CN102590416A - Method for extracting negative ions from extract medicine by means of matrix solid phase dispersion and ion chromatographic detection method - Google Patents

Abstract

The invention relates to a matrix solid-phase dispersion extraction method for anion in medicine and ion chromatographic detection method. The herbal drug raw material is ground and mixed together with the solid-phase extraction agent graphitized carbon and SPE carbon 18, and then subjected to solid-phase extraction. Rinse, and the eluent is the extract containing anions. Dilute the eluent with deionized water to the required concentration, and extract the solution and filter it with a 0.22 μm filter membrane, and the obtained filtrate is the solution to be tested. Tested with an ion chromatographic analyzer, the measured seven inorganic anions F ^- , Cl ^- , NO ₂ ^- , Br ^- , NO ₃ ^- , PO ₄ ^3- and SO ₄ ^2- have a good range of 0.4 to 12 mg/L Linear relationship, the detection limit is 1.20-5.00 μg/L, the recovery rate of the standard addition is 70%-98%, and the precision is 2.09%-5.87%.

Description

A method for detecting anions in a matrix solid-phase dispersion extraction drug and ion chromatography

1. Technical field

The invention relates to a matrix solid-phase dispersion extraction method for anions in medicine and an ion chromatography detection method, belonging to the technical field of chemical extraction and ion chromatography detection.

2. Background technology

Matrix solid-phase dispersion technology (MSPD, matrix solid-phase dispersion) is a rapid sample processing technology proposed by Professor Barker of Louisiana State University in the United States in 1989 and given a theoretical explanation. Its advantage is that it concentrates the sample homogenization, tissue cell lysis, extraction, purification and other processes in the traditional sample pretreatment, and does not need to perform tissue homogenization, precipitation, centrifugation, pH adjustment and sample transfer. loss. It is a simple, efficient and practical extraction and purification method, which is suitable for the extraction and purification of various molecular structures and polar pesticide residues, improves the analysis speed, reduces the amount of reagents, and is suitable for automatic analysis.

At present, the development of herbal medicines in our country is in the ascendant period, so the qualitative and quantitative analysis of chemical components is extremely important in the research process of herbal medicines. The combination of various chromatography and mass spectrometry is currently the mainstream of qualitative and quantitative analysis of organic chemical components. Wang Guangji et al. used high performance liquid chromatography-ion trap-time-of-flight tandem mass spectrometry to quickly screen and identify chemical components in complex systems of traditional Chinese medicine (public number : CN 101382525A), a quick and easy method is provided for the analysis of organic components in herbal medicines, but there are few reports on the analysis of inorganic components in herbal medicines.

Due to the wide variety of herbal medicines with obvious curative effects and low side effects, their clinical application has a long history. However, the complex organic and inorganic chemical components in herbal medicines have not yet been elucidated. This is the medicinal substance of herbal medicines. Major bottlenecks in basic, quality control, and pharmacological studies of action. At present, the development of herbal medicines in our country is still in the ascendant period. Therefore, the qualitative and quantitative analysis of chemical components is extremely important in the research process of herbal medicines. The combination of various chromatography and mass spectrometry is currently the mainstream of qualitative and quantitative analysis of organic chemical components. Wang Guangji et al. used high performance liquid phase-ion trap-time-of-flight tandem mass spectrometry to quickly screen and identify chemical components in complex systems of traditional Chinese medicine (published No.: CN 101382525A) provides a fast and easy method for the analysis of organic components in medicines, but there are few reports on the analysis of non-metallic elements in inorganic components in medicines.

The known matrix solid-phase dispersion technique (MSPD) is mainly used in pesticide residue detection (Analytical Chemistry, 2005, 33 (9): 1318-1320), has not yet been reported in ion chromatography analysis and determination, and its main principle is to combine adsorption filler with Put the sample into the mortar and grind at the same time to obtain a semi-dry mixture, then let it dry or dry it at low temperature, then pack the column, and rinse the column with a solvent to separate the analyte from the impurity substance. The patents of Zhao Haixiang etc. relate to the matrix solid-phase dispersion-high performance liquid chromatography determination method (publication number: CN101241114A) of sulfa drugs in animal-derived food, but what it uses is still the traditional matrix solid-phase dispersion technology.

