SOLAμ for Twenty-fold Sample Concentration Prior to Analysis Jon Bardsley and Ken Meadows Thermo Fisher Scientific, Runcorn, UK Introduction Despite advances in analytical detection technology achieving required limits of sensitivity, whilst maintaining reproducibility, can still be an issue for many bioanalytical laboratories. Results The assay gave a lin 0.998. The chromato above the acceptabl Traditional scale SPE helps to clean up the sample to minimize matrix effects, however in order to pre-concentrate the sample a lengthy dry down and reconstitution step needs to be employed. This process is not only time consuming but can have a detrimental effect on the recovery of the analyte due to volatility or non-specific binding. FIGURE 2. Linear 2.6 2.5 By using Thermo Scientific™ SOLAµ™ a twenty-fold concentration of sample can be achieved without the need for a lengthy dry-down step, increasing sensitivity without compromising assay performance. 2.4 2.3 2.2 2.1 2.0 1.9 FIGURE 1. Summary of workflow required to achieve a twenty-fold increase in sample concentration prior to analysis without altering the work-flow or compromising assay performance. 1.8 1.7 1.6 Area Ratio 1.5 1.4 1.3 1.2 1.1 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 5000 Low, mid and high 30000 pg/mL respe Assay is reproducib TABLE 1. Assay 2 SOLAμ for Twenty-fold Sample Concentration Prior to Analysis By loading 500 µL twenty-fold conce even with low elut cleanliness were a By loading 500 µL o twenty-fold concent even with low elutio cleanliness were ac FIGURE 3. Twentysample concentrat SOLAµ allows use allowing greater li analyte recovery, Thermo Scientific Poster Note • PN20950_HPLC_2014_E_05/14S 3 TABLE 2. Matr against pure r Results red limits of many bioanalytical The assay gave a linear dynamic range from 40 to 40000 pg/mL with an r2 coefficient of 0.998. The chromatography for the limit of quantitation sample at 40 pg/mL is significantly above the acceptable signal to noise limit. ix effects, however in itution step needs to a detrimental effect g. FIGURE 2. Linear calibration line for Niflumic acid from 40 to 40000 pg/mL Niflumic Acid Y = 0.00375684+5.93882e-005*X R^2 = 0.9975 W: 1/X^2 2.6 2.5 of sample can be ensitivity without 2.4 Analyte recov fortified blank against pure r less than 9% 2.3 2.2 2.1 2.0 1.9 old increase in k-flow or 1.8 1.7 TABLE 3. Ass against fortifi 1.6 Area Ratio 1.5 1.4 1.3 1.2 1.1 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 5000 10000 15000 20000 25000 30000 35000 40000 Low, mid and high QC samples were prepared at concentrations of 400, 20000 and 30000 pg/mL respectively. Good levels of accuracy at all QC levels were observed. Assay is reproducible for replicate extractions (n=18) at both high and low QC levels. TABLE 1. Assay accuracy and precision data Discussi This poster de analysis while cleanliness. By loading a s decrease the evaporation p SOLAμ provid utilizing the re technology. T ensures highl SOLAμ deliv By loading 500 µL of sample onto the SOLAµ plate and eluting in a total of 25 µL a twenty-fold concentration of the analyte was achieved. The results demonstrate that Concentration Prior to Analysis 4 SOLAμ for Twenty-fold Sample even with low elution volume high levels of accuracy, precision, recovery and sample cleanliness were achieved. • Lower sam • Increased • The ability • Improved By loading 500 µL of sample onto the SOLAµ plate and eluting in a total of 25 µL a twenty-fold concentration of the analyte was achieved. The results demonstrate that even with low elution volume high levels of accuracy, precision, recovery and sample cleanliness were achieved. FIGURE 3. Twenty-fold increase in response achieved by using SOLAµ plate for sample concentration © 2014 Thermo Fi Scientific and its s This information is intellectual propert SOLAµ allows users to pre-concentrate the sample up to 20-times prior to injection, allowing greater limits of sensitivity to be achieved whilst maintaining a high level of analyte recovery, accuracy and precision. Thermo Scientific Poster Note • PN20950_HPLC_2014_E_05/14S 5 TABLE 2. Matrix effects results calculated by comparing fortified blank samples against pure reference standards coefficient of L is significantly 2 0 pg/mL Analyte recovery was shown to be greater than 89.9% by comparison to post extraction fortified blank samples. Post extraction fortified blank samples were also compared against pure reference standards to demonstrate matrix effects which were calculated at less than 9% at both high and Low QC levels. TABLE 3. Assay recovery results calculated by comparing extracted samples against fortified blank samples 40000 20000 and observed. w QC levels. Discussion This poster demonstrates the advantages of SOLAµ for sample concentration prior to analysis while maintaining high levels of precision, accuracy, recovery and sample cleanliness. By loading a sample volume of 500 µL and eluting in a volume of 25 µL it is possible to decrease the lower limit of quantitation by a factor of twenty without the need for lengthy evaporation procedures that may compromise sample integrity. SOLAμ provides reproducibility, robustness and ease of use at low elution volumes by utilizing the revolutionary Thermo Scientific™ SOLA™ , Solid Phase Extraction (SPE) technology. This removes the need for frits delivering a robust, reproducible format which ensures highly consistent results at low elution volumes. SOLAμ delivers: • Lower sample failures due to high reproducibility at low elution volumes • Increased sensitivity due to lower elution volumes • The ability to process samples which are limited in volume • Improved stability of bio-molecules by reduction of adsorption and solvation issues l of 25 µL a onstrate 6that SOLAμ for Twenty-fold Sample Concentration Prior to Analysis ry and sample technology. This removes the need for frits delivering a robust, reproducible format which ensures highly consistent results at low elution volumes. SOLAμ delivers: • Lower sample failures due to high reproducibility at low elution volumes • Increased sensitivity due to lower elution volumes • The ability to process samples which are limited in volume • Improved stability of bio-molecules by reduction of adsorption and solvation issues of 25 µL a nstrate that and sample Aµ plate for © 2014 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others. to injection, high level of www.thermoscientific.com PO20950_E 04/14S ©2014 Thermo Fisher Scientific Inc. All rights reserved. ISO is a trademark of the International Standards Organization. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. This information is presented as an example of the capabilities of Thermo Fisher Scientific Inc. products. It is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others. Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details. 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