SOLA SPE Cartridges and Plates | Thermo Fisher Scientific - US
SOLA Solid-Phase Extraction (SPE) cartridges and plates Preventing sample failures for bioanalysis Prevent sample failure in bioanalytical workflows by using SOLA SPE Choose from Thermo ScientificTM SOLATM Solid-Phase Extraction (SPE) cartridges and plates available in a range of phases, formats, and bed weights to suit any bioanalysis application. SOLA and SOLA SPE products help prevent costly reanalysis of bioanalytical samples by preventing blocking, voiding, and channeling during the SPE sample preparation process. Award-winning fritless polymeric technology eliminates the issues encountered in conventional SPE. Experience cleaner, highly reproducible and robust sample extractions in high throughput workflows. SOLA products provide unparalleled performance characteristics compared to conventional SPE, phospholipid removal and protein precipitation products · Higher levels of reproducibility · Higher levels of extract cleanliness · Reduced solvent requirements · Increased sensitivity 2 2 The proprietary manufacturing process involved in the production of SOLA cartridges and plates, provides an SPE product which eliminates issues normally associated with conventional loose-packed SPE, by combining the polyethylene frit material and media components into a uniform sorbent bed, removing the need for frits. SOLA products eliminate common issues associated with conventional SPE The manufacturing process has the additional benefit of removing extractables from component parts, resulting in cleaner sample extracts. SOLA products provide reduced failure rates, higher analysis speeds and lower solvent requirements, which are critical in today's laboratory environment. The increased performance delivered by SOLA products provides higher confidence in analytical results and lowers cost without compromising ease of use or requiring complex method development. Conventional SPE cartridges and well plates are packed with a loose powder of silica or polymeric material positioned between two frits. These packed beds are potentially prone to settling and voiding in production or transportation. This creates phase channeling and packing irreproducibility, resulting in reduced recovery and reproducibility in analytical results. No voiding No voiding No channeling Reproducible packing time after time No Reproducible packing channeling time after time Examples of conventional SPE product issues Voiding Voiding Channeling Channeling Flow through Flow through Packing inconsistency Packing inconsistency 3 Cleanliness of extract SOLA SPE versus other forms of sample preparation SOLA products offer greater selectivity, reproducibility and cleanliness of sample extract, compared to other sample preparation technologies such as protein precipitation and phospholipid removal plates. This is exemplified below, which shows MS contour plots from these respective technologies. It can be seen that SOLA products provide cleaner sample extracts resulting in greater confidence in your analytical results. m/z m/z m/z 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 Protein precipitation 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 Phospholipid removal plate MS contour plots from protein precipitation, phospholipid removal plates and the SOLA SAX cartridge 4 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 SOLA SAX cartridge Significantly more interferences have been removed using the Thermo ScientificTM SOLATM SAX cartridge Failure to remove the matrix interferences in the primary sample preparation process can result in substantial carry over of phospholipids from injection-to-injection. The image below shows MS contour plots of subsequent blank injections. This shows that there is considerable carry over when using protein precipitation or phospholipid removal products when compared to SOLA products. Removal of phospholipids is key to reducing ion suppression, obtaining improved sensitivity in MS detection and providing confidence in analytical results. It also prevents the need for costly column and system maintenance. m/z m/z m/z 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 Protein precipitation 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 Phospholipid removal plate MS contour plots of the subsequent blank injections--protein precipitation, phospholipid removal and the SOLA SAX cartridge 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 Time (Minutes) 1050 1000 950 900 850 800 750 700 650 600 550 500 450 400 350 300 250 200 150 SOLA SAX cartridge The subsequent blank injection shows a clean MS contour plot with the SOLA SAX cartridge 5 SOLA SPE leachables and extractables SOLA products proprietary manufacturing process provides a cleaner product and, as a result, a cleaner sample extract. As shown below, SOLA products are compared against a competitor (i) conventional loose-packed SPE product, which have both been extracted with acetonitrile, dichloromethane and methanol, respectively. SOLA products are significantly cleaner than the equivalent loosepacked SPE product from competitor (i) Acetonitrile extract comparison: SOLA product versus competitor (i) Dichloromethane extract comparison: SOLA product versus competitor (i) Methanol extract comparison: SOLA product versus competitor (i) 6 Relative abundance Relative abundance Relative abundance 100 SOLA product Competitor (i) 80 60 40 20 0 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 Time (min) 100 SOLA product