HPLC-ICPMS detection method for arsenic in various forms in whole blood

Abstract

The invention relates to an HPLC-ICPMS (High Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry) detection method for arsenic in various forms in whole blood. The HPLC-ICPMS detection method comprises the following steps: step 1, preparing a standard solution for constructing calibration curves of six arsenic Step 2, constructing a calibration curve of six arsenic forms; 3, collecting a whole blood sample and collecting a sample blank; and 4, taking 0.30 mL of the whole blood sample collected in the step 3, and adding 0.30 mL of the extraction solution. According to the method, an AG7 guard column and an AS7 chromatographic column are selected, and a mobile phase A is 5mmol / L ammonium carbonate; the separation effect of six forms of arsenic is improved and the separation time is saved under the conditions that a mobile phase A is 100 mmol / L ammonium carbonate, a mobile phase B is 100 mmol / L ammonium carbonate, and a mobile phase A is 100% in 0-3 min, 0% in 3-6 min and 100% in 6-12 min, a methanol acetonitrile solution (V / V = 7: 3) is used as an extraction solution, the sample extraction effect is improved, and the processes of sample pretreatment, HPLC separation of the forms of arsenic, ICP-MS detection and the like are optimized by utilizing HPLC and HPLC-ICP MS technologies.

Description

Technical Field

[0001] The present invention relates to the technical field of methods for detecting arsenic forms, and in particular to a HPLC-ICPMS method for detecting various forms of arsenic in whole blood. Background Art

[0002] Arsenic (As) is a toxic nonmetallic element and a confirmed human carcinogen. Occupational exposure can cause lung and skin cancers in individuals. Common forms of arsenic found in nature include arsenite (As(III)), arsenate (As(V)), monomethylarsenic (MMA), dimethylarsenic (DMA), arsenobetaine (AsB), arsenocholine (AsC), and arsenosaccharide (AsS). Its toxicity is closely related to the chemical form present. With the development of modern industry and science and technology, arsenic mining and smelting, semiconductor material manufacturing, and the production of arsenic-containing pesticides have made individuals who are occupationally exposed to arsenic a vulnerable group. The primary routes of occupational arsenic exposure are respiratory and dermal absorption. Following absorption, arsenic is present in the human body in various chemical forms. Blood, as a vital component of the human body, can directly reflect internal exposure to the toxicant. Internal exposure can directly link arsenic to health effects. Integrating information on the route and timing of arsenic ingestion allows for a more scientific assessment of exposure risks and expected outcomes.

[0003] Arsenic speciation analysis in blood involves the separation and detection of various arsenic forms. Currently, reports both domestically and internationally often utilize high-performance liquid chromatography (HPLC) for separation and atomic fluorescence spectroscopy (AFS) for detection. Due to the complex matrix, small sample size, and low arsenic content of blood samples, the detection of arsenic speciation in blood samples places higher demands on the selection of arsenic extraction methods, separation, and detection. Compared to AFS, inductively coupled plasma mass spectrometry (ICP-MS) offers advantages such as low detection limits, wide dynamic linear range, rapid analysis, and excellent stability. The combination of HPLC and ICP-MS for elemental speciation analysis has become a growing trend. Currently, there are few reports on the use of HPLC-ICPMS for the detection of six arsenic forms in the blood of occupationally exposed individuals. Furthermore, to date, there are no established reference values ​​or biological exposure limits for arsenic forms in the blood of occupationally exposed individuals, nor are there any standardized assay methods.

[0004] In order to solve the above problems, the present invention proposes a HPLC-ICPMS detection method for various forms of arsenic in whole blood. Summary of the Invention

[0005] (1) Technical problems to be solved

[0006] The purpose of the present invention is to overcome the deficiencies of the prior art, meet practical needs, and provide an HPLC-ICPMS detection method for various forms of arsenic in whole blood to solve the above technical problems.

