CN112946056A - Method for detecting aluminum element in alanyl glutamine injection - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 50
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 238000002347 injection Methods 0.000 title claims abstract description 31
- 239000007924 injection Substances 0.000 title claims abstract description 31
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 title claims abstract description 23
- 108010044940 alanylglutamine Proteins 0.000 title claims abstract description 22
- 229960002648 alanylglutamine Drugs 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 47
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910017604 nitric acid Inorganic materials 0.000 claims abstract description 13
- 230000006920 protein precipitation Effects 0.000 claims abstract description 8
- 238000007865 diluting Methods 0.000 claims abstract description 6
- 238000001914 filtration Methods 0.000 claims abstract description 6
- 239000007789 gas Substances 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 10
- 238000009616 inductively coupled plasma Methods 0.000 claims description 6
- 238000010790 dilution Methods 0.000 claims description 5
- 239000012895 dilution Substances 0.000 claims description 5
- 239000003085 diluting agent Substances 0.000 claims description 3
- 239000001307 helium Substances 0.000 claims description 2
- 229910052734 helium Inorganic materials 0.000 claims description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims 2
- 229910052786 argon Inorganic materials 0.000 claims 1
- 239000012159 carrier gas Substances 0.000 claims 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 abstract description 10
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 abstract description 10
- 238000012795 verification Methods 0.000 abstract description 10
- 238000011084 recovery Methods 0.000 abstract description 5
- 239000002253 acid Substances 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 4
- 238000011002 quantification Methods 0.000 abstract description 3
- 238000002133 sample digestion Methods 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 28
- 239000000523 sample Substances 0.000 description 26
- 229910052782 aluminium Inorganic materials 0.000 description 16
- 239000012488 sample solution Substances 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 239000011550 stock solution Substances 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000003916 acid precipitation Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229910002804 graphite Inorganic materials 0.000 description 2
- 239000010439 graphite Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 108010016626 Dipeptides Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- -1 aluminum salt Chemical class 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000000120 microwave digestion Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 235000016236 parenteral nutrition Nutrition 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/626—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode using heat to ionise a gas
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- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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Abstract
The invention discloses a method for detecting aluminum element in alanyl glutamine injection, which is used for measuring the content of aluminum salt in alanyl glutamine injection by inductively coupled plasma mass spectrometry. And optimizing ICP-MS technical parameters by adopting a standard addition method according to an HMI-He mode, diluting a sample by using nitric acid, directly injecting a sample after protein precipitation and filtration, detecting the content of aluminum salt in alanyl glutamine injection, and carrying out methodology verification. The aluminum element is in a good linear relation within the range of 0-30 mu g/L, the correlation coefficient is 0.999, the detection limit is 0.88 mu g/L, the quantification limit is 2.9 mu g/L, the average recovery rate is 104.0%, and the relative standard deviation of two detection values of the stability experiment is 2.3%. The method has the advantages of simple and accurate operation, good precision, accuracy, repeatability and stability, direct sample injection after sample acid protein precipitation, no complicated steps such as sample digestion treatment and the like, and can be used for measuring the content of aluminum salt in alanyl glutamine injection.
Description
Technical Field
The invention relates to the technical field of chemical analysis and quantitative detection, in particular to a quantitative detection method for aluminum metal element in alanyl-glutamine injection.
Background
Glutamine (Gln) is an amino acid constituting a protein, is one of energy substances necessary for secretion, proliferation and function maintenance of lymphocytes, has an important immunoregulatory effect, and is very important for muscle strength, gastrointestinal mucosa and repair and protection of damaged tissues of a human body. However, it has many limitations in clinical application due to its poor water solubility and heat resistance. Therefore, the dipeptide alanyl glutamine containing Gln is mainly used for supplementing glutamine to patients clinically, can be decomposed into glutamine and alanine by organisms after injection, is used as an parenteral nutrient, and has high absorption and utilization rate and no toxic or side effect.
Research shows that most parenteral nutrient solutions have residual aluminum element, which has great influence on the health of human bodies, especially children of low age. FDA in the united states has required manufacturers to control the quality of aluminum in parenteral nutrition since 2004 and requires explicit specification of aluminum content on the packaging of products.
