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Precise Determination of Atropine using a simple gradient
No Ion Pair Reagent Necessary
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printable Application Sheet
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Note: After a simple sample clean up procedure, the method can be applied for monitoring atropine
concentrations in biological specimens in cases of drug poisoning. The recoveries of atropine added to drug-free
specimens which were analyzed using the described method were satisfactory with coefficients of variation of
4% or less.
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Method Conditions
| Column |
Cogent Bidentate C18™, 4µm, 100A |
| Catalog No. |
40018-75P |
| Dimensions |
4.6 x 75 mm |
| Mobile Phase |
A: DI water + 0.1 acetic acid + 0.005% TFA
B: Acetonitrile + 0.1% acetic acid + 0.005% TFA
Both solutions were vacuum filtered through a 0.45µm nylon filter |
| Flow Rate |
1 µL/min. |
| Gradient |
0 min - 10% B,
0 to 4 min to 30%B
4 – 6 min hold 30%B
6.01 min back to 10%B |
| Injection |
1 microL |
| Sample |
Prepared in 50% solution A/50% solution B, concentration 1 mg/mL and was filtered through a 0.45 µm nylon membrane |
| Peak |
Atropine
Injection 1: RT = 4.772 min, Injection 2: RT = 4.773 min,
Injection 3: RT = 4.772 min Injection 4: RT = 4.774 min |
| Detection |
UV 214 nm |
Discussion
Chromatographic separation and quantification methods of tropa alkaloids are often described in the literature and the method of choice
is usually ion-pair chromatography (IPC), which requires long equilibration times and it is not very robust. This method shows a
symmetrical peak for atropine using a simple gradient method that does not include any ion pair reagents which can cause damage to
columns and lack eproducibility. The retention times are extremely repeatable but one of the best advantages to this method is the time
savings between runs. Using a Cogent Bidentate C18 column the equilibration time was very short (5 min between every gradient run).
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