YY/T 0618-2017 English PDF (YYT0618-2017)
YY/T 0618-2017 English PDF (YYT0618-2017)
YY/T 0618-2017: Test methods for bacterial endotoxins of medical devices—Routine monitoring and alternatives to batch testing
PHARMACEUTICAL INDUSTRY STANDARD
OF THE PEOPLE’S REPUBLIC OF CHINA
Replacing YY/T 0618-2007
Test methods for bacterial endotoxins of medical devices -
Routine monitoring and alternatives to batch testing
ISSUED ON: FEBRUARY 28, 2017
IMPLEMENTED ON: JANUARY 01, 2018
Issued by: China Food and Drug Administration
Table of Contents
Foreword ... 3
Introduction ... 4
1 Scope ... 6
2 Normative references ... 6
3 Terms and definitions ... 6
4 Principles of quality management system ... 11
5 Non-pyrogenic label ... 13
6 Selection of product unit ... 14
7 Choice of technique ... 15
8 Methodological validation ... 15
9 Use of technique ... 20
10 Alternatives to batch testing ... 24
Appendix A (Informative) Background information on bacterial endotoxin testing .. 28 Appendix B (Informative) Guidelines for test methods, routine monitoring, alternatives to batch testing ... 32
Appendix C (Informative) Out-of-specification (OOS) and failure investigation guide ... 50
References ... 53
Test methods for bacterial endotoxins of medical devices -
Routine monitoring and alternatives to batch testing
This standard specifies basic guidelines for test methods for bacterial endotoxins, which are applicable to the determination of medical devices, components or raw materials. Note: Although the scope of this standard is limited to medical devices, the requirements specified in this standard and the guidelines given may also apply to other medical products. This standard does not apply to the evaluation of pyrogens, other than bacterial endotoxin.
2 Normative references
The following documents are essential to the application of this document. For the dated documents, only the versions with the dates indicated are applicable to this document; for the undated documents, only the latest version (including all the amendments) is applicable to this standard.
Pharmacopoeia of the People's Republic of China 2015 Edition
3 Terms and definitions
The following terms and definitions apply to this document.
Bacterial endotoxin test; BET
Tests for the determination of active bacterial endotoxins, by mixing a liquid test sample with Limulus amebocyte lysate, wherein the results of proportional responses are measured by visual inspection, turbidity, chromogenic or other validated test methods.
A specified quantity of raw material, intermediate or finished product, which is The abnormal phenomenon of bacterial endotoxin test, which is caused by non- endotoxin-related factors (usually caused by the characteristics of the test sample), making the test response higher than the actual total amount of endotoxin present. 3.10
The BET method for quantification or detection of endotoxin, which is based on the principle that the resulting gel-clot reaction is a proportional reaction -- between the Limulus amebocyte lysate and the endotoxin.
Geometric mean endpoint
The mean value, which is obtained from repeated serial dilutions, as converted to a base 10 endpoint logarithm value; it is used for determining the central tendency or mean value, from a test solution.
The abnormal phenomenon of bacterial endotoxin test, which is caused by non- endotoxin-related factors (usually caused by the characteristics of the test sample), so that the test response is lower than the actual total amount of endotoxin present. 3.13
A test, which is used to determine whether a specific bacterial endotoxin test sample contains factors that reduce the accuracy of the test. That is, whether it inhibits or enhances the test system.
Reanalysis of a sample extract or preparation, to verify the original test results. 3.15
LAL reactive material; LAL-RM
Any non-endotoxin component that activates the Limulus amebocyte lysate and causes enhancement.
LAL reagent water; LRW
Purified water or other identified solvents, diluents and/or extractants, for use in BET, which shall be non-reactive and non-interfering in the method used. 3.17
The nominal lambda of the LAL gel reagent, expressed in EU/mL. OR, for
chromogenic and turbidimetric methods, refer to the nadir of the standard curve (endotoxin concentration).