The detection of the aluminum element is a technical problem, because the aluminum element widely exists in nature, including air, water and the inner wall of a container, the detection of the aluminum element is easy to be interfered by the outside, and the requirements on the sensitivity of a detection instrument and an experimental environment method are high.
At present, the commonly used sample pretreatment methods for measuring the metal element aluminum in the organic sample mainly comprise a digestion method and an acid precipitation method, and because the residual amount of aluminum in the alanyl glutamine injection is very low, the digestion method has the disadvantages of tedious process, long time consumption and complex digestion equipment cleaning and maintenance, and meanwhile, the aluminum in the environment is easily introduced in the operation process of microwave digestion, so that the reproducibility of the aluminum content is poor.
The main component of the alanyl-glutamine injection is dipeptide, and when the aluminum residue of the alanyl-glutamine injection is measured by adopting an atomic absorption method, a sample can generate a plurality of impurities attached to a graphite tube after being combusted, so that the invention result is greatly influenced. Impurities are not easy to remove, a new graphite tube needs to be frequently replaced, and the loss of the invention is increased. At present, no method for determining aluminum element in alanyl-glutamine injection by utilizing ICP-MS is clear at home. The invention aims to establish an accurate and feasible HMI-ICP-MS detection method for measuring aluminum in alanyl glutamine injection and carrying out methodology verification.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides an accurate and feasible HMI-ICP-MS detection method for determining aluminum in alanyl glutamine injection and carrying out methodology verification.
The retrieval method comprises the following steps: and establishing an inductively coupled plasma mass spectrometry (ICP-MS) to determine the content of aluminum salt in alanyl glutamine injection. And optimizing ICP-MS technical parameters by adopting a standard addition method according to an HMI-He (atomizing gas dilution-helium collision) mode, diluting a sample by using nitric acid, directly feeding a sample after protein precipitation and filtration, and detecting the content of aluminum salt in alanyl glutamine injection.
(1) Determination of sample treatment method:
because the Agileng7800 inductively coupled plasma mass spectrometer can accept 100% liquid organic samples to be directly injected, the method combines an acid protein precipitation method, adopts nitric acid to dilute the samples, adopts alanyl glutamine injection to generate flocculent precipitates, filters the precipitates and directly injects the samples, and the detection result is closer to a real value, so the samples are pretreated by adopting an acid protein precipitation technology.
(2) Determination of tuning mode:
the Agileng7800 type inductively coupled plasma mass spectrometer is provided with a No Gas mode and a He collision mode, and generally, elements with small mass numbers are detected by the No Gas mode, but the result measured by the method adopting the No Gas mode is not ideal, and the reason is presumed to be that after the sample is atomized by an atomizer, the sample is diluted by a nitric acid precipitation mode, a solution contains a large amount of nitrogen elements, the nitrogen elements are just interfered with organic carbon by CN, and the nitrogen elements and the detected AL elements are not effectively distinguished and enter a detection pool together through a quadrupole rod to interfere the determination of the aluminum elements, so the He collision mode is adopted to eliminate the influence of the ions with small mass numbers on the AL ions through collision. Because the alanyl glutamine injection liquid matrix is complex and has certain influence on the determination of AL, the atomized sample is further diluted by adding a path of diluent gas (HMI-He mode), and the better detection effect is achieved by adjusting the ratio of the flow of the atomizer to the flow of the diluent gas.
The invention provides a method for detecting aluminum element in alanyl glutamine injection, which comprises the following steps:
(1) diluting the sample with nitric acid;
(2) directly feeding a sample after protein precipitation and filtration;
the inductively coupled plasma mass spectrometer adopts a He collision mode, adds one path of dilution gas to further dilute the atomized sample, and adjusts the flow ratio of the atomizer to the dilution gas.
Further, the detection method adopts a standard addition method.
Has the advantages that: the detection limit is 0.88 mug/L, the quantification limit is 2.9 mug/L, the average recovery rate is 104.0 percent, and the relative standard deviation of the two detection values of the stability experiment is 2.3 percent. The method has the advantages of simple and accurate operation, good precision, accuracy, repeatability and stability, direct sample injection after sample acid protein precipitation, no complicated steps such as sample digestion treatment and the like, and can be used for measuring the content of aluminum salt in alanyl glutamine injection.
