Mitra (RUO) Microsampling Device User Manual
Contents
1. Once the sampler tip is applied look for it to go fully red This will happen VERY fast Do not get surprised and yank the sampler tip away Once you see the sampler tip go fully red count an additional gt 2 seconds and then SLOWLY and SMOOTHLY YN e Total contact time for the the sampler tip to NY blood will typically not be longer than 6 seconds ALA IN SX unless hematocrit of the blood is gt 60 MO SY NX e You will not over sample if sampler tip is left in blood longer than 2 additional seconds as long as IY it is not immersed 3 or more seconds is better than stopping too early to ensure sampler tip is full remove sampler tip from the blood A WN C AS N clamshell back together and press closed You will hear it snap into place Closed clamshell can be dried under ambient conditions or immediately bag with 1 gram of desiccant per sampler e MPORTANT ensure that the desiccant is fresh or proper drying will not sample may be compromised analysis or stored for future use e See page 9 for extraction recommendations Repeat steps 3 4 for the remainder of the sampler tips in the clamshell and then fold placedina occur and Once tips are fully dry 2 hours the samplers can be transported processed for Drying Storage amp Transportation considerations or special couriers Fully dried tips are not a biohazard and can be shipped without dry ice hazar2. Collection Label sampler bodies and outer box according to your needs protocol Remove rack of 96 samplers from outer box by pinching the two raised tabs on the rack rocking gently to release and lifting out gripping samplers also aids in removal Remove individual sampler from rack with Mitra Sampling Tool e See page 7 of manual to see Sampling Tool use instructions Apply tip of sampler to surface of blood sample Ensure 15 uL of blood is available per sample to avoid under sampling NY e ry to sample from a 45 angle for best results not perpendicularly B e Do NOT fully immerse tip e Do NOT make contact with skin surface e Do NOT drip blood onto the tip if sampling from a cannula Once the sampler tip is applied look for it to go fully red This will happen VERY fast Do not get surprised and yank the sampler tip away Once you see the sampler tip go fully red count an additional 2 seconds and then SLOWLY and SMOOTHLY remove sampler tip from the blood e Total contact time for the sampler tip to blood will typically not be longer than 6 seconds unless hematocrit of the blood is gt 60 e You will not over sample if sampler tip is left in blood longer than 2 additional seconds as long as itis not aceon 3 or more seconds is nelle than stopping too early to ensure sampler tip is full Transfer full tip to drying rack Be careful not to touch it to the sides of the rack A sim
3. All extractions using more than 30 water will require centrifugation for the removal of solids e 2 step extractions such as water followed by acetonitrile can be used e Alternative methods of physical agitation such as longer vortexing steps sonication or heated solvents can be used to improve recoveries Example Methanol MeOH Extraction Add MeOH to extraction plate Acceptable volumes are of 200 500 uL We also recommend adding the internal standard at this point Add Mitra Samplers to the extraction plate Vortex extraction plate for 15 minutes at 1100 RPM using a platform mixer rotator Remove Mitra Samplers from the extraction plate If particulate is present centrifuge the Extraction Plate at 1500G for 5 minutes Remove supernatant from extraction plate and transfer to a suitable container e g another extraction plate Blow down to dryness using N Reconstitute sample with solvent suitable for dissolution of analytes Analyze extract IMPORTANT Extraction of dried blood from Mitra sampler should be optimized using your target analyte at a concentration of 5x your LLOQ doped into 65 hematocrit blood Absolute recoveries of gt 85 under these conditions will provide the best possible results across a range of analyte concentrations and hematocrit levels mitrasupportfaphenomenex com 44 www phenomenex com mitra ALANE eoteryx The Mitra Microsampler 96 Well
4. Plate and 4 Pack Clamshell formats should be stored in a dry location under ambient conditions The shelf life is 12 months Parts are labeled with the expiration date mitrasupportfaphenomenex com 44 www phenomenex com mitra ALANE eoteryx N N Issue Most Likely Cause Solution Optimize extraction method using recommendations found on p 9 of this manual or contact Phenomenex for assistance Low lt 75 or no recovery of A non optimized extraction target analyte method Mitra Sampler Tip is not absorbing a Sampler tip is expired a Use a fresh sampler tip or taking longer than 6 seconds to fully adsorb b Sampler tip was stored at a b Use a fresh sampler tip that temperature gt 110 F has been stored under ambient conditions c Blood has gt 65 hematocrit level c Hold sampler tip to blood pool longer than 6 seconds to allow more time for wicking Mitra Sampler Tip is not dry after a Ambient conditions were near a Move tips to dry in a low 2 hours 100 humidity humidity environment or dry under desiccated conditions b The samplers in the drying rack were returned to storage box b Remove 96 well dry rack from before they were dry outer box so air freely circulates around sampler tips Mitra Sampler Tip is not wicking a Tip was submerged into blood a Resample with a new tip and volumes within the specification pool apply it just to surface o