Limulus amebocyte lysate; LAL
Reagents which are extracted from the circulating blood amoeba of horseshoe crab (Limulus polyphemus or Tachypleus tridentatus). It interacts with endotoxin to produce gels, for bacterial endotoxin assays to determine bacterial endotoxin levels. 3.19
Gram-negative bacterial cell wall components, which are typically composed of lipid A, core polysaccharide, and O-side chains.
Maximum valid dilution; MVD
The maximum dilution of the sample, at which the specified test endotoxin limit can be detected, in relation to the lambda of the BET.
Out of specification; OOS
Samples with valid BET test results exceeding the specification for product endotoxin limits.
Note: The term OOS applies only to this document. It is different from other regulatory guidance that deals with OOS results, such as the U.S. Food and Drug Administration (FDA) "Studies of OOS Test Results for Drug Manufacturing".
Standard control series
Serial dilutions of RSE or CSE, which are used for validating endotoxin lambda. 3.30
Test endotoxin limit
The maximum endotoxin concentration is allowed in the extract. This determined limit is used to calculate the maximum effective dilution factor, for sample leaching (mix or one unit).
Note: This limit is determined by dividing the product endotoxin limit by the volume of LRW used per unit of sample extract.
The BET method, for quantification or determination of endotoxin, which is based on the principle that the measured color reaction is a proportional reaction -- between the Limulus amebocyte lysate and the endotoxin.
Establish documented procedures for obtaining, recording, interpreting results, to ensure that products conform to predetermined specifications.
4 Principles of quality management system
4.1 Document formation
4.1.1 The bacterial endotoxin test procedure shall be described in detail. 4.1.2 The documents and records which are required in this standard shall be reviewed and approved by designated personnel (see 4.2.1). Meanwhile, it shall be under the control of the specified quality management system requirements.
4.1.3 The retention of original data, calculated and derived data, final data records shall be specified, in the quality management system requirements. Records shall include information on those involved in sample preparation and testing.
The inspection technique steps and instructions used are approved. All relevant equipment for use and operation shall be available.
4.1.4 Calculations and data transfer shall be properly controlled. If electronic data is used, the software used shall be validated and a record of validation shall be kept. 4.2 Management responsibilities
4.2.1 Responsibilities and rights to complete and carry out the steps, which are described in this document, shall be specified. Responsibility shall be given to those who meet the requirements of the quality management system.
4.2.2 If an organization, which has a separate quality management system, undertakes the requirements of this standard, the responsibilities and rights of each individual shall be specified.
4.3 Product realization
The steps for procurement shall be detailed. These steps shall be consistent with the quality management system requirements.
4.4.1 The responsibilities of personnel, who are assigned to perform bacterial endotoxin testing, shall be specified in the quality management system requirements. 4.4.2 The training shall be carried out in accordance with the documented procedures. The records of the relevant identification, training, and experience of the technical personnel shall be kept.
4.4.3 The analyst shall qualify the bacterial endotoxin test (see 8.3.2), before undertaking it.
4.5.1 All equipment which is required for the proper conduct of the specified tests, shall be available. The planned maintenance and calibration shall be carried out, in accordance with documented procedures. Maintenance and calibration records shall be kept.
4.5.2 All equipment shall be operated in accordance with the document specifying the guidelines.
4.6 Reagents and materials
4.6.1 The material preparation method, which is used in the bacterial endotoxin test, shall be specified, including the corresponding identification test.
Note: The corresponding identification test includes the identification of the lambda of the 5.4 All components of the product claimed to be non-pyrogenic shall be included in the test procedure. Any item of product within an exempt package shall be documented (e.g., a handle or strap). Claims of "non-pyrogenic fluid circuits" shall be supported by appropriate evaluation of the associated components and surfaces of the fluid circuits used.
5.5 For multi-component kits, the labels and claims shall be consistent with the documented evaluation of the components, in the package in contact with the circulatory, lymphatic or cerebrospinal fluid system. This label should be consistent with the intended clinical use of the package and its components.
5.6 Product endotoxin limits shall be determined, in accordance with appropriate regulatory requirements.
Note: Endotoxin limits for medical infusion (blood) and injection equipment are recommended in GB/T 14233.2-2005.