[0007] (2) Technical solution

[0008] In order to achieve the purpose of the present invention, the technical solution adopted by the present invention is:

[0009] A HPLC-ICPMS method for detecting multiple forms of arsenic in whole blood and monitoring internal exposure levels comprises the following steps:

[0010] Step 1: Prepare standard solutions for constructing calibration curves for six arsenic species;

[0011] Step 2: construct calibration curves for six arsenic species;

[0012] Step 3: Collect a whole blood sample and a sample blank;

[0013] Step 4: Take 0.30 mL of the whole blood sample collected in step 3, add 0.30 mL of the extraction solution to obtain a blood sample solution;

[0014] Step 5: Vortex the blood sample solution in step 4 for 2 minutes, centrifuge at 12,000 rpm for 10 minutes, take 0.2 mL of the supernatant, blow nitrogen to near dryness at 60°C, add 0.2 mL of pure water to reconstitute, vortex for 2 minutes, centrifuge at 12,000 rpm for 10 minutes, and take the supernatant to obtain the test solution;

[0015] Step 6. Use an AG7 guard column and an AS7 chromatographic column, mobile phase A: 5 mmol / L ammonium carbonate; mobile phase B: 100 mmol / L ammonium carbonate, the ratio of mobile phase A is 100% for 0-3 min, 0% for 3-6 min, and 100% for 6-12 min, the injection volume is 50 μL, and the flow rate is 1.2 ml / min as a conditional gradient elution to separate the target compound in the test solution to obtain the target compound;

[0016] Step 7: Detect the target compound described in step 6 using an inductively coupled plasma mass spectrometer.

[0017] Furthermore, the step 1 specifically includes: preparing an arsenic form standard intermediate solution (1.00 ug / mL, calculated as As): accurately weighing a certain amount of arsenite ion (As(III)), arsenate ion (As(V)), monomethylarsenic (MMA), dimethylarsenic (DMA), arsenobetaine (AsB), and arsenocholine (AsC) in a 10 mL volumetric flask, and diluting the solution to the mark with water;

[0018] Accurately pipette a certain amount of 1.00 ug / mL mixed standard intermediate solution of As(III), As(V), MMA, DMA, AsB and AsC, and dilute with water to prepare a standard series with concentrations of 0.0 ng / mL, 0.5 ng / mL, 1.0 ng / mL, 2.0 ng / mL, 5.0 ng / mL, 10.0 ng / mL, 20.0 ng / mL and 50.0 ng / mL.

[0019] Furthermore, step 2 specifically includes: adjusting various instrument indicators with a tuning solution so that the instrument sensitivity, oxide, double charge, resolution, and other indicators meet the measurement requirements. After measuring a mixed standard solution of arsenic forms with a concentration of 10.0 ng / mL and confirming that the separation of each arsenic form meets the requirements, a series of mixed standard solutions of As(III), As(V), MMA, DMA, AsB, and AsC are injected into a liquid chromatography-inductively coupled plasma mass spectrometry instrument in ascending order of mass concentration to obtain a chromatogram and qualitatively analyze the retention time;

[0020] The standard curve was drawn with the concentration of arsenic species as the abscissa and the chromatographic peak area as the ordinate to obtain the calibration curves of the six arsenic species.

[0021] Furthermore, the step three specifically includes: collecting 2.00 mL of venous blood using a sodium heparin anticoagulant blood collection tube, placing the blood in a clean container for refrigerated transportation, and storing the blood at -70°C for later use;

[0022] Use a blood collection needle to draw 2.00 mL of ultrapure water into a blood collection tube as a sample blank, place it in a clean container for refrigerated transportation, and store it at -70°C until use.

[0023] Furthermore, the extraction solution in step 4 is a methanol-acetonitrile solution (V / V=7:3), and step 4 specifically includes: taking 0.30 mL of whole blood sample, adding 0.30 mL of methanol-acetonitrile solution (V / V=7:3) to form a blood sample solution.

[0024] (3) Beneficial effects:

[0025] The present invention uses an AG7 guard column and an AS7 chromatographic column, mobile phase A: 5mmol / L ammonium carbonate; mobile phase B: 100mmol / L ammonium carbonate, and the ratio of mobile phase A is 100% for 0-3 minutes, 0% for 3-6 minutes, and 100% for 6-12 minutes as conditions, thereby improving the separation effect of six forms of arsenic and saving separation time. By using a methanol-acetonitrile solution (V / V=7:3) as an extraction solution, the sample extraction effect is improved, and HPLC and HPLC-ICPMS technologies are used to optimize processes such as sample pretreatment, HPLC separation of form arsenic, and ICP-MS detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The effect of the mobile phase ratio and chromatographic column of the present invention on the separation effect of the mixed standard solution of six arsenic forms (10 ng / mL);

[0027] Figure 2 is the calibration curve of the six arsenic forms of the present invention;

[0028] Figure 3 The chromatograms of six arsenic forms at low, medium and high concentrations according to the present invention are shown (n=6). DETAILED DESCRIPTION

[0029] The following is a further explanation of the HPLC-ICPMS detection method of various forms of arsenic in whole blood of the present invention in the form of specific examples. However, it should be understood that the contents of the examples are intended to provide a clearer understanding of the present invention and are not intended to limit the scope of protection of the present invention.

