BIOANALYSIS CONSUMABLE SOLUTIONS Sample Preparation and Liquid Chromatography Solutions for Quantitative BIOANALYSIS
At Waters, we understand the unique challenges faced by the bioanalytical community, such as the need to maximize sensitivity, increase throughput, and develop new, robust methods that can be transferred to laboratories across the globe. As a result, we create sample preparation and liquid chromatography solutions for quantitative bioanalysis around a simple, logical workflow that provides clean samples and selective separations. And for the ultimate in bioanalytical data quality, laboratory productivity, and compliance, integrate these eading chemistries with the Waters Regulated Bioanalysis System Solution. It purposefully combines the new reduceddispersion ACQUITY UPLC I-Class now the best-in-class inlet to MS with the high-sensitivity Xevo TQ-S, under new software control using the fully compliant-ready UNIFI Scientific Information System. 10
BIOANALYSIS CONSUMABLE SOLUTIONS 11
SAMPLE PREPARATION SOLUTIONS Key Benefits: Unique design eliminates time-consuming steps for increased throughput Precipitate-free filtrate for maximum system uptime vs. classical methods Ideally suited for cleanliness, MS compatibility, and analyte recovery 10
SIROCCO PROTEIN PRECIPITATION PLATE SIMPLY CLEAR The Sirocco protein precipitation (PPT) plate eliminates additional, time-consuming sample-handling steps traditionally done with classical PPT. Unique membrane and valve-tip technology enable the analyst to carry out in-well protein precipitation followed by the final filtration step. This approach leads to efficient sample processing, resulting in clear filtrates from small sample volumes. Sirocco can be automated to process large numbers of samples efficiently. PROCESSING TIME COMPARISON With Sirocco plates, samples can be processed in significantly less time than with classical PPT, thus increasing laboratory throughput. RECOV ERY The Sirocco plate was tested for analyte recovery using 7 analytes spiked into rat plasma, using an internal standard. The results show excellent recoveries across all analytes listed. Classical PPT Sirocco PPT 100 45 minutes 15 minutes PPT recovery (%) 80 60 40 Load Sample Load Sample 20 Mix Mix Vacuum 0 Atenolol Nadolol Metoprolol Oxprenolol Labetalol Alprenolol Propranolol Centrifuge Direct Inject Transfer Supernatant 11
Sum of Area Counts of 5 Phospholipids SAMPLE PREPARATION SOLUTIONS THE FASTER WAY TO CLEANER Ostro sample preparation products provide a novel solution for the cleanup of phospholipids in plasma and serum. Requiring minimal to no method development and using a simple protocol, this technology can be quickly implemented to optimize your laboratory s workflow. Providing cleaner, more reproducible extracts than competitive phospholipid removal devices or techniques, the Ostro 96-well plate allows for more sensitive analyses, increased sample throughput, and reduced instrument downtime. PHOSPHOLIPID REMOVAL The level of 5 phospholipids remaining after sample preparation using Ostro is less than with both liquidliquid extraction and solid-supported liquid extraction. REPRODUCIBILITY Comparative % removal of total phospholipids from 6 different lots of plasma using Ostro (0.19% RSD), phospholipid removal plate from competitor A (24.5% RSD), and phospholipid removal plate from competitor B (40.9% RSD). 400000 350000 300000 250000 200000 150000 100000 % Phospholipid Removal 100 90 80 70 60 50 40 30 20 10 0 1 2 3 4 5 6 Ostro Competitor A Competitor B 50000 0 Ostro LLE SSLE Sum of 5 abundant phosphatidylcholine containing phospholipids with m/z 496, 524, 704, 758, and 806. 10
OSTRO SAMPLE PREPARATION PRODUCTS Key Benefits: Removes significantly more phospholipids than competitive techniques for cleaner extracts Improves reproducibility for more consistent, robust methods Increases throughput with easy-to-implement protocol 11
SAMPLE PREPARATION SOLUTIONS Key Benefits: Highest sensitivity for traditional pharmaceutical compounds and therapeutic peptides Minimal matrix effects for LC/MS/MS analyses Unmatched product quality, performance, and reproducibility 10
