HPLC Method for Acetaminophen (Paracetamol), Phenylephrine, Phenylephrine hydrochloride, Ascorbic Acid on Primesep S2 by SIELC Technologies
High Performance Liquid Chromatography (HPLC) Method for Analysis of Acetaminophen, Phenylephrine, and Ascorbic Acid
Acetaminophen (or paracetamol) is one of the most popular over-the-counter painkillers all over the world (in the US it is best known under the brand name Tylenol). Ascorbic acid (also known as Vitamin C) helps your body build up muscle, cartilage, and other important building blocks in your body. Phenylephrine is a common over-the-counter decongestant, but can also be used for pupil dilation and hemorrhoid treatment. While all of these compounds are available at your local pharmacy, Vitamin C intake can reduce the body’s ability to break down acetaminophen.
You can find detailed UV spectra of Acetaminophen and information about its various lambda maxima by visiting the following link.
You can find detailed UV spectra of Ascorbic Acid and information about its various lambda maxima by visiting the following link.
All three of these compounds can be measured at low UV. Using a Primsep S normal phase column and a mobile phase consisting of water and acetonitrile (MeCN) with an Ammonium acetate (AmAc) buffer, these three compounds can be separated and UV detected at 270 nm. Varying the buffer concentration changes the order in which phenylephrine and ascorbic acid are retained. This method is compatible with Mass Spectrometry.
| Column | Primesep S2, 4.6 x 150 mm, 5 µm, 100 A, dual ended |
| Mobile Phase | Gradient MeCN – 97 to 70% |
| Buffer | Gradient Ammonium Formate pH 3.0 – 10 to 50 mM |
| Flow Rate | 1.0 ml/min |
| Detection | UV 270 nm MS- compatible mobile phase |
| Class of Compounds | Drug, Acid |
| Analyzing Compounds | Acetaminophen (Paracetamol), Phenylephrine, Phenylephrine hydrochloride, Ascorbic Acid |
Application Column
Primesep S2
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended
Ascorbic Acid
Phenylephrine
Phenylephrine hydrochloride