[0030] Experimental instruments and reagents

[0031] iCAP RQplus ICP-MS (thermo scientific); Vanquish HPLC (thermoscientific); sodium heparin blood collection tubes (Jiangsu Kangjian Medical Products Co., Ltd.); HH-8 electric constant temperature water bath (Changzhou Shenguang Instrument Co., Ltd.); N-EVAP 112 nitrogen blowdown apparatus (Organomation, USA); H2100R refrigerated centrifuge (Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.); Milli- Reference water purifier (Millipore, USA); Dionex IonPac AS7 (4×250 mm, thermo scientific); Dionex IonPac AG7 (4×50 mm, thermo scientific).

[0032] Arsenite solution standard substance (GBW08666, mass concentration: as As: 75.5 μg / g, China National Institute of Metrology, batch number 21071, k = 2); arsenobetaine solution standard substance (GBW08670, mass concentration: as As: 38.8 μg / g, China National Institute of Metrology, batch number 21101, k = 2); monomethylarsenic solution standard substance (GBW08668, mass concentration: as As: 25.1 μg / g, China National Institute of Metrology, batch number 21101, k = 2 ); dimethylarsenic solution standard substance (GBW08669, mass concentration: as As: 52.9 μg / g, National Institute of Metrology, batch number 21071, k = 2); arsenate solution standard substance (GBW08667, mass concentration: as As: 17.5 μg / g, National Institute of Metrology, batch number 2104, k = 2); arsenic choline solution standard substance (GBW08671, mass concentration: as As: 28.0 μg / g, National Institute of Metrology, batch number 21071, k = 2). Ammonium carbonate (99.999% metals basis, Aladdin, batch number G1922003). All experimental water was ultrapure water (18.2 MΩ / cm).

[0033] Example

[0034] A HPLC-ICPMS method for detecting various forms of arsenic in whole blood comprises the following steps:

[0035] Step 1: Prepare standard solutions for constructing calibration curves for six arsenic species:

[0036] Prepare an intermediate standard solution of arsenic species (1.00 μg / mL, calculated as As): accurately weigh a certain amount of arsenite ion (As(III)), arsenate ion (As(V)), monomethylarsenic (MMA), dimethylarsenic (DMA), arsenobetaine (AsB), and arsenocholine (AsC) into a 10 mL volumetric flask and dilute to the mark with water.

[0037] Accurately pipette a certain amount of 1.00 ug / mL mixed standard intermediate solution of As(III), As(V), MMA, DMA, AsB and AsC, and dilute with water to prepare a standard series with concentrations of 0.0 ng / mL, 0.5 ng / mL, 1.0 ng / mL, 2.0 ng / mL, 5.0 ng / mL, 10.0 ng / mL, 20.0 ng / mL and 50.0 ng / mL.

[0038] Step 2: Construct calibration curves for six arsenic species:

[0039] The instrument was adjusted using a tuning solution to ensure that sensitivity, oxides, double charges, and resolution met the measurement requirements. A mixed standard solution of arsenic species at a concentration of 10.0 ng / mL was measured. After confirming that the separation of each arsenic species met the requirements, a series of mixed standard solutions of As(III), As(V), MMA, DMA, AsB, and AsC were injected into the liquid chromatography-inductively coupled plasma mass spectrometry instrument in ascending order of mass concentration. Chromatograms were obtained and qualitatively analyzed using retention times.

[0040] The standard curve was drawn with the concentration of arsenic species as the horizontal axis and the chromatographic peak area as the vertical axis. Figure 2 Calibration curves for six arsenic species.

[0041] Step 3: Collect whole blood sample and sample blank:

[0042] Collect 2.00 mL of venous blood using a sodium heparin anticoagulant blood collection tube, place it in a clean container for refrigerated transportation, and store it at -70°C until use;

[0043] Use a blood collection needle to draw 2.00 mL of ultrapure water into a blood collection tube as a sample blank, place it in a clean container for refrigerated transportation, and store it at -70°C until use.

