Analytical method use in a TDM lab

TDM lab can be classified into 3 gps

1. Chromatographic method
2. Immunological method
3. One step dry chemistry

1) CHROMATOGRAPHIC METHOD
HPLC & GC

Advantage
• Very sensitive method
• Multiple drugs/metabolites of the drg can be estimated in a
single run
• These methods use bulk chemicals not kits
Disadvantage
• Preparation of the sample may be a lengthy procedure &
technically demanding
• Very expensive
• Sequential inj to chk the linearity

HPLC
• Selection of column, mobile phase, pH req
• Quality attributes/validation requirement should be fulfilled
to get proper result

Validation parameter

Selectivity, specificity, sensitivity, linearity (dyanamic range),

precision/accuracy (% recovery), stabilty, rugudness,
robustness
Reason for validation
It provide reliability of a system or a procedure to meet the pre-
determined criteria/attributes providing high degree of
confidence

Selection of analytical method

• As simple as possible
• Most productive, convenient & economical
• As accurate & precise as required
• Most specific

1) Specificity/Selectivity
• To ensure selectivity & specificity, internal std is used
• Internal std may be any compound or drug
• Its purpose to give a constant peak throughout sampling
2) Sensitivity
Minimum quantity of drug can be determined by LOD or LOQ

3) Range
Range of drug is made through serial dilution that much range is
detectable by instrument

4) Linearity
Observe signal throughout concentrated range ( response of
instrument)

5) Accuracy (% recovery)
% recovery is determined by
I. Relative accuracy
II. Absolute accuracy
1 Relative accuracy
• Known concentration of range of drug in plasma is made
• Take pt sample then determine % recovery

11 Absolute accuracy
• Take mobile phase, make concentrated range, then determine pt
plasma drug concentration
• % recovery is found by formula
% recovery = mean area in tab x 100
Mean area in std

6) Precision
• A/f linearity, determine precision
• Known concentration is made then chk either the method is
reproducible using that particular % conc which indicates
authenticity of method
a) Inter-day precision
b) Intraday precision

7) Stabilty
• Chk the biological sample is how much stable, in which drug
concentration is to be determined
• If sample degrade actual Cp is not determined
• Provide storage of plasma @ -20⁰C
• When to determine the conc, defrost the sample
• Chk during defrost, drug is degraded or not
• Freeze & then method is perform in 3 cycles

8) Ruggedness
To chk the inter-instrumental variation
2) Immunological method
Many immuno assays depend on competetive protein binding
assays