Competitor (i) 80 60 40 20 0 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 Time (min) 100 SOLA product Competitor (i) 80 60 40 20 0 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 Time (min) SOLA SPE usability Your sample preparation robustness and reproducibility are affected by more than just the product; the operator performing the extraction, and how they carry it out, can also have a large effect. The method of applying a sample to the SPE sorbent can vary from a `neat' application directly onto it, to a `messy' application by pipetting down the walls of the wells. The vacuum speed applied to an SPE product can also influence the recovery and reproducibility of your extraction. We recommend applying liquid samples directly onto the sorbent bed and using a vacuum flow rate of approximately 1 drop per second. Sometimes, novice users of SPE could perform their extraction under sub-optimal conditions that go against this advice. Luckily, the robustness of SOLA products is very high, allowing reproducible extractions time-and-time again, as shown in the example below. 4-Pentylbenzoic acid 100 4-Pentylbenzoic Acid SOLA is a robust SPE platform that delivers high recoveries and high reproducibility no-matter the extraction conditions 80 Recovery (%) 60 40 SOLA product 20 Competitor (i) 0 Slow/Neat Slow/Messy Fast/Neat Vacuum speed/application method Fast/Messy Recovery (%) Diclofenac 100 80 60 40 20 0 Slow/Neat Dichlofenac Slow/Messy Fast/Neat Vacuum speed/application method Fast/Messy SOLA product Competitor (i) Methodology: 4-Pentylbenzoic acid and Diclofenac were used as test probes in solution (no matrix) and loaded onto Thermo ScientificTM SOLAµTM SAX 2 mg/1 mL 96-well plates and competitor microelution plates of equivalent phase chemistry (200 µL load volume and 2 × 25 µL elution volumes). Neat vs Messy application method--liquid placed with single channel pipette directly onto sorbent vs multichannel pipette down the walls of the wells. Slow vs Fast elution speed--a vacuum started at slow speed with a gradual increase until liquid slowly moves through the sorbent vs a vacuum started at the fastest speed possible until the sorbent is dry. 7 How to choose the chemistry phase Selectivity options for SOLA cartridges and plates Thermo ScientificTM SOLATM and SOLAµTM SPE products are manufactured using high-quality polymeric material which provides a wide range of selectivity options (see below) to meet all your analytical requirements. The use of polymeric sorbents in the design provides a robust high capacity bed which is stable over a wide range of pH (014) and does not lose sample capacity upon drying. SOLA selectivity options and compound applicability Chemistry HRP Hydrophobic ReversedPhase SCX Mixed-Mode Strong CationExchange SAX Mixed-Mode Strong AnionExchange WCX Mixed-Mode Weak Cation- Exchange WAX Mixed-Mode Weak Anion- Exchange Base polymer NO O S OO N+ NO O NH+ S ONH+ HO ON+ O-+HN+ O O NH+ S ONH+ HO ON+ O-+H+ NO NH+ O NH+ HS OO O ON-++H+ NO NH+ O NH+ HS OO O ON-++H+ Functional groups pKa Primary use Secondary use N N N O O OS O- S O- SO O- O O N+ N+ N+ O O O <1 NH+ NH+N H NNHH++ NH+ HO NH+ H S OO- O OO-+H+ O O-+H+ O-+H+ >18 O ~4.5 N+ Neutral compounds Weakly basic compounds (pKa 810) Weakly acidic compounds (pKa 24) Strongly basic compounds (pKa >10) Moderately polar compounds Neutral compounds Neutral compounds Neutral compounds NH+ NH+ H Strongly ~8.5 acidic compounds Neutral compounds (pKa <2) O O-+H+ Description Cat. no. Hydrophobic retention of compounds with complementary retention of moderately polar analytes. An all-purpose phase. Strong ion-exchange retention of basic compounds. Complementary reversedphase retention of neutral compounds. Strong ion-exchange retention of acidic compounds. Complementary reversedphase retention of neutral compounds. Weak ion-exchange retention of basic compounds. Sorbent charge can be activated or deactivated. Complementary reversed-phase retention of neutral compounds. Weak ion-exchange retention of acidic compounds. Sorbent charge can be activated or deactivated. Complementary reversed-phase retention of neutral compounds. 60109-001 60209-001 60309-001 60409-001 60509-001 60109-002 60209-002 60309-002 60409-002 60509-002 60109-003 60209-003 60309-003 60409-003 60509-003 60109-004 60209-004 60309-004 60409-004 60509-004 60109-005 60209-005 60309-005 60409-005 60509-005 NH+ NH+ H O O-+H+ 8 How to choose the bed weight Bed size options for SOLA and SOLA SPE products The choice of bed weight is an important point to consider when developing an SPE protocol and is dictated by the volume and complexity of the sample matrix, along with the amount of analyte to be extracted in your application. When compared to traditional silica-based media, the SOLA packing sorbent has approximately 23 times more reversed-phase mass capacity. This allows SOLA and SOLAµ SPE products to retain more analyte in your sample than the equivalently sized silica-based sorbent. Smaller bed weights, such as the SOLAµ microelution well plate, lend themselves to lower elution volumes, whereas larger bed weights, such as the SOLA 30 mg well plate, offer greater loading capacity for hard to retain analytes found in low concentrations. Both approaches can be used to increase extraction sensitivity. The deciding factor for bed size is often the sample concentration and volume of matrix used in your application. Bed size options for SOLA and SOLA SPE products SOLA bed size Typical sample volumes Elution