Technical specification of efficacy evaluation for removing of heavy metal by biological products

GB/T 39029-2020.Technical specification of efficacy evaluation for removing of heavy metal by biological products.
1 Scope
GB/T 39029 specifies the general principles, basic requirements, evaluation methods and evaluation report expressions
for the evaluation of the efficacy of biological products in removing heavy metals. GB/T 39029 is applicable to the evaluation of the effect of biological products in removing heavy metals from water.
2 Normative references
The following documents are indispensable for the application of this document. For any dated referenced document, only the dated version applies to this document. For any undated referenced document, its latest version (including all amendments) applies to this document.
GB/T 6682 Specifications and test methods for water for analytical laboratories
GB/T 5750.6 Standard test methods for drinking water Metal indicators
3 Terms and definitions
The following terms and definitions apply to this document.
3.1
Biological products
Products prepared by biotechnology.
Note: The biological products in this standard specifically refer to microorganisms or enzyme products with the function of removing heavy metals.
4 General principles
4.1 Biological product labels or instructions can only be evaluated if there is a clear name of the microorganism or enzyme product.
4.2 The efficacy evaluation work shall be carried out scientifically and impartially, and the evaluation process and evaluation methods shall be scientific.
4.3 Statistical methods shall be used for comparison, and the inspection report shall not be forged or altered.
5 Basic requirements
5.1 Samples shall be received, stored, handed over, prepared, recovered and returned/destroyed in accordance with the requirements of relevant laws, regulations, standards and product instructions, and corresponding management systems and procedures shall be formulated.
5.2 Laboratories conducting microbial product evaluation shall meet the requirements of biosafety level 1.
5.3 The types, quantity, performance, range and accuracy of the instruments and equipment used for evaluation shall meet the needs of efficacy evaluation. The utensils and materials required for the evaluation of the efficacy of microbial products should be sterilized.
5.4 The reagents used in the evaluation should be analytically pure, and the water should comply with the secondary water specified in GB/T 6682.
5.5 The heavy metals used in the evaluation are prepared from heavy metal standards with a purity greater than or equal to 98%, or standard substances certified by the state and awarded with standard substance certificates, into standard solutions of a certain mass concentration, and the culture medium should be selected according to the type of microorganism.
5.6 After the evaluation experiment is completed, the experimental materials should be harmlessly treated.
This standard specifies the general principles, basic requirements, evaluation methods, and evaluation report expressions for the evaluation of the efficacy of biological products in removing heavy metals. This standard applies to the evaluation of the effect of biological products in removing heavy metals in water.

ICS07.080
National Standard of the People's Republic of China
GB/T39029—2020
Technical specification of efficacy evaluation for removing ofheavymetalbybiologicalproducts2020-07-21Issued
State Administration for Market Regulation
National Administration of Standardization
Issued
2021-02-01Implemented
Foreword
This standard was drafted in accordance with the rules given in GB/T1.1-2009 GB/T39029—2020
Please note that some of the contents of this document may involve patents. The issuing agency of this document does not assume the responsibility for identifying these patents. This standard was proposed and managed by the China National Institute of Standardization. The drafting units of this standard are: Jiangnan University, China National Institute of Standardization, and Beijing Sambo Technology Co., Ltd. The main drafters of this standard are Wu Xiaoling, Xu Chuanlai, Kuang Hua, Ma Aijin, Lin Lu, Liu Liqiang, Xu Liguang, Ma Wei, Guo Lingling, and Hao Shuai. I
1 Scope
Technical specification for evaluation of heavy metal removal efficacy of biological products GB/T39029—2020
This standard specifies the general principles, basic requirements, evaluation methods and evaluation report expressions for the evaluation of heavy metal removal efficacy of biological products. This standard applies to the evaluation of the effect of biological products in removing heavy metals from water. Normative references
The following documents are indispensable for the application of this document. For all dated references, only the dated version applies to this document. For all undated references, the latest version (including all amendments) applies to this document GB/T6682 Specifications and test methods for water for analytical laboratories GB/T5750.6 Standard test methods for drinking water Metal index terms and definitions
The following terms and definitions apply to this document. 3.1
biologicalproducts
Biological products
Products prepared by biotechnology.
