KR20210007799A - Method and device for detecting inorganic arsenic of food - Google Patents

Abstract

The present invention provides a method for detecting inorganic arsenic from food regardless of a type of an object to be analyzed, and an apparatus thereof. According to one embodiment of the present invention, a method for detecting inorganic arsenic from food in an inorganic arsenic detection apparatus comprises the following steps: checking the type of an object to be analyzed input for detection of inorganic arsenic; determining whether food requires inorganic arsenic detection in accordance with the type of the object to be analyzed; controlling extraction of a test solution through a pretreatment method corresponding to the type of the object to be analyzed when it is determined that the food requires the inorganic arsenic detection; controlling the test solution to be separated through liquid chromatography (LC); controlling the test solution to be analyzed through an inductively coupled plasma/mass spectrometer (ICP/MS); and determining whether to detect the inorganic arsenic for the object to be analyzed based on an inorganic arsenic analysis result of the test solution.

Description

Method and device for detecting inorganic arsenic in food {METHOD AND DEVICE FOR DETECTING INORGANIC ARSENIC OF FOOD}

The present invention relates to a method and apparatus for detecting inorganic arsenic in food, and more particularly, to determine whether or not a food requiring inorganic arsenic detection according to the type of analyte, and if it is determined as a food requiring inorganic arsenic detection, a test solution It relates to an inorganic arsenic detection method and apparatus for detecting inorganic arsenic in food irrespective of the type of analysis object by automatically controlling the extraction, separation and analysis.

Arsenic is found naturally in soils, minerals and ores containing copper or lead. When these ores are heated, a lot of arsenic is released into the atmosphere in the form of fine dust. Contamination of arsenic in food is mainly contaminated with groundwater. In particular, grains, fruits, and vegetables such as rice grown under water are contaminated with arsenic by irrigation water. Arsenic is present in both natural and processed foods, and marine foods and some grain products are known to contain high amounts of arsenic.

Arsenic is an element with the element symbol As, atomic number 33, and atomic weight of 74.9216. It is an element belonging to the nitrogen group of Group 5 (N, P, As, Sb, Bi) of the periodic table, and is chemically classified as a metalloid. Metals and non-metals It has all the characteristics of, but is often regarded as a metal. Arsenic can exist in four valence states such as -3, 0, +3, +5. In general, trivalent arsenic (arsenite, AsIII) is in a reduced state, and pentavalent arsenic (arsenate, AsV) in an oxidized state is stable. to be. The chemical properties of arsenic are very complex and form various arsenic compounds, and are classified into inorganic arsenic compounds that bind to oxygen, chlorine, and sulfur, and organic arsenic that binds to carbon and hydrogen. .

Inorganic arsenic exists in various chemical forms in nature, but there are arsenic trioxide (AsIII) and arsenic pentoxide (AsV) in the environment and food. In general, trivalent inorganic arsenic compounds include arsenic trioxide, arsenenous acid, and sodium arsenite, and pentavalent inorganic arsenic compounds include arsenic salts such as arsenic pentoxide, arsenic acid, and lead arsenate. (arsenate). Organic arsenic includes monomethylarsinic acid (MMA), dimethylarsinic acid (DMA), trimethylarsine oxide, tetramethylarsonium ion, arsenobetaine (AsB), arsenocholine (AsC), and various arsenosugars and arsenolipids.

The International Agency for Research on Cancer (IARC) categorizes and manages arsenic as a human carcinogen, "Group 1" through rigorous epidemiological investigations, and the United States Environmental Protection Agency (EPA) manages the inorganic metal endocrine system. Arsenic as an impaired substance (ECD, dendocrine disruptor) is managed as "Group A", the first list of toxic chemicals.

Arsenic has a difference in toxicity depending on the type of compound, and inorganic arsenic is more toxic than organic arsenic. Among inorganic arsenic, AsIII is 10 times more toxic than AsV and 700 times more toxic than methylated organic arsenic species such as MMA and DMA. The degree of toxicity according to the form of arsenic is in the order of AsIII, AsV, MMA, DMA, AsC and AsB.

On the other hand, since most of the arsenic in foods is organic arsenic with low toxicity, only inorganic arsenic should be selectively identified and managed, but the test method for inorganic arsenic among the universal foods used internationally has not been suggested. Currently, there are many known LC-ICP/MS methods in which liquid chromatography (LC) is connected to an inductively coupled plasma mass spectrometer (ICP/MS) for analysis of inorganic arsenic, but because of the high skill level and high analysis cost. The investigation of the content of inorganic arsenic in food is insufficient.

In addition, although selective management of highly toxic inorganic arsenic is required, an inorganic arsenic analysis method has been suggested for some foods containing rice and seaweed in excluded countries, and most foods are analyzed as total arsenic instead of inorganic arsenic, which is difficult to analyze. . The safety management of arsenic in the future needs to be converted to research and management as inorganic arsenic that needs to be managed in practice, and accordingly, a technology to automatically detect inorganic arsenic in food is required.

The present invention is to solve the problems of the prior art described above, by determining whether or not a food requiring inorganic arsenic detection according to the type of analyte, and when it is determined as a food requiring inorganic arsenic detection, a test solution is extracted, separated and It is an object of the present invention to provide a method and apparatus for detecting inorganic arsenic for detecting inorganic arsenic in food irrespective of the type of an analysis target by automatically controlling the analysis to be analyzed.

The object of the present invention is not limited to the above-mentioned object, and other objects that are not mentioned will be clearly understood from the following description.

According to an embodiment of the present invention for achieving the above object, there is provided a method for detecting inorganic arsenic in food in an inorganic arsenic detection device, the method comprising the steps of: checking the type of analyte input for detection of inorganic arsenic; Determining whether or not the food is required to detect inorganic arsenic according to the type of the analysis target; Controlling the test solution to be extracted through a pretreatment method corresponding to the type of the analysis target when it is determined that the target for analysis is a food requiring detection of inorganic arsenic; Controlling the test solution to be separated through liquid chromatography (LC); Controlling the test solution to be analyzed through an inductively coupled plasma/mass spectrometer (ICP/MS); And determining whether or not to detect inorganic arsenic in the analysis target, based on the result of the analysis of inorganic arsenic in the test solution, there is provided a method for detecting inorganic arsenic in food.

The step of confirming the type of the analysis target may include checking the type of the analysis target through at least one of the viscosity of the analysis target and an image of the analysis target.

The inorganic arsenic detection method comprises the steps of controlling a standard solution to be separated through the liquid chromatography (LC); And setting the separation time of the test solution based on the peak detection time of the substance separated most recently from the standard solution.

The inorganic arsenic detection method comprises the steps of controlling the standard solution to be analyzed through the inductively coupled plasma/mass spectrometer (ICP/MS); And when it is confirmed that the peaks for some substances of inorganic arsenic and the peaks of some substances of organic arsenic are adjacent based on the results of inorganic arsenic analysis of the standard solution, the pH of the mobile phase used for separation of the test solution is preset. It may further comprise the step of controlling to be adjusted to the pH of the range.

The inorganic arsenic detection method may include determining whether or not a food requiring removal of an analyte interfering substance according to the type of the analysis target; And if it is determined that the analysis object is a food that needs to be removed from the analysis object, controlling the analysis object to be removed from the analysis object through at least one of a chemical removal method and a physical removal method.

According to another embodiment of the present invention for achieving the above object, in an inorganic arsenic detection device for detecting inorganic arsenic in food, the type of an analysis object injected for detection of inorganic arsenic is identified, and the type of the analysis object is Accordingly, an analysis target confirmation unit that determines whether or not the food is required to detect inorganic arsenic; A solution extracting unit for extracting a test solution through a pretreatment method corresponding to the type of the analysis target when it is determined that the analysis target is a food requiring detection of inorganic arsenic; A solution separation unit separating the test solution through liquid chromatography (LC); A solution analyzer for analyzing the test solution through an inductively coupled plasma/mass spectrometer (ICP/MS); And an inorganic arsenic detection unit configured to determine whether or not to detect inorganic arsenic in the analysis target based on the result of the analysis of inorganic arsenic in the test solution.

The analysis target identification unit may check the type of the analysis target through at least one of a viscosity of the analysis target and an image photographed of the analysis target.