volumes 2 mg (SOLAµ) Up to 500 µL 25 µL Benefits of use · Low volume samples · R educed cost and increased throughput by removal of evaporation and reconstitution steps · Increased signal sensitivity by up to 20 fold 10 mg Up to 1 mL 150 µL · Good option for most analyses · Lower elution volumes than silica based products thus reducing time for evaporation and increasing throughput 30 mg Up to 2 mL 250 µL · When high loading volumes are required to reach lower limits-of-quantitation · For difficult to retain analytes · When experiencing analyte breakthrough on a smaller bed weight Want to know more about how SOLA products can revolutionize your analysis? thermofisher.com/solaspe 9 How to choose the format Format options for SOLA and SOLA SPE products SPE devices come in various formats, and two common designs are the cartridge and the 96-well plate. SOLA products are available in both formats, while SOLA is available in a 96-well plate format with individually removable wells, which are especially convenient for method development. Cartridges SOLA SPE product 10 mg/1 mL 30 mg/3 mL HRP 60109-001 60409-001 SCX 60109-002 60409-002 When to select cartridges or plates · 1 mL cartridges are typically used for routine or method development purposes · 3 mL cartridges can be used in method development and in analyses where the larger cartridge volume is required, (e.g., analysis from urine) · 96-well plates are typically used in high throughput analyses where many samples are needed to be processed in parallel SAX 60109-003 60409-003 WCX 60109-004 60409-004 WAX 60109-005 60409-005 96-well plates SOLA SPE product 2 mg/1 mL (SOLAµ) 10 mg/2 mL 30 mg/2 mL HRP 60209-001 60309-001 60509-001 SCX 60209-002 60309-002 60509-002 SAX 60209-003 60309-003 60509-003 WCX 60209-004 60309-004 60509-004 WAX 60209-005 60309-005 60509-005 SOLA SPE products are available in 5 polymeric phases-- micro-elution, 10 mg and 30 mg bed weights, cartridges, and 96-well plates--which fit the method development and throughput processes in bioanalysis laboratories 10 SOLA SPE 10 mg cartridges and plates Reproducibility--What benefits do SOLA products have over conventional loose-packed SPE products? Improved reproducibility and recovery The reproducibility and recovery levels of SOLA products for three test probes; caffeine, hydrocortisone and carbamazepine when compared to two equivalent, loose-packed, low bed weight competitor products. The data below shows that SOLA products outperform competitor products, even when utilizing the recommended generic competitor methodology. Error bars illustrate significantly lower variability sample-to-sample for SOLAproducts compared to conventional SPE products. This shows that you will achieve the correct result time after time. SOLA product precision (%RSD) 2.7 Competitor (i) precision (%RSD) 21.8 Competitor (ii) precision (%RSD) 7.8 SOLA products show significantly higher reproducibility and recovery levels Recovery (%) 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Ca eine SOLA product Competitor (i) Competitor (ii) Hydrocortisone Carbamazepine SO Co Co HSyOdLrAopcroodrutcistsoonuetperform competitor products for reproducibilitCy arbamazepine Caffeine Hydrocortisone Carbamazepine Method SPE SOLA product precision (%RSD) 4.4 3.3 2.7 Condition Equilibrate Competitor (i) precision (%RSD) 23.9 20.5 21.8 Load Wash Competitor (ii) precision (%RSD) 12.1 10.4 Elute 7.8 200 µL methanol 200 µL water 1 mL sample 200 µL 5% methanol in water 200 L methanol 11 Improved reproducibility The top figure highlights the reproducibility of SOLA products with three test probes; caffeine, hydrocortisone and carbamazepine when compared to an equivalent, loose-packed, low bed weight competitor product. The data shows that SOLA products have consistent recoveries across all thirty test samples. The conventional loose-packed SPE product from competitor (i) shows that on average one in every four samples gives a significantly lower recovery. The results delivered are inconsistent. In comparison, SOLA products provide significantly higher levels of reproducibility, which is vitally important for high-throughput studies. This improved reproducibility is shown in the lower figure which demonstrates that SOLA products have more uniform flow-through characteristics compared to the equivalent, loose-packed, low bed weight competitor product. Inconsistency of loose-packed competitor (i) product compared to SOLA product % Recovery % Recovery % Recovery Caffeine 120.00% 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% 0 5 Hydrocortisone 120.00% 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% 0 5 Carbamazepine 120.00% 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% 0 5 SOLA product Competitor (i) 10 15 20 25 30 35 Sample number 10 15 20 25 30 35 Sample number 10 15 20 25 30 35 Sample number Air flow mL/min 25.00 20.00 15.00 10.00 5.00 0.00 0 SOLA Competitor (i) Competitor (ii) 5 10 15 20 25 30 35 Sample number The consistent flow rate of SOLA products compared to equivalent loose-packed competitor (i) and (ii) products 12 Reproducibility in plasma Due to their nature, biological matrices such as plasma present a difficult challenge in obtaining reproducible results. The excellent performance characteristics of SOLA products provide high levels of reproducibility, even when dealing with these difficult matrices. This has been demonstrated by the extraction of rosuvastatin from human plasma using the Thermo ScientificTM SOLATM 96-Well Plate. Precision