6 Selection of product unit
6.1 The sampling criteria for the selection of product units, in the bacterial endotoxin test, are based on the premise that the production process is controlled and the requirements of the quality management system are met.
6.2 The selection of test product units shall be based on the criteria, as specified in the sampling plan, which may be based on regulatory requirements, published statistical programs or guidelines, or validation of production runs.
6.3 The sampling parent (group) is generally defined as the production batch. If data support different selection bases or alternatives for batch testing is selected, sample selection can be based on the sampling parent being not a production batch. If alternatives for batch testing is selected, see Chapter 10, B.6 ~ B.8, B.10. Note: Sampling specifications, where the sampling matrix is not a production batch, should include a risk assessment, to evaluate the suitability of the criteria for establishing the sampling matrix (see Appendix B).
6.4 Test samples should be selected from the finished product, which includes all factors that may affect or increase endotoxin levels (e.g., packaging).
Note: Samples for endotoxin testing may be selected from products, which are rejected in other production quality items, provided that the rejected samples represent non-rejected products. 6.5 Samples may include pre-sterilized and post-sterilized products. The sterilized sample contains all the factors that could affect the product or endotoxin testing. If sample testing before sterilization is selected, the acceptability of the sample to represent the endotoxin level of the final product shall be documented. The inspection procedures performed should consistently reflect pre-sterilized or post-sterilized samples. See B.6.5 for guidance on validation of equivalence.
Note: For products that promote microbial growth, it may not be appropriate to select samples prior to sterilization.
6.6 In the inspection of multi-component kits (program packages) or complete sets of products, depending on the use of the product, sometimes each component can be evaluated individually, sometimes all the contents can be regarded as a whole for evaluation. Standard test procedures should be applied to each component case. When using a product kit or kit as a stand-alone unit, consideration shall be given to sample preparation and applicable product endotoxin limits, without changing the method. See B.6.6, for more guidance.
7 Choice of technique
7.1 Testing laboratories currently have three bacterial endotoxin testing techniques to choose from. The selection of each technique should be based on adequate evaluation of laboratory skills, experience, sample size requirements, data handling requirements, test sample properties. Current techniques and methods include:
a) Gel-clot technique: Limit test and determination method;
b) Chromogenic techniques: Kinetic and endpoint methods;
c) Turbidity techniques: Kinetic and endpoint methods.
See Appendix B, for information on the three methods.
7.2 The selected method shall be confirmed, in accordance with Chapter 8. If the test method or technique is changed, validation/verification shall be carried out (see 8.6). 8 Methodological validation
8.1 Identification of test endotoxin limits
8.1.1 The test endotoxin limit is defined as the maximum allowable concentration of endotoxin, which may exist in the extract of a product, which is related to the product endotoxin limit. See Appendix B, to determine product endotoxin limits. 8.1.2 Once the product endotoxin limit is determined, the test endotoxin limit can be calculated, according to formula (1):
Medical device test endotoxin limit (EU/mL) = KN/V ………………………… (1)
a) BET testing laboratory changes;
b) Changes to BET materials, equipment that may affect the test; and
c) Changes to lambda of Limulus amebocyte lysate.
Note: Product positive controls (PPCs) are available for validation in some cases. PPC may adequately detect that a change has occurred.
9 Use of technique
9.1 Key test parameters
The general incubation temperature for bacterial endotoxin test methods is (37 ± 1) °C. Refer to the reagent manufacturer's instructions for selection of appropriate parameters. 9.1.2 Time
Limulus amebocyte lysate manufacturer's product instructions give the reagent addition and incubation time length. The general incubation time of gel bacterial endotoxin test method is (60 ± 2) min.
