ANNEX 7 OF THE OMCL NETWORK GUIDELINE “QUALIFICATION OF EQUIPMENT” QUALIFICATION OF MASS SPECTROMETERS

Note: Mandatory requirements in this guideline and its annexes are defined using the terms «shall» or «must». The use of «should» indicates a recommendation. For these parts of the text other appropriately justified approaches are acceptable. The term «can» indicates a possibility or an example with non-binding character.

Introduction

The present document is the 7th Annex of the core document “Qualification of Equipment” and it shall be used in combination with it when planning, performing and documenting the qualification process of Mass Spectrometers coupled with Chromatographic equipment.

The core document “Qualification of Equipment” contains the introduction and general forms for Level I and II of qualification, which are common to all type of instruments.

The present Annex 7 contains a general introduction and requirements for GC-EI-MS and LC-ESI-MS.

Level III and IV qualifications must be carried out being an ISO 17025 requirement.

Requirements and (if applicable) corresponding typical acceptance limits given in bold should be applied, however other appropriately justified deviations are acceptable provided they are traceable.

Exemplary procedures provided in Annexes have non-binding character. They can be helpful to carry out the required qualification. Nevertheless, other procedures can be applied depending on the model of the MS equipment.

7.1: QUALIFICATION OF GAS CHROMATOGRAPHY-MASS SPECTROMETERS WITH ELECTRON IMPACT IONIZATION (GC-EI-MS)

TABLE I

Requirements and related typical acceptance limits are indicated in bold.

*Other figures given under column of “typical acceptance limits” are typical values obtained when applying the exemplary procedures provided in annexes, therefore these values are not binding. 
** PFTBA (FC-43): Perfluoro-tributyl-amine (CAS NO.: 311-89-7). 
*** To be checked when quantification is requested. 

TABLE II

Requirements and related typical acceptance limits are indicated in bold.

ANNEX I

This Annex contains practical exemplary procedures for several parameters related to the qualification of GC-EI-MS.

These examples can be considered by the OMCLs as possible approaches to perform the Level III of the equipment qualification process: “Periodic and motivated instrument checks”.

However, alternative procedures can be applied.

GENERAL CONSIDERATION: GC-MS is mainly used for the identification of unknown substances or quantification of low concentrated substances where high specificity is needed.

MASS ACCURACY

Materials:

• PFTBA (FC-43) or internal calibration gas

Method:

• Internal instrument check or spectrum of PFTBA (FC-43) in full scan mode

Limits:

• As defined in Table I

LINEARITY

Materials:

• Stock solutions: 1-Octanol in methanol at concentrations of 0.2, 0.4, 0.6, 0.8, 1.0 µL/mL

Method:

• Injection volume: 1.0 µL (2 injections of each level)

Limits:

• Limit to be set up by OMCL based on experience and type of regression mode chosen

NOTE: Linearity is typically performed in SIM/MRM, since this mode is normally applied for quantification of analytes in low concentration.

SYSTEM/INSTRUMENT PRECISION

Materials:

• Stock solution: 1-Octanol in methanol at concentration of 1.0 µL/mL

Method:

• Injection volume: 1.0 µL (6 injections)

Limits:

• RSD ≤ 10.0 % on peak area (without internal standard)

ANNEX II

IDENTIFICATION (by mass spectral library)

Materials:

• Papaverine 20.0 µg/mL in methanol, Caffeine 10.0 μg/mL in methanol or another compound chosen according to the specific method

Method:

• Identification by mass spectral library

Limits:

• Match reference spectra

SYSTEM/INSTRUMENT PRECISION **

Materials:

• See identification

Method:

• Injection volume: 1.0 µL (6 injections)

Limits:

• RSD ≤ 10.0 % on peak area

** to be checked when performing quantification testing

7.2 QUALIFICATION OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETERS (LC-MS)

Ionisation sources of Mass Spectrometry include:

• ESI: Electrospray Ionisation

• APCI: Atmospheric Pressure Chemical Ionisation

• APPI: Atmospheric Pressure Photo-Ionisation

TABLE III

Requirements and related typical acceptance limits are indicated in bold.
IDENTIFICATION - Positive mode: 

** only for instruments with MS/MS capabilities.

IDENTIFICATION - Negative mode:

* Other figures given under column of “typical acceptance limits” are typical values obtained when applying the exemplary procedures provided in annexes, therefore these values are not binding.

** Chloramphenicol, CAS NO.: 56-75-7.

*** only for instruments with MS/MS capabilities.

QUANTIFICATION: 
Check following parameters both in positive and negative modes:

TABLE IV

Level IV. In-use instrument checks 
Requirements for LC-ESI-MS

ANNEX III

This Annex contains practical exemplary procedures for several parameters related to the performance of LC-ESI-MS.

These examples can be considered by the OMCLs as possible approaches to perform the Level III of the equipment qualification process: “Periodic and motivated instrument checks”.

However, alternative procedures can be applied.

GENERAL CONSIDERATION: LC-MS is widely used for the identification of unknown substances or quantification of low concentrated substances where high specificity is needed.

MASS ACCURACY

Materials:

• ESI positive: Reserpine at concentration of 0.01 mg/mL in methanol/water (60/40 V/V)*

• ESI negative: Chloramphenicol at concentration of 0.01 mg/mL in methanol containing 0.1% of formic acid*.

• *Note: concentrations should be selected depending on the instrument and experimental conditions to be applied

Method:

• Direct infusion or flow injection

Limits:

• As defined in Table III.

RESOLUTION:

• Procedure to be followed depends on the instrument. Instructions to check resolution can be provided by service supplier/ instrument manual.

LINEARITY

Materials:

• Solutions as mixture of betamethasone-17,21-dipropionate and betamethasone-17-valerate in methanol at concentrations of 0.002, 0.004, 0.006, 0.008, 0.01 mg/mL

Method:

• Column: BEH-C18 1.7 µm 50 x 2.1 mm or equivalent

• Suitable gradient of acetonitrile/water containing 0.1 % formic acid

• Injection volume: 1.0 µL (2 injections at each concentration)

Limits:

• Limit to be set up based on OMCL experience/service provider specifications and type of regression mode chosen

SYSTEM/INSTRUMENT PRECISION

Materials:

• Solution as mixture of betamethasone-17, 21-dipropionate and betamethasone-17-valerate in methanol at concentration of 0.006 mg/mL.

Method:

• Injection volume: 1.0 µL (6 injections)

Limits:

• RSD ≤ 5.0 % on peak area

CARRY OVER

Materials:

• - Solution as mixture of betamethasone 17, 21-dipropionate and betamethasone 17- valerate in methanol at concentration of 0.002 mg/mL.

• - Methanol (blank)

Method:

• Injection volume: 1.0 µL

Limits:

• The percentage of the peaks corresponding to betamethasone-17,21-dipropionate and betamethasone 17-valerate in the blank (injected after 0.002 mg/mL solution) does not exceed 1.0 % of the area of said peaks in the chromatogram obtained injecting 0.002 mg/mL solution of betamethasone-17,21-dipropionate and betamethasone 17-valerate.

REFERENCES
(For all references, the latest version applies)
1) Ph. Eur. Chapter 2.2.43. MASS SPECTROMETRY.