Katsutoshi Shimizu*1, Yushi Harada*2, Atsuki Morisaki*2, Minling Pei*2, Masaru Tomioka*2
Fig.1 The Chromaster (including the 6310 column oven)
We have recently made two new additions to the lineup of modules for the Chromaster series of instruments: the 5280 autosampler and the 6310 column oven.
The 5280 autosampler uses the direct-injection method and supports operation at a pressure tolerance of 60 MPa. The direct-injection method allows microscopic sample volumes to be injected without loss. In addition, support for operation at a pressure tolerance of 60 MPa enables the use of columns with particle sizes under 2.0 µm and core shell columns; this not only allows conventional HPLC analysis but also improves separation performance and reduces analysis times.
In the 6310 column oven, we have taken the column oven from the ChromasterUltra Rs lineup and made it controllable within the Chromaster system. Compared to its predecessor, the 5310 column oven, the 6310 boasts an expanded temperature control range and improved heating and cooling capability, making it a good choice for a wide variety of applications.
The use of the direct-injection method, in which the sampling needle is incorporated into the analytical flow path, ensures that samples may be gathered via suction and injected without waste (no sample loss), making the method a powerful technique for measuring samples with microscopic volumes.
By synchronizing the timing of sample injection with the operation of the liquid transport system, the instrument achieves reproducibility with peak retention times, ensuring the acquisition of highly reliable analytical results.
The pressure tolerance has been improved to 60 MPa. This extends the choice of columns, enabling support for a wide variety of analytical applications.
Because the structure of the flow path is similar to that of its predecessor, the L-2200, appropriate operating conditions are easily adapted from those for the L-2200.
The standard instrument comes equipped with a Peltier-based temperature-control system that allows not only heating but also cooling. The temperature may be controlled over the wide range of [room temperature - 15°C] - [room temperature + 75°C] (temperature control range 4-90°C).
The temperature is controlled to a high degree of accuracy: within ±0.1°C over the entire range.
Heating and cooling capabilities have been significantly improved, and system startup times have been reduced. The temperature rises from room temperature to 40°C in 4 minutes.
The addition of a low-capacity preheater ensures stable peak retention times even with varying environment temperatures.and dramatically reduces the internal volume associated with peak dispersion. (Capacity of preheater unit: 1 µL).
A waste-liquid tank may be placed beneath the 6310 column oven to store waste liquids expelled from the system (capacity: 3 L).
The Hitachi-developed Moment-Enhancing Mechanism allows pressure-tolerance performance of 140 MPa via hand-tightening and reduces internal volume.
In Supplement II to the Japanese Pharmacopoeia, 16th edition, enacted in February 2014, the test items for D-mannitol were revised, and analysis via HPLC was added. Purity (4) related substances and assay for D-mannitol involve analysis at 85°C using a column with a length of 300 mm, requiring an instrument with enough capacity to accommodate a wide column, and temperature control to ensure stable operation at high temperature. Here we present the results of analyses of D-mannitol using a system confi gured with the 5280 autosampler and the 6310 column oven in accordance with the assay of Supplement II to the Japanese Pharmacopoeia, 16th edition.
D-mannitol assay: Confi rmation of system suitability
Standard solution: Dissolve 0.5 g of D-mannitol in pure water and adjust to yield a total volume of 10 mL (5 %).
System suitability test solution (1): Dissolve 0.25 g each of D-mannitol and D-sorbitol in pure water and adjust to yield a total volume of 10 mL (2.5% each)
System suitability test solution (2): Dissolve 0.5 g each of maltitol and isomalt in pure water and adjust to yield a total volume of 100 mL (0.5% each).
FIg.2 Chromatogram of system compliance test solution 1
FIg.3 Chromatogram of system compliance test solution 2
Fig.4 Chromatogram of standard solution
Measurement conditions
Moving phase : H2O
Flow volume : 0.5 mL/min
Column : CARBOsep COREGEL-87C 7.8 mmI.D.×300 mmL (Transgenomic)
Column temperature : 85°C
Detection : RI
Injection volume : 20 µL
| Item | Specification | Results of measurement | |
|---|---|---|---|
| System performance (relative retention time) (Figures 2, 3) |
D-mannitol - isomalt (1) D-mannitol - maltitol D-mannitol - isomalt (2) D-mannitol - D-sorbitol |
Approximately 0.6 Approximately 0.69 Approximately 0.73 Approximately 1.2 |
0.63 0.69 0.73 1.23 |
| System performance (resolution) (Figure 1) |
D-mannitol - D-sorbitol | 2.0 or higher | 5.74 |
| System repeatability (Figure 4) |
D-mannitol Relative standard deviation (n=6) in peak area (%) |
1.0 or lower | 0.21 |
Table 1 shows the results for D-mannitol assay and system suitability. Measured results for all test items were in compliance with specifications.
In this article we presented the key features of two new modules for the Chromaster series: the 5280 autosampler and the 6310 column oven. In future work we plan to continue our efforts to support new applications of these new modules.
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