AU2019100873A4 - Analytical method for simultaneously determining content of harmful heavy metals in edible fungi cultivation matrix by using combined digestion - Google Patents
Analytical method for simultaneously determining content of harmful heavy metals in edible fungi cultivation matrix by using combined digestion Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/4044—Concentrating samples by chemical techniques; Digestion; Chemical decomposition
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
An analytical method for simultaneously determining a content of harmful heavy metals in an edible fungi cultivation matrix by using combined digestion, including weighing 0.35 g (accurate to 0.0001 g) of a matrix sample which has been air-dried, ground and sieved through a C0.15 mm; placing the matrix sample in a Poly tetra fluoroethylene (PTFE) microwave digestion tank; adding 6 mL of nitric acid and soaking overnight, and then adding 2 mL of hydrogen peroxide to perform microwave digestion. After the digestion is complete and cooled, performing the acid-driven to 0.5-1.0 mL at 130 °C, and taking the matrix sample down for cooling. The digestion liquid is transferred to a 25 mL volumetric flask with ultra pure water to perform a constant volume. The measuring method has high accuracy, simple and quick operation, is suitable for batch measurement, and improves work efficiency.
Description
ANALYTICAL METHOD FOR SIMULTANEOUSLY DETERMINING
CONTENT OF HARMFUL HEAVY METALS IN EDIBLE FUNGI CULTIVATION MATRIX BY USING COMBINED DIGESTION
TECHNICAL FIELD [0001] The present invention relates to the field of agricultural product quality and safety, particularly to an analytical method for the content of harmful heavy metals (arsenic, mercury, lead, cadmium) in the edible fungi cultivation matrix, more specifically, relating to an analytical method for simultaneously determining a content of harmful heavy metals in edible fungi cultivation matrix by using combined digestion.
BACKGROUND [0002] As a sunrise industry in the 21st century, edible fungi have the advantages of high protein, low fat, no pollution, no pollution, which can act as food and medicine both, is recommended by the UN Food and Agriculture Organization as healthy food. The edible fungi industry is an short-adaptable-fast economic development project that integrates economic benefits, ecological benefits and social benefits. However, with the rapid increase in the number of edible fungi cultivation, the main raw materials used for artificial cultivation of edible fungi have shifted from simple wood chips or logs to agricultural and sideline products and even industrial waste. Because edible fungi have a certain absorption and enrichment effect on heavy metals, in addition, the cultivation environment (air, water, soil, etc.) of edible fungi has been polluted to varying degrees in recent years, resulting in the occurrence of heavy metals exceeding the standard in edible fungi. Studies have shown that because heavy metals can not be degraded by microorganisms, and will be enriched and transformed in living organisms, and finally enter the human body through the food chain, when the heavy metals in the human body exceed a certain limit, it will cause serious harm to the body. Therefore, strengthening the quality control of the edible fungi production environment is of great significance to ensure the quality and safety of edible fungi. At present, China has not issued a national standard method for determining the content of harmful heavy metals in the edible fungi cultivation matrix.
2019100873 08 Aug 2019
The method for determining the content of harmful heavy metals in the the edible fungi cultivation matrix at home and abroad mainly refers to the test method for determining total arsenic, total mercury, lead and cadmium in the soil. Due to the large differences in the formation mechanism, conditions and composition of the cultivation matrix and the soil, it is not appropriate to determine the harmful heavy metals in the cultivation matrix completely according to the soil measurement method. In this paper, the determination method of harmful heavy metals (arsenic, mercury, lead, cadmium) in edible fungi cultivation matrix is designed and verified by methods. At present, there is no public report in China.
SUMMARY [0003] In order to accurately evaluate the safety of edible fungi, the present invention establishes a method for determining harmful heavy metals such as arsenic, mercury, lead and cadmium in edible fungus matrix by using combined digestion. The method has high accuracy of determining the content of arsenic, mercury, lead and cadmium in the edible fungus matrix, and the operation is simple and quick, which is suitable for batch measurement and improving work efficiency.
Experimental materials [0004] 1.1 Reagents: nitric acid (MOS); 30% hydrogen peroxide; multi-element mixed standard stock solution Part# 5183-4688 (Agilent; USA): 10 μ g/mL As, Cd, Pb; the internal standard solution Part #5188-6525 (Agilent; USA): Rh concentration is 100 μ g/mL, diluted to 1 μ g/mL with (1+19) HNO3 solution; tuning solution Part# 5188 -6545 (Agilent; USA): Li, Co, Y, Ce, T1 mixed standard solution with a concentration of 10 ng/mL, the medium is (1+49) HNO3 solution; the mercury standard solution GBW(E) 080392 (National Standards Research Center); the experimental water is ultrapure water with a resistivity greater than 18.2 ΜΩ prepared by Milli-Q.