OASIS SAMPLE EXTRACTION PRODUCTS SENSITIVITY IN ITS PUREST FORM Sensitivity is one of the key requirements for a successful bioanalytical assay. Achieving optimum sensitivity requires the isolation of compounds of interest from interferences present in biological samples. The patented Oasis solid-phase extraction (SPE) products are available in a wide array of formats and sorbents, allowing scientists to develop methodologies that are selective for the compounds of interest. Oasis products enable scientists to create highly accurate and reproducible SPE methods that are unsurpassed in sensitivity and selectivity. SENSITIVITY The Oasis mixed-mode sorbents, all built upon the unique water-wettable Oasis HLB copolymer, provide dual modes of retention, enabling greater cleanup selectivity and sensitivity for both acidic and/or basic compounds even if the sorbent in the wells runs dry. Achieve unsurpassed sensitivity with the Oasis μelution plate which can concentrate a sample up to 15x without requiring evaporation or reconstitution. LOQ of Benzodiapine Alprazolam 1 pg/ml from 375 μl of Plasma LOQ of Fluticasone Propionate 5 pg/ml from 375 μl of Plasma 100 0% 100 100 0% 1.61 1.52 1.66 1.55 A. Spiked sample 1.73 1.54 1.58 1.62 1.66 1.70 1.74 1.78 B. Matrix blank 1.54 1.58 1.62 1.66 1.70 1.74 1.78 min 100 1.19 0% 0% A. Spiked sample 0.8 1.0 1.2 1.4 1.6 1.8 2.0 B. Matrix blank 1.01 1.18 1.50 1.73 2.03 0.88 1.06 1.28 1.38 1.82 1.92 0.8 1.0 1.2 1.4 1.6 1.8 2.0 min MATRIX EFFECTS Following extraction with an Oasis mixed-mode sorbent, the matrix effects for a panel of 5 antidepressant drugs were evaluated in multiple lots of urine. The RSDs of the matrix factors were determined to be between 5.5 and 8.4%, which are well within the 15% limit required by regulatory agencies. Analyte % RSD of Matrix Factors Amitriptyline 6.03 Nortriptyline 5.53 Imipramine 8.36 Desipramine 7.15 Doxepin 5.96 QUALITY Waters careful process design and stringent quality controls have set a new standard in batch-to batch and lot-to-lot reproducibility for SPE sorbents. This enables scientists to develop robust, accurate, and precise bioanalytical methods. Recovery % 105 95 No Batch Reservations Needed: Consistently high recoveries over more than 15 years of production. 85 0 5 10 15 years RSDs of recovery for: Procanamide: 1.90% Ranitidine: 1.68% Acetominophen: 1.65% 11
LIQUID CHROMATOGRAPHY SOLUTIONS COLUMN TECHNOLOGY Due to the increased sensitivity, improved throughput, and exceptional assay reliability that UPLC technology provides, the ACQUITY UPLC system has been established as the LC platform of choice for bioanalytical assays. With 5 particle substrates and 20 sub-2 µm column chemistries available for both small molecule and biopharmaceutical applications, Waters has a UPLC column solution to meet your application needs. ACQUITY UPLC BEH C 18 O O Si O Providing unprecedented levels of peak shape, efficiency and chemical stability, the 1.7 µm BEH Technology [Ethylene Bridged Hybrid] C 18 column is a versatile, ultra-performance separation solution for a diverse set of analytes. With the ability to operate between ph 1 12, the BEH C 18 column employs the power of ph to impact the retention, selectivity, and sensitivity of ionizable compounds, resulting in sensitivity improvements for LC/MS assays. ACQUITY UPLC HSS T3 O O Si O Specifically designed for the enhanced retention of polar compounds and metabolites, the 1.8 µm HSS [High Strength Silica] T3 column is a low-ligand density C 18 column that exhibits exceptional retentivity without the need for ion-pairing reagents. This enables lower limits of detection and quantitation to be achieved for LC/MS methods. ACQUITY UPLC BEH HILIC Employing an acetonitrile-rich mobile phase to improve the retention, ionization efficiency, and mass spectrometry response for extremely polar compounds and metabolites, the 1.7 µm BEH HILIC [Hydrophilic Interaction Chromatography] column provides lower limits of detection compared to conventional reversed-phase methods. Additionally, high organic extracts from PPT, LLE, or SPE can be directly injected, dramatically improving sample throughput, robustness, and sensitivity. ACQUITY UPLC BEH300 C 18 O O Si O Specifically designed and QC tested for the rapid, high resolution analysis of peptides, the 300Å BEH C 18 column assures the successful separation of a diverse physiochemical range of peptides. A single column can now be used to retain and separate peptides of a wide range of hydrophobicity, isoelectric point, and molecular weight. 10
ACQUITY UPLC COLUMNS Key Benefits: Maximize laboratory productivity by reducing analysis time while improving data quality Reach lower limits of detection by enabling higher MS response with narrow chromatographic peaks Improve sample characterization by reducing MS suppression due to residual matrix components 11
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