The solution separation unit may separate the standard solution through the liquid chromatography (LC), and set the separation time of the test solution based on the peak detection time of the material separated from the standard solution last.

The solution analysis unit analyzes the standard solution through the inductively coupled plasma/mass spectrometer (ICP/MS), and the solution separation unit, based on the result of the analysis of inorganic arsenic in the standard solution, for some substances of inorganic arsenic. When it is confirmed that the peak and the peak for some substances of organic arsenic are adjacent, the pH of the mobile phase used for separation of the test solution can be adjusted to a pH within a preset range.

The analysis target identification unit determines whether or not a food requiring removal of an analysis interfering substance according to the type of the analysis target, and when it is determined that the analysis target is a food requiring removal of an analysis interfering substance, one of a chemical removal method and a physical removal method Through at least one, an analysis interfering substance may be removed from the analysis target.

According to an embodiment of the present invention, it is determined whether or not a food needs to be detected for inorganic arsenic according to the type of analyte, and when it is determined as a food requiring detection of inorganic arsenic, the test solution is automatically controlled to be extracted, separated, and analyzed. However, it has the advantage of being able to easily detect inorganic arsenic in food regardless of the type of analysis target.

The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a block diagram showing the configuration of an inorganic arsenic detection apparatus according to an embodiment of the present invention.
2 is a flowchart illustrating a process of detecting inorganic arsenic in food according to an embodiment of the present invention.
3 is a flowchart illustrating a process of adjusting the pH of a mobile phase according to an embodiment of the present invention.
4 is a flowchart illustrating a process of removing an analyte interfering substance according to an embodiment of the present invention.
5 is a view showing a standard material of arsenic species according to an embodiment of the present invention.
6 is a view showing a schematic diagram of a pre-treatment of food containing rice, tot or maternal and half according to an embodiment of the present invention.
7 is a view showing an experimental method for pretreatment of food containing rice, tot or maternal and half according to an embodiment of the present invention.
8 is a diagram showing a schematic diagram of pretreatment of fish oil (krill oil) according to an embodiment of the present invention.
9 is a view showing an experimental method for pretreatment of fish oil (krill oil) according to an embodiment of the present invention.
10 is a view showing a reverse phase column used for separation of arsenic species according to an embodiment of the present invention.
11 is a view showing a liquid chromatography-inductively coupled plasma/mass spectrometer according to an embodiment of the present invention.
12 is a view showing tubing between a column and an inductively coupled plasma-mass spectrometer according to an embodiment of the present invention.
13 is a view showing the analysis results of inorganic and organic arsenic according to an embodiment of the present invention.
14 is a view showing a calibration curve for inorganic and organic arsenic according to an embodiment of the present invention.
15 is a diagram showing a chromatogram of another standard solution according to an embodiment of the present invention.
16 is a view showing a blank chromatogram of a mobile phase preparation reagent according to an embodiment of the present invention.
17 is a diagram showing a blank chromatogram according to the nitric acid concentration of a test solution according to an embodiment of the present invention.
18 is a diagram showing a standard solution chromatogram according to the nitric acid concentration of a test solution according to an embodiment of the present invention.
19 is a diagram showing a co-experiment chromatogram of a standard solution according to a mobile phase pH according to an embodiment of the present invention.
20 is a view showing a chromatogram and a recovery rate (Rice CRM) comparison according to an extraction solution and an extraction method according to an embodiment of the present invention.
21 is a view showing a chromatogram and a recovery rate (Tot CRM) comparison according to an extraction solution and an extraction method according to an embodiment of the present invention.
22 is a diagram showing a chromatogram of a mixed standard solution according to an embodiment of the present invention.

Prior to the detailed description of the present invention, terms or words used in the present specification and claims described below should not be construed as being limited to their usual or dictionary meanings, and the inventors shall use their own invention in the best way. For explanation, based on the principle that it can be appropriately defined as a concept of terms, it should be interpreted as a meaning and concept consistent with the technical idea of the present invention. Therefore, the embodiments described in the present specification and the configurations shown in the drawings are only the most preferred embodiments of the present invention, and do not represent all the technical spirit of the present invention, and various equivalents that can replace them at the time of application It should be understood that there may be water and variations.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. In this case, it should be noted that the same components in the accompanying drawings are indicated by the same reference numerals as possible. In addition, detailed descriptions of known functions and configurations that may obscure the subject matter of the present invention will be omitted. For the same reason, some components in the accompanying drawings are exaggerated, omitted, or schematically illustrated, and the size of each component does not entirely reflect the actual size.

First, an apparatus for detecting inorganic arsenic in food according to an embodiment of the present invention will be described. 1 is a block diagram showing the configuration of an inorganic arsenic detection apparatus according to an embodiment of the present invention.

Referring to FIG. 1, the inorganic arsenic detection device 100 according to an embodiment of the present invention includes an analysis target identification unit 110, a solution extraction unit 120, a solution separation unit 130, and a solution analysis unit 140. And it may include an inorganic arsenic detection unit 150.

The analysis target identification unit 110, the solution extraction unit 120, the solution separation unit 130, the solution analysis unit 140, and the inorganic arsenic detection unit 150 may be program modules or hardware capable of communicating with external devices. . Such a program module or hardware may be included in the inorganic arsenic detection device 100 or another device capable of communicating with the inorganic arsenic detection device 100 in the form of an operating system, an application program module, and other program modules, and may be physically stored on various known storage devices. I can. Meanwhile, these program modules or hardware include routines, subroutines, programs, objects, components, data structures, etc. that perform specific tasks to be described later or execute specific abstract data types according to the present invention, but are not limited thereto and various It can also be implemented in a form.

The analysis target identification unit 110 may check the type of the analysis target injected for detection of inorganic arsenic.

Specifically, the analysis object confirmation unit 110 includes an input unit (not shown) for inputting the analysis object, or an input unit (not shown) implemented as an independent device is connected to the analysis object confirmation unit 110 Thus, when an analysis object is input through an input unit (not shown), the analysis object confirmation unit 110 can check what kind of the input analysis object.

When checking the analysis target, the analysis target identification unit 110 may check the type of the analysis target through at least one of the viscosity of the analysis target and an image of the analysis target.

Specifically, the analysis object confirmation unit 110 includes a viscosity measuring device (not shown) for measuring the viscosity of the analysis object, or a viscosity measuring device (not shown) implemented as an independent device is the analysis object confirmation unit 110 Since it is connected to, the analysis target identification unit 110 may measure the viscosity of the analysis target through a viscosity measuring device (not shown), and check the type of the analysis target according to the viscosity measurement result.

For example, when the analysis target is input while the category of the analysis target is set to rice, the analysis target verification unit 110 measures the viscosity of the analysis target through a viscosity meter (not shown), and the viscosity measurement value of the analysis target If it is measured to be higher than the reference value set for each category, the type of the analysis object can be identified as glutinous rice, and when the viscosity measurement value of the analysis object is measured to be lower than the reference value set for each category, the type of analysis object can be confirmed as hot rice.

In addition, the analysis object confirmation unit 110 includes a camera (not shown) for photographing the analysis object, or is connected to a camera (not shown) implemented as an independent device, so that the analysis object confirmation unit 110 An image photographed of an analysis target may be acquired through a camera (not shown), and the type of the analysis target may be identified according to the image analysis result.

For example, the analysis target identification unit 110 analyzes the image of the analysis target acquired through a camera (not shown) and classifies it into rice, barley, beans, etc. according to the color of the image, thereby confirming the type of analysis target. I can.

When the type of the analysis target is confirmed, the analysis target identification unit 110 may determine whether or not a food requiring inorganic arsenic detection according to the type of the analysis target.

For example, when rice is registered as a food that requires inorganic arsenic detection, the analysis target identification unit 110 may determine that the type of the analysis target is a food that requires inorganic arsenic detection for the input analysis target. And, if the type of the analysis object is confirmed as coffee, it can be determined as a food that does not require detection of inorganic arsenic for the input analysis object.

When it is determined that the analysis target is a food that needs inorganic arsenic detection, the analysis target identification unit 110 may further determine whether or not a food requiring removal of an analysis interfering substance according to the type of the analysis target.