[0005] 1.2 Standard Product: National first-class standard reference material soil GBW07427, poplar leaves GBW07604.
[0006] 1.3 instruments: Agilent 7500a inductively coupled plasma mass
2019100873 08 Aug 2019 spectrometer (Agilent Technologies Co. Ltd., USA), with a Babington high salt atomizer; spray chamber: quartz double-channel and Piltier semiconductor with the temperature controlled at (2 ± 0.1)°C; Rectangular tube: quartz integrated, 215mm center channel; sample cone: Ni material cone. MARS5 Microwave Digestion Apparatus (CEM, USA) with poly tetra fluoroethylene digestion tank. DKQ-3C intelligent temperature control electric heater (Shanghai Yiyao Analytical Instrument Co., Ltd.).
Experimental method
2.1 Preparation of sample solution:
[0007] 0.35 g (accurate to 0.0001 g) of a matrix sample which has been air-dried, ground and sieved through a Φ0.15 mm is weighed, and then the matrix sample is placed in a poly tetra fluoroethylene microwave digestion tank, 6 mL of nitric acid is added, soaking overnight. After then, 2 mL of hydrogen peroxide is added to perform microwave digestion, the microwave digestion procedure is shown as Table 1. After the microwave digestion is complete and cooled, an acid-driven is performed to 0.5-1.0 mL at 130 °C and taken the matrix sample down for cooling. Then, the digestion liquid is transferred to a 25 mL volumetric flask with ultra pure water to perform a constant volume for use. A blank test is set at the same time, wherein standard material soil and poplar leaves are treated in the same way as the matrix sample, and treatment liquid is further diluted according to the actual sample content.
Table 1 Microwave Digestion Procedure
| Step | Power (W) | Power Percentage (%) | Rise Time (min) | Temperature (°C) | Holding time (min) |
| 1 | 1200 | 100 | 6 | 120 | 2 |
| 2 | 1200 | 100 | 4 | 140 | 5 |
2019100873 08 Aug 2019
| 3 | 1200 | 100 | 3 | 160 | 20 |
2.2 Sample determination
2.2.1 Optimization of instrument conditions [0008] The instrument is tuned with tuning fluids (the content of 7Li, 89Y, 59Co, 140Ce, and 2O5T1 is 10 ng/mL) by regulating the parameters such as RF power, carrier gas flow, torch horizontal and vertical position, and sampling depth, etc. so that the sensitivity and stability is optimized to allow the instrument to meet detection requirements. The optimized instrument operating parameters are shown in Table 2. According to the principle of large abundance, small interference and high sensitivity to select the isotope is shown in Table 3.
Table 2 ICP-MS operating parameters
| Item | Parameters |
| Power | 1200 w |
| Coolant Gas Flow | 15.0 1/min |
| Auxiliary Gas Flow | 1.0 1/min |
| Carrier Gas Flow | 1.10 1/min |
| Sample Lifting Rate | 1.0 ml/min |
| Sampling Depth | 7.0 mm |
| Analysis Model | Quantitative Model |
| Acquisition Points Of Unit Mass Number | 3 |
| Data Acquisition Model | Peak Jumping Acquisition Model |
| Integration Time | 0.3 s/isotope |
| Data Acquisition Repetition Number | 3 |
Table 3 Isotope Selection
| Element | As | Cd | Hg | Pb |
| Isotope | 75 | 111 | 202 | 208 |
2019100873 08 Aug 2019
2.2.2 Standard curve drawing [0009] Diluting multi-element mixed standard solution Part#5183-4688 with (1+19) HNO3 solution to dilute standard stock solution into 5, 10, 20, 50, 100, 200 ng/mL of 6 concentration gradients.
[0010] Diluting the mercury standard solution GBW(E)080392 with (1+19) HNO3 solution to dilute standard stock solution into 0.2, 0.5, 1.0, 1.5, 2.0 ng/mL of 5 concentration gradients.
[0011] Reagent blank: (1 + 19) HNO3 solution.
2.2.3 Sample Measurement [0012] The measuring method is edited under optimized instrument conditions, the on-line internal standard calibration instrument drift is introduced to improve matrix influence, and after the instrument is stable, the reagent blank, standard curve solution, standard solution and matrix sample solution are introduced in sequence. A calibration file is edited to select an appropriate internal standard calibration element, and the concentration of each element in the sample is calculated based on the calibration equation.