[0044] Step 4: Take 0.30 mL of the whole blood sample collected in step 3 and add 0.30 mL of extraction solution to obtain a blood sample solution:

[0045] Take 0.30 mL of whole blood sample and add 0.30 mL of methanol-acetonitrile solution (V / V=7:3) to form a blood sample solution.

[0046] Step 5: Vortex the blood sample solution described in step 4 for 2 minutes, centrifuge it at 12000 r / min for 10 minutes, take 0.2 mL of the supernatant, blow it to near dryness with nitrogen at 60°C, add 0.2 mL of pure water to reconstitute it, vortex it for 2 minutes, centrifuge it at 12000 r / min for 10 minutes, and take the supernatant to obtain the test solution.

[0047] Step 6. Select an AG7 guard column and an AS7 chromatographic column, mobile phase A: 5 mmol / L ammonium carbonate; mobile phase B: 100 mmol / L ammonium carbonate, the ratio of mobile phase A is 100% for 0-3 min, 0% for 3-6 min, and 100% for 6-12 min, the injection volume is 50 μL, and the flow rate is 1.2 ml / min as conditional gradient elution to separate the target compound in the test solution to obtain the target compound. The elution program is shown in Table 1.

[0048] Table 1 Gradient elution program

[0049]

[0050] Step 7: Using Table 2 as the reference conditions for the inductively coupled plasma mass spectrometer, the target compound described in step 6 was detected using an inductively coupled plasma mass spectrometer.

[0051] Table 2 Reference conditions for inductively coupled plasma mass spectrometry

[0052]

[0053]

[0054] Comparison of experimental conditions

[0055] 1. Influence of mobile phase ratio and chromatographic column

[0056] 1. Using ammonium carbonate as the mobile phase, the effects of different mobile phase ratios on the separation of six forms of arsenic using Hamilton PRP-X 100 column, AG7 guard column and AS7 column were investigated. Figure 1 A and Figure 1 B:

[0057] Figure 1 A shows the separation results of a Hamilton PRP-X 100 column. Mobile phase A consisted of 1 mmol / L ammonium carbonate; mobile phase B consisted of 100 mmol / L ammonium carbonate. The proportions of phase A were: 80% for 0-5 min, 50% for 6-20 min (a); 95% for 0-4 min, 20% for 5-10 min, and 95% for 11-15 min (b); and 90% for 0-4 min, 20% for 5-10 min, and 90% for 11-15 min (c). Figure 1 Among a, b and c in A, c has a higher separation degree and a shorter separation time.

[0058] Figure 1 B is the separation effect of AG7 guard column and AS7 column on 6 forms of arsenic at different mobile phase ratios. Mobile phase A: 5mmol / L ammonium carbonate; mobile phase B: 100mmol / L ammonium carbonate, mobile phase A is: 40% at 0-10min (a); 80% at 0-3min, 0% at 3-5min, 80% at 5-12min (b); 100% at 0-3.5min, 0% at 3.6-6.5min, 100% at 6.5-10min (c); 100% at 0-3min, 0% at 3-6min, 100% at 6-12min (d), from Figure 1 B shows that isocratic elution (a) is not effective for separation; b, c and d, three different mobile phase ratios can all achieve effective separation. Considering the matrix of the blood sample, the interference of other forms of arsenic besides the six forms of arsenic, and the analysis time, the condition d is better.

[0059] 2. Comparison of the separation effects of Hamilton PRP-X 100 column, AG7 guard column and AS7 column on the six forms of arsenic. Figure 1 C. It can be seen that under the ammonium carbonate mobile phase condition, the elution order of the six forms of arsenic on different columns is inconsistent, and the separation effect and separation time of the AG7 guard column and AS7 column (Figure C, Bd) for the six forms of arsenic are significantly better than those of the PRP-X 100 column (Figure C, Ac).

[0060] Therefore, the detection method uses an AG7 guard column and an AS7 chromatographic column, mobile phase A: 5mmol / L ammonium carbonate; mobile phase B: 100mmol / L ammonium carbonate, and the proportion of mobile phase A is 100% for 0-3min, 0% for 3-6min, and 100% for 6-12min as the conditions.

