purpose
Using the Waters ACQUITYUPC2 system, the normal phase HPLC content determination method of anthraline in the Pharmacopoeia was successfully converted to supercritical fluid chromatography.
background
At present, the determination method of the USP for the drug anthraline, (9 (10H) -anthrone, 1,8-dihydroxy-9-anthrone) [CAS # 1143-38-0] is the normal phase HPLC method . Isocratic separation was performed using a 4.6x250mm silica gel column (L3). The mobile phase was 82: 12: 6 n-hexane: dichloromethane: glacial acetic acid mixed eluent with a flow rate of 2mL / min, as shown in Figure 1. The running time of the current measurement method is about 10 minutes (2 times the peak time of the last main peak). Although this method is feasible and reliable, due to health, safety, environmental and cost considerations, many laboratories want to reduce the use of typical normal phase chromatography solvents (such as n-hexane and methylene chloride). Supercritical liquid chromatography (SFC) is a normal phase chromatography separation technique that uses CO2 as the main mobile phase
Use polar solvents (such as methanol) as modifiers. Since the principle of SFC is similar to that of HPLC, the current method should be able to be converted into an SFC method, thereby reducing the use and handling of solvents, reducing the cost of each analysis, and enhancing health, safety and environmental protection.
Successfully converted the HPLC method in the United States Pharmacopoeia to a high-quality UPC2 method at a cost of .05 per analysis (compared to .90 in Pharmacopoeia) and at 1.6 times the speed of Pharmacopoeia.
The conversion of these methods to SFC methods must maintain the quality of the analysis data (reproducibility of retention time, resolution between the target compound and other components in the sample), and must obtain analysis results consistent with current normal phase chromatography methods.
solution
An anthraline sample was prepared and analyzed using the current USP method (this sample was also used for UPC2 analysis). The analysis results were compared with those obtained using the ACQUITY UPC2 method. The conditions of the Ultra Performance Convergence Chromatography (UPC2) method are as follows:
The comparison of main system adaptability parameters is shown in Table 1. In all analysis conditions, the results obtained by the converted UPC2 method can easily reach the system suitability values ​​required by the USP, and the results are ideal compared with the values ​​obtained by the normal phase chromatography method. Interestingly, the selectivity of the adaptive mixture (anthraline and danthron) has changed, but it does not adversely affect the method conversion. The two methods were used to determine anthraline samples of unknown purity, and the analysis results were consistent. When using normal phase HPLC analysis, the content of anthraline sample was 94.3%; when using UPC2, the content was 94.6%.
In this example, a single normal phase HPLC analysis consumes 16.4 mL of n-hexane and 1.2 mL of methylene chloride. In contrast, the UPC2 method consumes only 1.05 mL of methanol. This shows that the conversion of normal phase chromatography method to UPC2 method can greatly reduce the use of organic solutions. Based on current solution prices, the cost of a single normal phase HPLC analysis is approximately US $ 0.90, compared to US $ 0.05 for UPC2.
to sum up
Using the ACQUITY UPC2 system, the HPLC method of the United States Pharmacopeia can be successfully converted to the UPC2 method. The data obtained by this new UPC2 method is comparable to the current HPLC method, or even better, at 1.6 times the speed of the current HPLC method, and consumes less solvent. When we obtain high-quality analysis data at a faster rate, laboratory productivity increases, while the analysis cost per sample decreases. The ACQUITY UPC2 system is an ideal solution for laboratories to convert the current normal-phase HPLC method to a more efficient and cost-effective UPC2 method, while also enhancing health, safety and environmental protection.
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