Application Note

Analysis of Paracetamol in Different Formulationsusing Thin-Layer Chromatography & Compact Mass Spectrometry (TLC/CMS)

Introduction

ExperimentalMass spectrometer: Advion expression CMS

Interface: CAMAG TLC-MS

Plate: Merck HPTLC Silica gel 60 F

254 MS-grade, 20x10 cm

Application volume: 0.3 - 1 μl

Detection: UV 254 nm ESI (+) mode MS (m/z 100-300)

Migration distance: 5 cm

Migration time: 9 min

Mobile phase: Acetone/toluene 1:1 + 0.1% acetic acid

Extraction solvent: Acetonitrile/water 95:5 + 0.1% formic acid

Extraction flow: 0.1 ml/min

Sample preparation: Suppository: One suppository containing 125 mg Paracetamol is diluted in 125 ml ethanol at 36°C and applied directly Pain syrup: 0.2 ml Paracetamol pain syrup containing 4% Paracetamol is diluted in 7.8 ml methanol and applied directly Pill: One pill containing 500 mg Paracetamol is diluted in 500 ml methanol, filtered with a 0.45 μm Millex® syringe filter and applied directly

Sample application: Using the ATS4 sample applicator (CAMAG) 6 mm bandwise

Paracetamol 151.16 g/mol

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The analysis of Paracetamol is performed using the elution based TLC – MS Interface from CAMAG coupled with the expression compact masss spectrometer (CMS) from Advion. Because of the high matrix tolerance of TLC, very easy sample preparation methods can be applied for suppositories, pain syrup and pills.

Summary

Results

Normal-Phase/CMS

TLC is a widespread, routine, reliable tool used in nearly every chemist’s laboratory. Coupling the CAMAG TLC-MS interface with Advion’s expression CMS brings the specificity and sensitivity of mass spectrometry to TLC plate analysis. Now chemists can quickly, simply and unambiguously identify and quantify their compounds, even in complex mixtures, using TLC/CMS.

reaction monitoring

Mass Spectrum

1 2 3 4 5 6 7 8 9 1110 12 13 14

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[M+H]+ [M+ACN]+

152.12 m/z 193.18 m/zIntensity

m/z

Chromatographic Data

Track Compound Concentration Application volume

hRf Detected mass([M+H]+/[M+ACN]+)

1, 8 Paracetamol Standard 1 mg/ml 0.3 μl 452, 9 Paracetamol Standard 1 mg/ml 0.5 μl 453, 10 Paracetamol syrup 1 mg/ml 0.5 μl 45 152.1 / 193.2 m/z4, 11 Paracetamol pill 1 mg/ml 0.5 μl 45 152.1 / 193.2 m/z5, 12 Paracetamol suppository 1 mg/ml 0.5 μl 45 152.1 / 193.2 m/z6, 13 Paracetamol Standard 1 mg/ml 0.7 μl 457, 14 Paracetamol Standard 1 mg/ml 1.0 μl 45