For example, if seaweed is registered as a food with a lot of analysis interfering substances, the analysis target identification unit 110 determines that the type of analysis target is a food that requires removal of the analysis interfering substance when the type of analysis target is confirmed as seaweed. can do.

When it is determined that the analysis target is a food that needs to be removed from the analysis target, the analysis target identification unit 110 may process the analysis target to be removed from the analysis target through at least one of a chemical removal method and a physical removal method.

Specifically, the analysis target identification unit 110 includes an analysis interfering substance remover (not shown) for removing an analysis interfering substance, or an analysis interference remover (not shown) implemented as an independent device is an analysis target identification unit Since it is connected to 110, the analysis target identification unit 110 may process the analysis interfering material adhered to the analysis target to be removed through an analysis interfering material remover (not shown).

For example, the analysis target identification unit 110 may remove the analysis interfering material from the analysis target through a chemical removal method through active gases such as ammonia, oxygen, methane, and hydrogen, and physically through an inert gas such as helium. Through the removal method, an analyte interfering substance can be removed from the analyte.

When the analysis target identification unit 110 determines that the analysis target is a food that requires inorganic arsenic detection, the solution extraction unit 120 may extract the test solution through a pretreatment method corresponding to the type of the analysis target.

Specifically, the solution extraction unit 120 includes one or more extractors (not shown) for extracting the test solution through sample pretreatment, or at least one extractor (not shown) implemented as an independent device is an analysis target identification unit Since it is connected to 110, the analysis target identification unit 110 controls the extractor (not shown) corresponding to the type of the analysis target to be operated, so that the test solution can be extracted in a different pretreatment method according to the type of the analysis target. have.

For example, when the type of the analysis object is rice, the solution extraction unit 120 can control the rice extractor to operate to extract the test solution, and when the type of the analysis object is fish oil, the fish oil extractor is operated. The test solution can be extracted under control.

The solution separation unit 130 may separate the test solution extracted from the solution extraction unit 120 through liquid chromatography (LC).

Specifically, the solution separation unit 130 includes a liquid chromatography (LC) device for separating the mixed material into each component by using the difference in the interaction between the mobile phase and the material, or a liquid chromatography implemented as an independent device. Since the (LC) device is connected to the solution separation unit 130, the solution separation unit 130 can separate the test solution into respective components through a liquid chromatography (LC) device.

The solution analysis unit 140 may analyze the test solution separated by the solution separation unit 130 through an inductively coupled plasma/mass spectrometer (ICP/MS).

Specifically, the solution analysis unit 140 includes an inductively coupled plasma/mass spectrometer (ICP/MS) that ionizes an element through an inductively coupled plasma and separates the ions through a mass spectrometer to analyze the element, or is an independent device. The inductively coupled plasma/mass spectrometer (ICP/MS) implemented as is connected to the solution analysis unit 140, so the solution separation unit 140 is used in each of the test solutions through an inductively coupled plasma/mass spectrometer (ICP/MS). The components of can be analyzed.

The inorganic arsenic detection unit 150 may determine whether to detect inorganic arsenic for an analysis target based on the inorganic arsenic analysis result of the test solution analyzed by the solution analysis unit 140.

For example, the inorganic arsenic detection unit 150 checks the content of inorganic arsenic contained in the analysis target based on the inorganic arsenic analysis result of the test solution, and if the content of inorganic arsenic is higher than the reference value, the It can be determined that arsenic has been detected, and when the content of inorganic arsenic is lower than the reference value, it can be determined that inorganic arsenic has not been detected in the analysis target.

According to an embodiment, the solution extraction unit 120 may additionally extract various solutions such as a background solution and a standard solution in addition to the test solution.

The solution separation unit 130 may separate the standard solution through liquid chromatography (LC), and may set the separation time of the test solution based on the peak detection time of the material separated most recently from the standard solution.

For example, when the AsC non-small species is separated at the latest in the process of separating the standard solution, the solution separation unit 130 can check the peak detection time of AsC and set the AsC peak detection time as the separation time of the test solution. .

When it is confirmed that AsC is not separated from the standard solution, the solution separation unit 130 may set a preset time (eg, 20 minutes) as the separation time of the test solution.

According to an embodiment, the solution separation unit 130 may adjust the pH of the mobile phase used for separation of the test solution so that the test solution can be separated through the mobile phase set to a pH suitable for separation of non-small species.

Specifically, the solution analysis unit 140 may analyze the standard solution through an inductively coupled plasma/mass spectrometer (ICP/MS), and the solution separation unit 130 is based on the inorganic arsenic analysis result of the standard solution, If the peak for some substances of arsenic and the peaks for some substances of organic arsenic are confirmed to be adjacent, the pH of the mobile phase used for separation of the test solution can be adjusted to a pH within a preset range.

For example, as a result of component analysis of the standard solution, if it is confirmed that the peaks for AsIII of inorganic arsenic and MMA of organic arsenic are adjacent, the organic arsenic may be mistaken as inorganic arsenic, so that the solution separation unit 130 By adjusting the pH to a pH in the range of 2.5 to 2.8, it is possible to treat the peaks of AsIII and MMA to be separated when analyzing the components of the test solution.

Next, a process of detecting inorganic arsenic in food according to an embodiment of the present invention will be described in more detail. 2 is a flowchart illustrating a process of detecting inorganic arsenic in food according to an embodiment of the present invention.

First, when an analysis target is input into the inorganic arsenic detection device 100, in step S201, the inorganic arsenic detection device 100 may check the type of the analysis target.

Specifically, the inorganic arsenic detection device 100 can check the type of the analysis target injected for the detection of inorganic arsenic, and check which type of the analysis target is through at least one of the viscosity of the analysis target and an image of the analysis target. I can.

Thereafter, in step S202, the inorganic arsenic detection apparatus 100 may determine whether inorganic arsenic detection is necessary.

Specifically, the inorganic arsenic detection apparatus 100 may determine whether or not a food for which inorganic arsenic detection is required for the analysis target according to the type of the analysis target identified in step S201.

If it is determined in step S202 as a food that requires detection of inorganic arsenic for the analysis target, in step S203, the inorganic arsenic detection device 100 may extract the test solution, and detection of inorganic arsenic is not required for the target to be analyzed in step S202. If it is determined that there is no problem with the food, in step S208, the inorganic arsenic detection device 100 may output a pass message to inform that there is no problem with the food.

In extracting the test solution, the inorganic arsenic detection device 100 may control the test solution to be extracted through a pretreatment method corresponding to the type of the analysis object identified in step S201.

For example, the inorganic arsenic detection device 100 can control the extraction of the test solution through the first pretreatment method when the type of the analysis target is identified as rice, tot, or food containing mother and child, and the type of the analysis target is If it is identified as fish oil, krill oil, etc., the test solution can be controlled to be extracted through the second pretreatment method.

Thereafter, in step S204, the inorganic arsenic detection apparatus 100 may control the test solution extracted in step S203 to be separated through liquid chromatography (LC).

Thereafter, in step S205, the inorganic arsenic detection device 100 may control the test solution separated in step S204 to be analyzed through an inductively coupled plasma/mass spectrometer (ICP/MS).

Thereafter, in step S206, the inorganic arsenic detection apparatus 100 may determine whether inorganic arsenic is detected in the test solution.

Specifically, the inorganic arsenic detection apparatus 100 may determine whether to detect inorganic arsenic for an analysis target based on the inorganic arsenic analysis result of the test solution analyzed in step S205.

If it is determined that inorganic arsenic has been detected for the analysis target in step S206, in step S207, the inorganic arsenic detection device 100 may output a warning message to inform that there is a problem with the food, and the analysis target in step S206 If it is determined that inorganic arsenic has not been detected, in step S208, the inorganic arsenic detection apparatus 100 may output a pass message to inform that there is no problem with the food.

Next, a process of adjusting the pH of the mobile phase to prepare for separation of the test solution according to an embodiment of the present invention will be described in more detail. 3 is a flowchart illustrating a process of adjusting the pH of a mobile phase according to an embodiment of the present invention.

First, a standard solution may be directly extracted from the inorganic arsenic detection device 100, or a separately prepared standard solution may be added to the inorganic arsenic detection device 100.