data for extractions of a fixed concentration of analyte across the entire plate 100 The table shows the precision data for extractions of a 1.59 fixed concentration of analyte across the entire plate. This can be visually observed in the figure, which shows randomly selected overlaid chromatograms of 75 rosuvastatin. Relative abundance (%) Precision (%RSD) data for rosuvastatin Rosuvastatin (area of 96 replicates) d6-Rosuvastatin (area of 96 replicates) Response ratio (96 replicates) Precision (%RSD) 5.4 3.9 2.7 50 25 0 1.52 1.54 1.56 1.58 1.60 1.62 1.64 Time (min) Overlaid chromatograms of rosuvastatin Elution volumes Higher sensitivity and lower solvent consumption SOLA products achieve excellent recovery levels even with low volumes of extract solvents, resulting in a more concentrated analyte and increased sensitivity. Additional cost and time saving benefits can be achieved from reduced sample dry-down time and solvent usage. % Recovery 140.00% 120.00% 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% Ca eine Hydrocortisone High recovery levels are achieved with SOLA products at low elution volumes, resulting in increased sample concentrations and sensitivity Carbamazepine 100 µL 150 µL 200 µL 250 µL 300 µL 500 µL 13 These low-volume extractions would be significantly compromised when using a conventional loose-packed, low bed weight, SPE product. SOLA products exhibit recovery and reproducibility levels at low extraction volumes which are significantly better than conventional loose-packed, low bed weight competitor products. SOLA products recovery and reproducibility levels at lower extraction volumes SOLA product Competitor (i) Competitor (ii) SOLA product Competitor (i) Competitor (ii) % Recovery Caffeine 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% -20.00% 150 µL 200 µL 250 µL Elution volume SOLA Competitor (i) Competitor (ii) % Recovery Hydrocortisone 100.00% 80.00% 60.00% 40.00% 20.00% 0.00% -20.00% 150 µL 200 µL 250 µL SOLA Elution volume Competitor (i) Competitor (ii) The error bars illustrate significantly lower variability sample-to-sample for SOLA products compared to conventional SPE products. Correct results are achieved with SOLA products time after time, even at low elution volumes. Conventional loose-packed SPE products are unable to compete with the reproducibility or recovery levels of SOLA products at these low elution volumes. Significantly higher recovery levels are achieved with SOLA products at an elution volume of 150 µL for caffeine, hydrocortisone and carbamazepine compared to competitor loose-packed SPE products Carbamazepine 100.00% 80.00% 60.00% % Recovery 40.00% 20.00% 0.00% -20.00% 150 µL 200 µL 250 µL Elution volume 14 SOLA product Competitor (i) Competitor (ii) SOLA SPE 30 mg cartridges and plates Relative abundance (%) Reproducibility SOLA products, such as the SOLA 30 mg series, demonstrate exceedingly high levels of reproducibility when dealing with biological matrices, whilst maintaining excellent recovery and minimal matrix effects, even at high loading volumes. 100 counts 75 50 OH-Vit D3 The example below shows three full 96-well plate extractions of 25-hydroxyvitamin D3 from 1 mL human plasma using the Thermo ScientificTM SOLATM HRP 30 mg phase. The precision of each full-plate extraction is high, delivering reproducible batch-to-batch results time and time again. 25 0 0.06 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.06 Time (min) Chromatogram and extraction reproducibility data for 25-hydroxyvitamin D3 (IS-corrected with 13C5-OH-Vitamin D3) at 150 ng/mL (mid-QC) in 1 mL human plasma Batch 1 Batch 2 Batch 3 Mean Inter-batch reproducibility (%RSD) Average peak response factor 3.37 3.27 3.31 3.32 1.60 Precision of 96-well plate extraction (%RSD) 3.61 3.33 2.45 3.13 Also evident is the high, reproducible recoveries of extraction against competitors, as well as minimal matrix effects. Low quality control (LQC) and high quality control (HQC) samples were extracted (n = 12) and their recoveries and matrix effects calculated and averaged. The summary data is presented below. SOLA Competitor (i) Competitor (ii) Recoveries to 2 dp 95.23 102.15 118.12 Precision (CV%) 1.78 2.85 10.28 Matrix effects (%) -0.40 -3.99 -9.36 140.00 0.00 OH-Vit D3 120.00 100.00 -1.00 -2.00 -3.00 Recovery (%) Matrix e ects (%) 80.00 60.00 40.00 20.00 SOLA product Competitor (i) Competitor (ii) -4.00 -5.00 -6.00 -7.00 -8.00 -9.00 SOLA product Competitor (i) Competitor (ii) 0.00 OH-Vit D3 -10.00 Recovery comparison of 25-hydroxyvitamin D3 Matrix effect comparison of hydroxyvitamin D3 15 Even when extracting analytes from large volumes of urine, SOLA 30 mg can maintain high levels of recovery against the competition. In the example below, an extraction of 11 opioids and opiate-derivatives was conducted from 1 mL of urine using SOLA SCX 30 mg 96-well plates. We can see that not only is SOLA able to carry out a reproducible extraction, but also delivers high recoveries time-after-time. Recovery (%) 120.00 100.00 80.00 60.00 40.00 20.00 0.00 Morphine Oxymorphone Hydromorphone Noroxycodone Hydrocodone NorhydrocOod-doensemethyl-cis-tramadol Norfentanyl cis-tramadol Comparison of SPE recovery between SOLA SCX 30 mg and competitor SPE products Fentanyl Methadone SOLA SCX 30 mg Competitor (i) Competitor (ii) Competitor (iii) Morphine Oxymorphone Hydromorphone Noroxycodone Hydrocodone Norhydrocodone O-desmethyl-cis-tramadol Norfentanyl cis-tramadol Fentanyl Methadone SOLA SCX 106.8 86.9 81.2 92.6 87.6 94.8 102.1 