(1)

Point 1.2.1. is replaced by the following:

‘1.2.1.   General requirements for confirmatory methods

For prohibited or unauthorised substances, the CCα shall be as low as reasonably achievable. For prohibited or unauthorised substances, for which an RPA is established under Regulation (EU) 2019/1871 the CCα shall be lower than or equal to the reference point for action.

For authorised substances, the CCα shall be higher than, but as close as possible to, the MRL or ML.

For confirmation purposes, only analytical methods for which it can be demonstrated in a documented traceable manner that they are validated and have a false non-compliant rate (α error) which is less or equal to 1 % for prohibited or unauthorised substances, or which is less or equal to 5 % for authorised substances, shall be used.

Confirmatory methods shall provide information on the structural chemical composition of the analyte. Consequently, confirmatory methods based only on chromatographic analysis without the use of mass spectrometric detection are not suitable on their own for use as confirmatory methods for prohibited or unauthorised pharmacologically active substances. In the case of mass spectrometry not being suitable for authorised substances, other methods such as HPLC-DAD and HPLC-FLD, or a combination of them, can be used.

When required according to the confirmatory method, a suitable internal standard shall be added to the test portion at the beginning of the extraction procedure. Depending on availability, either stable isotope-labelled forms of the analyte, which are particularly suited for mass spectrometric detection, or analogue compounds that are structurally closely related to the analyte, shall be used.’;

(2)

Point 1.2.1a. is inserted between Points 1.2.1. and 1.2.2.:

‘1.2.1a.   Specific use of Co-chromatography when no internal standard available

When no suitable internal standard can be used, the identification of the analyte shall preferably be confirmed by co-chromatography (*). In this case, only one peak shall be obtained, the enhanced peak height (or area) being equivalent to the amount of added analyte. If this is not practicable, matrix-matched or matrix-fortified standards shall be used.

(*)  Co-chromatography is a procedure in which the sample extract prior to the chromatographic step(s) is divided into two parts. Part one is chromatographed as such. Part two is mixed with the standard analyte that is to be measured. Then this mixture is also chromatographed. The amount of added standard analyte has to be similar to the estimated amount of the analyte in the extract. Co-chromatography is used to improve the identification of an analyte when chromatographic methods are used, especially when no suitable internal standard can be used.’;"

(3)

in Point 1.2.2.2., the sentence after Table 2 is replaced by the following:

‘For analyses carried out under repeatability conditions, the coefficient of variation under repeatability conditions is usually below two thirds of the values listed in Table 2 and shall be lower than or equal to the coefficient of variation under reproducibility conditions.’;

(4)

Point 1.2.3. is replaced by the following:

‘1.2.3.   Requirements for chromatographic separation

1.2.3.1.   Minimum acceptable retention time

For liquid (LC) or gas chromatography (GC), the minimum acceptable retention time for the analyte(s) under examination shall be twice the retention time corresponding to the void volume of the column.

1.2.3.2.   Retention time of the analyte in the extract

The retention time of the analyte in the extract shall correspond to that of the calibration standard, a matrix-matched standard or a matrix-fortified standard with a tolerance of ± 0,1 minute. For fast chromatography, where the retention time is below 2 minutes, a deviation of less than 5 % of the retention time is acceptable.

1.2.3.3.   Retention time in case of using of internal standard

In case an internal standard is used, the ratio of the chromatographic retention time of the analyte to that of the internal standard, that means the relative retention time of the analyte, shall correspond to that of the calibration standard, matrix-matched standard or matrix-fortified standard with a maximum deviation 0,5 % for gas chromatography and 1 % for liquid chromatography for methods validated from the date of entry into force of this Regulation.’;

(5)

in Point 2.1., Table 5 is replaced by the following:

‘Table 5

Classification of analytical methods by the performance characteristics that have to be determined

(6)

Point 2.2.1. is replaced by the following:

‘2.2.1.   Conventional validation

The calculation of the parameters in accordance with conventional methods requires the performance of several individual experiments (see Table 5 in this Annex). For major changes the ongoing validity of the performance characteristics has to be verified. For multi-analyte methods, several analytes can be analysed simultaneously, as long as possibly relevant interferences have been ruled out. Several performance characteristics can be determined in a similar way. Therefore, in order to minimise the workload, it is advised to combine experiments as much as possible (e.g., repeatability and within-laboratory reproducibility with specificity, analysis of blank samples to determine the decision limit for confirmation and testing for specificity).’;

(7)

in Point 2.2.1.2., in point 1., subpoint (a) is replaced by the following:

(**)  Where, for a non-allowed pharmacologically active substance validation of a concentration of 0,5 times the RPA is not reasonably achievable, the concentration of 0,5 times the RPA can be replaced by the lowest concentration between 0,5 times and 1,0 times the RPA, which is reasonably achievable or by the LCL in case that it is lower than 0,5 times the RPA.’;"

(8)

Point 2.2.1.3. is amended as follows:

(9)

Point 2.2.1.4. is amended as follows:

(10)

in Point 2.2.2., the sentence after Table 6 is replaced by the following:

‘The calculation of the method characteristics shall be performed as described by Jülicher et al. (*****) or by ISO/TS 23471:2022 (******).

(*****)  Jülicher, B., Gowik, P. and Uhlig, S. (1998) Assessment of detection methods in trace analysis by means of a statistically based in-house validation concept. Analyst, 123, 173."

(******)  ISO/TS 23471:2022 Experimental designs for evaluation of uncertainty – Use of factorial designs for determining uncertainty functions.’;"

(11)

in Point 2.5., the third paragraph is replaced by the following:

‘In case the required stability data are not available, the following approaches should be used. In addition, also the application of an isochronous approach (*******) with a storage temperature scheme similar to the one in Table 7 of this Annex allows a determination of potential analyte instabilities and also an estimation of appropriate storage periods, and may also be used.

(*******)  Lamberty, A., Schimmel, H. and Pauwels, J. (1998) The study of the stability of reference materials by isochronous measurements. Fres. J. Anal. Chem. 360, 359.’;"

(12)

Point 2.5.2. is replaced by the following:

‘2.5.2.   Determination of the stability of analyte(s) in matrix

(13)

Point 2.7. is amended as follows:

(14)

in Point 2.9., the first paragraph is replaced by the following:

‘The absolute recovery of the method does not need to be determined if an internal standard, a matrix-fortified calibration or both mentioned are available. In all other cases, the absolute recovery of the method shall be determined.’;

(15)

in Point 2.10., the last paragraph is replaced by the following:

‘The coefficient of variation shall not be greater than the values listed in Table 2 of this Annex for the MF (standard normalised for IS).’;

(16)

in Chapter 3, the third paragraph is replaced by the following:

‘During routine analysis, the analysis of certified reference materials (CRMs) is the preferable option to provide evidence of method performance. Since CRMs that contain the relevant analytes at the required concentration levels are seldom available, also reference materials provided and characterised by the EURLs or laboratories that hold an ISO/IEC 17043:2023 (********) accreditation may be used as an alternative. As another alternative in-house reference materials, which are controlled regularly, may be used.

(********)  ISO/IEC 17043:2023 Conformity assessment – General requirements for proficiency testing.’."