CN112649552A - Method for measuring selenium form by using high performance liquid inductively coupled plasma mass spectrometry - Google Patents
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- 125000003748 selenium group Chemical group *[Se]* 0.000 title claims abstract description 29
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 title claims abstract description 11
- 239000007788 liquid Substances 0.000 title claims abstract description 8
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims abstract description 43
- 239000011669 selenium Substances 0.000 claims abstract description 36
- 229910052711 selenium Inorganic materials 0.000 claims abstract description 33
- RJFAYQIBOAGBLC-BYPYZUCNSA-N Selenium-L-methionine Chemical compound C[Se]CC[C@H](N)C(O)=O RJFAYQIBOAGBLC-BYPYZUCNSA-N 0.000 claims abstract description 18
- RJFAYQIBOAGBLC-UHFFFAOYSA-N Selenomethionine Natural products C[Se]CCC(N)C(O)=O RJFAYQIBOAGBLC-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229960002718 selenomethionine Drugs 0.000 claims abstract description 18
- XDSSPSLGNGIIHP-VKHMYHEASA-N Se-methyl-L-selenocysteine Chemical compound C[Se]C[C@H]([NH3+])C([O-])=O XDSSPSLGNGIIHP-VKHMYHEASA-N 0.000 claims abstract description 17
- FDKWRPBBCBCIGA-REOHCLBHSA-N (2r)-2-azaniumyl-3-$l^{1}-selanylpropanoate Chemical compound [Se]C[C@H](N)C(O)=O FDKWRPBBCBCIGA-REOHCLBHSA-N 0.000 claims abstract description 15
- FDKWRPBBCBCIGA-UWTATZPHSA-N D-Selenocysteine Natural products [Se]C[C@@H](N)C(O)=O FDKWRPBBCBCIGA-UWTATZPHSA-N 0.000 claims abstract description 15
- ZKZBPNGNEQAJSX-UHFFFAOYSA-N selenocysteine Natural products [SeH]CC(N)C(O)=O ZKZBPNGNEQAJSX-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229940055619 selenocysteine Drugs 0.000 claims abstract description 15
- 235000016491 selenocysteine Nutrition 0.000 claims abstract description 15
- 238000001927 high performance liquid chromatography-inductively coupled plasma mass spectrometry Methods 0.000 claims abstract description 10
- 238000010606 normalization Methods 0.000 claims abstract description 10
- 229940082569 selenite Drugs 0.000 claims abstract description 6
- MCAHWIHFGHIESP-UHFFFAOYSA-L selenite(2-) Chemical compound [O-][Se]([O-])=O MCAHWIHFGHIESP-UHFFFAOYSA-L 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 10
- 150000003863 ammonium salts Chemical class 0.000 claims description 6
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 6
- 238000007872 degassing Methods 0.000 claims description 5
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims description 5
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims description 5
- 235000019838 diammonium phosphate Nutrition 0.000 claims description 5
- 239000012266 salt solution Substances 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 3
- -1 methanol-ammonium citrate Chemical compound 0.000 claims description 3
- 239000005696 Diammonium phosphate Substances 0.000 claims description 2
- 239000007979 citrate buffer Substances 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- MCAHWIHFGHIESP-UHFFFAOYSA-N selenous acid Chemical compound O[Se](O)=O MCAHWIHFGHIESP-UHFFFAOYSA-N 0.000 abstract description 10
- 238000004458 analytical method Methods 0.000 abstract description 5
- 238000004445 quantitative analysis Methods 0.000 abstract description 3
- 238000000926 separation method Methods 0.000 abstract 1
- 230000014759 maintenance of location Effects 0.000 description 14
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 8
- 238000001514 detection method Methods 0.000 description 6
- 239000007789 gas Substances 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 4
- 229940000207 selenious acid Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 229910000831 Steel Inorganic materials 0.000 description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N argon Substances [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000010959 steel Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 101000666730 Homo sapiens T-complex protein 1 subunit alpha Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 102100038410 T-complex protein 1 subunit alpha Human genes 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000021329 brown rice Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012806 monitoring device Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds
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- 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)
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Abstract
The invention discloses a method for measuring selenium form by using high performance liquid inductively coupled plasma mass spectrometry, which is characterized by comprising the following steps: (1) preparing a mobile phase; (2) measuring the standard selenium form solution and the extract to be detected by adopting high performance liquid chromatography-inductively coupled plasma mass spectrometry; (3) and quantitatively analyzing the forms of five selenium, namely selenocysteine, selenomethionine, methyl selenocysteine, selenite and selenate, in the sample by adopting a normalization method. The method for detecting the selenium form of the plants has the advantages of complete form separation, convenient preparation of the mobile phase, accurate and reliable result obtained by quantitative analysis, and scientific analysis of the selenocysteine (SeCys) in the plants2) Selenomethionine (SeMet), methylselenocysteine (MeSeCys), and selenous acid (SeO)3 2‑) And selenate (SeO)4 2‑) Five forms of selenium.
