Hair Cotinine Extraction
Hair samples weighing 30 mg from both case and control groups washed with distilled water, and extracted with 2 ml of hair extraction buffer (Immunalysis, USA) for 30 minutes using an ultrasonic water bath at 23 °C. Following the extraction process, 200 µl of neutralization buffer (Immunalysis, USA) were added to finalize the extraction procedure, followed by initiation of the ELISA process\sout.