CN110988154A - Method for detecting glycine content in hemodialysis solution - Google Patents
Method for detecting glycine content in hemodialysis solution Download PDFInfo
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- CN110988154A CN110988154A CN201911122124.6A CN201911122124A CN110988154A CN 110988154 A CN110988154 A CN 110988154A CN 201911122124 A CN201911122124 A CN 201911122124A CN 110988154 A CN110988154 A CN 110988154A
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- G—PHYSICS
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- 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
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Abstract
The invention provides a method for detecting glycine content in hemodialysis solution, which comprises the step of using 90-98% by volume of 0.05mol/L potassium dihydrogen phosphate solution and 2-10% by volume of methanol as mobile phases, and measuring glycine in an Agilent 1260 high performance liquid chromatograph with C18 as a chromatographic column. The invention adopts the conventional C18 chromatographic columns in all laboratories, and the preparation process of the mobile phase is extremely simple, the acquisition time is short, and the analysis cost is saved.
Description
Technical Field
The invention belongs to the field of methods for detecting glycine content in hemodialysis solution, and particularly relates to a method for detecting glycine content in hemodialysis solution.
Background
Glycine is the simplest amino acid in the 20 members of the amino acid series, is also called aminoacetic acid, is an essential amino acid for human body, has both acidic and basic functional groups in the molecule, and is a strong electrolyte in aqueous solution. The glycine has high solubility in a strong polar solvent, is basically insoluble in a non-polar solvent, has high boiling point and melting point, and can be made to present different molecular forms through the adjustment of the acidity and the alkalinity of an aqueous solution.
Since the side bond of glycine is a hydrogen atom, it can occupy space that other amino acids cannot occupy, for example, as an amino acid in a collagen helix. At normal temperature, the amino acid is white crystal or light yellow crystalline powder, and has unique sweet taste, and can alleviate acid and alkali taste, mask bitter taste of saccharin added in food, and enhance sweet taste. The synthesis process of the glycine is more, three methods of chloroacetic acid ammonolysis, Scherrer's method and hydantoin method are needed, and the components of the glycine mother liquor synthesized by the three methods are complex, so that certain difficulty is brought to accurate detection of the glycine content in the mother liquor.
In the prior art, various methods for analyzing amino acids have been disclosed, such as liquid chromatography, amino acid analyzer method, photometry, formaldehyde titration method and kjeldahl method. The liquid chromatography generally uses a scheme of pre-column derivatization, namely firstly derivatizing glycine, then separating reactants on an analytical column for detection, and indirectly reacting the content of the glycine. Although pre-column derivatization allows a relatively free choice of reaction conditions, there are no reaction kinetic limitations; moreover, the derivatized by-products can be pretreated to reduce or eliminate interference, easily allow multi-step reactions, have more derivatization reagents to be selected, and do not require complicated instrumentation, but the by-products formed by derivatization can cause great difficulties in chromatographic separation, and impurities or interfering peaks are easily introduced during derivatization, resulting in sample loss. In addition, derivatization reagents are expensive, complicated to operate and not suitable for determining one amino acid.
The Kjeldahl method has the greatest advantages of high accuracy, but because the reagent consumption is large, the measurement period is long, and other nitrides cannot be contained in the sample, otherwise, the result is easy to be inaccurate.
In summary, a glycine detection method with simple operation is needed.
Disclosure of Invention
In order to solve the technical problem, the invention provides a method for detecting the content of glycine in hemodialysis solution, which comprises the steps of using 90-98% by volume of 0.05mol/L potassium dihydrogen phosphate solution and 2-10% by volume of methanol as mobile phases, and measuring the glycine in an Agilent 1260 high performance liquid chromatograph with C18 as a chromatographic column.
Preferably, the potassium dihydrogen phosphate solution with the concentration of 0.05mol/L comprises sodium heptane sulfonate with the concentration of 0.05mol/L, and phosphoric acid is used for adjusting the pH value to 3.
Preferably, the concentration of 0.05mol/L potassium dihydrogen phosphate solution accounts for 98% by volume, the methanol accounts for 2% by volume, and the mobile phase is eluted at equal intervals for 10 min.
Preferably, the column temperature of the hplc at the time of measuring glycine is set to 25 ℃.
Preferably, the wavelength of the HPLC at the time of measuring glycine is 214 nm.
Preferably, the sample amount of the high performance liquid chromatograph for measuring glycine is set to 20 μ L.