In step S301, the inorganic arsenic detection device 100 may control the standard solution to be separated through the liquid chromatography (LC).

Thereafter, in step S302, the inorganic arsenic detection apparatus 100 may determine whether or not it is necessary to set the separation time of the test solution.

Specifically, the inorganic arsenic detection device 100 can check the peak detection time of the substance separated most recently from the standard solution, and determine whether it is necessary to set the separation time of the test solution through the peak detection time of the substance. have.

For example, the inorganic arsenic detection device 100 may determine that it is necessary to set the separation time of the test solution when the peak detection time of the substance separated most recently from the standard solution is greater than the reference value. If the peak detection time of the material separated late is less than the reference value, it can be determined that the setting of the separation time of the test solution is not necessary.

If it is determined that it is necessary to set the separation time of the test solution in step S302, the inorganic arsenic detection device 100 may set the separation time of the test solution in step S303, and set the separation time of the test solution in step S302. If it is determined that this is not necessary, step S304 may be performed without performing step S303.

In setting the separation time, the inorganic arsenic detection apparatus 100 may set the separation time of the test solution based on the peak detection time of the substance separated most recently from the standard solution.

For example, in the process of separating the standard solution, when the AsC arsenic species was separated last and the peak detection time of AsC is 15 minutes, the inorganic arsenic detection device 100 uses the AsC peak detection time of 15 minutes as the separation time of the test solution. It can be set as the AsC peak detection time of 15 minutes and the preset spare time of 5 minutes, combined with 20 minutes as the separation time of the test solution.

Thereafter, in step S304, the inorganic arsenic detection device 100 may control the standard solution to be analyzed through an inductively coupled plasma/mass spectrometer (ICP/MS).

Thereafter, in step S305, the inorganic arsenic detection apparatus 100 may determine whether a peak for some substances of inorganic arsenic and a peak for some substances of organic arsenic are adjacent to each other.

Specifically, the inorganic arsenic detection device 100 may compare and confirm the peak for each material of inorganic arsenic and the peak for each material of organic arsenic based on the result of analysis of inorganic arsenic in a standard solution. It is possible to determine whether or not the organic arsenic is in a range that can be mistaken for inorganic arsenic because the peak for the substance and the peak for some substances of the organic arsenic are adjacent.

If it is determined that the peaks for some substances of inorganic arsenic and the peaks of some substances of organic arsenic are adjacent in step S305, in step S306, the inorganic arsenic detection device 100 adjusts the pH of the mobile phase used for separation of the test solution. The preparation for the separation of the test solution can be completed by controlling so that it is possible to complete the preparation for the separation of the test solution, and if it is determined that the peaks for some substances of inorganic arsenic and the peaks of some substances of organic arsenic are not adjacent in step S305, step S306 is not performed. Preparation for separation of the test solution can be completed.

In adjusting the pH of the mobile phase, the inorganic arsenic detection device 100 controls the pH of the mobile phase used for separating the test solution to be adjusted to a pH within a preset range (for example, in the range of 2.5 to 2.8). When analyzing the components of, the peaks for some substances of inorganic arsenic and the peaks of some substances of organic arsenic can be treated to be separated.

Next, a process of removing an analyte interfering substance in preparation for extraction of a test solution according to an embodiment of the present invention will be described in more detail. 4 is a flowchart illustrating a process of removing an analyte interfering substance according to an embodiment of the present invention.

First, when an analysis target is input to the inorganic arsenic detection device 100, in step S401, the inorganic arsenic detection device 100 may check the type of the analysis target.

Thereafter, in step S402, the inorganic arsenic detection device 100 may determine whether or not the analysis interfering material needs to be removed.

Specifically, the inorganic arsenic detection apparatus 100 may determine whether or not a food requiring removal of an analysis interfering substance from the analysis target is based on the type of the analysis target identified in step S401.

If it is determined in step S402 as a food that needs to be removed from the analysis target, in step S403, the inorganic arsenic detection device 100 controls to remove the analysis target from the analysis target, thereby completing preparation for extraction of the test solution. If it is determined in step S402 as a food that does not require removal of an analysis interfering substance from the analysis target, preparation for extraction of the test solution may be completed without performing step S403.

In removing the analyte interfering substance, the inorganic arsenic detection apparatus 100 may control the analyte to be removed from the analysis target through at least one of a chemical removal method and a physical removal method.

Hereinafter, the process of analyzing inorganic arsenic in food will be described in more detail.

First, looking at the measurement principle, inorganic arsenic can be separated and analyzed by extracting inorganic arsenic from food using 1% nitric acid solution, separating it into an LC column using the ionicity of the arsenic compound, and quantifying it by ICP/MS. I can.

Next, looking at reagents and instruments, the reagents include distilled water (resistance 18 MΩ or higher), Arsenite (AsⅢ) standard, Arsenate (AsⅤ) standard, Monomethyl arsonic acid (MMA) standard, nitric acid, C ₃ H ₄ O ₄ , 60-62%), hexane (Hexane, CH ₃ (CH ₂ ) ₄ CH ₃ , 95% or more, MW 86.18), chloroform (Chloroform, CHCl ₃ , 99.999%, MW 119.38), methanol (Methanol, CH ₃ OH) , 99.999%, MW 32.04), 1-butane sulfonate sodium salt (Sodium 1-butane sulfonate, C ₄ H ₉ NaO ₃ S, 98% or more, MW 160.17), Malonic acid C ₃ H ₄ O ₄ , 98% or more, MW 104.06), tetramethylammonium hydroxide pentahydrate ((CH ₃ ) ₄ N(OH) 5H ₂ O, 97% or more, MW 181.23), etc. can be used, and a precision balance ( Those with an accuracy of at least 3 decimal places), micro pipette (1000, 5000 µl), volumetric flask (25, 50, 100, 1000 mL), reagent spoon, grinder, sealed container for sample storage, 15, 50 mL centrifuge tube , Agitator (Vortex mixer), weighing dish, scented water tank (to be kept above 80℃), centrifuge, syringe (10 ㎖), 0.45 ㎛ membrane filter (diameter 25-30 mm), pH meter , A reciprocating shaker or the like may be used.

The glassware directly affects the accuracy and precision in the process of the experiment, and to obtain the result of minimizing the contamination of inorganic arsenic from the glassware or decomposition vessel, a container made of polymer materials such as PP, PE, Teflon can be used. If a material is used, it can be used after washing with HCl solution (1:1).

CAPCELL PAK C18 MG (4.6 × 250 mm, 5 µm) or equivalent, liquid chromatography (LC), inductively coupled plasma/mass spectrometry (ICP/MS), etc. may be used as the analytical instrument.

Next, looking at the standard substances and preparation, the standard substances and preparation methods used in the analysis are as follows.

Inorganic arsenic and organic arsenic are standard substances of arsenic species, and inorganic arsenic is Arsenite (AsⅢ) standard (Sodium arsenite, NaAsO ₂ , FW129.91, 98% or more, Fisher chemical), Arsenate (AsV) standard (Sodium arsenate dibasic). heptahydrate, Na ₂ HAsO ₄ 7H ₂ O, FW312.01, 98% or more, Sigma-aldrich), etc., and organic arsenic includes Monomethyl arsonic acid (MMA) standard (Disodium methyl arsonate hexahydrate, CH ₃ AsNa ₂ O ₃ 6H ₂ O, FW291.90, 98% or higher, Chem service), Arsenocholine (AsC) standard (Arsenocholine bromide, C ₅ H ₁₄ AsBrO, FW244.99, 95% or higher, Wako).

Next, a picture related to the standard substance of the arsenic species is shown in FIG. 5. 5 is a view showing a standard material of arsenic species according to an embodiment of the present invention.

Looking at the standard stock solution and mixed standard solution, in preparing the standard stock solution, reagents and equipment include distilled water (resistance 18 MΩ or more), precision balance (three decimal places or more), weighing dish, reagent spoon, volumetric flask (100 mL) and the like can be used.

In the preparation of the standard stock solution, the standard form of arsenic species can be dissolved in distilled water to make it 1,000 ppm (100 mg / 100 mL) as arsenic (As), which can be calculated by considering the purity of the standard form. .