Description
Technical Field
The invention relates to a method for determining selenium form by using high performance liquid inductively coupled plasma mass spectrometry, and belongs to the field of analysis and test.
Background
The current research shows that selenium is mainly SeO in the environmental medium3 2-、SeO4 2-、Se0、Se2-Exists in the form of (1); while selenium mainly comprises SeMet, SeCysCysSe, MeSeCys and SeO in crops3 2-、SeO4 2-Etc., and is generally analyzed by LC-AFS, HPLC-ICP-MS, etc. The detection method recommended in the national supply and marketing industry standard 'GH/T1135-2017 for selenium-rich agricultural products' is LC-AFS, and the low-selenium, selenium-containing or natural selenium-rich agricultural products can not reach the instrument detection limit of the AFS after sample pretreatment.
Disclosure of Invention
In view of the above, the main objective of the present invention is to provide a method for measuring selenium form by using high performance liquid inductively coupled plasma mass spectrometry, which not only detects the total amount of elements, but also optimizes the quantitative method, eliminates experimental interference, and improves detection efficiency and accuracy.
In order to achieve the above object, the present invention provides a method for measuring selenium form by high performance liquid inductively coupled plasma mass spectrometry, comprising the steps of:
(1) preparation of a mobile phase: preparing a methanol-ammonium citrate buffer solution from the ammonium salt solution and chromatographic pure methanol and adjusting the pH value; performing suction filtration on the mobile phase by using a 0.22-micron filter membrane, and performing ultrasonic degassing to obtain the mobile phase;
(2) measuring the standard selenium form solution and the extract to be detected by adopting high performance liquid chromatography-inductively coupled plasma mass spectrometry (ICP-MS); ICP-MS has the advantages of high sensitivity, good selectivity and the like when analyzing and detecting the selenium form, has the capability of simultaneously detecting multiple elements and isotopes, but has relatively low selenium ionization efficiency, and the dimer Ar of argon2Serious ion interference exists in selenium detection, so that the CCT collision pool technology is adopted to eliminate the ion interference;
(3) normalization method is adopted to measure selenocysteine (SeCys) in sample2) Selenomethionine (SeMet), methylselenocysteine (MeSeCys), and selenous acid (SeO)3 2-) And selenate (SeO)4 2-) Five forms of selenium were quantitatively analyzed. Compared with other methods, the normalization method has the advantages that: the method is simple, convenient and accurate, the quantitative result is independent of the repeatability of the sample volume (within the range of no overload of a chromatographic column), the result is slightly influenced when the operating conditions are slightly changed, and the method can be used for quantitatively analyzing unknown substances.
Further, in the step (1), the ammonium salt is selected from one of diammonium citrate, diammonium phosphate and ammonium citrate, and ammonium citrate is preferred.
Further, in the step (1), the chromatographically pure methanol is the chromatographically pure methanol frozen at the temperature of-20 ℃ so as to ensure the stability of the physical and chemical properties of the chromatographically pure methanol and save the experimental cost to the utmost extent.
Further, in the step (1), the volume concentration of the methanol in the mobile phase is 2-10%, preferably 2%, so that the effect of completely separating five selenium forms is preferably achieved.
Further, in the step (1), the concentration of the ammonium salt solution in the mobile phase is 2-10mmol/L, preferably 6mmol/L, so that the effect of completely separating five selenium forms is preferably achieved.
Further, in step (1), the pH value of the mobile phase is 5-6, preferably 5.7, so that the effect of completely separating five selenium forms is preferably achieved.
