Inhalation Toxicology Support Optimisation David Howie April 2014

Department organisation  Inhalation Analysis section is part of the Pharmacy, Formulation & Inhalation Analysis Department (FIA)  Split into Formulation & Inhalation analysis sections  Made up of: Team Leaders Study Managers Technical Specialists Analysts Laboratory support

The Role of Inhalation Analysis  To analyse test atmosphere samples taken from the exposure system for determination of test substance concentration and particle size distribution  We support: Inhalation Toxicology Pharmacology (PK, respiratory) Pharmacy

Test items  Physical form solids (powder blend) liquids gases  Pharmaceuticals for inhalation administration drugs for treating diseases of the respiratory tract drugs for treating non-respiratory diseases (MS, Alzheimer’s etc)  Non pharmaceuticals that may be inhaled accidentally industrial chemicals agrochemicals air pollutants

4 key parts  Method development  Method Validation  Preliminary trials  Main study analysis

T0T0 T end T -5 weeks T -10 weeks T end + 8 weeks Start of Tox Phase Last Tox Samples Taken Chamber Characterisation Stability Method Set-up, development and validation Firm enquiry Expert Report = 1 week Timelines T end + 5 weeks

METHOD DEVELOPMENT  Conducted by the Technical Specialist  Aim: To establish a working method of analysis  Extraction techniques (Sonication, dissolution etc)  Quantification range  Extraction solvent  Collection media (Filter, bubbler)  Method of detection

 UPLC, HPLC & GC

Other techniques: LCMS/MS, UV & IR (gases) Atomic Absorption (AA) (Metal containing excipients, industrial chemicals)

METHOD VALIDATION Used to demonstrate that we can accurately quantify the test item  Validation considerations Target dose levels in protocol Quantitation range limits determined during method development Sample volumes agreed with ATS Extraction volumes

Validation  Specificity (control & blank assessed)  Carryover (<10x LOQ)  Linearity of detector response  Precision of injection (top & bottom std)  Standard stability  Recovery & Stability from the collection media & solution  PSD plate compatibility test  LOQ & LOD

 The coal face

PRELIMINARY TRIALS  Used to ensure the delivery of the test item meets target dose levels  Establish M/T ratio in dog or cyno studies  MDI studies Pack test (Delivery & consistency between cans) Bench test (Distribution of TI in the dosing apparatus) Drug recovery test (Recovery from dose apparatus during in-life phase)

Preliminary trials  Only conducted following the initial validation stage  Timings of trials arranged in consultation with ATS  Any changes to the confirmed schedule agreed in advance  Analysis performed the day after receipt unless specifically agreed (Received before 15.30hrs)

MAIN STUDY (in-life)  Data provides indication of “dose available” to the animal  Particle size data provides an indication of the respirability of the test material  Results used in conjunction with the gravimetric data

Sampling & analysis Frequency determined by: Study duration & design (DRF, staggered, carc) Dose duration (10min or 6hr for whole body) Sponsor requirements (daily throughout) May be reduced dependent on: Sponsor & SD Variability of results from exposure system Stability of test item on collection media

Sampling from the exposure system  A known volume of test atmosphere is withdrawn from the system Filter paper is sent for analysis Bubbler trap Gas bag or syringe (gas analysis)  Headspace Analysis Samples of blood (ca. 1 ml) are sent for analysis in sealed headspace vials

 Samples are taken from a vacant port Inhalation Dosing - Rodents

Inhalation Dosing - Dogs

Particle Size Distribution the test material

Sample extraction  Automated sample booking in system  Quick and simple extraction techniques (sonication, dissolution, gas analysis)  Use of wide selection of controlled forms during analysis (minimises manual input)  Use of controlled methods (test item and study specific)

Instrument analysis & processing  Wide use of UPLC Generic methods Short run times / fast turn around  Automated data capture  Validated results spreadsheets

Data review  Analyst & SM discuss any issues, OOS or atypical result QC check sheet  Study Manager issues results (SD, ATS, HOD Inh tox)  Any further actions i.d by SD/ATS are dealt with by SM/TL Main aim to reduce unnecessary re-inj’s)  Daily department meeting to discuss any issues

Reporting of results  Results & reporting Results available by 9.30am Action limits set on results (80% – 120%) Report issue date in timeplan (2 weeks from last exposure) Expert or standalone

Questions?