Note: The biological products in this standard specifically refer to microorganisms or enzyme products with the function of removing heavy metals. 4 General principles
4.1 Biological product labels or instructions can only be evaluated if there is a clear name of the microorganism or enzyme product. The efficacy evaluation work shall be carried out scientifically and impartially, and the evaluation process and evaluation methods shall be scientific. 4.2
4.3 Statistical methods shall be used for comparison, and the inspection report shall not be forged or altered. 5 Basic requirements
5.1 Samples shall be received, stored, handed over, prepared, recovered and returned/destroyed in accordance with the requirements of relevant laws, regulations, standards and product instructions, and corresponding management systems and procedures shall be formulated. 5.2 Laboratories conducting microbial product evaluation shall meet the requirements of biosafety level 1. 5.3 The types, quantity, performance, range and accuracy of the instruments and equipment used in the evaluation shall meet the needs of efficacy evaluation. The blood materials required for the efficacy evaluation of microbial products shall be sterile. 5.4 The reagents used in the evaluation should be analytically pure, and the water should comply with the secondary water specified in GB/T6682. 5.5 The heavy metals used in the evaluation are prepared from heavy metal standards with a purity greater than or equal to 98%, or standard substances certified by the state and awarded with standard substance certificates, into standard solutions of a certain mass concentration, and the culture medium should be selected according to the type of microorganism. 5.6 After the evaluation experiment is completed, the experimental materials should be harmlessly treated. SAG
GB/T39029—2020
6 Evaluation method
Experimental design
The experimental design should comply with the requirements of Table 1.
Project
Treatment design
Heavy metal selection
Number of repetitions
Reaction system
Table 1 Experimental design
Microbial products
Product type
Test samples, inactivated samples, blank control enzyme products
Select one or more according to the evaluation objectives and evaluation needs for no less than 3 times
Determine according to the results of orthogonal experiments
If the reaction system is clearly specified in the instructions of microbial or enzyme products, proceed in accordance with the requirements of the instructions. For biological products without a clear reaction system, the best reaction system for removing heavy metals shall be determined by orthogonal test design. The removal rate shall be used as the investigation index, and five factors shall be selected as the investigation objects (substrate mass concentration, biological product mass concentration, pH, temperature and time). Four levels shall be selected for each factor, and an orthogonal test with five factors and four levels shall be conducted. 6.2
Preparation of heavy metal solution
The heavy metal standard solution shall be diluted with water to prepare a stock solution with a mass concentration of 100μg/mL. During the test, it shall be diluted with water to a working solution with a certain mass concentration, and prepared immediately before use.
Sample treatment
Test sample
If the biological product to be tested is a block solid product, it shall be crushed or fully ground and passed through a 0.25mm aperture sieve. Accurately weigh a certain weight of solid test sample, dilute it with different pH buffers to different mass concentrations (g/mL) as required; if the test sample is liquid, dilute it with different pH buffers to different mass concentrations (g/mL) as required 6.3.2 Inactivated sample
Take part of the test sample and treat it in a sterilizer at 121℃ for 30min, and set it aside 6.3.3 Blank sample
Replace the biological product to be tested with buffer, and set it aside. 4 Determination of reaction system
Design an orthogonal experiment, and select factors that affect the action of microorganisms or enzyme products, such as substrate mass concentration, sample mass concentration, pH, temperature and time, to determine the reaction system.
Heavy metal removal treatment
6.5.1 Microbial product removal treatment
Take 9 250mL Erlenmeyer flasks, including 3 blank samples, 3 test samples and 3 inactivated samples. According to the substrate mass concentration and the amount of microbial product added determined in the optimal reaction system, a culture solution with a reaction system of 100 mL was prepared, and the removal reaction was carried out under the conditions of the optimal reaction system.
6.5.2 Enzyme product removal treatment
Take 9 50mL centrifuge tubes, including 3 blank samples, 3 test samples and 3 inactivated samples. According to the amount of enzyme product added and the substrate mass concentration determined in the optimal reaction system, a reaction solution with a reaction system of 15mL was prepared, and the removal reaction was carried out under the conditions of the optimal reaction system.
6.6 Determination
After the removal treatment is completed, put it on the centrifuge and continue to centrifuge at a speed of 6000r/min for (20±1) min, and take the supernatant. Determine the mass concentration of heavy metals according to the method specified in GB/T5750.6. 6.7 Calculation of heavy metal removal rate
Heavy metal removal rate should be calculated according to formula (1): R = (po -pi) -(po -p2) -(p. -p)po
Where:
R——removal rate,%;
Heavy metal mass concentration in the initial substrate at the beginning of the reaction, in micrograms per liter (g/L); Q
Heavy metal mass concentration in the residual substrate after the test sample reaction, in micrograms per liter (μg/L); e
Heavy metal mass concentration in the residual substrate after the inactivated sample reaction, in micrograms per liter (ug/L); o
—Heavy metal mass concentration in the residual substrate after the blank sample reaction, in micrograms per liter (ug/L). The average value of the parallel samples is taken as the final removal rate value, and the calculation result is retained to two decimal places. 7 Evaluation report presentationbzxZ.net
7.1 The efficacy evaluation results are expressed in removal rate, see Table 2. Table 2 Efficacy evaluation results
Category
Removal rate (R)
R≥80%
80%>R≥60%
Result
General
60%>R≥40%
40%>R≥20%
（1）
20%>R
7.2 The name of the evaluated product, the name of the production organization, the name of the heavy metal, the mass concentration of the heavy metal, the removal rate and reaction system parameters (including pH value, temperature and time, etc.), and the removal effect (good, good, good, general and poor) should be stated. SAG
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