COMMISSION IMPLEMENTING REGULATION (EU) 2021/808
of 22 March 2021
on the performance of analytical methods for residues of pharmacologically active substances used in food-producing animals and on the interpretation of results as well as on the methods to be used for sampling and repealing Decisions 2002/657/EC and 98/179/EC
(Text with EEA relevance)
Article 1
Subject matter and scope
Article 2
Definitions
Article 3
Methods of analysis
Article 4
Quality control
Article 5
Interpretation of results
Article 6
Methods for sampling
Article 7
Repeals and transitional measures
Article 8
Entry into force
ANNEX I
CHAPTER 1
PERFORMANCE CRITERIA AND OTHER REQUIREMENTS FOR ANALYTICAL METHODS
1.1.
Requirements of screening methods
1.1.1.
Categories of suitable screening methods
1.1.2.
Requirements for biological, biochemical or physico-chemical screening
methods
1.2.
Requirements of confirmatory methods
1.2.1.
General requirements for confirmatory methods
1.2.2.
General performance criteria for confirmatory methods
1.2.2.1. Trueness by recovery
Mass Fraction |
Range |
≤ 1 μg/kg |
–50 % to +20 % |
> 1 μg/kg to 10 μg/kg |
–30 % to +20 % |
≥ 10 μg/kg |
–20 % to +20 % |
1.2.2.2. Precision
1.2.3.
Requirements for chromatographic separation
1.2.4.
Specific performance criteria for mass spectrometry
1.2.4.1. Mass spectrometric detection
1.2.4.2. Identification
Technique |
Identification Points |
Separation (mode GC, LC, SFC, CE) |
1 |
LR-MS ion |
1 |
Precursor ion selection at <±0,5 Da mass range |
1 (indirect) |
LR-MSn product ion |
1,5 |
HR-MS ion |
1,5 |
HR-MSn product ion |
2,5 |
Technique(s) |
Separation |
Number of ions |
Identification points |
GC-MS (EI or CI) |
GC |
n |
1 + n |
GC-MS (EI and CI) |
GC |
2 (EI) + 2 (CI) |
1 + 4 = 5 |
GC-MS (EI or CI) 2 derivates |
GC |
2 (Derivate A) + 2 (Derivate B) |
1 + 4 = 5 |
LC-MS |
LC |
n (MS) |
1 + n |
GC- or LC-MS/MS |
GC or LC |
1 precursor + 2 products |
1 + 1 + 2 × 1,5 = 5 |
GC- or LC-MS/MS |
GC or LC |
2 precursor + 2 products |
1 + 2 + 2 × 1,5 = 6 |
GC- or LC-MS3 |
GC or LC |
1 precursor + 1 MS2 product + 1 MS3 product |
1 + 1 + 1,5 + 1,5 = 5 |
GC- or LC-HRMS |
GC or LC |
n |
1 + n × 1,5 |
GC- or LC-HRMS/MS |
GC or LC |
1 precursor (<±0,5 Da mass range) + 1 product |
1 + 1 + 2,5 = 4,5 |
GC- or LC-HRMS and HRMS/MS |
GC or LC |
1 full scan ion + 1 HRMS product ion (a) |
1 + 1,5 + 2,5 = 5 |
GC- and LC-MS |
GC and LC |
2 ions (GCMS) + 1 ion (LCMS) |
1 + 1 + 2 + 1 + 1 = 6 |
1.2.5.
Specific performance criteria for the determination of an analyte using liquid chromatography with detection techniques other than mass spectrometry
1.2.5.1. Performance criteria for full-scan diode array spectrophotometry
1.2.5.2. Performance criteria for fluorescence detection spectrophotometry
CHAPTER 2
VALIDATION
2.1.
Performance characteristics to be determined for analytical methods
Method |
Confirmation |
Screening |
|||
Qualitative |
Quantitative |
Qualitative |
Semi-quantitative |
Quantitative |
|
Substances |
A |
A, B |
A, B |
A, B |
A, B |
Identification in accordance with 1.2 |
x |
x |
|
|
|
CCα |
x |
x |
|
|
|
CCβ |
- |
|
x |
x |
x |
Trueness |
x |
|
|
x |
|
Precision |
x |
|
(x) |
x |
|
Relative matrix effect/absolute recovery (*) |
x |
|
|
x |
|
Selectivity/Specificity |
x |
x |
x |
x |
|
Stability (#) |
x |
x |
x |
x |
|
Ruggedness |
x |
x |
x |
x |
|
x: It is required to prove by means of the validation that the requirements for the performance characteristic are met. (x) The precision requirements of Chapter 1.2.2.2 do not need to be met for semi-quantitative screening methods. However, the precision shall be determined to prove the suitability of the method for avoiding false compliant analytical results. A: prohibited or unauthorised substances B: authorised substances |
2.2.
Trueness, repeatability and within-laboratory reproducibility
2.2.1.
Conventional validation
2.2.1.1. Trueness on the basis of a certified reference material
2.2.1.2. Trueness on basis of fortified samples
2.2.1.3. Repeatability
2.2.1.4. Within-laboratory reproducibility
2.2.2.
Validation according to alternative models
Factor |
I |
II |
III |
IV |
V |
VI |
VII |
Run 01 |
A |
A |
A |
A |
A |
A |
A |
Run 02 |
A |
A |
B |
A |
B |
B |
B |
Run 03 |
A |
B |
A |
B |
A |
B |
B |
Run 04 |
A |
B |
B |
B |
B |
A |
A |
Run 05 |
B |
A |
A |
B |
B |
A |
B |
Run 06 |
B |
A |
B |
B |
A |
B |
A |
Run 07 |
B |
B |
A |
A |
B |
B |
A |
Run 08 |
B |
B |
B |
A |
A |
A |
B |
2.2.3.
Other validation approaches
2.3.
Selectivity/Specificity
2.4.
Ruggedness
2.5.
Stability
2.5.1.
Determination of the stability of the analyte in solution
|
–20 °C |
+4 °C |
+20 °C |
Dark |
10 aliquots |
10 aliquots |
10 aliquots |
Light |
|
|
10 aliquots |