Preferably, the flow rate of the HPLC chromatograph for measuring glycine is set to 1 mL/min.
The invention has the beneficial effects that: the invention adopts the conventional C18 chromatographic columns in all laboratories, and the preparation process of the mobile phase is extremely simple, the acquisition time is short, and the analysis cost is saved.
Detailed Description
The invention is further described below:
example (b):
a method for detecting glycine content in hemodialysis solution comprises measuring glycine in Agilent 1260 high performance liquid chromatograph with C18 as chromatographic column by using 90-98% potassium dihydrogen phosphate solution with concentration of 0.05mol/L and 2-10% methanol as mobile phase.
Specifically, the potassium dihydrogen phosphate solution having a concentration of 0.05mol/L contains sodium heptanesulfonate having a concentration of 0.05mol/L, and the pH is adjusted to 3 using phosphoric acid.
Specifically, the volume of the potassium dihydrogen phosphate solution with the concentration of 0.05mol/L accounts for 98 percent, the volume of the methanol accounts for 2 percent, and the mobile phase is eluted with the same degree, and the running time is 10 min.
Specifically, the column temperature of the hplc during the determination of glycine was set to 25 ℃.
Specifically, the wavelength of the HPLC at the time of measuring glycine is 214 nm.
Specifically, the sample size of the HPLC chromatograph for glycine determination is set to 20. mu.L.
Specifically, the flow rate of the HPLC chromatograph for glycine determination was set to 1 mL/min.
Example 1
Selecting 98% by volume of 0.05mol/L potassium dihydrogen phosphate solution and 2% by volume of methanol as mobile phases, eluting 98% by volume of 0.05mol/L potassium dihydrogen phosphate solution and 2% by volume of methanol at the same flowing temperature, operating for 10min, and measuring glycine in an Agilent 1260 high performance liquid chromatograph with C18 as a chromatographic column, the column temperature of 25 ℃, the wavelength of 214nm, the sample injection amount of 20 mu L and the flow rate of 1 mL/min.
It should be noted that, in this document, moreover, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A method for detecting the content of glycine in hemodialysis solution is characterized by using 90-98% by volume of 0.05mol/L potassium dihydrogen phosphate solution and 2-10% by volume of methanol as mobile phases and measuring the glycine in an Agilent 1260 high performance liquid chromatograph with C18 as a chromatographic column.
2. The method according to claim 1, wherein the 0.05mol/L potassium dihydrogen phosphate solution contains sodium heptane sulfonate with a concentration of 0.05mol/L, and the pH is adjusted to 3 by using phosphoric acid.
3. The method according to claim 1, wherein the concentration of potassium dihydrogen phosphate solution is 98% by volume and the concentration of methanol is 2% by volume, and the elution is carried out at a constant flow rate for 10 min.
4. The method for detecting glycine content in hemodialysis solution according to claim 1, wherein the HPLC column temperature for measuring glycine is set to 25 ℃.
5. The method for detecting glycine content in hemodialysis solution according to claim 1, wherein the wavelength of HPLC for measuring glycine is 214 nm.
6. The method for detecting glycine content in hemodialysis solution of claim 1, wherein the HPLC sample size for glycine determination is set to 20 μ L.
7. The method for detecting glycine content in hemodialysis solution of claim 1, wherein the flow rate of HPLC for measuring glycine is set to 1 mL/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201911122124.6A CN110988154A (en) | 2019-11-15 | 2019-11-15 | Method for detecting glycine content in hemodialysis solution |
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| CN201911122124.6A CN110988154A (en) | 2019-11-15 | 2019-11-15 | Method for detecting glycine content in hemodialysis solution |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106596799A (en) * | 2017-01-20 | 2017-04-26 | 阳泉煤业（集团）有限责任公司 | Method for detecting glycine and impurities thereof by high performance liquid chromatography |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106596799A (en) * | 2017-01-20 | 2017-04-26 | 阳泉煤业（集团）有限责任公司 | Method for detecting glycine and impurities thereof by high performance liquid chromatography |
Non-Patent Citations (2)
| Title |
|---|
| 孙宝丹等: "HPLC法研究还原型谷胱甘肽降解产物", 《沈阳药科大学学报》 * |
| 薛晓等: "HPLC法同时测定复方甘草酸苷注射液中甘氨酸与盐酸半胱氨酸的含量", 《中国新药杂志》 * |
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Application publication date: 20200410 |