In step ①, Arsenite (AsⅢ) standard: As arsenic in Sodium arsenite (NaAsO2, MW 129.91), 0.173 g (value calculated as 100% purity) is dissolved in 100 mL of distilled water to make 1,000 ppm, 100 mL. I can.

In step ②, Arsenate (AsⅤ) standard: Sodium arsenate dibasic heptahydrate (Na ₂ HAsO ₄ 7H ₂ O, MW 312.01), 1,000 ppm as arsenic, to make 100 mL, 0.416 g (calculated as 100% purity) in distilled water. It can be prepared by dissolving in 100 mL.

In step ③, Monomethyl arsonic acid (MMA) standard: Disodium methyl arsonate hexahydrate (CH ₃ AsNa ₂ O ₃ 6H ₂ O, FW291.90) as arsenic in 1,000 ppm, 0.390 g (calculated as 100% purity) to make 100 mL One value) can be prepared by dissolving in 100 mL of distilled water.

In step ④, Arsenocholine (AsC): Arsenocholine bromide (C ₅ H ₁₄ AsBrO, FW244.99) as arsenic in 1,000 ppm, to make 100 mL 0.326 g (value calculated as 100% purity) 100 mL of tertiary distilled water It can be dissolved and prepared.

In MMA analysis, since AsIII and MMA peaks are adjacent to each other, there is a possibility that the MMA peak may be mistaken as AsIII. For accurate analysis, AsV, AsIII and MMA are analyzed together to confirm that they are separated.

In analyzing AsC, AsC is the latest material to be separated among non-small species, so the analysis time can be determined after confirming the AsC peak detection.

In preparing a mixed standard solution, reagents and instruments include distilled water (resistance 18 MΩ or higher), nitric acid (Nitric acid, C ₃ H ₄ O ₄ , 60-62%), micro pipette (1000, 5000 µl), volumetric flask (50 mL), 15, 50 mL centrifuge tubes, etc. may be used.

In the case of the standard stock solution, it can be stored in a refrigerator with little influence of light and temperature in order to minimize the change of arsenic species.Before preparing the mixed standard solution, take it out before a preset time (for example, 1 hour) and bring it to room temperature. Can be left unattended.

In the preparation of a mixed standard solution, a 100 ppm mixed standard solution can be prepared by taking 5 mL of each standard solution prepared at 1,000 ppm as arsenic (As) and dissolving 50 mL with 0.2% nitric acid solution, and 0.2% nitric acid solution It can be diluted with and prepared at the concentration of the calibration curve for the target food to be analyzed.

Mixed standard solutions can be prepared and used on the day of analysis because conversion between non-small species occurs when stored for a long time.

According to an embodiment, in the case of white rice with an inorganic arsenic standard of 0.2 mg/kg or less, calibration curves 1, 2, 5, 10, 20 ppb, etc. may be used, and if the test solution is 8 ppb or more based on a sample amount of 1 g, it will be judged as inappropriate. I can.

According to another embodiment, in the case of foods using brown rice, fusiformes, etc. of 0.1 mg/kg or less based on inorganic arsenic, calibration curves of 0.5, 1, 2, 5, 10 ppb, etc. may be used, and 4 ppb of test solution based on 1 g of sample amount In case of abnormality, it may be judged as inappropriate.

According to another embodiment, in the case of foods using brown rice, Japanese hemp, etc., which are 0.1 mg/kg or less based on inorganic arsenic, calibration curves 1, 10, 20, 50, 100 ppb, etc. may be used, and test solution 40 based on 1 g of sample amount. If ppb or more, it may be judged as inappropriate.

According to another embodiment, in the case of fish oil (krill oil) of 0.1 mg/kg or less based on inorganic arsenic, calibration curves of 0.5, 1, 2, 5, 10 ppb, etc. may be used, and test solution 2 ppb or more based on 1 g of sample amount It may be judged as nonconforming.

Next, referring to the sample pretreatment, the contents related to the schematic diagram of the pretreatment of the food containing rice, tot, or mother and child are shown in FIG. 6. 6 is a view showing a schematic diagram of a pre-treatment of food containing rice, tot or maternal and half according to an embodiment of the present invention.

In the experimental process related to the pretreatment of foods containing rice, soybean paste, or mother-and-child banquet, reagents and instruments include distilled water (resistance 18MΩ or higher), nitric acid (C ₃ H ₄ O ₄ , 60-62%), precision balance (third decimal point). One with more than one place accuracy), reagent spoon, micro pipette (1000, 5000 µl), volumetric flask (25 mL), shaker (Vortex mixer), 15, 50 mL centrifuge tube, scented water bath (maintain above 80℃) One), a centrifuge, a syringe (Plastic syringe, 10 mL), a 0.45 μm membrane filter (diameter 25-30 mm), and the like can be used.

Next, an experimental method for the pretreatment of food containing rice, tot, or maternal and half will be described with reference to FIG. 7. 7 is a view showing an experimental method for pretreatment of food containing rice, tot or maternal and half according to an embodiment of the present invention.

As shown in (a) of FIG. 7, in step ①, 1 g (0.2 to 1 g in the case of dried seaweed) can be precisely taken in a 50 mL centrifuge tube with a reagent spoon for the homogenized sample as a powder, After adding 5 mL of 1% nitric acid as an extraction solvent, capping the cap, you can sufficiently shake until homogenized using a Vortex mixer.

When shaking, using a Vortex mixer can be more effectively homogenized, and if the sample is taken in a 15 mL centrifuge tube, a 50 mL centrifuge tube can be used because there is a difficulty in the heating extraction process.

As shown in (b) of FIG. 7, in step ②, extraction can be performed by heating for 90 minutes in a constant temperature water bath set at 90°C, and at this time, 5 to 10 so that the sample and the extraction solvent are sufficiently mixed during the initial 30 minutes. You can shake it vigorously every minute.

During the extraction time, 90°C can be maintained, and heating and shaking can be repeated so that inorganic arsenic can be sufficiently extracted for the initial 30 minutes before starch is gelatinized.

In step ③, after the 90-minute heating extraction process is over, after cooling, 25 mL of distilled water can be distilled, and after distillation, it can be shaken once more using a Vortex mixer before centrifugation.

As shown in (c) of FIG. 7, in step ④, the first centrifugation may be performed through a process of centrifuging (3000G) for 10 minutes.

As shown in (d) of FIG. 7, in step ⑤, the second centrifugation may be performed through a process of appropriately taking the supernatant after the first centrifugation and centrifuging for 10 minutes (3000G).

As shown in (e) of FIG. 7, in step ⑥, the supernatant after the second centrifugation may be filtered through a membrane filter (nylon, 0.45 μm) and extracted as a test solution.

The two centrifugation and filtration processes are to minimize impurities injected into the analysis equipment (LC-ICP/MS). If the test solution is not clearer than the standard value, steps ⑤ and ⑥ may be additionally performed.

Next, the contents related to the schematic diagram of the pretreatment of fish oil (krill oil) are shown in FIG. 8. 8 is a diagram showing a schematic diagram of pretreatment of fish oil (krill oil) according to an embodiment of the present invention.

In the experimental process related to the pretreatment of fish oil (krill oil), reagents and equipment include distilled water (resistance 18 MΩ or higher), nitric acid (Nitric acid, C3H4O4, 60-62%), %), hexane (Hexane, CH ₃ (CH ₂ ) ₄ CH ₃ , 95% or more, MW 86.18), methanol (Methanol, CH ₃ OH, 99.999%, MW 32.04), chloroform (Chloroform, CHCl ₃ , 99.999%, MW 119.38), precision balance (accuracy of more than 3 decimal places) With), micro pipette (1000, 5000 µl), volumetric flask (25, 50 mL), 15, 50 mL centrifuge tube, reciprocating shaker, centrifuge, plastic syringe (10 mL), 0.45 A µm membrane filter (diameter 25-30 mm) or the like can be used.

Next, an experimental method for pretreatment of fish oil (krill oil) will be described with reference to FIG. 9. 9 is a view showing an experimental method for pretreatment of fish oil (krill oil) according to an embodiment of the present invention.

