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Merck

TLC Analysis of Caffeine in Coffee Samples, Quantification of Analyte Concentration

TLC Analysis of Caffeine in Coffee Samples, Quantification of Analyte Concentration application for TLC

Materials

accessory

Product No.
Description
Pricing

Bottle-type sprayer

capacity 240 mL

Rectangular TLC developing tanks, complete

L × H × W 27.0 cm × 26.5 cm × 7.0 cm

Sigma TLC spray box

Spectroline® E-Series UV lamp

output8 W, AC/DC input 115 V AC

Rectangular TLC developing tanks, complete

L × H × W 7.5 cm × 15.5 cm × 8.0 cm

Hamilton® TLC syringes

1702, volume 25 μL, needle size 22s ga, needle L 51 mm (2 in.),

coated tip

Spectroline® E-Series UV lamp

output6 W, AC/DC input 120 V AC

Hamilton® TLC syringes

1705, volume 50 μL, needle size 22 ga, needle L 51 mm (2 in.),

coated tip

Hamilton® TLC syringes

1701, volume 10 μL, needle size 26s ga, needle type, fixed, needle L 51 mm (2 in.),

coated tip

Rectangular TLC developing tanks, complete

L × H × W 17.5 cm × 16.0 cm × 8.2 cm

Spectroline® E-Series UV lamp

output6 W, AC/DC input 230 V AC

Spectroline® E-Series UV lamp

output4 W, AC/DC input 230 V AC

Spectroline® E-Series UV lamp

output8 W, AC/DC input 230 V AC

Latch-lid TLC developing chambers

For use with 10 cm × 10 cm plates

Hamilton® TLC syringes

1710, volume 100 μL, needle size 22 ga, needle L 51 mm (2 in.),

coated tip

Latch-lid TLC developing chambers

For use with 20 cm × 20 cm plates

Spectroline® E-Series UV lamp

output4 W, AC/DC input 115 V AC

Kimble® microcapillary pipettes

Kimble® microcapillary pipettes

Kimble® microcapillary pipettes

capacity 20 μL, accuracy: ±0.5%

Reagent sprayer for TLC plates

volume 250 mL, Bottle-Type

CONDITIONS

sample preparation

The boiled coffee samples were extracted and filtered with a 0.45 μm Millex syringe filter.

plate

HPTLC Silica gel 60 F254, 20 cm x 10 cm (1.05642.0001)

eluent

[A] acetonitrile; [B] methanol; [C] n-Heptane; (6:4:1, A:B:C)

migration distance

5 cm

sample

0.1 μL to 2.0 μL

detector

UV, 254 nm

Description

Analysis Note

Abstract: HPTLC (High Performance Thin Layer Chromatography) is a very useful analytical technique with very low sample preparation. Shown in the example: the quantification of caffeine in different coffee samples. The boiled coffee samples
were extracted and filtered with a 0.45 μm syringe filter. The plate was pre-conditioned with the mobile phase. After chromatographic separation the plate was scanned with a TLC-Scanner and subsequently quantified.

Results: Caffeine was detected under UV light (254 nm, hRf = 56, Fig. 1), (quantification information Fig. 2). A triple determination of each sample is possible in parallel.

Conclusion: These results demonstrate that caffeine can be easily quantified by TLC with a three-fold determination and minimal sample preparation.