In order to investigate minute quantities of impurities in such areas as pharmaceuticals and pesticide residues in food products, both the ability to separate and detect trace compounds more reliably and increased analysis efficiency are required.
Utilizing features of the Prominence UFLC, which offered high speed as well as high analysis accuracy and reliability, thereby increasing analysis cost-efficiency, we have developed the Prominence UFLCXR, which achieves greater data quality by means of higher separation performance.
While maintaining the high analysis accuracy and reliability cultivated with the Prominence series by using a high-speed separation column and optimizing the system, we have increased separation performance to a level greater than that of conventional LC.
The Ultimate in High Separation Performance
The ability to reliably perform the high-separation, high-sensitivity detection of trace compounds is required. Improving separation performance by simply increasing the column length also increases the analysis time, which reduces analysis efficiency.
With the Prominence UFLC, high-speed analysis while maintaining a high degree of analysis accuracy and reliability is possible and with the Prominence UFLCXR, greater separation performance is attained. In order to achieve the ultimate in high separation performance, column efficiency has been improved by optimizing the balance between the particle diameter of the XR-ODS and XRII-ODS high-speed separation column and the column length, and the system has been optimized to attain the highest possible level of column performance.
The results obtained in the analysis of seven alkylphenones are shown below. The Prominence UFLCXR demonstrated a level of separation performance equivalent to that of at least a 250-mm (5-µm) column. The analysis time was a quarter of that required by a general-purpose LC, indicating that both high separation performance and high speed were achieved.
High-Sensitivity Detection of Trace Components
The combination of the Prominence UFLCXR and a Shim-pack XR-ODS II column (length: 150 mm) achieves a level of separation performance equivalent to that of at least a 250-mm column containing packing material with a particle diameter of 5 µm. The wide dynamic range of the SPD-20A detector enables the high-separation, high-sensitivity detection of fine peaks, such as those obtained for minute quantities of impurities in pharmaceuticals, and thereby enables highly reliable analysis in which the slightest presence of impurities is not missed. Also, the low-carryover SIL-20AXR autosampler supports high-accuracy analysis.
The results obtained in the analysis of a minute quantity of an impurity in cefazolin are shown below. This impurity was sufficiently separated from other impurities, and although its peak was only one thousandth the height of the peak obtained for the main component, the wide dynamic range ensured that it was detected properly.
Evaluation of Minute Quantities of Impurities in Cefazolin
Column: Shim-pack XR-ODS II (150 mm (L) x 3 mm (i.d.))
Column temperature: 40 °C
Mobile phase: Phosphate buffer solution / acetonitrile, gradient elution
Flow rate: 0.9 mL/min
Detection: 245 nm
Attaining Higher Separation Performance than the Prominence UFLC
In general, the aldehydes and ketones found in indoor locations and the atmosphere are subjected to HPLC analysis with pre-column derivatization performed using a 2,4-DNPH (2,4-dinitrophenylhydrazine) reagent. In this case, 13 aldehydes and ketones were analyzed with the Prominence UFLCXR. In comparison with general-purpose LC, relatively high speed and separation performance were attained. Also, separation was achieved in roughly the same analysis time required for the high-speed Prominence UFLC with greater reliability.
Analytical Conditions
Mobile phases: A: Water/tetrahydrofuran, B: Acetonitrile, gradient elution
Column temperature: 40°C
Flow rate: 1.0 mL/min
Detection: 360 nm
Excellent Reproducibility and Low Sample Carryover
Of the basic performance specifications required of analytical instruments, high reproducibility and low sample carryover are particularly important, and are essential for high-separation, high-sensitivity analysis.
With Prominence, these needs were addressed at an early stage, and superior technology was cultivated for all the fundamental elements. This included a high-performance pump, an autosampler capable of a high level of injection reproducibility, even for minute quantities, and an injection mechanism that prevents sample carryover. These technologies have all been inherited by the Prominence UFLCXR, and support high-separation, high-sensitivity analysis.