Further, in the step (1), the ultrasonic degassing time is 15 +/-5 min.
Further, wherein in step (2), the standard selenium form solution comprises selenocysteine (SeCys)2) Selenomethionine (SeMet), methylselenocysteine (MeSeCys), and selenous acid (SeO)3 2-) And selenate (SeO)4 2-) Five selenium form standard sample stocks, wherein each selenium form standard sample stock has a concentration of 1000mg/mL (as selenium).
Further, the step (3) specifically comprises: according to the characteristic that the quantity of each component in a sample is in direct proportion to the peak area of the component detected by HPLC-ICP-MS, all components in the sample can generate signals to obtain corresponding chromatographic peaks, and the corresponding forms of the components can be known by the retention time of each component, so that the content of each component can be calculated by using a normalization formula (1):
wherein: setting the sample groups as n, wherein the content of each group is m1，m2，……，mn。
Further, in the step (3), when the sample is quantitatively analyzed by the normalization method, all standard sample configurations need to be calculated in selenium concentration, and it is satisfied that all selenium form components are eluted and the peak areas are the same in the case that the detector responds and all selenium form components have the same selenium concentration.
Compared with the prior art, the invention has the following beneficial effects:
the invention can respectively detect the selenocysteine (SeCys) in the plant by using HPLC-ICP-MS combined technology and adopting a normalization method to carry out quantitative analysis on the sample2) Methyl selenocysteine (SeMeCy), selenomethionine (SeMet), and selenous acid (SeO)3 2-) And selenate (SeO)4 2-) Five selenium formsThe state and the content. The method is simple and rapid, the mobile phase is simple and convenient to prepare, and the analysis result is accurate and reliable. Compared with the existing methods such as spectrometry, mass spectrometry, chromatography and the like, the method has better selectivity and stronger stability on element forms.
Drawings
FIG. 1 is a Se element standard curve of the present invention without CCT collision pool technology;
fig. 2 is a Se element standard curve of the CCT collision cell technology employed in the present invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings.
The following materials or reagents, unless otherwise specified, are commercially available.
The embodiment of the invention provides a method for measuring selenium form by using high performance liquid inductively coupled plasma mass spectrometry, which comprises the following steps:
(1) preparation of a mobile phase: weighing diammonium citrate, diammonium hydrogen phosphate or ammonium citrate, adding ultrapure water, and uniformly stirring by using a glass rod; after addition of refrigerated (-20 ℃) chromatographically pure methanol (2%), the pH was adjusted to 5.7 using 10% by weight of dilute hydrochloric acid; filtering the mobile phase with 0.22 μm filter membrane, and ultrasonic degassing the prepared mobile phase for 15 min; the selenium forms of samples are respectively subjected to chromatographic analysis by using the diammonium citrate, diammonium hydrogen phosphate and ammonium citrate to prepare mobile phase, and the retention time and peak area of each form show that the mobile phase prepared by using the diammonium hydrogen phosphate and the diammonium citrate cannot effectively separate five forms, so that the experimental result is influenced, and the five forms can be completely separated by using the ammonium citrate as mobile phase salt.
(2) The standard selenium form solution and the extract to be detected are measured by using high performance liquid chromatography-inductively coupled plasma mass spectrometry, and the CCT collision cell technology is adopted to eliminate ion interference (see the figure 1 and the figure 2),
the CCT collision cell technology is specifically operated as follows (taking 7% hydrogen + 93% helium mixture as an example).
Opening a He gas steel cylinder (the purity of the He gas steel cylinder is required to be more than 99.999 percent), and regulating the gas pressure to be 0.1 MPa.
② selecting standard mode in Instrument → Configurations, and calling out a set of parameters.
Thirdly, igniting the plasma according to a normal mode and entering an operation state.
Fourthly, inserting a sample injection capillary into the 1ppb regulating solution, and checking the signal conditions under the standard conditions, such as sensitivity, RSD and the like.
Fifthly, opening the Gas with CCT1 in the Instrument → Tune-added Gas, adjusting the flow rate to 7ml/min, and waiting for about 10 min.
Sixthly, inserting the sample injection capillary into the 1ppb In solution.
Seventhly, check V CCTCH1 in Instrument → Configurations → check V, then check V again in the lower box.
