Determination of caffeine using HPLC

1. STEP 1

1.1. Preparation of stock standard (A) solution

1.1.1. Standard compound (5 mg of caffeine in 10 ml HPLC grade methanol

2. STEP 2

2.1. Preparation of series of standard (B) solution

2.1.1. 4 dilutions of standard (20 ppm, 50 ppm, 100 ppm and 150 ppm)

3. STEP 3

3.1. Preparation of mobile phase

3.1.1. 40:60 of methanol:water

4. STEP 4

4.1. Preparation of sample

4.1.1. Dilute 3 in 1 coffee as sample

5. STEP 5

5.1. Preparation of instrument

5.1.1. Setting up HPLC, load standard and sample, run HPLC