5. 6 Well Plate Assembly 1 ea 9R K002 CD Mitra RUO 10uL Microsampling Device 96 Well Plate Assembly 12 pk Typical Uses For high throughput environments e g discovery labs For animal studies in drug development For method development 9R K002 BF Mitra RUO 10uL Microsampling Device 4 Pack Clamshell 60 pk For sample collection in clinical trials including Direct Sampling At home Sampling Remote Locations AK 9268 A Mitra Drying Rack 96 Well Plate 1ea To organize sampling for high throughput environments AK 9269 A AK 9270 A Mitra SBS Deck Adapter 96 Well Plate Mitra Sampling Tool 1ea 1ea For mounting 96 well plate on deck of liquid handlers To easily manipulate and transfer samplers in 96 well plate assembly AH0 8636 96 Well Collection Plate 2 mL round well with round bottom 50 pk To perform extractions prior to analysis mitrasupportfaphenomenex com www phenomenex com mitra ry Asneoteryx breaking barriers N IN e Mitra RUO 10 uL Microsampling Device e Mitra Drying Rack recommended for sample organization e Mitra Sampling Tool recommended for sampler handling e Mitra SBS Deck Adapter recommended for automation e 96 Well Collection Plate recommended for extractions
6. Mitra RUO Microsampling Device User Manual OC Mitra RUO is developed and nanufactured by Neoteryx Mitra RUO is distributed and supported mitrasupportfaphenomenex com ry www phenomenex com mitra 44 ryx breaking tery oe es nN pe 2014 Neoteryx LLC All rights reserved La A Overview Formats and Accessories General Use Protocols e 96 Well Plate e 4 Pack Clamshell Sampling Tool Use Automation Guidance for 96 Well Plate Extraction Recommendations Storage and Stability Troubleshooting Frequently Asked Questions mitrasupportfaphenomenex com 44 www phenomenex com mitra Ad n rea oteryx ng barriers A The patent pending Mitra RUO Microsampling Device Is based on revolutionary yet simple Volumetric Absorptive Microsampling VAMS technology for the collection transport storage and analysis of biological fluids In seconds the Mitra Microsampler collects 10 uL resulting in accurate and precise quantitative data while reducing or eliminating volumetric blood hematocrit bias The Mitra Microsampler Is dried stored and transported under ambient conditions Once ready for analysis a simple and automatable extraction procedure is performed mitrasupportfaphenomenex co 44 M www phenomenex E Md neoteryx SN K SZ DY Part No 9R K002 CA Description Mitra RUO 10uL Microsampling Device 9
7. all percentages of hematocrit blood No The sampler tip will wick more slowly with higher percentages of blood hematocrit Q Are there any precautions I need to take to make sure the clamshell doesn t open during shipment No The clamshell has been designed with a tight fit and has been tested to not open during shipment Q What happens if contaminate the sampler body with blood This should not negatively affect the results as only the sampler tips will be immersed in the extraction solvent mitrasupportfaphenomenex com VV i www phenomenex com mitra 44 neoteryx breaking barriers
8. cal purpose It should not be used with the intent or in the course of clinical diagnosis treatment or care of humans or animals 2014 Neoteryx LLC All rights reserved mitrasupportfaphenomenex com VV i www phenomenex com mitra 44 neoteryx breaking barriers amp Mitra MICROSAMPLING DEVICE FREQU ENTLY Q Is there a specific contact angle that the Mitra Sampler Tip should be applied to the blood pool with ASKED QUESTIONS For best sampling performance we recommend sampling from 45 angle Q Can other fluids be collected using the Mitra Microsampler Yes Other biological fluid urine ocular cerebral spinal etc can be collected as long as there is at least a 20 ul pool to collect from Q Can the Mitra Microsampler only be used to collect samples from a live host No Samples can be collected directly from a live host or from blood collected in another device e g Vacutainer Hematocrit tube etc Q Is there a correct way to add an internal standard We have had much success with adding the internal standard as part of the extraction solvent Q What happens if samples are transported before they are fully dry If the desiccant that they are transported with is functioning the samples will dry during transport however samples need to be dried completely before transport or they may be considered a biohazard Q What is the best way to label the 4 pack clamshell and the individual samplers For t
9. dous mitrasupportfaphenomenex com 44 www phenomenex com mitra 44 neoter breaking barriers yx N N The Mitra RUO Sampling Tool assists in picking up the Mitra Samplers from the 96 well drying rack and can also be used to take the sample There are several ways to hold the sampling tool according to what is most comfortable for you Below are two recommended holds Hold Recommendation 1 Place thumb on top of plunger and grasp the body of the tool between middle and ring finger Hold Recommendation 2 Grasp body of tool between thumb and middle finder and place index finder on top of plunger Guide tip of Sampling Tool into distal end of Mitra Sampler and press down for a tight fit Collect blood sample Press down on plunger to eject Mitra Sampler back into drying rack mitrasupportfaphenomenex com 44 www phenomenex com mitra ALANE eoteryx N IN The Mitra SBS Deck Adapter aligns and holds the Mitra 96 well plate assembly to the correct SBS positioning of the sampler bodies to allow automation of sample extraction Once samples are dry replace the drying rack holding the dried samples into the bottom portion of the storage box Place the adapter platels in the desired locations on the liquid handler deck Place the Mitra 96 well assembly drying rack inside storage box on the adapter plate in the desired orientat