The manufacturer's instructions for the Limulus amebocyte lysate will give the optimum pH range, for endotoxin response. A pH outside this range in the test may cause interference. If the pH of the product positive control is acceptable, in the validation test, subsequent pH measurements may not be required. However, if the pH must be adjusted in validation, the pH shall be measured, in subsequent routine tests. 9.2 Equipment and reagents
9.2.1 Since the bacterial endotoxin test requires a limited temperature range, the heating box or water bath, which is used for the incubation of the gel-clot test, shall record the temperature during the incubation. Mechanical pipettes (including fixed, adjustable, reusable units) shall be calibrated regularly. If the laboratory uses chromogenic or turbidity techniques, the hardware and software shall be identified, in accordance with the manufacturer's instructions for use and validation requirements. Materials that are not non-pyrogenic supplies (e.g., microplates), shall be carefully evaluated before use (see 4.6.4), to ensure that they do not interfere with the determination in the testing laboratory.
9.2.2 All limulus amebocyte lysates used in the test shall be certified products. The activity of the control standard endotoxin shall be calibrated, by the national endotoxin standard, which shall be carried out by the testing laboratory OR the Limulus amebocyte lysate manufacturer provides the calibration documents to the testing laboratory.
9.2.3 The product label of the Limulus amebocyte lysate manufacturer will give storage requirements for lyophilized and reconstituted reagents. If the storage conditions in the laboratory are different from those recommended by the manufacturer, the storage conditions shall be confirmed.
9.3 Sample preparation
220.127.116.11 Samples for testing should be collected and stored, in a manner that prevents changes in endotoxin levels.
18.104.22.168 The routine bacterial endotoxin test shall adopt the sample
preparation/extraction method, which is used for validation. The product can be rinsed or soaked, to prepare a test rinse or leaching solution. The leaching method used depends on the product labelled as non-pyrogenic.
Note: For a long time, there has been no recognized extraction method for bacterial endotoxin test. However, endotoxin limits have been established with a safety factor, to ensure patient safety (see Appendix A.8).
9.3.2 Medical devices
22.214.171.124 The instruments are usually leached together, for routine endotoxin testing. When the test determines the endotoxin level per device or the variability between devices, the device needs to be tested separately. See Appendix B or relevant regulations, for guidelines on the appropriate sample size.
126.96.36.199 In order to ensure the safety of the patient's pyrogenic threshold, usually the endotoxin limit of a batch refers to the measured value of a maximum of 10 devices, which are pooled together. If a device unit is tested, the endotoxin limit of the product can be evaluated. Additional information is given in B.8.1.1.
188.8.131.52 The instruments are usually immersed in LRW for sample extraction. Depyrogenation equipment can be used to separate or disassemble the instruments for extraction. The shortest extraction time is 15 min at 37 °C ~ 40 °C, OR not less than 1 h at controlled room temperature (generally 18 °C ~ 25 °C), OR other proven equivalent conditions. It is recommended to supplement the leaching process with agitation. 184.108.40.206 For the instruments marked with "non-pyrogenic liquid circuit", inject the endotoxin test water, which has been heated to 37 °C ± 1 °C, into the liquid circuit, so that the extract is in contact with the liquid circuit, at a controlled room temperature (usually 18 °C ~ 25 °C) for not less than 1 h, OR other equivalent conditions. When appropriate, the extracts of each sample are mixed, to obtain a pooled sample test product positive control is 50% ~ 200% of the known concentration of added endotoxin.
9.5.2 For the gel limit test, it is acceptable, if the validity of each parameter of the test product complies with 9.5.1 AND both test tubes of the sample solution are negative. If a positive result appears in any sample tube, see Appendix C.
9.5.3 For gel-clot determination, determine the endotoxin concentration in the sample solution, by calculating the end point concentration of each series of parallel tubes AND multiplying the dilution factor of each end point by λ. The endotoxin concentration of the sample is the geometric mean end point concentration of each parallel tube dilution series. When necessary, the endotoxin concentration of the sample solution can be used to calculate the total endotoxin per unit of product, using appropriate mathematical factors (i.e., sample/extract volume, product weight, sample to product ratio, etc.). If the diluted sample solution is used in the test, the endotoxin concentration is calculated, by multiplying the leaching stock solution by the dilution factor. It is acceptable, if the measured endotoxin level of the test sample is l...