First, in step ①, 2.5 g of fish oil (krill oil) can be precisely taken into a 50 mL centrifuge tube, and 5 mL of hexane, 5 mL of methanol, and 0.5 mL of 1% nitric acid can be added as the extraction solution and the stopper can be closed.

If the sample is taken in a 15 mL centrifuge tube, it may be recommended to use a 50 mL centrifuge tube because the extraction process is difficult.

As shown in (a) of FIG. 9, in step ②, the first extraction may be performed through a process of shaking for 30 minutes using a shaker.

In step ③, centrifugation (3000G) can be performed for 10 minutes, and oil (fish oil) may be present in the supernatant (hexane), and inorganic arsenic may be present in the lower layer (methanol + 1% nitric acid).

As shown in (b) of FIG. 9, in step ④, the separated lower layer can be taken and collected in another centrifuge tube, and inorganic arsenic is extracted in the lower layer (methanol + 1% nitric acid). You can take it.

In step ⑤, the second extraction can be performed by adding 5 mL of chloroform, 3.75 mL of methanol, and 1.25 mL of 1% nitric acid to the remaining filtrate, and shaking for 30 minutes using a shaker.

In step ⑥, centrifugation (3000G) can be performed for 10 minutes, oil (fish oil) may be present in the lower layer (hexane + chloroform), and inorganic arsenic may be present in the supernatant (methanol + 1% nitric acid).

As shown in (c) of FIG. 9, in step ⑦, the separated supernatant can be taken and combined with the primary extract, and since inorganic arsenic is extracted in the supernatant (methanol + 1% nitric acid), pay attention to You can take it to the fullest.

In step ⑧, it can be adjusted to 50 mL with 0.2% nitric acid in the first and second extracts.

In step ⑨, centrifugation (3000G) can be performed for 10 minutes, and suspension may occur during normalization, so a clear liquid can be taken through centrifugation.

In step ⑩, the supernatant can be taken and diluted 2.5 times with 0.2% nitric acid, and dilution can be proceeded to reduce the methanol content while obtaining a clear test solution without suspension.

As shown in (d) of FIG. 9, in step ⑪, it can be filtered with a membrane filter (nylon, 0.45 ㎛) and extracted as a test solution, and the pressing force is adjusted so that the pressure is not applied strongly during filtration with the membrane filter. You can get a clear liquid.

Next, looking at the device analysis, all devices and conditions used for the analysis are as follows.

Regarding the test equipment, a column (CAPCELL PAK C18 MG (4.6×250 mm, 5 μm) or equivalent), liquid chromatography (LC), inductively coupled plasma/mass spectrometry (ICP/MS), etc. can be used. .

Regarding the mobile phase, reagents and instruments include water (resistance 18 MΩ or higher), nitric acid (C ₃ H ₄ O ₄ , 60-62%), methanol (Methanol, CH ₃ OH, 99.999%, MW 32.04), 1 -Butane sulfonate sodium salt (Sodium 1-butane sulfonate, C ₄ H ₉ NaO ₃ S, 98% or more, MW 160.17), Malonic acid C ₃ H ₄ O ₄ , 98% or more, MW 104.06), hydroxide Tetramethylammonium hydroxide pentahydrate ((CH ₃ ) ₄ N(OH)5H ₂ O, more than 97%, MW 181.23), precision balance (three-digit precision), micro pipette (1000, 5000 [Mu]l), volumetric flask (1000 mL), reagent spoon, pH meter, and the like can be used.

In the mobile phase preparation and method, the mobile phase is 0.05% (v/v) methanol, 10 mM sodium 1-butane sulfonate, 4 mM malonic acid, 4 mM tetramethylammonium hydroxide pentahydrate (TMAH) (pH 2.7), that is, 0.5 mL of methanol, 1 -Butanesulfonate sodium salt 1,634 g, malonic acid 0.425 g, tetramethylammonium hydroxide pentahydrate 0.747 g are dissolved in 990 mL distilled water, and the pH 2.7 is accurately adjusted with 10% nitric acid solution (about 1.8 mL/L), and the final 1000 It can be quantified in mL.

Since the separation of non-small species is very sensitive to pH, the pH must be accurately adjusted. If not, the pH meter can be calibrated, or a mobile phase in the range of pH 2.5-2.8 can be prepared and analyzed after confirming the separation.

Regarding the analysis method, the principle of analysis is that after making polarity using an ion exchange reagent in the reversed-phase column, the arsenic species is separated by liquid chromatography (LC) and qualitatively performed by an inductively coupled plasma-mass spectrometer (ICP/MS). And a method of quantifying.

Analysis sequence can be analyzed in the order of blank solution (0.2% nitric acid), mixed standard solution, and test solution. When analyzing a large number of samples, a mixed standard solution with a concentration of 10 ㎍/L at intervals of 15 test solutions to check the condition of the device If the recovery rate is less than 70% or exceeds 125% by analysis, it can be analyzed after rewriting the standard calibration curve.

LC instrument analysis conditions are shown in the following table.

LC conditions column CAPCELL PAK C18 MG (4.6×250 mm, 5 μm) or equivalent, Mobile phase 0.05%(v/v) methanol, 10 mM sodium 1-butane sulfonate, 4 mM malonic acid, 4 mM tetramethylammonium hydroxide pentahydrate (TMAH) (pH 2.7) Column temperature 25℃ Flow rate 0.75 mL/min Injection volume 10-50 μl Total analysis time 13 to 20 minutes

In the total analysis time, AsC is the latest material to be separated among non-small species, so the analysis time can be determined after confirming the detection of the AsC peak in the standard solution.If AsC is not added to the standard solution and analyzed, the next sample will not be affected. In order to be sufficiently eluted, 20 minutes may be set as the analysis time. Next, a picture related to the reverse phase column used for separating arsenic species is shown in FIG. 10. 10 is a view showing a reverse phase column used for separation of arsenic species according to an embodiment of the present invention.

Referring to FIG. 10, the reverse phase column is CAPCELL PAK C18 MG (4.6×250 mm, 5 μm), and the MG column among CAPCELL PAK C18 may be most suitable for analysis.

ICP/MS instrument analysis conditions are shown in the following table.

ICP/MS conditions RF power 1550 W Argon gas flow rates
Carrier
Makeup
0.80 L/min
0.35 L/min He gas flow rate 3.0 mL/min Analyte ion (m/z) As(75)

In the case of measuring foods with a lot of salt or many interfering substances (salt, seaweed, etc.), use a chemical interference removal method through active gas (ammonia, oxygen, methane, hydrogen, etc.), or use an inert gas (helium, etc.). The physical interference cancellation method used can be used, for example, it can be analyzed with AsO 91 in the DRC mode. Next, a picture related to liquid chromatography-inductively coupled plasma/mass spectrometer is shown in FIG. . 11 is a view showing a liquid chromatography-inductively coupled plasma/mass spectrometer according to an embodiment of the present invention.

Referring to FIG. 11, the liquid chromatography device and the inductively coupled plasma/mass spectrometer may be implemented as one integrated device, and the liquid chromatography device and the inductively coupled plasma/mass spectrometer may be implemented as separate devices and connected to each other. have.

Next, a picture related to the tubing between the column and the inductively coupled plasma-mass spectrometer is shown in FIG. 12. 12 is a view showing tubing between a column and an inductively coupled plasma-mass spectrometer according to an embodiment of the present invention.

Referring to FIG. 12, the length of the tubing entering from the column to the inductively coupled plasma-quality analyzer may be set to about 40 cm.

Next, a picture related to the analysis results of inorganic arsenic (AsIII, AsV) and organic arsenic (MMA, AsC) is shown in FIG. 13. 13 is a view showing the analysis results of inorganic and organic arsenic according to an embodiment of the present invention.

Referring to FIG. 13, it is possible to check the peak separation of AsIII and MMA, check the AsC peak elution, or set an analysis time of 20 minutes.

Next, a figure related to the calibration curves of inorganic arsenic (AsIII, AsV) and organic arsenic (MMA, AsC) is shown in FIG. 14. 14 is a view showing a calibration curve for inorganic and organic arsenic according to an embodiment of the present invention.