Adjusting the flow rate of the collision gas and instrument parameters to enable the count of Ar (80) In pure water to be less than 200 and the sensitivity of In to be more than 20000/ppb.
Ninthly, editing the analysis method, and selecting the database as Default CCT.
Sample for analysis in r.
As can be seen from comparison between fig. 1 and fig. 2, the fitting coefficient of the standard curve of the CCT collision cell technology is 0.999583, which is significantly higher than the fitting coefficient 0.986383 that is not used, which indicates that there is substantially no interference of other ions when the CCT collision cell technology is used, and the measured value is more accurate; the standard selenium form solution comprises selenocysteine (SeCys)2) Selenomethionine (SeMet), methylselenocysteine (MeSeCys), and selenous acid (SeO)3 2-) And selenate (SeO)4 2-) Five selenium form standard sample stocks, wherein each selenium form standard sample stock has a concentration of 1000mg/mL (as selenium). Storing at-20 deg.C after preparation, preparing 5 kinds of selenium form mixed standard solutions before machine detection, and gradually diluting the mixed standard to 100 μ g/kg;
(3) normalization method is adopted to measure selenocysteine (SeCys) in sample2) Selenomethionine (SeMet), methylselenocysteine (MeSeCys), and selenous acid (SeO)3 2-) And selenate (SeO)4 2-) Five forms of selenium were quantitatively analyzed. Detecting the amount of each component in the sample and the peak area thereof according to the combination of HPLC-ICP-MSProportional characteristic, all components in the sample can generate signals to obtain corresponding chromatographic peaks, the corresponding forms of the components can be known by the retention time of the components, and the content of the components can be calculated by the following normalization formula. Setting the sample groups as n, wherein the content of each group is m1，m2，……，mnThen the percentage content of the i components is as follows:
respectively detecting a sample 1 (tea), a sample 2 (rice leaf) and a sample 3 (brown rice) by using HPLC-ICP-MS (high performance liquid chromatography (HPLC, Shimadzu LC-20A, Japan) and X series 2 type ICP-MS (ThermoFisher)) as an online HPLC monitoring device, and establishing an HPLC-ICP-MS combined detection system to obtain peak areas of all forms, which are shown in Table 1; then, the peak areas of the samples 1 to 3 are respectively substituted into the formula (1), and the percentage contents of the forms of the samples 1 to 3 are respectively obtained, which is shown in table 1.
TABLE 1
The present invention is further illustrated by the following specific examples.
Example 1 salt concentration comparison of mobile phase
Table 2 retention times for five forms of mobile phase salt at different concentrations
Preparing three mobile phases with the ammonium citrate content of 2mmol/L, 6mmol/L and 10mmol/L respectively (the three mobile phases have no other variables except the ammonium citrate concentration), and measuring by using three selenium form mixed standard solutions with the flow ratio of 100 mu g/kg (the concentration of each form standard solution) according to the instrument instruction, so as to obtain the retention time of each form and analyze.
As can be seen from the results in Table 2, when the concentration of ammonium citrate is 2mmol/L, the retention time of selenite is increased, which is superimposed with the retention time of selenomethionine and cannot be separated; when the concentration of the ammonium citrate is 10mmol/L, the retention time of selenious acid root is reduced, so that the selenious acid root and the methylselenocysteine are overlapped and cannot be separated; therefore, ammonium citrate with the concentration of 6mmol/L is selected to configure the mobile phase.
Example 2 pH comparison of the mobile phase
Three mobile phases with pH values of 5.5, 5.7 and 5.9 (no other variables except the pH value of the three mobile phases) are prepared, and the three mobile phases are measured by using three selenium form mixed standard solutions with the flow rate of 100 mu g/kg (the concentration of each form of standard solution) according to the instrument instruction, so that the retention time of each form is obtained and analyzed.
TABLE 3 Retention time of the mobile phase at different pH values for five morphologies
As can be seen from the results in table 3, the retention times of selenite and selenate are extremely sensitive to changes in the pH of the mobile phase, with greater pH leading to smaller retention times. When the pH value is 5.5, the mobile phase can elute the form to be detected within 7min, but the selenite and the methylselenocysteine cannot be separated; and when the pH value is 5.9, the selenious acid root and selenomethionine produce peaks which are overlapped, the sample to be detected cannot be separated, and the retention time of the selenious acid root is over 10 min. Therefore, the optimum condition was that the mobile phase pH was 5.7, and all of the 5 forms of selenium to be tested could be eluted within 10min with no overlap in retention time.