Due to the traditional MSPD technology, due to the small amount of adsorption at the bottom, when using a solvent to elute the ions to be measured into the solvent, some impurity ions and organic matter are eluted together with the eluting solvent, and the peak shape of the obtained ion chromatogram is disordered, which interferes with the determination. ; At the same time, it is easy to cause damage to the ion chromatography column system. These all limit the wide application of this method in ion chromatography analysis and determination.

3. Contents of the invention

The object of the present invention is to provide a kind of matrix solid-phase dispersion to extract the anion in the medicine and ion chromatographic detection method, the botanical medicine raw material and the solid-phase extraction agent graphitized carbon and SPE carbon 18 are ground and mixed together and then removed by solid-phase extraction. Ionized water rinses, and the rinse solution is the extract solution containing anions. Dilute the eluent with deionized water to the required concentration, and extract the solution and filter it with a 0.22 μm filter membrane, and the obtained filtrate is the solution to be analyzed. Ion Chromatography Analyzer Test

The present invention comprises the following steps:

1. Extraction of anion-containing liquid in herbal raw materials

Wash the botanical raw materials with water and then rinse with deionized water for 3 to 5 times, dry in the sun or at no more than 60°C until the water content is 3-5%, crush to a particle size of 0.15 to 0.25mm, and put it in a dryer in spare.

Weigh 0.05g-0.5g of the herbal drug raw material with the above particle size, grind and mix them together for 3-6min with the solid-phase extractant graphitized carbon 0.04-0.8g and carbon 18 0.3-3g, then dry naturally until the water content is between 3- 5%, to obtain the mixture of raw material drug and solid phase extractant, take 0.4～0.5g of the mixture and put it into the solid phase extraction column, fill in 0.02～0.03g of carbon eighteen at the bottom of the solid phase extraction column, add 8 ~9mL of deionized water is used to wash the mixed raw material, and then it is separated and purified in the solid phase extraction column. The eluent collected from the lower part of the column is the extract containing anions in the herbal drug raw material. The extract was diluted with deionized water to the required concentration, and 3 mL of the solution was extracted and filtered through a 0.22 μm filter membrane, and the obtained filtrate was the extract to be tested.

The structure of the solid-phase extraction column is shown in Figure 1, which includes a column tube 1, a sieve plate 2, a mixture 3 of a solid-phase extraction agent and a raw material drug, and SPE carbon 18-4.

2. Simultaneous determination and analysis of inorganic anion components

(1) Test conditions of ion chromatography analyzer

The separation column of the ion chromatograph is 250mm×4mm, the guard column is 50mm×4mm, the current of the automatic regeneration suppressor is 24mA; the column temperature of the chromatographic column is 30°C, and the cell temperature of the temperature-controlled conductivity detector is 35°C.

The analysis parameters set in the present invention: sample size is 25 μ L (extract to be detected), and flow rate is 1.5mL/min, and eluent is the sodium bicarbonate of concentration 2.8mmol/L and concentration is 3.5mmol/L mixture of sodium carbonate.

Take 1 mL of the extract and perform anion detection according to the ion chromatography analysis flow chart in Figure 2 under the parameters set above. That is: the eluent in the eluent bottle 1 is transported to the injection valve 3 through the pump 2, and together with the extract to be analyzed, it is transported to the guard column 4 and the separation column 5 to separate different anions and pass through the large suppressor Reduce the background conduction, improve the sensitivity, and use the detection pool 7 and the detector 8 to detect the negative ions.