Regarding the preparation of the calibration curve, the peaks of the mass value (As, m/z 75) of the target element from the chromatogram of the test solution obtained under the above-described test operating conditions should be consistent with the retention time of the peak of the mixed standard solution. A calibration curve is prepared for each concentration of the standard solution, and the correlation coefficient (R ² ) of the standard calibration curve must have a linearity of 0.99 or more, and the result value of the analyzed test solution can be included within the range of the standard calibration curve.

Regarding the quantification of the test solution, a standard calibration curve can be drawn with the height or area of the peak obtained from each solution tested using the standard solution as the Y-axis and the concentration of arsenic species (㎍/L) as the X-axis. By applying the area of the arsenic species obtained from the test solution to the standard calibration curve, the concentration A (㎍/L) of the arsenic species in the test solution is calculated through [Equation 2], and the content of each arsenic species in the sample (mg/kg ) Can be calculated through [Equation 3].

Hereinafter, contents related to improvement of the problem of the inorganic arsenic test method will be described in detail.

Regarding the inorganic arsenic test method in rice, there are problems with the extraction solution for glutinous rice, and the problem of quantification of excess AsIII due to the change of the extraction solvent.

Regarding the problem of the extraction solution for glutinous rice, the existing malonic acid warm extraction method for glutinous rice produces a viscous extract and is difficult to filtration.To solve this problem, the extraction solvent can be improved using 1% nitric acid.

In the problem of quantifying the excess AsIII due to the change of the extraction solvent, the viscosity problem of glutinous rice is solved with the improved extraction solution (1% nitric acid), but the excessive quantification phenomenon of AsIII specifically occurs when analyzing the device. To solve this problem In addition, arsenic-detecting reagents (bicarbonate, phosphte) can be excluded and the mobile phase can be changed and replaced with a C18 column.

An isocratic analysis can be performed through a mobile phase with 0.05% (v/v) methanol, 10 mM sodium 1-butane sulfonate, 4 mM malonic acid, 4 mM tetramethylammonium hydroxide pentahydrate (TMAH), pH 3.0 (nitric acid), and CAPCELL Columns that are PAK C18 MG can be used.

Next, a picture related to the chromatogram of the 6 standard arsenic solution is shown in FIG. 15. 15 is a diagram showing a chromatogram of another standard solution according to an embodiment of the present invention.

Referring to FIG. 15, as a graph for chromatograms (0.5, 1, 2.5, 5, 10 (ug/L) of 6 arsenic standard solutions, the resolution for 6 arsenic species was confirmed, and the existing 4 Analysis of AsIII, AsV, MMA, DMA may be possible, and analysis of trace concentrations of non-small species with a low baseline may also be possible.

Regarding the inorganic arsenic test method for seaweed and rice and seaweed processed foods, there is a problem of generating a ghost peak in inorganic arsenic AsV, and a ghost peak occurs in inorganic arsenic AsV by mobile phase manufacturing reagent (Ammonium phosphate). I can.

Next, a picture related to a blank chromatogram with a mobile phase preparation reagent (Ammonium phosphate) is shown in FIG. 16. 16 is a view showing a blank chromatogram of a mobile phase preparation reagent according to an embodiment of the present invention.

Referring to FIG. 16, a ghost peak may be detected at a concentration of 0.2 ppb to 2 ppb in terms of AsV.

In confirming the ghost peak, as a result of analysis with the mobile phase prepared using a reagent with a high arsenic count, the concentration of the AsV ghost peak can be confirmed as high as about 10 times, and 0.2 ppb (minimum As count) ~ 2 ppb as As V It can be checked within the (Max As count) range.

To solve this problem, it is possible to change the mobile phase and replace it with a C18 column by excluding reagents (bicarbonate, phosphte) that detect arsenic.

An isocratic analysis can be performed through a mobile phase with 0.05% (v/v) methanol, 10 mM sodium 1-butane sulfonate, 4 mM malonic acid, 4 mM tetramethylammonium hydroxide pentahydrate (TMAH), pH 3.0 (nitric acid), and CAPCELL Columns that are PAK C18 MG can be used.

Next, in the problem of deterioration of the resolution according to the nitric acid concentration of the test solution, when the instrument is analyzed with 1% nitric acid as the extraction solution, the AsV peak turns off and the AsV peak spreads. As the concentration of nitric acid decreases (0.2% nitric acid) ) It can be seen that the base of the chromatogram is stable and the efficiency of the AsV peak is high.

In order to solve this problem, as shown in Figs. 17 and 18, the nitric acid concentration of the final test solution and standard solution can be adjusted to 0.2%.

17 is a view showing a blank chromatogram according to the nitric acid concentration of the test solution according to an embodiment of the present invention, Figure 18 is a standard solution chromatogram according to the nitric acid concentration of the test solution according to an embodiment of the present invention It is a figure shown.

In addition, regarding the problem of examining the peak resolution of AsIII and MMA, if the resolution is low because the peaks of AsIII and MMA are adjacent, there is a possibility that MMA is misrecognized as AsIII.

To solve this problem, as shown in Fig. 19, the mobile phase pH can be adjusted from 3.0 to 2.7.

19 is a diagram showing a co-experiment chromatogram of a standard solution according to a mobile phase pH according to an embodiment of the present invention.

Next, regarding the problem of unification of pretreatment methods, improvement is necessary to an objective and universal test method applicable to a wide range of food media, and the current test method divided into rice and seaweed and processed foods containing rice and seaweed is the scope of the standard. In the same way, it can be unified by pre-treatment for foods containing rice, tot or maternal.

In order to solve this problem, as shown in FIGS. 20 and 21, the same pretreatment method may be applied by heating extraction at 90° C. with 1% nitric acid.

20 is a view showing a chromatogram and a recovery rate (Rice CRM) comparison according to an extraction solution and an extraction method according to an embodiment of the present invention, and FIG. 21 is a view showing an extraction solution and an extraction method according to an embodiment of the present invention. It is a diagram showing the comparison of the chromatogram and the recovery rate (Tot CRM).

Accordingly, a unified test method can be prepared by integrating the inorganic arsenic test method in rice and the inorganic arsenic test method in seaweed and processed rice and seaweed foods, and the related conditions are as follows.

Foods that contain rice, tot or half hat Pretreatment 90℃ hot extraction in 1% nitric acid Standard solution 0.2% nitric acid dilution Mobile phase 0.05% (v/v) methanol, 10 mM sodium 1-butane sulfonate, 4 mM malonic acid, 4 mM tetramethylammonium hydroxide pentahydrate (TMAH), pH 2.7 (nitric acid) → isocratic analysis column Reversed phase column (C18)

Hereinafter, the contents related to the test method for inorganic arsenic in food will be described in detail. First, with respect to the scope of the test method for inorganic arsenic, it is applied to foods for which the standard standard for inorganic arsenic has been established ( Fish oil).

Regarding the analysis principle, inorganic arsenic in the sample can be extracted with 1% nitric acid solution, and arsenic species can be separated by liquid chromatography and analyzed by inductively coupled plasma/mass spectrometry.

Regarding the device, a liquid chromatograph-inductively coupled plasma/mass spectrometer (LC-ICP/MS) can be used, and when the liquid chromatograph and inductively coupled plasma/mass spectrometer are connected, the tube length can be set within 40 cm.

Regarding reagents and reagents, the standard stock solution contains sodium arsenite (AsIII, Sodium arsenite, NaAsO ₂ ) and sodium arsenate (AsV, Sodium arsenate dibasic heptahydrate, Na ₂ HAsO ₄ 7H ₂ O) standard foam in distilled water, and the concentration is as It can be set to 1,000mg/L, and the mixed standard solution can be used as a standard solution for preparing a calibration curve by diluting each standard stock solution with 0.2% nitric acid solution, and a mixed standard solution can be prepared and used on the day of analysis.