[0013] Presentation of analysis results:
2019100873 08 Aug 2019 x (C-Co)xv mx(l-Ao)xlOOO
X—the element content in the sample (mg/kg); C—the concentration of the digestion liquid of the sample (ng/mL); V—the total volume of the digestion liquid of the sample (mL); m—the mass of the sample (g); X0— The water content of the air drying test.
2.3 Controlled experiments and precision test [0014] According to the above method, the standard substance soil (GBW07427) and poplar leaves (GBW07604) are used as controlled experiments to determine the contents of arsenic, mercury, lead and cadmium, and the repeated measurements are carried out for six times, the results are shown in Table 4. The results show that the above determination method can satisfy the determination of each element in this experiment, and the precision of the instrument is good.
Table 4 Controlled experiments and precision test (n=6)
| Element | Solid (GBW07427) | Poplar Leaves (GBW07604) | ||||
| Measured value (mg/kg) | RSD (%) | Certificate value (mg/kg) | Measured value (mg/kg) | RSD (%) | Certificate value (mg/kg) | |
| As | 11.2 | 2.04 | 10.6±0.8 | 0.418 | 3.75 | 0.37±0.09 |
| Cd | 0.135 | 3.28 | 0.13±0.01 | 0.295 | 3.02 | 0.32±0.07 |
| Hg | 0.049 | 2.93 | 0.052±0.006 | 0.030 | 5.41 | 0.026±0.003 |
| Pb | 20.8 | 1.26 | 21.6M.2 | 1.34 | 1.59 | 1.5±0.3 |
2019100873 08 Aug 2019
2.4 Repeatability test [0015] Under the same conditions, 6 edible fungi cultivation matrices of different varieties are weighed and made into 6 parallels test. The parallel samples are prepared and determined according to the above method, and the results are calculated and expressed by RSD. The results are shown in Table 5 and show that the method has good repeatability.
Table 5 Repeatability test (n = 6)
2019100873 08 Aug 2019
2.5 Recovery test [0016] In order to verify the accuracy of the method, six different varieties of edible fungi cultivation matrix samples are performed spike-and-recovery testing. The analysis results are shown in Table 6-9. It can be seen from the results that the recovery of arsenic is between 95.3% and 99.0%, the recovery of cadmium is between 97.4% and 101.5%, the recovery of mercury is between 91.7% and 102.0%, and the recovery of lead is 97.7%~101.9%. It shows that the method can accurately determine the content of each element of the cultivation matrix, and the method is accurate and reliable.
Table 6 Arsenic spike-and-recovery testing
| Sample item | Measured value (mg/kg) | Spike amount (mg/kg) | Spike measured value (mg/kg) | Recovery rate (%) |
| oyster mushroom matrix | 0.072 | 1.00 | 1.05 | 97.9 |
| shiitake mushroom matrix | 0.170 | 1.00 | 1.13 | 96.6 |
| agrocybe cylindracea matrix | 0.081 | 1.00 | 1.07 | 99.0 |
| pleurotus nebrodensis matrix | 0.144 | 1.00 | 1.09 | 95.3 |
| pleurotus eryngii matrix | 0.058 | 1.00 | 1.03 | 97.4 |
| ganoderma lucidum matrix | 0.053 | 1.00 | 1.02 | 96.9 |
Table 7 Cadmium spike-and-recovery testing
2019100873 08 Aug 2019
| Sample item | Measured value (mg/kg) | Spike amount (mg/kg) | Spike measured value (mg/kg) | Recovery rate (%) |
| oyster mushroom matrix | 0.049 | 1.00 | 1.03 | 98.2 |
| shiitake mushroom matrix | 0.034 | 1.00 | 1.05 | 101.5 |
| agrocybe cylindracea matrix | 0.037 | 1.00 | 1.04 | 100.3 |
| pleurotus nebrodensis matrix | 0.041 | 1.00 | 1.02 | 98.0 |
| pleurotus eryngii matrix | 0.047 | 1.00 | 1.02 | 97.4 |
| ganoderma lucidum matrix | 0.179 | 1.00 | 1.18 | 100.1 |
Table 8 Mercury spike-and-recovery testing
| Sample item | Measured value (mg/kg) | Spike amount (mg/kg) | Spike measured value (mg/kg) | Recovery rate (%) |
| oyster mushroom matrix | 0.030 | 0.20 | 0.221 | 91.7 |
| shiitake mushroom matrix | 0.047 | 0.20 | 0.238 | 96.4 |
2019100873 08 Aug 2019
| agrocybe cylindracea matrix | 0.029 | 0.20 | 0.217 | 94.8 |
| pleurotus nebrodensis matrix | 0.061 | 0.20 | 0.252 | 96.6 |
| pleurotus eryngii matrix | 0.036 | 0.20 | 0.230 | 97.5 |
| ganoderma lucidum matrix | 0.105 | 0.20 | 0.311 | 102.0 |
Table 9 Lead spike-and-recovery testing
| Sample item | Measured value (mg/kg) | Spike amount (mg/kg) | Spike measured value (mg/kg) | Recovery rate (%) |
| oyster mushroom matrix | 0.422 | 1.00 | 1.39 | 97.7 |
| shiitake mushroom matrix | 1.15 | 1.00 | 2.13 | 99.1 |
| agrocybe cylindracea matrix | 0.214 | 1.00 | 1.22 | 100.5 |
| pleurotus nebrodensis matrix | 0.998 | 1.00 | 1.99 | 99.6 |
| pleurotus eryngii matrix | 0.203 | 1.00 | 1.19 | 98.9 |
| ganoderma lucidum matrix | 1.65 | 1.00 | 2.70 | 101.9 |
2019100873 08 Aug 2019 [0017] Throughout the specification and the claims that follow, unless the context requires otherwise, the words “comprise” and “include” and variations such as “comprising” and “including” will be understood to imply the inclusion of a stated integer or group of integers, but not the exclusion of any other integer or group of integers.
