like, a high-proportion organic phase (methanol, acetonitrile and the like) is mostly adopted to wash the chromatographic column and elute impurities adsorbed on the chromatographic column, thereby achieving the purpose of recovering the column efficiency or directly replacing a new chromatographic column. However, in practice, methanol is used to wash the columnThe recovery degree of column efficiency is limited, and Hg after washing 2+ The chromatographic peak can not reach the column efficiency of the new chromatographic column. Therefore, after the chromatographic column is analyzed to a certain amount of sample, only the chromatographic column can be replaced in most cases, so that the measurement cost is greatly increased.
Therefore, a method for effectively recovering Hg in the method for analyzing the form and valence state of mercury element by HPLC-ICP-MS is developed 2+ The processing method of chromatographic peak column efficiency and peak shape is very important for improving the accuracy of the method and reducing the analysis cost, and becomes one of the problems which need to be solved urgently in the analysis of mercury form.
Disclosure of Invention
Aiming at the existing method for analyzing the form and valence state of mercury element by HPLC-ICP-MS, inorganic mercury (Hg) is used 2+ ) The invention provides a method for effectively recovering Hg, which solves the problems of reduced chromatographic peak column efficiency, poor peak deformation and influence on accurate quantification 2+ Chromatographic peak shape and column efficiency.
In a first aspect of the invention, an inorganic mercury (Hg) for use in elemental mercury morphological and valence state analysis is provided 2+ ) The regenerated reagent for recovering the chromatographic peak with reduced column efficiency consists of silver nitrate aqua and methanol in certain proportion.
Preferably, the concentration of the silver nitrate substance in the silver nitrate aqueous solution is 0.1 to 0.5mmol/L.
Preferably, the concentration of the amount of silver nitrate substance of the silver nitrate aqueous solution is 0.1mmol/L.
Preferably, the regeneration reagent comprises methanol with the mass concentration percentage of 2-8%.
In a second aspect, the present invention provides an inorganic mercury (Hg) in the form and valence state analysis of Hg element 2+ ) The recovery processing method after the chromatographic peak column efficiency is reduced, the chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, and when the liquid chromatograph is a binary pump, the method comprises the following steps:
step S1, chromatographic column pretreatment: the connection between the outlet end of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet end of the chromatographic column is connected with a waste liquid collecting device;
step S2A, a chromatographic column pre-cleaning step: mobile phase: the phase A is water, the phase B is methanol, and the volume ratio of the mobile phase A to the mobile phase B is 95:5, the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, and the cleaning time is 30-60 min;
step S3A, a chromatographic column regeneration treatment step: placing the silver nitrate aqueous solution into a mobile phase A, wherein the volume ratio of the mobile phase A to the mobile phase B is 95:5, the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, and the regeneration treatment time is 30-60 min;
step S4, replacing a chromatographic column regeneration reagent: and (3) changing the mobile phase A into water, wherein the volume ratio of the mobile phase A to the mobile phase B is 95: and 5, the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, the replacement time is 30-60 min, and the outlet end of the chromatographic column is connected to the inlet of the ICP-MS atomizer after replacement is finished.
Preferably, the preparation process of the silver nitrate aqueous solution comprises the following steps: precisely taking 1-5 mL of 0.1mol/L silver nitrate solution, adding about 700-900 mL of water, adding water to 1000mL, and shaking up to obtain the silver nitrate solution.
Further preferably, the preparation process of the silver nitrate aqueous solution is as follows: precisely taking 1mL of 0.1mol/L silver nitrate solution, adding about 900mL of water, adding water to 1000mL, and shaking up to obtain the silver nitrate solution.