Regarding the preparation of the test solution, 1 g of a homogenized sample (0.2 g to 1 g in the case of dried seaweed) is precisely weighed and placed in a container (excluding glass containers) with 5 ml of 1% nitric acid solution for foods containing rice, rice, fusiformes, or mackerel. Hot water extraction can be performed at 90° C. for 90 minutes by adding. At this time, the sample and 1% nitric acid solution can be shaken vigorously every 5 to 10 minutes so that the sample and 1% nitric acid solution are sufficiently mixed during the initial 30 minutes.After extraction, add water to make 25 mL, mix well, and centrifuge for 10 minutes (3,000 G) After centrifugation, the supernatant is appropriately taken, centrifuged again for 10 minutes (3,000G), and the supernatant is filtered through a membrane filter (nylon, 0.45 ㎛) to extract the test solution.

In fish oil, 2.5 g of a sample was precisely weighed in a tube for a centrifuge, 5 mL of hexane, 5 mL of methanol, and 0.5 mL of 1% nitric acid were added, shaken for 30 minutes on a shaker, and centrifuged for 10 minutes (3,000 G). Extract) can be completely taken and transferred to a centrifuge tube, add 5 mL of chloroform, 3.75 mL of methanol, and 1.25 mL of 1% nitric acid to the remaining filtrate, shake again with a shaker for 30 minutes, and centrifuge for 10 minutes to separate the layered supernatant (Secondary extract) can be completely taken and combined with the first extract taken earlier. Add 0.2% nitric acid to make 50 mL, mix well, and centrifuge (3,000 G) for 10 minutes.After that, take 4 mL of the supernatant and adjust it to 10 mL with 0.2% nitric acid, and filter the membrane (nylon, 0.45 ㎛). The test solution can be extracted by filtering a part with a furnace.

Regarding the test operation, according to an embodiment of the measurement conditions of the liquid chromatograph, the column is C18 MG (4.6 × 250 mm, 5 μm) or equivalent, and the mobile phase is 0.05% (v/v) methanol, 10 A mixed solution of mM sodium 1-butane sulfonate, 4 mM malonic acid, and 4 mM tetramethylammonium hydroxide pentahydrate (TMAH) is prepared, and the pH of the solution is adjusted to exactly 2.7 using 10% nitric acid solution (about 1.8 mL/L). The column temperature is 25°C, the flow rate is 0.75 mL/min, the injection amount is 10-50 ul, and the analysis time can be set to a total of 13 minutes.

According to an embodiment of the measurement conditions of the inductively coupled plasma/mass spectrometer, the RF power is 1300 to 1500 W, the nebulizer argon gas is 0.95 to 1.15 L/min, and the auxillary argon gas flow is 1.2 to 1.5 L/min. , Plasma argon gas flow is 15~20 L/min, Lens voltage is 5~10 V, Ion monitored is As(m/z 75).When using DRC mode, it can be analyzed as AsO(m/z 91). have.

According to an example of preparing a calibration curve, a calibration curve can be prepared by taking a mixed standard solution by concentration (1-50 ppb) and analyzing the peak areas of AsV and AsIII on a chromatogram.

Next, a picture related to the chromatogram of the mixed standard solution (10 ppb each) is shown in FIG. 22. 22 is a diagram showing a chromatogram of a mixed standard solution according to an embodiment of the present invention.

According to an embodiment of the qualitative test, peaks of the mass value (m/z 75) of the target element from the chromatogram of the test solution obtained under the above-described test operating conditions may coincide with the retention time of the peak of the mixed standard solution.

According to an embodiment of the quantitative test, when the peak of the test solution obtained under the same conditions as the qualitative test coincides with the peak of the mixed standard solution, the peak area is substituted into the calibration curve to quantify AsIII and AsV, and the respective quantitative values are summed to It can be calculated from the content of arsenic.

All the above-described processes can be automatically performed by the control of the inorganic arsenic detection device 100, the inorganic arsenic detection device 100 can perform all the processes by itself, a device connected to the inorganic arsenic detection device 100 You can also do the whole process through.

As described above, according to an embodiment of the present invention, it is determined whether or not the food needs inorganic arsenic detection according to the type of the analysis target, and when it is determined that the food needs inorganic arsenic detection, the test solution is automatically extracted, separated, and analyzed. By controlling it as, there is an advantage in that it is possible to easily detect inorganic arsenic in food regardless of the type of analysis object.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not limiting. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as being distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the claims to be described later, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

100: inorganic arsenic detection device
110: Analysis target confirmation unit
120: solution extraction unit
130: solution separation unit
140: solution analysis unit
150: inorganic arsenic detection unit

Claims (10)

In the method for detecting inorganic arsenic in food in an inorganic arsenic detection device,
Confirming the type of the analysis object input for detection of inorganic arsenic;
Determining whether or not the food is required to detect inorganic arsenic according to the type of the analysis target;
Controlling the test solution to be extracted through a pretreatment method corresponding to the type of the analysis target, if it is determined that the target for analysis is a food requiring detection of inorganic arsenic;
Controlling the test solution to be separated through liquid chromatography (LC);
Controlling the test solution to be analyzed through an inductively coupled plasma/mass spectrometer (ICP/MS); And
A method for detecting inorganic arsenic in food comprising the step of determining whether or not to detect inorganic arsenic in the analysis target based on the result of inorganic arsenic analysis of the test solution. The method of claim 1,
The step of confirming the type of the analysis object,
A method for detecting inorganic arsenic in food comprising the step of confirming the type of the analysis target through at least one of the viscosity of the analysis target and an image photographed of the analysis target. The method of claim 1,
Controlling the standard solution to be separated through the liquid chromatography (LC); And
The method for detecting inorganic arsenic in food, further comprising the step of setting the separation time of the test solution based on the peak detection time of the material separated most recently from the standard solution. The method of claim 3,
Controlling the standard solution to be analyzed through the inductively coupled plasma/mass spectrometer (ICP/MS); And
Based on the results of the analysis of inorganic arsenic in the standard solution, if it is confirmed that the peaks for some substances of inorganic arsenic and the peaks for some substances of organic arsenic are adjacent, the pH of the mobile phase used for separation of the test solution is within a preset range. The method further comprising the step of controlling to be adjusted to the pH of the food, inorganic arsenic detection. The method of claim 1,
Determining whether or not a food requiring removal of an analysis interfering substance according to the type of the analysis target; And
Inorganic arsenic in food further comprising the step of controlling to remove the analysis interfering substance from the analysis target through at least one of a chemical removal method and a physical removal method when it is determined that the analysis target is a food requiring removal of the analysis interfering material. Detection method. In the inorganic arsenic detection device for detecting inorganic arsenic in food,
An analysis target identification unit that checks the type of an analysis target input for detection of inorganic arsenic, and determines whether or not a food requiring inorganic arsenic detection is determined according to the type of the analysis target;
A solution extracting unit for extracting a test solution through a pretreatment method corresponding to the type of the analysis target when it is determined that the target is to be a food that requires detection of inorganic arsenic;
A solution separation unit separating the test solution through liquid chromatography (LC);
A solution analyzer for analyzing the test solution through an inductively coupled plasma/mass spectrometer (ICP/MS); And
Inorganic arsenic detection apparatus comprising an inorganic arsenic detection unit for determining whether or not to detect the inorganic arsenic in the analysis target based on the inorganic arsenic analysis result of the test solution. The method of claim 6,
The analysis target confirmation unit,
Inorganic arsenic detection device for confirming the type of the analysis target through at least one of the viscosity of the analysis target and an image photographed of the analysis target. The method of claim 6,
The solution separation unit,
Separating the standard solution through liquid chromatography (LC), and setting the separation time of the test solution based on the peak detection time of the material separated last from the standard solution, inorganic arsenic detection device. The method of claim 8,
The solution analysis unit,
Analyze the standard solution through the inductively coupled plasma/mass spectrometer (ICP/MS),
The solution separation unit,
Based on the result of the analysis of inorganic arsenic in the standard solution, when it is confirmed that the peaks for some substances of inorganic arsenic and the peaks for some substances of organic arsenic are adjacent, the pH of the mobile phase used for separation of the test solution is set within a preset range. Inorganic arsenic detection device to adjust the pH of. The method of claim 6,
The analysis target confirmation unit,
Depending on the type of the analysis target, it is determined whether or not the food needs to be removed from the analysis target, and if it is determined that the target is a food requiring removal of the analysis target, the analysis is performed through at least one of a chemical removal method and a physical removal method. Inorganic arsenic detection device that processes to remove analyte interfering substances from the target.

Images