108.8 88.8 102.8 80.4 Recovery (%) Competitor Competitor Competitor (i) (ii) (iii) MCX PCX CX 82.8 93.2 78.4 75.5 81.0 66.9 73.8 72.4 62.1 71.4 79.7 64.7 71.8 79.4 63.0 72.9 77.6 61.4 81.8 93.1 80.8 80.3 87.7 66.9 68.5 79.2 65.0 63.2 101.8 70.6 56.1 76.7 55.0 SOLA SCX 1.39 4.46 10.33 3.44 5.82 5.02 1.82 1.46 2.68 2.94 5.61 Precision (%RSD) Competitor Competitor Competitor (i) (ii) (iii) MCX PCX CX 3.09 2.52 4.17 3.31 5.30 5.28 3.24 2.13 6.00 2.78 4.31 4.36 2.48 4.49 3.96 3.54 4.34 5.05 2.20 2.16 3.77 2.59 2.18 4.98 4.15 1.78 6.59 5.39 5.18 7.56 7.29 3.28 9.32 Compound recovery and precision details between SOLA SCX 30 mg and competitor SPE products 16 Loading capacity When scaling up an assay, the larger loading capacity of SOLA 30 mg allows it to retain far more analyte, even in the presence of more matrix interferences associated with the higher loading volume. Higher sample loading volumes can also be used to boost analyte signal, as shown below. This benefit could be used to reach lower quantifiable levels, if required. 50.000 40.000 30.000 Intensity (counts) 20.000 10.000 0 50.000 40.000 SOLA 10 mg in1c.6r1et.aim6sexesin reinrsecpsrepoaonsnseseein 30.000 20.000 10.000 SOLA 30 mg 0 0.00 0.50 1.00 1.50 2.00 2.50 Time (min) The high loading capacity of SOLA HRP 30 mg also improves signal response when compared to smaller bed weights. The chromatograms show an improved signal response of OH-Vit D3 for SOLA HRP 30 mg after 1 mL of spiked human plasma (150 ng/mL) was loaded onto each bed weight. A higher SPE bed weight, such as SOLA 30 mg, can also prevent analyte breakthrough, which can happen if the loading capacity of the sorbent is insufficient. This figure is a comparison between SOLA 30 mg and smaller bed weights and demonstrates a higher recovery of analyte when loading the same volume of 1 mL spiked human plasma. Greater recovery of analyte when loading 1 mL onto SOLA 30 mg vs smaller bed weights Recovery (%) 100 75 50 25 0 SOLA 10 mg SOLA 30 mg OH-Vit D3 OH-Vit D2 17 SOLA 30 mg products are designed to efficiently extract analytes from high matrix loading volumes, including plasma and urine Loading caLpoaacditiyn:gacnaaplyatceitrye:caonvaelryytefrroemcopvlearsymfaroSmOpLlAasHmRaPS3O0LmAgHRP 30 mg plasma SOLA HRP 30 mg ticosterone 17-OH-progesterone 1150 µL 1050 µL 950 µL 850 µL 750 µL 650 µL 550 µL 450 µL Recovery (%) Recovery (%) 100.00 100.00 80.00 80.00 60.00 60.00 40.00 40.00 20.00 20.00 1150 µL 1050 µL 950 µL 850 µL 750 µL 650 µL 550 µL 450 µL 0.00 0.00 Ca eine Ca eine CorticosteronCe orticostero1ne7-OH-proge1st7e-rOonHe-progesterone Plasma loading volume 450 µL 550 µL 650 µL 750 µL 850 µL 950 µL 1050 µL 1150 µL Caffeine 1.64 0.22 1.52 1.06 1.64 3.15 1.81 2.18 Precision (%RSD) Corticosterone 1.54 4.92 0.51 0.69 3.69 2.08 3.15 2.14 17-OH-progesterone 0.52 2.05 0.95 0.39 1.74 0.86 1.35 0.33 SOLA 30 mg has been shown able to extract analytes from large plasma loading volumes whilst maintaining very high levels of reproducibility Method Sample pre-treatment Dilute spiked matrix samples, as per graph, 1:1 with 0.1 % formic acid in water 18 SPE Condition 1 mL methanol Equilibrate 1 mL water Load Pretreated samples Wash 1 mL 5% methanol in water Elute 2 × 400 L methanol Evaporate to dryness under nitrogen, no heat Reconstitute in × L mobile phase, where × = 50% loading volume R 40.00 20.00 0.00 Ca eine Corticosterone 17-OH-progesterone 1150 µL 1100 µL 1050 µL 1000 µL Analyte recovery from urine by matrix volume using SOLA HRP 30 mg Recovery (%) 1150 µL 100.00 1050 µL 950 µL 80.00 850 µL 750 µL 60.00 650 µL 550 µL 40.00 450 µL 20.00 0.00 Ca eine Corticosterone 17-OH-progesterone Urine loading volume 1000 µL 1050 µL 1100 µL 1150 µL 1200 µL 1250 µL 1300 µL 1350 µL 1400 µL 1450 µL 1500 µL Corticosterone 0.74 2.50 4.06 2.98 2.12 3.18 0.85 1.82 0.69 3.45 3.63 Precision (%RSD) Caffeine 1.11 1.33 5.79 2.03 3.06 1.11 0.49 0.90 1.25 0.61 1.86 Corticosterone 1.21 0.45 0.86 0.59 0.29 0.50 0.82 0.60 0.71 1.46 0.13 SOLA 30 mg has been shown able to extract analytes from large urine loading volumes whilst maintaining very high levels of reproducibility. This high loading capacity means that SOLA can be used to concentrate very large volumes of matrix such as urine, to reach the lowest of LOQs. 1500 µL 1450 µL 1400 µL 1350 µL 1300 µL 1250 µL 1200 µL 1150 µL 1100 µL 1050 µL 1000 µL Method Sample pre-treatment Dilute spiked matrix samples, as per graph, 1:1 with 1% formic acid in water SPE Condition 1 mL methanol Equilibrate 1 mL water Load Pretreated samples Wash 1 mL 5% methanol in water Elute 2 × 400 L methanol Evaporate to dryness under nitrogen, no heat Reconstitute in × L mobile phase, where × = 20% loading volume 19 Elution volumes With SOLA 30 mg, compounds can be extracted from biological matrices such as plasma and urine and eluted from the SPE sorbent with only 250 µL. Elution volumes from plasma Ca eine recovery from 140 plasma by elution volume 120 100 Recovery (%) 80 60 40 20 0 SOLA product Corticosterone recovery from plasma by elution volume Recovery (%) 120 100 80 60 40 20 0 SOLA product 17-OH-progesterone recovery 120 from plasma by elution volume 100 80 60 40 20 0 20 Recovery (%) SOLA product Competitor (i) Competitor (i) Competitor (i) SOLA 30 mg products outperform the competition--good recovery can be maintained with minimum elution volumes in plasma and urine matrices Competitor (ii) Competitor (iii) 150 µL 200 µL 250 µL 300 µL 350 µL 400 µL 450 µL 500 µL 150 µL 200 µL 250 µL 300 µL 350 µL 400 µL 450 µL 500 µL Competitor (ii) Competitor (iii) Competitor (ii) Competitor (iii) 150 µL 200 µL 250 µL 300 µL 350 µL 400 µL 450 µL 500 µL Method Sample pre-treatment Dilute spiked matrix samples, as per graph, 1:1 with 0.1% formic acid in water (plasma samples) or 1:1 with 1% formic acid in water (urine samples) SPE Condition 1 mL methanol Equilibrate 1 mL water Load 1 mL pretreated sample Wash 500 L 5% methanol in water Elute (150500) L methanol, see graph Evaporate to dryness under nitrogen, no heat Reconstitute in 500 L mobile phase Elution volumes from urine Ca eine recovery from 120 urine by elution volume 100 Recovery (%) 80 60 40 20 0 SOLA product Competitor (i) Competitor (ii) Competitor (iii) 150 µL 200 µL 250 µL 300 µL 350 µL 400 µL 450 µL 500 µL Corticosterone recovery from urine by elution volume Recovery (%) 120 100 80 60 40 20 0 SOLA product Competitor (i) Competitor (ii) Competitor (iii) 150 µL 200 µL 250 µL 300 µL 350 µL 400 µL 450 µL 500 µL 17-OH-progesterone recovery 140 from urine by elution volume 120 Recovery (%) 100 150 µL 200 µL 80 250 µL 300 µL 60 350 µL 400 µL 40 450 µL 20 500 µL 0 SOLA product Competitor (i) Competitor (ii) Competitor (iii) 21 SOLAµ SPE products Pharmaceutical and biopharmaceutical analytical challenges The modern bioanalytical and clinical research laboratory must provide high quality analytical results from complex biological samples in a high throughput environment while complying with strict legislation. These demands are compounded by the continued drive to higher efficacy drugs and long-acting formulations which continue to push the required quantification limits to lower levels. There is also the desire to take advantage of the replacement, refinement and reduction policy. The growth of biopharmaceuticals also brings into consideration additional analytical challenges such as solvation and non-specific binding. What is required of the bioanalytical method to meet these demands? · Robustness--low analytical failure rates · Ability to process low sample volumes · High sensitivity · High reproducibility · Ease of use · High throughput processing · Efficient and fast The micro elution SPE format is uniquely positioned to deliver on these requirements Delivering reproducible, low volume extractions Reproducible extraction with Reproducible extraction with 25 uL elution v2o5luµmLeesluutsiionng SOLA SPE well plates are designed for bioanalytical SOLAu WAX volumes using the and clinical research analysts who require cleaner, SOLAµ WAX 96-well highly reproducible, and robust sample extraction at SPE plate very low sample and solvent volumes in high throughput 120% workflows. SOLA well plates achieve this with unique and innovative fritless SPE technology. 100% SOLA well plates are the first micro elution product to truly meet the requirements of the bioanalyst. The example below demonstrates the excellent reproducibility of SOLAµ at low elution volumes. Three compounds were extracted from matrix across three Thermo ScientificTM SOLAµTM WAX 96-well SPE plates and successfully eluted with only 25 µL of elution solvent. High recoveries and low imprecision were maintained across each SPE plate, whilst inter-batch variation between plates was also less than 3.9%. 80% 60% 40% 20% 0% 4-Propylbenzoic acid Niflumic acid 4-Pentylbenzoic acid Plate 1 Plate 2 Plate 3 Combined 22 Maintaining excellent loading capacity The utilization of our advanced polymeric technologies in the SOLA plate provides an SPE phase with excellent loading capacity. This ensures that good retention of analyte and removal of matrix interferences is achieved when a larger range of sample volumes are applied. In the following example, incremental volumes of human plasma spiked at 200 ng/mL with a polar (atenolol) and non polar (imipramine) analyte were extracted. Recovery and matrix effects were monitored across the loading range to demonstrate acceptable assay performance. Loading capacity: recovery and reproducibility 120% 100% 80% 120% 60% 100% 40% 80% 20% 60% 0% 40% 50 20% 100 150 250 350 450 500 Undiluted plasma load volume (µL) 0% 40% 50 100 150 250 350 450 500 30% Undiluted plasma lead volume (µL) L2o0w%matrix effects: enhancement and suppression 1400%% 300%% -1200%% -2100%% -300%% --4100%% 50 -20% -30% -40% 50 100 150 250 350 450 500 Atenolol Imipramine 100 150 250 350 450 500 Atenolol Imipramine Loading capacity maintained over incremental volumes of human plasma spiked at 200 ng/mL with atenolol and imipramine analytes 23 Providing reproducible sensitivity By combining the impressive loading capacity and very low elution volume capability of SOLAµ, you can improve the sensitivity of your bioanalytical assays by up to 20 times the original starting concentration. In the following example, 500 µL human plasma was loaded onto the SOLAµ plate for the analysis of niflumic acid. The compound was eluted in 25 µL providing a 20 times increase in concentration whilst maintaining excellent precision. The problem: improvement in assay sensitivity required 1:1 dilution with 4% H3PO4 40 pg/mL 500 L sample 20 pg/mL SOLA well plate solution: up to 20 times increase in concentration of sample without changes to workflow 1:1 dilution with 4% H3PO4 40 pg/mL 500 L sample 20 pg/mL Conventional scale SPE 500 L elution 40 pg/mL 20× concentration SOLA well plate 25 L elution 800 pg/mL Sample enrichment (20 times concentration) 20 times increase in sensitivity Sample preparation method Sample pre-treatment 500 µL of human plasma diluted 1:1 with 4% phosphoric acid Sample preparation