Preferably, the silver nitrate in the 0.1mol/L silver nitrate solution is one or two of analytically pure and chromatographically pure.
More preferably, the silver nitrate in the 0.1mol/L silver nitrate solution is chromatographically pure;
in a third aspect, the present invention provides an inorganic mercury (Hg) in the form and valence state analysis of Hg element 2+ ) The recovery processing method after the chromatographic peak column efficiency is reduced, the chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, and when the liquid chromatograph is a unit pump, the recovery processing method comprises the following steps:
step S1, chromatographic column pretreatment: the connection between the outlet end of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet end of the chromatographic column is connected with a waste liquid collecting device;
step S2B, pre-cleaning of the chromatographic column: placing the aqueous solution containing 5% of methanol in a mobile phase, wherein the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, and the cleaning time is 30-60 min;
step S3B, a chromatographic column regeneration treatment step: placing the regeneration solution into a mobile phase, wherein the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, and the regeneration treatment time is 30-60 min;
step S4, a chromatographic column regeneration reagent replacement step: and (3) placing the aqueous solution containing 5% of methanol into a mobile phase, setting the flow rate at 0.5-1.5 mL/min, the column temperature at room temperature, and the replacement time at 30-60 min, and connecting the outlet end of the chromatographic column to the inlet of the ICP-MS atomizer after replacement is finished.
Preferably, the method further comprises a checking step of: according to the operation steps of analyzing the form and valence state of mercury element by normal high performance liquid-inductively coupled plasma mass spectrometry, 10-50 mul Hg is analyzed by sample injection 2+ 、CH 3 Hg、CH 2 CH 3 Hg mix control solution.
Preferably, in step S1 to step S4, the water is one or more of HPLC grade water, ultrapure water and deionized water.
More preferably, in step S1 to step S4, the methanol is HPLC grade.
Preferably, in step S1 to step S4, the methanol is one or more of HPLC grade, chromatographic purity or mass spectrum purity.
More preferably, in step S1 to step S4, the methanol is HPLC grade or chromatographically pure.
The invention provides Hg in the analysis of the form and valence state of mercury element 2+ The invention discloses a regeneration recovery processing method after column efficiency of chromatographic peaks is reduced or invalid, which has the beneficial effects that:
residual silicon hydroxyl of octadecylsilane chemically bonded silica chromatographic column in mercury element form and valence state analysis is to Hg 2+ Has certain adsorption effect, and Hg is adsorbed due to the influence of matrix after multiple sample injection analysis 2+ The chromatographic peak is gradually widened, and the separation degree from the methyl mercury is reduced, even Hg is generated 2+ The disappearance of chromatographic peaks leads to the failure of accurate quantitative analysis. The invention is directed to Hg 2 + The chromatographic peak column effect is reduced and the peak shape is deteriorated, a specially-made regeneration treatment reagent is provided, silver nitrate with a certain concentration is used as a water phase and is mixed with methanol in a certain proportion, and the problems of Hg in the existing mercury morphological analysis are solved by sequentially adopting the steps of pre-cleaning, regeneration treatment, replacement cleaning and the like 2+ The problems of poor and widened chromatographic peak shape, reduced chromatographic column efficiency and the like are not solved, and the pertinence is strong.
The invention also determines the concentration, the cleaning flow rate and the cleaning time of the regeneration reagent, and can effectively improve Hg under the specific concentration, flow rate and cleaning time 2+ The chromatographic peak-shaped chromatographic column has long service life and thus wide application foreground.
Drawings