The aluminum element is in the concentration range of 0-30 mug/L, the correlation coefficient is larger than 0.995, the verification requirement is met, and the aluminum element response value and the concentration form a good linear relationship.
The method can measure the quantitative limit and the detection limit of the aluminum element, and the limit requirement can be met. The RSD% of the repeatability detection value measured by the method is 1.8%, the method meets the verification requirement, and the method is good in precision.
The recovery rate of the aluminum element in the accuracy experiment is 100.8-107.3%, the average value is 104.06%, and the RSD value is 2.2%, which shows that the accuracy of the method is good.
6 parts of standard sample solution is prepared in parallel, the RSD% value of the aluminum element detection value is 0.4%, the verification requirement is met, and the repeatability of the method is good.
6 parts of standard-added test sample solution are prepared by the same method on different analysts and different dates, the RSD percent of 12 detection values is calculated to be 1.3 percent, and the method meets the verification requirement, and the intermediate precision of the method is good.
The relative standard deviation of the two detection values of the stability experiment is 2.3 percent, which shows that the method detects the aluminum element and has better stability.
Detailed Description
The present invention will be further described with reference to the following examples for facilitating understanding of those skilled in the art, and the description of the embodiments is not intended to limit the present invention.
Example 1
Setting of the Agileng7800 inductively coupled plasma mass spectrometer:
TABLE 1 ICP-MS Instrument parameters
Instrument and reagent
Ultra pure water machine (millipore corporation, usa) model: Milli-Q A10; inductively coupled plasma mass spectrometer (agilent) model number: 7800; standard solution of aluminum single element (China institute of metrology science, 100. mu.g/mL, lot number: 14125); high-purity nitric acid: suzhou Jingrui chemical Co., Ltd
Preparation of the solution
25mL of nitric acid (68% -70%) is measured and quantitatively diluted to 500mL with water to obtain a 5% nitric acid solution.
Aluminum standard stock solution
0.5mL of an aluminum single element standard solution (concentration: 100.0. mu.g/mL) was precisely measured, placed in a 50mL measuring flask, graduated with a 5% nitric acid solution, and shaken up to give a standard stock solution (concentration: 1.0. mu.g/mL).
Standard addition method test solution
Precisely measuring 6 parts of the product (7.5 mL), respectively placing in 15mL plastic centrifuge tubes, precisely adding 0, 75 μ L, 150 μ L, 225 μ L, 300 μ L and 450 μ L of aluminum standard stock solution (1.0 μ g/mL), respectively fixing the volume to the scale with 5% nitric acid, shaking, and filtering with 0.22 μm water film to obtain the concentration values of the added standard aluminum element of 0, 5 μ g/L, 10 μ g/L, 15 μ g/L, 20 μ g/L and 30 μ g/L, respectively.
Determination of L in test sample for aluminum salt
Taking a test sample solution, and carrying out sample injection detection. And drawing a standard curve solution by taking the CPS value of the element to be detected as a vertical coordinate and the detection concentration of the element to be detected as a horizontal coordinate. And extending and intersecting the standard curve on the abscissa, wherein the content corresponding to the distance between the intersection point and the origin is the content of the element to be detected in the sample. And calculating the content of the element to be detected.
Content of aluminum salt (. mu.g/L) × dilution factor of test solution
Verification of aluminum salt inspection methodology
The methodology verification is carried out on the aluminum salt inspection method in the alanyl glutamine injection according to the verification guiding principle of the quality standard analysis method of the four-part general rule 9101 in 2015 edition of Chinese pharmacopoeia.
The method detects the content of aluminum in alanyl glutamine injection by an ICP-He mode through an ICP-MS technical means and a standard addition method, and verifies the detection method, including linearity and range, detection limit, quantitative limit, precision, accuracy, repeatability, intermediate precision and stability.
Linear and range test
Taking a sample solution, carrying out sample injection analysis, carrying out linear regression by taking CPS of the aluminum element as a vertical coordinate and the concentration (mu g/L) of the detected aluminum element as a horizontal coordinate, and obtaining test results shown in Table 2.