Compound(s) Niflumic acid, niflumic acid d5 (IS) Matrix Human plasma SOLAµ WAX 96-well plate (60209-005) Condition 200 µL methanol Equilibrate 200 µL 4% phosphoric acid Load Apply sample at 0.5 mL/min Wash 200 µL 25 mM ammonium acetate (pH4) 200 µL 70% methanol (pH4) Elute 2 × 12.5 µL 50/50 methanol/acetonitrile with 2% ammonia Direct injection of eluent HPLC system Thermo ScientificTM DionexTM UltiMateTM 3000 RSLC System Column Thermo ScientificTM AccucoreTM RP-MS HPLC Column 50 mm × 2.1 mm 2.6 µm (17626-052130) Guard column Thermo ScientificTM AccucoreTM RP-MS DefenderTM Guard Cartridge (17626-012105) Thermo ScientificTM UniguardTM Drop-in Guard Holder (852-00) Mass spec system Thermo ScientificTM TSQ VantageTM Triple-Stage Quadruple Mass Spectrometer Relative abundance 100 90 80 70 60 50 40 30 20 10 100 90 80 70 60 50 40 30 20 10 0 24 1.33 400 pg/mL aqueous solution of niflumic acid taken through the extraction procedure (20 times increase in sensitivity) 400 pg/mL aqueous solution of niflumic acid injected neat (no concentration factor) 0.5 1.0 1.33 1.5 Time (min) Peak area 144300 counts Peak area 7215 counts 2.0 2.5 Low QC (0.4 ng/mL) High QC (30 ng/mL) Precision data for niflumic acid peak area ratio (%RSD) n = 18 1.31 1.06 Recovery of niflumic acid (%) 89.9 94.0 Matrix effects (%) 8.63 3.21 Precision, recovery and matrix effects data for niflumic acid at Low QC 0.4 ng/mL and High QC 30 ng/mL (n=18) 3.0 SOLAµ SPE methods: Perfect for sample limited assays or scaled-down conventional SPE methods SOLAµ products can maintain equivalency when scaling products the ideal choice when processing low sample down conventional, large-scale SPE methods. That said, volumes in your bioanalytical assays. you can directly scale down the volumes used in their analytical methods. This means a reduction in sample usage--there is less need for excessive sampling from animal and human models, and in turn, stress endured by the hosts is alleviated. By using less sample to begin with, you can also reduce your solvent costs by using smaller wash and elution volumes, making SOLAµ SOLA allows you to achieve the same results even when sample volumes are low The following example shows that by loading 25 µL of niflumic acid sample onto the SOLAµ plate and eluting in a total of 25 µL, a ten-fold decrease in sample volume was achieved when compared to a traditional scale higher bed weight product. Equivalent method performance and high levels of reproducibility provided by SOLA technology were still maintained. Sample preparation method Sample pre-treatment Human plasma diluted 1:1 with 4% phosphoric acid Sample preparation The problem: Sample volume requires reduction without compromising sensitivity 1:1 dilution with 4% H3PO4 250 L sample 500 L diluted Traditional scale SPE 250 L elution 250 L ready to inject Compound(s) Matrix Condition Equilibrate Load Wash Niflumic acid, niflumic acid d5 (IS) Human plasma SOLAµ WAX 96-well plate (60209-005) 200 µL methanol 200 µL water Apply 25 µL sample at 0.5 mL/min 200 µL 25 mM ammonium acetate (pH4) 200 µL methanol SOLA well plate solution: 10 fold reduction in sample volume with no additional step 1:1 dilution with 4% H3PO4 25 L sample 50 L diluted SOLA well plate 25 L elution 25 L ready to inject Elute 2 × 12.5 µL methanol with 2% ammonia Direct injection of eluent HPLC system Dionex UltiMate 3000 RSLC system ColumnAccucore RP-MS HPLC column 50 mm × 2.1 mm 2.6 µm (17626-052130) Guard columnAccucore RP-MS Defender guard cartridge (17626-012105) Uniguard drop-in guard holder (852-00) Equivalency of results obtained with niflumic acid (500 ng/mL) extracted with 10 mg SOLA WAX using 250 µL of sample and SOLAµ WAX using 25 µL of sample Mass spec systemTSQ Vantage triple-stage quadruple mass spectrometer Intensity 500,000 450,000 Precision data for niflumic acid 400,000 350,000 300,000 250,000 200,000 SOLA well plate Low QC Analyte peak area (%RSD) 7.32 Peak area ratio (%RSD) 0.356 150,000 100,000 High QC 5.33 0.195 50,000 0 500,000 Precision data niflumic acid at low QC 0.4 ng/mL and high QC 30 ng/mL (n=18) 450,000 400,000 350,000 300,000 250,000 SOLAµ well plate 200,000 150,000 100,000 50,000 0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 Time (min) 25 SOLA and SOLAµ method scaling With traditional SPE the eluted sample is typically blown down to increase the concentration of the sample and thus improve the sensitivity. This causes an issue for certain compound types which can be lost during this step, resulting in reduced sensitivity. SOLA well plates allow the sample to be extracted without the need for dry down and reconstitution. Not only does this maximize recovery of the analytes it also improves workflow efficiency and increases productivity. 