(2) Results: The measured seven inorganic anions F ^- , Cl ^- , NO ₂ ^- , Br ^- , NO ₃ ^- , PO ₄ ^3- and SO ₄ ^2- have a good linear relationship in the range of 0.4 to 12 mg/L , the detection limit is 1.20～5.00μg/L. The recovery rate of the standard addition was 70%-98%, and the precision was 2.09%-5.87%.

Advantages over known techniques

1. The separation and purification of anions in the herbal medicine extract can be completed at the same time. Improve the packing layer, use double-layer dispersant (the upper layer is a mixture of raw material medicine and solid phase extraction agent, and the lower layer is SPE carbon 18) technology to extract and purify, that is, put a certain amount of adsorbent on the bottom sieve plate, Avoiding incomplete purification during elution from damaging the ion chromatography separation column and interfering with the determination;

2. The ion chromatography detection method of matrix solid-phase dispersion extraction of 7 common inorganic anions can be accurately and comprehensively analyzed under the best chromatographic conditions;

3. This method has no strict restrictions on the types of herbal medicines.

4. Description of drawings

Figure 1 is a structural diagram of a solid phase extraction column, in which 1 is a column tube, 2 is a sieve plate, 3 is a mixture of a solid phase extraction agent and a raw material drug, and 4 is SPE carbon 18.

Fig. 2 is the ion chromatographic analysis flowchart of inorganic anion, among the figure 1 is eluent, 2 is pump, 3 is injection valve, 4 is guard column, 5 is separation column, 6 is suppressor, 7 is detection cell, 8 is a detector.

5. Specific implementation

Embodiment one:

1 Extraction and Detection of Inorganic Anions in Kingsberry Raw Materials

Rinse the raw material of Goldenseal Plum with water and then rinse it with deionized water three times, dry it at 60°C until the water content is 3%, grind it with a pulverizer until the particle size is 0.15-0.25mm, and put it in a desiccator for later use.

After weighing 0.1g of the above-mentioned particle size of the Kingsberry bulk drug, 0.8g of graphitized carbon and 0.3g of SPE carbon eighteen 0.3g, they are jointly ground and mixed for 3min, and then naturally dried to a water content of 3%, to obtain the bulk drug and solid phase extractant 0.5 g of the mixture is packed into a solid-phase extraction column, the bottom of which has been filled with 0.02 g of carbon eighteen, and 8 mL of deionized water is added from the upper part of the extraction column to rinse the raw material, and the eluent is collected from the lower part of the extraction column to obtain Anion-containing extract. Dilute the eluent with deionized water to 100 mL, and extract 3 mL of the solution and filter it with a 0.22 μm filter membrane, and the obtained filtrate is the extract to be tested.

2. Ion chromatography detection

(1) Test conditions of ion chromatography analyzer

Using Dionex ICS1500 ion chromatograph from American Diane Company, Chromeleon6.0 chromatographic workstation, IonPac AS14 separation column 250mm×4mm, IonPac AG14 guard column 50mm×4mm, ASRS-300 automatic regeneration suppressor current 24mA, column temperature 30°C, The detection temperature of the DS6 temperature-controlled conductivity detector is 35°C, the injection volume is 25 μL, the flow rate is 1.5mL/min, and the eluent is a mixture of 2.8mmol/L sodium bicarbonate and 3.5mmol/L sodium carbonate.

Take 1 mL of the extract to be analyzed and perform anion analysis according to the ion chromatography analysis flow chart in Figure 2 under the analysis parameters set above.

(2) Measurement results: the measured seven inorganic anions F ^- , Cl ^- , NO ₂ ^- , Br ^- , NO ₃ ^- , PO ₄ ^3- and SO ₄ ^2- have good linearity in the range of 0.4 to 12 mg/L The detection limit is 2 μg/L. Its spike recovery was 80% with a precision of 3%.