10. f blood stated on the Certificate of pool Conformance b Did not add the additional 2 seconds once tip went fully red b Allow tip to have an additional two seconds of contact time with c Blood has gt 65 hematocrit samples level c Hold sampler tip to blood pool NOTE if you are determining longer than 6 seconds to allow the volume by using a density more time for wicking conversion ensure that the conversion is correct Variation in signal is observed Mitra Sampler Tip is not fully dry Tips may appear dry long before they are actually dry Dry tips longer and try again Extractions are showing a Samples were not completely a Remove 96 well dry rack from unacceptable variation in response dry outer box so air freely circulate i e gt 15 around sampler tips b Samples did not absorb completely b Allow tip to have an additional two seconds of contact time with c Ambient conditions were near samples 100 humidity c Move tips to dry in a low humidity environment or dry under desiccated conditions Trademarks Neoteryx Mitra and VAMS are trademarks of Neoteryx LLC Vacutainer is a registered trademark of Becton Dickinson and Company Disclaimer Mitra is patent pending Mitra RUO is developed and manufactured by Neoteryx Mitra RUO is distributed and supported by Phenomenex The Mitra RUO Microsampling Device is for research use only RUO and should serve no medi
11. he clamshell we recommend applying a label to the large flat square area on the front or back For the sampler we suggest applying a label to the distal end Q Can I take a sampler out of the 4 pack clamshell to sample 12 Yes Individual samplers can be removed from clamshell to sample if required To replace into clamshell after sampling snap back into place Q Is there a limit to how long can store the tip after the sample has been collected This will depend on your analyte method of storage and method of extraction You will need to determine the conditions required for keeping your assay within validation criterion Q Will the tip s color change after extraction Is the change in the color of the tip an indication of the recovery The color of the tip will change depending on the extraction solvents used It is an indicator that an extraction has occurred However it is not indicative of the percent recovery If the tip is extracted with aqueous it becomes whiter If the tip is extracted with aqueous and a modifier it will become completely white If the tip is extracted using an organic solvent such as methanol the tip will become brownish Q When collecting blood from a live host should I wipe the first drop of blood before sampling It depends on your sampling protocol However it is typically best practice to wipe the first drop of blood before taking the sample Q Will the sampler tip wick at the same speed for
12. ion e The Mitra drying rack has a raised corner for rapid identification of rack plate orientation IMPORTANT The Mitra drying rack should be contained within its storage box when used with a liquid handler and placed on top of the adapter The storage box itself Is NOT SBS compatible and should not be used directly on a liquid handler mitrasupportfaphenomenex com 44 www phenomenex com mitra ALANE eoteryx N N Extraction of the Mitra Sampler is quite simple and very similar to common methods used with dried blood spotting cards The recommended extraction plate P N AHO 8636 is designed to specifically hold the Mitra Samplers If desired the Mitra Sampler fits a standard 20 200 uL pipette head and a liquid handler robot properly equipped to perform the protocol can be used see page 8 of manual for automation guidance Common extraction solvents recommended include water methanol acetonitrile and ethyl acetate or mixtures thereof Below are tips to aid you in extraction method development e Neutralizing ionizable analytes i e addition of 1 formic acid for extraction of acidic analytes or 1 of ammonium hydroxide for bases can improve recoveries e For high log P analytes the use of lower polarity solvents such as ethyl acetate is advantageous e Follow water extractions with an appropriate liquid liquid extraction e g ethyl acetate or dichloromethane DCM
13. ple way to do this is to place the sampler almost vertical to the hole you want to place it in and guide straight down Repeat steps 3 6 for the remainder of the sampler tips Drying Storage amp Transportation Allow tips to dry in 96 sampler dry rack at ambient temperate for 2 hours e Air should freely circulate around tips to aid in drying Once tips are fully dry 2 hours the 96 sampler rack can be inserted back into outer box and stored for future use or can be processed for analysis e See page 9 for extraction recommendations Fully dried tips are not a biohazard and can be shipped without dry ice hazardous considerations or special couriers mitrasupportfaphenomenex com 44 www phenomenex com mitra ALANE eoteryx N IN e Mitra RUO 4 pack Clamshell e 96 Well Collection Plate recommended for extractions Label samplers and clamshell according to your needs protocol Uncover sampler bodies by pulling apart clamshell and fold over cover to create a handle for easy sample collection Apply sampler tip to surface of blood sample Ensure 15 uL of blood is available per sample to avoid under sampling Try to sample from a 45 angle for best results not perpendicularly Do NOT fully immerse tip Do NOT make contact with skin surface Do NOT drip blood onto the tip if sampling from a cannula
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