FIG. 1 is a chromatogram of a mixed control solution when the new octadecylsilane-bonded silica gel column of example 1 was activated;
FIG. 2 is a chromatogram of a sample solution of corrugated seeds when column I in example 1 is activated;
FIG. 3 is a chromatogram of a sample solution of Pheretima as obtained when column I of example 1 is activated;
FIG. 4 is a chromatogram of a mixed control solution after about 70 batches of sample solutions in example 1 are injected;
FIG. 5 is a chromatogram of a mixed control solution after a chromatographic column I has been subjected to the regeneration method of the present invention;
FIG. 6 is a chromatogram of a mixed control solution after regeneration by washing column I with conventional methanol in comparative example 1;
FIG. 7 is a chromatogram of a mixed control solution when column II in example 2 is activated;
FIG. 8 is a chromatogram of a mixed control solution after injecting sample solutions of animal and plant substrates such as Pheretima and astragali radix for 60 batches in example 2;
FIG. 9 shows that in example 2, after injecting sample solution of animal and plant substrates such as Lumbricus, radix astragali, etc. for about 60 batches, bivalent mercury (Hg) is added 2+ ) A control solution chromatogram;
FIG. 10 is a chromatogram of a bivalent mercury control solution after about 40 times of sample injection of the cinnabar sample solution of the mineral drug in example 2;
FIG. 11 is a chromatogram of a bivalent mercury control solution after the regeneration method of the present invention is applied to a chromatographic column II;
FIG. 12 is a chromatogram of the mixed control solution when column III of example 3 is active;
FIG. 13 is a chromatogram of a mixed reference solution after multiple batches of samples of mercury form and valence state have been injected in example 3;
FIG. 14 is a chromatogram of a mixed control solution after the regeneration method of the present invention is applied to the column III.
Detailed Description
The invention will be further described with reference to specific embodiments, the advantages and features of which will become apparent from the description, but the invention is not limited to the embodiments below.
In the following examples 1-3, the main apparatus used included:
model Agilent 8800 inductively coupled plasma mass spectrometer (Agilent, usa);
model Agilent 7700 inductively coupled plasma mass spectrometer (Agilent corporation, usa);
model 1260 high performance liquid chromatograph (Agilent, usa, with dual pump and on-line degasser);
a water bath (SW 22, jumbo);
scales (Sartorius, germany);
centrifuge (Eppendorf, germany);
in the following examples 1-3, the main materials and reagents used included:
mercury standard solution (1000 μ g/mL, fluka, also bivalent mercury standard solution);
ethyl mercury chloride (97.0%, lot80430, dr. Ehrenstorfer GmbH);
methylmercury chloride (97.0%, lot90917, dr. Ehrenstorfer GmbH);
the methyl mercury standard substance is Tuna (Tuna fish BCR463, european Standard office);
silver nitrate (super pure, shanghai national drug group);
pepsin (P7125, sigma);
nitric acid, hydrochloric acid, ammonium dihydrogen phosphate and phosphoric acid are all superior grade purities;
the water was Milli-Q treated deionized water (18.2. Omega. M);
high purity liquid argon (purity > 99.99%) and helium (purity 99.999%) are provided by Shanghai Power gas industries, inc.
Example one
In this embodiment, taking chromatographic column I, referring to appendix 2322 of 2020 edition of chinese pharmacopeia, conducting the study on the mercury element form and valence state of earthworm and concha arcae, sequentially injecting sample according to a predetermined condition to analyze the reference solution and the sample solution, respectively recording the chromatogram immediately after sample injection, the chromatogram after column efficiency is decreased after sample injection for several times, and the chromatogram after washing the chromatographic column according to the method of the present invention, and comparing them.
1.1 preparation of solution:
preparation of mixed control solution:
respectively taking a mercury standard solution and appropriate amount of methyl mercury chloride and ethyl mercury chloride reference substances, precisely weighing, and adding 5% methanol to prepare solutions each containing 100ng (calculated by mercury) per 1mL to obtain reference substance mixed stock solution;