Example 2: Extraction and Detection of Inorganic Anions in Bamboo Leopard Orchid Bulk Drugs

1. Dry the bulk drug until the water content is 5%, crush it to a particle size of 0.15-0.25 mm, put it in a desiccator for standby; weigh 0.12 g of the bulk drug with the above particle size and 0.1 g of solid-phase extractant graphitized carbon and SPE C18 0.7g, grind and mix together for 6min, then dry naturally to obtain a mixture of raw material drug and solid phase extraction agent, take 0.4g of the mixture and put it into a solid phase extraction column (the bottom has been filled with 0.03g C18) , add 9mL deionized water from the upper part of the solid phase extraction column to rinse the raw material, collect the eluent from the lower part of the column to obtain an anion-containing extract; dilute the eluent to 100m with deionized water, and extract 3mL of the solution for use Filter through a 0.22 μm filter membrane to obtain the extract to be tested.

2. Ion chromatography analysis

(1) Ion chromatographic analyzer test conditions: the same as Example 1. Take 1mL of the extract and perform anion analysis according to the ion chromatography analysis flow chart in Figure 2 under the analysis parameters set above.

(2) Measurement results: the measured seven inorganic anions F ^- , Cl ^- , NO ₂ ^- , Br ^- , NO ₃ ^- , PO ₄ ^3- and SO ₄ ^2- have good linearity in the range of 0.4 to 12 mg/L The detection limit is 5.μg/L. Its spiked recovery was 91% with a precision of 4.6%.

Claims (3)

1. Anion and ion chromatography detection method in a matrix solid phase dispersion extraction medicine, it is characterized in that: it comprises the following steps: 1) Extraction of anion-containing liquid in herbal medicine raw materials Wash the botanical raw materials with water and then rinse with deionized water for 3 to 5 times, dry in the sun or at no more than 60°C until the water content is 3-5%, crush to a particle size of 0.15 to 0.25mm, and put it in a dryer Standby; weigh 0.05g-0.5g of the herbal drug raw material with the above particle size, grind and mix with the solid-phase extractant graphitized carbon 0.04-0.8g and carbon eighteen 0.3-3g together for 3-6min, then dry naturally to the water content At 3-5%, the mixture of botanical drug raw material and solid phase extractant is obtained, 0.4～0.5g of the mixture is loaded into the solid phase extraction column, and 0.02～0.03g of carbon eighteen is filled in the bottom of the solid phase extraction column, and the extraction column Add 8-9mL deionized water to the upper part of the column to wash the mixed raw material, and then separate and purify it in the solid phase extraction column. Dilute the extract with water to the required concentration, and extract 3mL of the solution and filter it with a 0.22μm filter membrane, and the obtained filtrate is the extract to be tested; The structure of the solid-phase extraction column includes a column tube 1, a sieve plate 2, a mixture 3 of a solid-phase extraction agent and a raw material drug, and SPE carbon octave 4; 2) Simultaneous determination and analysis of inorganic anion components (1), ion chromatography analyzer test conditions Ion chromatography separation column 250mm×4mm, guard column 50mm×4mm, automatic regeneration suppressor current 24mA, column temperature 30°C, temperature-controlled conductivity detector cell temperature 35°C; injection volume 25μL, flow rate 1.5mL/ min, the eluent is a mixture of sodium bicarbonate with a concentration of 2.8mmol/L and sodium carbonate with a concentration of 3.5mmol/L; (2), test process: the eluent in the eluent bottle (1) is delivered to the injection valve (3) through the pump (2), and is delivered to the guard column (4) and the separation column (5) together with the extract to be tested ), different anions are separated, the background conduction is reduced by the suppressor (6), the sensitivity is improved, and the anions are detected by the detection cell (7) and the detector (8). 2. The method for detecting anions and ion chromatographic detection in matrix solid-phase dispersion extraction medicine according to claim 1, characterized in that: said botanical medicine is Hypericum or Phyllostachys japonica. 3. The method for detecting anions and ion chromatograms in solid-phase dispersion extraction of the matrix according to claim 1, characterized in that: the ion chromatograph is an ion chromatograph of Diane Corporation in the United States.

Images