[0061] 2. Influence of Sample Pretreatment Methods

[0062] Accurately pipette 0.30 mL of whole blood sample and add 0.3 mL of different extraction solutions:

[0063] Method 1: Add 0.30 mL of acetonitrile;

[0064] Method 2: Add 0.30 mL of methanol;

[0065] Method 3: Add 0.30 mL of methanol-acetonitrile solution (V / V = 7:3);

[0066] Method 4: Add 0.30 mL of methanol-acetonitrile solution (V / V = 5:5);

[0067] The mixed solutions prepared by the above four methods were vortexed for 2 minutes, centrifuged at 12000 r / min for 10 minutes, and 0.2 mL of the supernatant was taken. The mixture was blown to near dryness under nitrogen at 60°C, and 0.2 mL of pure water was added for reconstitution. The mixture was vortexed for 2 minutes, centrifuged at 12000 r / min for 10 minutes, and the supernatant was taken for testing. The test results are shown in Table 3.

[0068] Table 3 Spiked recoveries of six arsenic species

[0069]

[0070] After adding 0.30mL of methanol, vortexing, and centrifuging in Method 2, the mixture remained in a paste state and the supernatant could not be removed. The As(III) recovery rates of Methods 1, 3, and 4 were low, while the As(V) recovery rates were high. This is presumably due to the transformation of different forms of arsenic in whole blood. Comparing the sum of the recovery rates of the six forms of arsenic, the spiked recovery rate of Method 3 was higher than that of Method 4. Due to the particularity of whole blood samples and the differences in blood samples from different people, the transformation between forms of arsenic is also different. This time, there was transformation between the six forms of arsenic in the whole blood, and there was less transformation outside the six forms of arsenic. Therefore, Method 3 was selected as the sample pretreatment method.

[0071] Detection limit

[0072] According to the method for establishing the calibration curve, the solution close to the blank was measured 10 times continuously, and 3 times the standard deviation was used as the detection limit and 10 times the standard deviation was used as the quantification limit to obtain Table 4.

[0073] Table 4 Detection limit and quantification limit

[0074]

[0075] When the sampling volume was 0.30 mL and the whole blood sample was diluted 2-fold, the detection limits of arsenobetaine, dimethylarsenic, arsenite, arsenocholine, monomethylarsenic, and arsenate were 0.096 ng / mL, 0.077 ng / mL, 0.106 ng / mL, 0.092 ng / mL, 0.090 ng / mL, and 0.493 ng / mL, respectively; the quantification limits of arsenobetaine, dimethylarsenic, arsenite, arsenocholine, monomethylarsenic, and arsenate were 0.318 ng / mL, 0.256 ng / mL, 0.352 ng / mL, 0.308 ng / mL, 0.300 ng / mL, and 1.643 ng / mL, respectively.

[0076] Precision

[0077] Three concentration levels of standard solution were selected and the measurement was repeated 6 times at each concentration level. Figure 3 The corresponding test concentrations and precision test results are shown in Table 5. Table 5 shows that the precision of the six forms of arsenic in this method is in the range of 0.58% to 7.66%.

[0078] Table 5 Corresponding test concentration method precision (n=6)

[0079]

[0080] accuracy

[0081] Blood samples below the detection limit were mixed thoroughly, and 2.50 mL was pipetted into each sample. 50 μL (50 ng / mL), 150 μL (50 ng / mL), and 37.5 μL (1 μg / mL) of the mixed standard solution were added, respectively. Three spiked samples (high, medium, and low) were prepared to validate the accuracy of the method. The three spiked blood concentrations were 1 ng / mL, 3 ng / mL, and 15 ng / mL, respectively. 0.30 mL of the spiked samples were accurately pipetted and processed according to the sample preparation method before instrumental analysis. The results are shown in Table 6. The spiked recoveries for the sum of the six arsenic species ranged from 90.2% to 105.5%, demonstrating the high accuracy of the method.

[0082] Table 6 Spiked recovery of six forms of arsenic

[0083]

[0084] Actual sample testing and internal exposure level monitoring

[0085] An HPLC-ICPMS method was established for the detection of six forms of arsenic in blood, namely As(III), As(V), MMA, DMA, AsB and AsC. The established method was used to detect 71 actual blood samples. The test results are shown in Table 7.

[0086] Table 7 Detection results of arsenic forms in whole blood (in As)

[0087]

[0088]

[0089]

[0090] The measurement results of 71 blood samples were as follows: AsB: <LOD ~ 0.568ng / mL; DMA: <LOD ~ 0.794ng / mL; As(Ⅲ): <LOD ~ 0.270ng / mL; AsC: <LOD ~ 4.248ng / mL; MMA: <LOD ~ 0.268ng / mL; As(Ⅴ): <LOD ~ 0.878ng / mL.