Claims (4)
1. An analytical method for simultaneously determining a content of harmful heavy metals in an edible fungi cultivation matrix by using combined digestion, comprising (1) weighing 0.35 g (accurate to 0.0001 g) of a matrix sample which has been air-dried, ground and sieved through a Φ 0.15 mm, placing the matrix sample in a poly tetra fluoroethylene microwave digestion tank, adding 6 mL of nitric acid and soaking overnight, and then adding 2 mL of hydrogen peroxide to perform microwave digestion; after the microwave digestion is complete and cooled, performing an acid-driven to 0.5-1.0 mL at 130 °C, and taking the matrix sample down for cooling; transferring digestion liquid to a 25 mL volumetric flask with ultra pure water to perform a constant volume for use; setting a blank test at the same time, wherein standard material soil and poplar leaves are treated in the same way as the matrix sample, and treatment liquid is further diluted according to an actual sample content;
2. The method according to claim 1, wherein a sample to be tested is samples of edible fungi matrix, including various samples of the edible fungi matrix such as oyster mushroom, shiitake mushroom, enoki mushroom, and pleurotus eryngii.
(2) standard curve drawing: diluting multi-element mixed standard solution Part#5183-4688 with (1+19) HNO3 solution to dilute standard stock solution into 5, 10, 20, 50, 100, 200 ng / mL of 6 concentration gradients;
diluting mercury standard solution GBW(E)080392 with (1+19) HNO3 solution to dilute standard stock solution into 0.2, 0.5, 1.0, 1.5, 2.0 ng / mL of 5 concentration gradients;
reagent blank: (1 + 19) HNO3 solution;
3. The method according to claim 1, wherein the instrument is an inductively coupled plasma mass spectrometer (ICP-MS).
(3) sample measurement: editing the measuring method under optimized instrument conditions, introducing on-line internal standard calibration instrument drift to improve matrix influence, and after the instrument is stable, introducing the reagent blank, standard curve solution, standard solution and matrix sample solution in sequence; editing a calibration file, selecting an appropriate internal standard
2019100873 08 Aug 2019 calibration element, and calculating concentration of each element in the sample based on the calibration equation.
4. The method according to claim 1, wherein the microwave digestion procedure has a power of 1200 W and a power percentage of 100%, a temperature rise time is set to 6 min at first, a temperature is raised to 120°C and hold for 2 min, and then the temperature rise time is set to 4 min, the temperature is raised to 140°C and hold for 5 min, a final temperature rise time is set to 3 min, and the temperature is raised to 160°C and hold for 20 min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112014174A (en) * | 2020-07-14 | 2020-12-01 | 中国科学院城市环境研究所 | Method for measuring phosphorus content in livestock and poultry manure and biochar thereof |
| CN112080436A (en) * | 2020-09-29 | 2020-12-15 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Large fungus strain with high cadmium resistance and application thereof |
-
2019
- 2019-08-08 AU AU2019100873A patent/AU2019100873A4/en not_active Ceased
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112014174A (en) * | 2020-07-14 | 2020-12-01 | 中国科学院城市环境研究所 | Method for measuring phosphorus content in livestock and poultry manure and biochar thereof |
| CN112080436A (en) * | 2020-09-29 | 2020-12-15 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Large fungus strain with high cadmium resistance and application thereof |
| CN112080436B (en) * | 2020-09-29 | 2022-01-04 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Large fungus strain with high cadmium resistance and application thereof |
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