precisely sucking a proper amount of the reference substance mixed stock solution, and respectively preparing into series of standard solutions containing 1ng, 2ng, 5ng, 10ng and 20ng of mercury per 1mL with 5% methanol to obtain mixed reference substance solution.
Preparation of sample solution (prepared by using a method of ultrasonic extraction of artificial gastric juice prepared by nitric acid):
respectively taking earthworm and concha arcae samples, adding a solution containing 3.28 percent of dilute nitric acid and 10 percent of pepsin as a digestion solution, carrying out closed oscillation in a water bath at 45 ℃ for overnight (20-24 hours), taking out, filtering by a membrane, and taking a subsequent filtrate to obtain the earthworm and concha arcae sample solution.
Preparing a silver nitrate solution: precisely taking 1mL of 0.1mol/L silver nitrate solution, adding about 900mL of water, uniformly mixing, adding water to 1000mL, shaking up, and ultrasonically degassing to obtain 0.1mmol/L silver nitrate solution.
1.2 injecting a reference substance solution and a sample solution:
chromatographic conditions are as follows:
a chromatographic column: octadecyl silica gel bonded phase chromatography column (Hydro-RP 80A 150X 4.6mm, 4 μm, phenomenex);
mobile phase: methanol-0.1% of L-cysteine and 0.1% of L-cysteine hydrochloride (L-cysteine, HCl, H) 2 O) mixed solution (5;
flow rate: 1.0mL/min.
ICP-MS conditions:
adopting a platinum intercepting cone and a sampling cone, and adopting normal mode analysis to tune the sensitivity, precision and specificity of the liquid adjusting instrument; the mass number of the mass spectrum parameter is 202 for detection, and the time resolution mode is selected as the acquisition mode, and the acquisition time is 0.5s respectively.
When the column I is just analyzed, the mercury morphological chromatographic peaks of the mixed control solution, the earthworm sample solution and the concha arcae sample solution on the column all show sharper and more symmetrical peak shapes (see fig. 1 to 3).
When about 100 times of injection is carried out, hg is found 2+ The peak shape of the chromatographic peak is widened, and the separation and accurate quantification of the chromatographic peak are influenced if the broadening is serious (see figure 4).
Comparing FIGS. 1 to 3 and 4, it can be seen that the matrix and Hg are added as the number of times of injecting Lumbricus sample and corrugate sample increases 2+ The unavoidable effect of residual silicon hydroxyl groups on the chromatographic column leads to Hg 2+ The column efficiency of chromatographic peaks is reduced.
1.3 chromatographic column regeneration
Step S1: the connection between the outlet of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet of the chromatographic column is connected with waste liquid;
step S2: setting a mobile phase A as water and a mobile phase B as methanol, setting the flow rate to be 1.0mL/min and the column temperature to be room temperature, and setting the mobile phase composition to be A: b =95:5; the cleaning time is 60min;
and step S3: placing silver nitrate solution in phase A of high performance liquid chromatograph, setting flow rate at 1.0mL/min, column temperature at room temperature, and performing regeneration treatment for 60min;
and step S4: after the regeneration treatment, the flow rate is set to zero, the silver nitrate solution is taken off, A is exchanged for ultrapure water, and the ratio of A: b =95:5; performing chromatographic column regeneration reagent replacement for 30min.
After the above operation is completed, the mixed reference solution is taken, sample injection analysis is performed according to the sample injection condition in 1.2, and a chromatogram is recorded, as shown in fig. 5. The results show that Hg is generated after the regenerated reagent is washed by the method 2+ The chromatographic peak broadening phenomenon is obviously improved, and the column efficiency is recovered.
Comparative example 1
Comparative example 1 differs from example 1 in the method for regenerating the column, and the method for washing the column with conventional methanol in this comparative example specifically comprises the following steps:
step A1: the chromatographic column pretreatment step: the connection between the outlet of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet of the chromatographic column is connected with the waste liquid;
step A2: setting the mobile phase A as water and the mobile phase B as methanol, the flow rate to be 1.0mL/min and the column temperature to be room temperature, and setting the mobile phase composition to be A: b =95:5; the cleaning time is 60min;
step A3: setting the mobile phase composition as A: b =5: the flow rate of 95 is 1.0mL/min, the column temperature is room temperature, the cleaning time is 60min, and then the mobile phase composition is set as follows: b =95:5, cleaning for 30min.
After the above operation is completed, the mixed reference solution is sampled and analyzed according to the sampling conditions of 1.2, and a chromatogram is recorded, as shown in fig. 6.
Comparing fig. 5 and fig. 6, it can be seen that in the process of analyzing the mercury element form and valence states of the marine medicine and the animal medicine, the matrix and the Hg are added with the increase of the sample injection times 2+ The unavoidable effect of residual silicon hydroxyl groups on the chromatographic column leads to Hg 2+ The chromatographic peak column efficiency is reduced, and the chromatographic column is not improved greatly by using an organic solvent methanol to wash the chromatographic column; hg is regenerated using the reagent of the present invention and using the rinsing step of the present invention 2+ The chromatographic peak is obviously recovered and can be reused for mercury elementThe quantitative analysis of the form prolongs the service life of the chromatographic column.
Example 2
In this embodiment, taking chromatographic column II, referring to appendix 2322 of 2020 edition of chinese pharmacopeia, performing the research on the forms and valence states of mercury elements in the earthworm and astragalus membranaceus samples, sequentially injecting sample according to a predetermined condition to analyze the reference solution and the sample solution, and respectively recording the chromatogram immediately after sample injection, the chromatogram after column efficiency is decreased after sample injection for a plurality of times, and the chromatogram after washing the chromatographic column according to the method of the present invention, and comparing.
2.1 preparation of solution:
preparation of mixed control solution, same as 1.1.
The preparation of a divalent mercury control solution,
precisely absorbing a proper amount of mercury element standard solution (1 g/mL, the medium type is nitric acid), and adding water to prepare solution containing l00ng per 1mL, so as to obtain bivalent mercury reference substance stock solution;
precisely sucking a proper amount of the bivalent mercury reference substance stock solution, and respectively preparing a series of standard solutions containing 1ng, 2ng, 5ng, 10ng and 20ng of mercury per 1mL by using 8% methanol to obtain bivalent mercury reference substance solution.
Preparation of sample solution:
earthworm and astragalus root samples are prepared by a method of ultrasonic extraction of artificial gastric juice prepared by nitric acid, the same as 1.1;
preparing a cinnabar sample solution: weighing cinnabar powder (passing through a five-mesh sieve) about 30mg, precisely weighing, placing the cinnabar powder into a 250mL plastic measuring flask in a mode of 2 copies, adding artificial gastric juice about 200mL into one copy, adding artificial intestinal juice about 200mL into the other copy, shaking up, placing the mixture into a water bath at 37 ℃ for ultrasonic treatment (power 300W and frequency 45 kHz) for 2 hours (fully shaking up once every 15 minutes), cooling the mixture, respectively diluting the mixture to a scale with corresponding solutions, shaking up, taking a proper amount of the mixture into a 50mL plastic centrifuge tube, standing the mixture for 20-24 hours, gently blowing off the upper layer surface solution by using an ear washing ball, sucking the middle layer solution by about 15mL (avoiding particles from being brought in during sucking), filtering the mixture by using a microporous filter membrane (10 mu m), precisely weighing 2mL of subsequent filtrate, placing the subsequent filtrate into a 10mL plastic salt bottle, adding water to dilute the mixture to the scale, and shaking up to obtain the cinnabar powder.