[0091] The embodiments disclosed in the present invention are preferred embodiments, but are not limited thereto. Ordinary technicians in this field can easily understand the spirit of the present invention based on the above embodiments and make different extensions and changes. As long as they do not deviate from the spirit of the present invention, they are all within the scope of protection of the present invention.

Claims

1. A HPLC-ICPMS method for detecting various forms of arsenic in whole blood, characterized in that: The following steps are involved: Step 1: prepare standard solutions for constructing calibration curves of six arsenic forms; Step 2: construct calibration curves for six arsenic forms; Step 3, collecting a whole blood sample and a sample blank; Step 4: Take 0.30 mL of the whole blood sample collected in step 3, add 0.30 mL of the extraction solution to obtain a blood sample solution; Step 5, vortex the blood sample solution in step 4 for 2 minutes, centrifuge at 12000 r / min for 10 minutes, take 0.2 mL of supernatant, blow nitrogen to nearly dryness at 60°C, add 0.2 mL of pure water to reconstitute, vortex for 2 minutes, centrifuge at 12000 r / min for 10 minutes, take the supernatant to obtain the test solution; Step 6, select AG7 guard column and AS7 chromatographic column, mobile phase A: 5mmol / L ammonium carbonate; mobile phase B: 100mmol / L ammonium carbonate, the ratio of mobile phase A is 100% for 0-3min, 0% for 3-6min, and 100% for 6-12min, the injection volume is 50μL, and the flow rate is 1.2ml / min as conditional gradient elution to separate the target compound in the test solution to obtain the target compound; Step 7: Detect the target compound described in step 6 using an inductively coupled plasma mass spectrometer.

2. The HPLC-ICPMS detection method for various forms of arsenic in whole blood according to claim 1, characterized in that: The step 1 specifically includes: Prepare the standard intermediate solution of arsenic forms (1.00 ug / mL, in terms of As): accurately weigh a certain amount of arsenite ion (As(III)), arsenate ion (As(V)), monomethylarsenic (MMA), dimethylarsenic (DMA), arsenobetaine (AsB), and arsenocholine (AsC) in a 10 mL volumetric flask, and dilute to the mark with water; Accurately pipette a certain amount of 1.00ug / mL mixed standard intermediate solution of As(III), As(V), MMA, DMA, AsB and AsC respectively, and dilute with water to prepare a standard series with concentrations of 0.0ng / mL, 0.5ng / mL, 1.0ng / mL, 2.0ng / mL, 5.0ng / mL, 10.0ng / mL, 20.0ng / mL and 50.0ng / mL.

3. The HPLC-ICPMS detection method for various forms of arsenic in whole blood according to claim 1, characterized in that: The step 2 specifically includes: The instrument parameters were adjusted with tuning solution to make the instrument sensitivity, oxide, double charge, resolution and other parameters meet the measurement requirements. The arsenic form mixed standard solution with a concentration of 10.0 ng / mL was measured. After confirming that the separation degree of each arsenic form meets the requirements, As(III), As(V), MMA, DMA, AsB and AsC mixed standard series solutions were injected into the liquid chromatography-inductively coupled plasma mass spectrometry instrument in order of mass concentration from low to high, and the chromatogram was obtained, and the retention time was used for qualitative analysis. The standard curve was drawn with the concentration of arsenic species as the abscissa and the chromatographic peak area as the ordinate to obtain the calibration curves of the six arsenic species.

4. The HPLC-ICPMS detection method for various forms of arsenic in whole blood according to claim 1, characterized in that: The step three specifically includes: Collect 2.00 mL of venous blood using a sodium heparin anticoagulation blood collection tube, place in a clean container for refrigerated transportation, and store at -70°C for future use; Use a blood collection needle to draw 2.00 mL of ultrapure water into a blood collection tube as a sample blank, place it in a clean container for refrigerated transportation, and store it at -70°C for future use.

5. The HPLC-ICPMS detection method for various forms of arsenic in whole blood according to claim 1, characterized in that: The extraction solution in step 4 is a methanol-acetonitrile solution (V / V=7:3), and step 4 specifically includes: Take 0.30 mL of whole blood sample and add 0.30 mL of methanol-acetonitrile solution (V / V=7:3) to form a blood sample solution.

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