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Application Ultra fast Purification Liquid
Chromatography
News
Isolation and identification of
No. LC-15-ADI-036 Atorvastatin degradation impurities by
UFPLC
Introduction:
Experimental:
Atorvastatin is an antilipemic drug belonging to Acid Degradation
the statins class, whose reference drug is Pfizer’s
Lipitor® (shown in Figure 1). It is used to reduce 200 mg of Atorvastatin API sample was dissolved in
the levels of lipoproteins rich in cholesterol and 10 mL of methanol and added 10 mL of 0.1 N
reduce the risk of coronary artery disease. The 0
drug in question is commonly sought after by hydrochloric acid and kept at 80 C for 1 hr. After
pharmaceutical industries that produce generic degradation, added a few mL of methanol to
drugs, due to the fact that the drug has a high dissolve residue and diluted to 10 mL. This solution
value price, it is consumed globally, and its patent
expired in late 2010. Atorvastatin has been found was used for analysis on UFPLC for fraction
to degrade under acid and basic conditions. collection. 10 µL of the solution was taken and
diluted with 1mL of acetonitrile/water (1:1) to make
Prominence UFPLC, Ultra Fast Preparative and
Purification Liquid Chromatograph (Shown in 200 mg/L and then injected into HPLC
Figure 2.), enables fast recovery of highly purified Analytical Conditions
target compounds from complex samples such as
organic synthesis reaction mixtures and natural Mobile phase A : 0.1% TFA in water
products. Preparative LC (Prep LC) is a widely used Mobile phase B : Acetonitrile
technique in many research developments and
manufacturing applications, including the Gradient program : (0.01/ 40, 10.00/50, 15.00/70
synthesis of new drug compounds and the 20.00/90, 25.00/90, 30.00/40
discovery of active components in natural 35.00/40) (Time in mins /B%
products. It is mostly used to collect large
amounts of unknown compounds in foods and Column : Shim-pack GIS C-18
drugs for subsequent structural analysis. (250X10mm, 5µ)
Flow Rate : 5.0 ml/min
Wavelength : 245 nm
Preparation for Analysis
The degradation sample was diluted with methanol
to make the clear solution. After dilution the sample
concentration was 10 g/L. Before UFPLC analysis
Figure 1. Atorvastatin Figure 2. UFPLC diluted samples were analyzed on Nexera system to
Features check the extent of degradation. The fast method
i. Comprehensive Automation of Preparative LC, was developed on Shim-pack XR-ODS-II on Nexera
Concentration, Purification, Elution, Collection X2 system to check the purity of degradation
and Powderization Only in 1.5 Hours samples and fractions collected by UFPLC.
Dedicated automation software to assist Results and Discussion
chemists in preparative procedure through Automation of preparative LC, concentration,
collection purification, elution and collection controlled by
The time of evaporation can be reduced dedicated automation software (see page 3) assists
by up to 90% because of collection with chemists in clearly identifying the peaks which are
organic solvent. trapped and collected in specific color code. 1D
ii. High Purity as a Free Base chromatogram is shown in Figure 3 and
Removal of counter ions derived from corresponding area percentages are given in Table
preparative mobile phase 1. Table 1:Area Percentage
De-salting and conversion to free base
with ammonia/water Peak# Name Ret. Time Area Area%
iii. Small Footprint and Low-initial-cost 1 Atorvastatin 4.421 14932410 27.214
Your lab space can be kept with high
2 Impurity H 5.449 17169678 31.292
functionality by small footprint
3 Unknown imp 6.032 22767800 41.494
Available in two standard configurations
to match your requirements The UFPLC system is capable of trapping maximum
5 peaks in one injection run on 5 different trap
Standard system with one trapping columns. It also rinses the individual trap columns
column by different rinsing solution to remove salts. It
Advanced system with five trapping ensures that the compounds are transferred into
free bases before they are eluted.
columns