Preparing a silver nitrate solution: precisely taking 2mL of 0.1mol/L silver nitrate solution, adding about 700-900 mL of the silver nitrate solution, adding water to 1000mL of the silver nitrate solution, and shaking up to obtain 0.2mmol/L silver nitrate solution.
2.2 control and sample injection
The chromatographic conditions and ICP-MS conditions were the same as in example 1.
When the chromatographic column is just analyzed, the peak shape of the chromatographic peak of the three valence states on the chromatogram of the mixed reference solution is good, as shown in fig. 7.
After about 60 times of sample injection of Lumbricus and radix astragali samples, hg in chromatogram of mixed reference solution 2+ The chromatographic peak was slightly tailing but did not affect the separation and detection accuracy, as shown in fig. 8. Hg 2+ The chromatographic peak on the control solution chromatogram smears but does not affect the integral, as shown in fig. 9.
Continuously injecting sample to analyze Cinnabaris sample for about 40 times, mixing Hg on chromatogram of control solution 2+ The chromatographic peak broadening gradually worsened, and the peak width became 2min due to the chromatographic peak tailing, which has not been accurately quantified, as shown in fig. 10.
2.3 chromatographic column regeneration
The specific procedure was the same as in example 1.
After the above operation is completed, taking the bivalent mercury reference solution, performing sample injection analysis according to the sample injection condition in 1.2, and recording a chromatogram, as shown in fig. 11.
The results show that Hg is treated by the regeneration method of the invention 2+ The chromatographic peak-peak broadening and the peak shape are obviously improved.
Example 3
In this embodiment, a chromatographic column III is taken, a reference solution and a sample solution are sequentially subjected to sample injection analysis according to a predetermined condition, and a chromatogram immediately after sample injection, a chromatogram after column efficiency is decreased after sample injection for several times, and a chromatogram after washing the chromatographic column according to the method of the present invention are respectively recorded and compared.
3.1 preparation of solution:
the control solution and sample solution were mixed and prepared as 1.1 and 2.1.
Preparing a silver nitrate solution: precisely taking 5mL of 0.1mol/L silver nitrate solution, adding about 700-900 mL of the silver nitrate solution, adding water to 1000mL of the silver nitrate solution, and shaking up to obtain 0.5mmol/L silver nitrate solution.
3.2 reference injection
Chromatographic conditions are as follows:
the column was an Xselect HSS T3 (150 × 4.6mm, 5 μm, waters) column;
other chromatographic conditions and ICP-MS conditions were the same as in example 1.
Hg in the column immediately after analysis 2+ The peak shape of the chromatographic peak is sharp and symmetrical, and is shown in figure 12.
The chromatographic column is used for analyzing the Hg element form and valence state of hundreds of samples 2+ The peak shape of the chromatographic peak is widened, the separation degree of the chromatographic peak from methyl mercury is deteriorated, and the accurate quantification is influenced, which is shown in figure 13.
3.3 chromatographic column regeneration
The specific procedure was the same as in example 1.
After the above operation is completed, the mixed reference solution is sampled and analyzed according to the sampling conditions of 3.1, and chromatogram is recorded, as shown in FIG. 14, hg 2+ The chromatographic peak shape is restored to be sharp and symmetrical, the separation degree with the methyl mercury is obviously improved, and the chromatographic column can be continuously used for quantitative analysis of the form and valence state of the mercury element after being restored.
The embodiments of the present invention have been described in detail, but the embodiments are only examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.