TABLE 2 Linear and Range test results
Table 2 Linearity and range test results
Note: instrument automatic calculation linear equation and correlation coefficient
Limit of detection (LOD), limit of quantitation (LOQ) tests
Taking a sample solution (not added with a standard solution) as a blank sample, and continuously injecting the sample into 11 needles. The 3-fold standard deviation 3SD of the detected concentration of the element to be detected is calculated as the detection Limit (LOD), and the 10-fold standard deviation 10SD of the detected concentration is calculated as the quantification Limit (LOQ). The results are shown in Table 3.
TABLE 3 detection limit, quantitation limit test results (μ g/L)
Table 3 Results of detection limit and quantitative limit(μg/L)
Precision test
The sample solution with standard aluminum element concentration of 5.0 mug/L is taken and continuously measured for 6 times, and the RSD percent of the aluminum element detection value is calculated. The test results are shown in Table 4.
TABLE 4 results of precision test
Table 4 Prec i sion test results
Accuracy test
Precisely measuring 7.5mL (9 parts) of a sample, respectively placing the sample into a 15mL plastic centrifuge tube, respectively precisely adding 150, 300 and 450 μ L of aluminum standard stock solutions (1.0 μ g/mL), respectively diluting to a scale with 5% nitric acid, shaking up, and filtering with a 0.22 μm water film to obtain the concentration values of the added standard aluminum element of 10, 20 and 30 μ g/L. And (5) sample injection detection and calculation of recovery rate, and the method is used for inspecting the accuracy of the method. The test results are shown in Table 5.
TABLE 5 accuracy test results
Table 5 Accuracy test results
The calculation formula is that percent recovery is (detection concentration-test article concentration) ÷ addition concentration is multiplied by 100
Repeatability test
According to the preparation method under the accuracy item, 6 parts of sample solution with the standard concentration of 20 mug/mL is prepared, sample injection detection is carried out, RSD% of 6 detection values is counted, the RSD% is used for inspecting the repeatability of the method, and the test result is shown in table 6.
TABLE 6 results of the repeatability tests
Table 6 Repeatability test results
Intermediate precision test
6 parts of sample solution with the standard concentration of 20 mug/mL are prepared by different analysts and different dates according to the same method of repeatability test, sample introduction detection is carried out, RSD% of 6 detection values is counted, RSD% of 12 detection values is counted by combining the result of the repeatability test, the RSD% is used for investigating the intermediate precision of the method, and the test result is shown in Table 7.
TABLE 7 results of intermediate precision test
Table 7 Intermediate precision test results
Stability test
And taking a sample solution to be tested by a standard addition method, carrying out sample injection detection, then standing at room temperature for 7 hours, and carrying out sample injection detection again. The two detection values are respectively as follows: 20.23. mu.g/L, 19.52. mu.g/L, relative standard deviation 2.3%, indicating that the detection method is good.
It will be understood by those skilled in the art that, unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the prior art and will not be interpreted in an idealized or overly formal sense unless expressly so defined herein.
It should be understood that the above detailed description of the technical solution of the present invention with the help of preferred embodiments is illustrative and not restrictive. On the basis of reading the description of the invention, a person skilled in the art can modify the technical solutions described in the embodiments, or make equivalent substitutions for some technical features; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (7)
1. A method for detecting aluminum element in alanyl glutamine injection is characterized in that: the method comprises the following steps:
(1) diluting the sample with nitric acid;
(2) directly feeding a sample after protein precipitation and filtration;
the inductively coupled plasma mass spectrometer adopts a He collision mode, adds one path of dilution gas to further dilute the atomized sample, and adjusts the flow ratio of the atomizer to the dilution gas.
2. The detection method of claim 1, characterized in that: the detection method adopts a standard addition method.
3. The detection method of claim 1, characterized in that: the carrier gas and the diluent gas are argon.
4. The detection method of claim 1, characterized in that: collision cell parameters helium gas flow rate: 3.7 mL/min.
5. The detection method of claim 1, characterized in that: the nitric acid solution was 5%.
6. The detection method of claim 1, characterized in that: atomizing: 0.60L/min.
7. The detection method of claim 1, characterized in that: diluting gas: 0.40L/min.
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Application publication date: 20210611 |