100% 80% % Recovery 60% 40% 20% 0% SOLAµ SCX SOLA SCX 10 mg SOLA SCX 30 mg cis-Tramadol 99.4% 97.6% 88.8% Fentanyl 95.8% 96.8% 102.8% Hydrocodone 97.3% 96.3% 87.6% Morphine 100.1% 97.0% 106.8% Recovery comparison following method scaling of opioids extraction from urine Norhydrococone 96.6% 93.9% 94.8% Noroxycodone 98.5% 95.7% 92.6% O-Desmethyltramadol 97.3% 98.1% 102.1% This chart shows an established method using SOLA SCX (10 mg SPE) compared to a scaled-down method using SOLAµ SCX (2 mg SPE) and a scaled-up method using SOLA SCX 30 mg. By removing the sample blow down and reconstitution step and using microelution with SOLAµ instead, there is a time savings of almost 30 minutes per plate, whilst maintaining reproducibility and recovery when compared to the 10 mg SPE extraction. When utilizing the higher loading capacity of SOLA 30 mg, the matrix volume by was increased over 5 times (1 mL with SOLA 30 mg as opposed to 200 µL with SOLAµ and SOLA 10 mg), again whilst maintaining high recoveries and high precision. No matter which SOLA bed size you chose, SOLA will always deliver reproducible results. SOLA SCX 30 mg SOLA SCX 10 mg SOLAµ SCX 2 mg Vol (µL) Time (min) Vol (µL) Time (min) Vol (µL) Time (min) 1000 5 Condition with methanol 500 5 Condition with methanol 200 5 1000 5 Equilibrate with water 500 5 Equilibrate with water 200 5 2000 10 Load pre-treated sample 1000 5 Load pre-treated sample 1000 10 1000 5 Wash with 0.1% formic acid (aq) 500 5 Wash with 0.1% formic acid (aq) 200 5 1000 5 Wash with 0.1% formic acid (methanol) 500 5 Wash with 0.1% formic acid (methanol) 200 5 Place a collection plate under the SPE device to capture the extract 2 × 400 5 Elute with MeOH/ACN/TEA(45/45/10) 2 × 200 5 Elute with MeOH/ACN/TEA(45/45/10) 2 × 25 5 -- -- Post-extraction processing requirPemosetn-etsxtraction processing requirements Dilute with water -- -- Dilute with water 50 1 -- 45 Evaporate under nitrogen -- 30 Evaporate under nitrogen -- -- 100 5 Reconstitute with mobile phase 100 5 Reconstitute with mobile phase -- -- Method details for SOLA SCX and SOLA SCX showing each step, volume of solvent required, and length of time in minutes for each step 26 The problem: Workflow requires dry down to achieve sensitivity 1:1 dilution with 4% H3PO4 Conventional scale SPE 10 ng/mL 5 ng/mL 500 L elution 4 ng/mL Re-con in 50 L 40 ng/mL SOLA well plate solution: Low elution volume allows removal of dry down 1:1 dilution with 4% H3PO4 10 ng/mL 5 ng/mL SOLA well plate 50 L elution 40 ng/mL Example chromatogram at quantitation limit of 10 ng/mL for ibuprofen Sample preparation method Sample pre-treatment 200 L of rat plasma diluted 1:1 with 4% phosphoric acid Sample preparation Compound(s) Ibuprofen, ibuprofen d3 (IS) Matrix Rat plasma SOLA SAX 1 mL 96 well plate (60109-002) Condition 200 µL methanol Application Load sample at 0.5 mL/min Washing 200 L water with 1% NH4 200 L methanol with 1% NH4 Elution 2 × 25 L 50/50 methanol/acetonitrile with 2% formic acid Dilution Add 50 L water to each sample Direct injection of eluent HPLC system Dionex UltiMate 3000 RSLC system ColumnAccucore RP-MS HPLC column 50 mm × 2.1 mm 2.6 µm (17626-052130) Guard columnAccucore RP-MS Defender guard cartridge (17626-012105) Uniguard drop-in guard holder (852-00) Mass spec systemTSQ Vantage triple-stage quadruple mass spectrometer Conclusion: Key benefits of each format Whether it is scaling up an assay to reach lower limits of quantitation with SOLA 30 mg, or scaling down with SOLAµ to resolve solvation issues and improve sample enrichment, you can rest assured that reproducible, robust results are always available with the innovative, fritless design of SOLA cartridges and well plates. SOLAµ 2 mg · A robust low sample volume preparation platform · Reproducibility at low sample and solvent levels · Processing of low volume samples · Sample enrichment (20 times) · Mitigates against solvation and non-specific binding issues SOLA 10 mg · Good option for most analyses · Lower elution volumes than silica-based products thus reducing time for evaporation and increasing throughput SOLA 30 mg · When high loading volumes are required to reach lower limits-of-quantitation · For difficult to retain analytes · When experiencing analyte breakthrough on a smaller bed weight 27 Ordering information Product Bed weight (mg) Cartridge/well plates volume (mL) Thermo ScientificTM SOLAµTM Solid-Phase Extraction (SPE) 96-well plates SOLA HRP 96-well plate 2 1 SOLA SCX 96-well plate 2 1 SOLA SAX 96-well plate 2 1 SOLA WCX 96-well plate 2 1 SOLA WAX 96-well plate 2 1 Thermo ScientificTM SOLATM Solid-Phase Extraction (SPE) cartridges SOLA HRP SPE cartridge 10 1 SOLA SCX SPE cartridge 10 1 SOLA SAX SPE cartridge 10 1 SOLA WCX SPE cartridge 10 1 SOLA WAX SPE cartridge 10 1 SOLA HRP SPE cartridge 30 3 SOLA SCX SPE cartridge 30 3 SOLA SAX SPE cartridge 30 3 SOLA WCX SPE cartridge 30 3 SOLA WAX SPE cartridge 30 3 Thermo ScientificTM SOLATM Solid-Phase Extraction (SPE) 96-well plates SOLA HRP 96-well plate 10 2 SOLA SCX 96-well plate 10 2 SOLA SAX 96-well plate 10 2 SOLA WCX 96-well plate 10 2 SOLA WAX 96-well plate 10 2 SOLA HRP 96-well plate 30 2 SOLA SCX 96-well plate 30 2 SOLA SAX 96-well plate 30 2 SOLA WCX 96-well plate 30 2 SOLA WAX 96-well plate 30 2 Quantity 1 each 1 each 1 each 1 each 1 each 100 pack 100 pack 100 pack 100 pack 100 pack 50 pack 50 pack 50 pack 50 pack 50 pack 1 each 1 each 1 each 1 each 1 each 1 each 1 each 1 each 1 each 1 each Expect reproducible results with sample prep, columns and vials Cat. no. 60209-001 60209-002 60209-003 60209-004 60209-005 60109-001 60109-002 60109-003 60109-004 60109-005 60409-001 60409-002 60409-003 60409-004 60409-005 60309-001 60309-002 60309-003 60309-004 60309-005 60509-001 60509-002 60509-003 60509-004 60509-005 Don't see what you need? We would be happy to discuss your specific requirements. Please contact your local sales representative for custom orders. Find out more at thermofisher.com/solaspe For Research Use Only. Not for use in diagnostic procedures. © 2020 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. 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 locations. Please consult your local sales representative for details. BR22044-EN 1120MAdobe InDesign CC 13.1 (Macintosh) Adobe PDF Library 15.0