Claims (6)

1. A method for recovering and treating reduced column efficiency of inorganic mercury chromatographic peak in mercury element form and valence state analysis is characterized in that a chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, a liquid chromatograph is a binary pump, and the method comprises the following steps:
step S1, chromatographic column pretreatment: the connection between the outlet end of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet end of the chromatographic column is connected with a waste liquid collecting device;
step S2A, a chromatographic column pre-cleaning step: mobile phase: the phase A is water, the phase B is methanol, and the volume ratio of the mobile phase A to the mobile phase B is 95:5, the flow rate is 0.5 to 1.5mL/min, the column temperature is room temperature, and the cleaning time is 30 to 60min;
step S3A, a chromatographic column regeneration treatment step: placing a silver nitrate aqueous solution into a mobile phase A, wherein the volume ratio of the mobile phase A to the mobile phase B is 95:5, the flow rate is 0.5 to 1.5mL/min, the column temperature is room temperature, and the regeneration treatment time is 30 to 60min; the concentration of the silver nitrate substance in the silver nitrate aqueous solution is 0.1 to 0.5mmol/L;
step S4, replacing a chromatographic column regeneration reagent: and (3) changing the mobile phase A into water, wherein the volume ratio of the mobile phase A to the mobile phase B is 95: and 5, connecting the outlet end of the chromatographic column to the inlet of the ICP-MS atomizer after replacement, wherein the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, and the replacement time is 30-60min.
2. The recovery processing method as set forth in claim 1, wherein the silver nitrate aqueous solution is prepared by: precisely taking 1-5mL of 0.1mol/L silver nitrate solution, adding about 700-900mL of water, adding water to 1000mL, and shaking uniformly to obtain the silver nitrate solution.
3. The recovery processing method as set forth in claim 2, wherein the silver nitrate aqueous solution is prepared by: precisely taking 1mL of 0.1mol/L silver nitrate solution, adding about 900mL of water, adding water to 1000mL, and shaking up to obtain the silver nitrate solution.
4. The method of claim 1, wherein the silver nitrate in the aqueous silver nitrate solution is one or both of analytically pure and chromatographically pure.
5. A method for recovering and processing inorganic mercury chromatographic peak column effect after reduction in mercury element form and valence state analysis is characterized in that an octadecylsilane chemically bonded silica chromatographic column is used as a chromatographic column, a liquid chromatograph is a unit pump, and the method comprises the following steps:
step S1, chromatographic column pretreatment: the connection between the outlet end of the chromatographic column and the inlet of the ICP-MS atomizer is removed, the inlet end of the chromatographic column is connected with a high performance liquid chromatography mobile phase outlet valve, and the outlet end of the chromatographic column is connected with a waste liquid collecting device;
step S2B, pre-cleaning of the chromatographic column: placing an aqueous solution containing 5% of methanol in a mobile phase at a flow rate of 0.5-1.5 mL/min, a column temperature of room temperature and a cleaning time of 30-60min;
step S3B, a chromatographic column regeneration treatment step: placing a regeneration solution consisting of silver nitrate aqueous solution and methanol in a mobile phase at a flow rate of 0.5-1.5 mL/min and a column temperature of room temperature, wherein the regeneration treatment time is 30-60min; the concentration of the silver nitrate substance in the silver nitrate aqueous solution is 0.1 to 0.5mmol/L, and the mass concentration percentage of the methanol in the regeneration reagent is 2 to 8 percent;
step S4, replacing a chromatographic column regeneration reagent: and (3) placing an aqueous solution containing 5% methanol in a mobile phase, wherein the flow rate is 0.5-1.5 mL/min, the column temperature is room temperature, the replacement time is 30-60min, and after the replacement is finished, connecting the outlet end of the chromatographic column to the inlet of the ICP-MS atomizer.
6. A method of reprocessing according to any of claims 1-5 and also comprising a checking step of: according to the operation steps of analyzing the form and valence state of mercury element by normal high performance liquid-inductively coupled plasma mass spectrometry, sample introduction analysis is carried out for 10 to 50 mu l Hg 2+ 、CH 3 Hg、CH 2 CH 3 Hg mixes with the control solution.
CN202110358321.9A 2021-04-01 2021-04-01 Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis Active CN113219087B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110358321.9A CN113219087B (en) 2021-04-01 2021-04-01 Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110358321.9A CN113219087B (en) 2021-04-01 2021-04-01 Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis

Publications (2)

Publication Number Publication Date
CN113219087A CN113219087A (en) 2021-08-06
CN113219087B true CN113219087B (en) 2022-11-04

Family

ID=77086358

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110358321.9A Active CN113219087B (en) 2021-04-01 2021-04-01 Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis

Country Status (1)

Country Link
CN (1) CN113219087B (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006297244A (en) * 2005-04-19 2006-11-02 Japan Organo Co Ltd Pretreatment column for ion chromatography unit and its regeneration method, and ion chromatography unit
CN105866232A (en) * 2015-01-19 2016-08-17 上海市食品药品检验所 Methods for determining total amount mercury and valence mercury content in biological tissues and organs

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006297244A (en) * 2005-04-19 2006-11-02 Japan Organo Co Ltd Pretreatment column for ion chromatography unit and its regeneration method, and ion chromatography unit
CN105866232A (en) * 2015-01-19 2016-08-17 上海市食品药品检验所 Methods for determining total amount mercury and valence mercury content in biological tissues and organs

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
HPLC色谱柱性能异常的判别及再生;黄骏雄;《现代仪器》;19961231(第02期);16-18 *
糖醇高效液相色谱柱的再生;李俊平等;《天津化工》;20070331;第21卷(第02期);49-51 *
色谱柱的维护与保养在高压液相色谱中的应用;刘育红等;《中小企业管理与科技(下旬刊)》;20101231(第02期);205 *
高效液相色谱柱清洗与再生方法;刘翻等;《分析测试技术与仪器》;20100630;第16卷(第02期);71-77 *

Also Published As

Publication number Publication date
CN113219087A (en) 2021-08-06

Similar Documents

Publication Publication Date Title
Lee et al. Preconcentration of palladium, platinum and rhodium by on-line sorbent extraction for graphite furnace atomic absorption spectrometry and inductively coupled plasma atomic emission spectrometry
CN101871920A (en) Multistage improvement column for quickly pre-processing and purifying polychlorinated biphenyl in biological sample
McDowall et al. Sample preparation for the HPLC analysis of drugs in biological fluids
Yamamoto et al. Methylcellulose-immobilized reversed-phase precolumn for direct analysis of drugs in plasma by HPLC
CN113219087B (en) Method for recovering and treating reduced inorganic mercury chromatographic peak column efficiency in mercury element form and valence state analysis
CN111537647A (en) Ultra-high performance liquid chromatography analysis method for crimson-cleft-camaldine in flos farfarae
CN109884199B (en) Method for measuring content of flavonoid components in honey
CN111504754A (en) One-step sample purifying tube
CN112903836B (en) Method for determining isopropyl-beta-D-thiogalactopyranoside in-vitro cultured bear gall powder
CN110161169A (en) The rapid detection method of a variety of pharmaceutically active substances in a kind of water environment
CN109030668B (en) High performance liquid phase analysis method of gadoxetic acid disodium intermediate
CN106970178B (en) It is a kind of based on hydrophobic nonionic exchange Solid Phase Extraction cigarette mainstream flue gas in the peculiar N- nitrosamine of tobacco extraction and assay method
JP2021128005A (en) Blood pretreatment method and examination method
CN110687224A (en) Method for measuring triptolide A in tripterygium wilfordii medicinal material and tripterygium wilfordii multi-glycoside tablet prepared from tripterygium wilfordii medicinal material
CN116858978B (en) Method for simultaneously detecting insulin aspart and insulin deglutition and plasma sample processing method thereof
CN113640401B (en) Method for detecting aristolochic acid in soil
CN114878739B (en) Thin-layer identification detection method for determining liquorice and bitter apricot seeds in ephedra, apricot, shigan granules
CN116953128B (en) Analysis method, device and system for common antipyretic, antitussive and pharyngitis treatment medicine components in water body
CN115536542B (en) Preparation method of colchicine
CN110715996B (en) Method for full-automatic online detection of free-state small molecular compound in serum
CN111855833B (en) Analysis method for determining tetracyclic triterpenoid saponin in compound salvia miltiorrhiza preparation based on polymer-coated silica gel bonded filler
CN113533563B (en) Method for simultaneously detecting contents of four components of liver-soothing, stomach-harmonizing and pain-relieving traditional Chinese medicine
CN117288870B (en) Method for establishing fingerprint of Yiling oral liquid
CN108061761A (en) A kind of method for measuring alkaloid component content in blood-nourishing and brain-refreshing alcohol-extracted extract
CN118883742A (en) Method for enriching, separating and measuring content of 5 effective components in liquorice

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant