Page 3 - Shimadzu Nexera Mikros

M i cr o: I m p r o v e d D e t e c t i o n L i m i t s w i t h M i c r o f l o w

A b o v e a n d B e y o n d N a n o Nexera Mikros ™ Micro -flow LC /MS System

LC/MS/MS methods are now widely recognized as gold standard for the accurate quantitation of small molecules in biological fluids and results more suitable
and reliable compared to immunoassay based approaches. For metabolites in biological fluids, the concentration level of the target component is often low,
therefore more sensitive analysis is required than semimicro-flow LC/MS analysis, which is superior in analytical throughput, stability and robustness. Generally,
in LC/MS analysis, improvement of sensitivity can be expected by reducing the analytical scale which is defined by the total flow rate and column size. On the
other hand, by reducing the analytical scale to nano-flow, a well skilled analysts who have special technics in nano-flow analysis is required, and throughput,
stability and robustness are reduced by trade-offs. LC/MS analysis on a micro-flow scale using 0.1 to 1 mm I.D. column with 1 to several L/min has the
excellent balance of sensitivity, throughput, and robustness. Nexera Mikros, a Shimadzu microflow LC/MS system provide the stable and sensitive quantitative
analysis by not only improved sensitivity through stable performance of micro-flow solvent delivery and optimized ion source design, but also utilization of the
The High Sensitivity You Expect from a Low Flow unique UF-Link column connection and system variety that support a wide range of applications.

System with the Ruggedness of HPLC Covering the High Sensitivit y Analysis or the Bioanalysis
f
Complete Range from Micro to Semi-Micro While the results of quantification of vitamin D metabolites using LC/MS/MS are more reliable than other methods, in metabolite measurement, the vitamin D
is lack of easily ionizable functional groups and pose assay sensitivity challenges mainly for quantification of low-abundance metabolites such as 1,25(OH) 2
di-hydroxyvitamin D and 24,25(OH) 2 di-hydroxyvitamin D. To overcome this sensitivity problem, It is effective to perform analysis with micro-flow LC/MS/MS.
The comparison between micro-flow LC/MS/MS and semimicro-flow LC/MS/MS for the analysis of standard solutions of Vitamin D metabolites shown a 3 to 5
fold improvement in the ratios of signal/noise.

5 µL/min 225 µL/min 5 µL/min 225 µL/min
30000 30000 30000 30000
411.35 > 105.15 (+) 411.35 > 105.15 (+) 399.35 > 91.15 (+) 399.35 > 91.15 (+)
1,25(OH) 2
1,25(OH) 2 Vitamin D 3
Vitamin D 2
20000 20000 20000 20000

×6 24,25(OH) 2 ×4
10000 10000 10000 Vitamin D 3 10000
1,25(OH) 2 1,25(OH) 2
×3 Vitamin D 3
Vitamin D 2 24,25(OH) 2 Vitamin D 3
0 0 0 0
17 18 19 20 min 17 18 19 20 min 16 17 18 19 min 16 17 18 19 min

The micro-flow LC/MS/MS system resulted effective in providing better signal intensity also for the quantitation of antiarrhythmic agents that are usually
monitored in plasma, due to their narrow therapeutic window and inter individual variability. The increase of the sensitivity when using micro-flow rates was 5
fold for Verapamil and 4 fold for Nor-Verapamil in the ratios of signal/noise.

5 µL/min 225 µL/min 5 µL/min 225 µL/min
455.10 > 303.30 (+) 455.10 > 303.30 (+) 440.95 > 165.00 (+) 440.95 > 165.00 (+)
30000 Verapamil 30000
800000 Nor-verapamil 800000

20000 20000 600000 600000
×5 ×4
400000 400000
10000 10000 Verapamil Nor-verapamil
200000 200000

0 0 0 0
3.5 4.0 4.5 5.0 min 2.0 2.5 3.0 3.5 min 3.5 4.0 4.5 5.0 min 2.0 2.5 3.0 3.5 min

Nexera Mikros
Microflow Liquid Chromatography Mass Spectrometry System 3