The above-mentioned embodiments are merely examples provided to fully illustrate the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (10)
1. A method for measuring selenium form by using high performance liquid inductively coupled plasma mass spectrometry is characterized by comprising the following steps:
(1) preparation of a mobile phase: preparing a methanol-ammonium citrate buffer solution from the ammonium salt solution and chromatographic pure methanol and adjusting the pH value; performing suction filtration on the mobile phase by using a 0.22-micron filter membrane, and performing ultrasonic degassing to obtain the mobile phase;
(2) measuring the standard selenium form solution and the extract to be detected by adopting high performance liquid chromatography-inductively coupled plasma mass spectrometry;
(3) and quantitatively analyzing the forms of five selenium, namely selenocysteine, selenomethionine, methyl selenocysteine, selenite and selenate, in the sample by adopting a normalization method.
2. The method of determining selenium speciation of claim 1 wherein in step (1) the ammonium salt is selected from one of diammonium citrate, diammonium phosphate and ammonium citrate.
3. The method of determining selenium speciation of claim 1 wherein in step (1) the chromatographically pure methanol is chromatographically pure methanol frozen at-20 ℃.
4. The method of determining selenium speciation of claim 1 where in step (1) the mobile phase has a methanol concentration of 2-10% by volume.
5. The method of claim 1, wherein in step (1), the concentration of the ammonium salt solution in the mobile phase is 2 to 10 mmol/L.
6. The method of determining selenium speciation of claim 1 where in step (1) the mobile phase has a pH of 5-6.
7. The method of determining selenium morphology as claimed in claim 1, wherein in step (1) the time of ultrasonic degassing is 15 ± 5 min.
8. The method of determining selenium morphology according to claim 1, wherein in step (2), the standard selenium morphology solution comprises five selenium morphology standard sample stocks of selenocysteine, selenomethionine, methylselenocysteine, selenite, and selenate, wherein each selenium morphology standard sample stock has a concentration of 1000mg/mL in terms of selenium.
9. The method of claim 1, wherein step (3) comprises: according to the characteristic that the quantity of each component in a sample is in direct proportion to the peak area of the component detected by HPLC-ICP-MS, and all components in the sample can generate signals to obtain corresponding chromatographic peaks, the content of each component is calculated by using a normalized formula (1):
wherein: setting the sample groups as n, wherein the content of each group is m1，m2，……，mn。
10. The method of claim 9, wherein in step (3), the sample is quantitatively analyzed by normalization, all standard sample configurations are calculated as selenium concentrations, and all selenium species are eluted and have the same peak area in the presence of a detector response and the same selenium concentration for all selenium species.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114460197A (en) * | 2022-01-29 | 2022-05-10 | 南京恒宝田功能农业产业研究院有限公司 | Method for quantitatively detecting content of organic selenium in agricultural products |
| CN114705767A (en) * | 2022-03-01 | 2022-07-05 | 海南省食品检验检测中心(海南省实验动物中心) | Analysis and detection method for selenium form in rice |
| CN114924001A (en) * | 2022-05-12 | 2022-08-19 | 中国计量科学研究院 | Phosphorus-containing or selenium-containing compound standard solution value setting method based on independent calibration |
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| CN102721779A (en) * | 2012-06-19 | 2012-10-10 | 浙江农林大学 | Detection method of selenium forms in plants |
| CN102928500A (en) * | 2012-11-15 | 2013-02-13 | 中华人民共和国浙江出入境检验检疫局 | Method for detecting organic selenium, protein selenium or polysaccharide selenium in marine products through microwave digestion-ICP-MS (Inductively Coupled Plasma Mass Spectrometry) way |
| CN106033081A (en) * | 2015-03-16 | 2016-10-19 | 中国科学技术大学苏州研究院 | Determination method of selenium form in grain sample |
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| CN114924001A (en) * | 2022-05-12 | 2022-08-19 | 中国计量科学研究院 | Phosphorus-containing or selenium-containing compound standard solution value setting method based on independent calibration |
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