Fast, Convenient All-in-One Efficient Protein Removal Filtration for HPLC Sample Prep for Different Sample Types O Simple, one step, single tube method–Eliminates
centrifugation plus aspiration issues and transfer steps
O Process samples 3 times faster–Purify 6 samples
in 3 minutes, 48 samples in <30 minutes
O >99% protein removal for plasma samples– % Reduction in [pr
Enables compound quantitation and extends
Sample Type O Self-sealing septa allows for repeated
The effectiveness of removing protein from different sample
injections–Prevents evaporation and
types was evaluated. Plasma, serum, and whole blood (diluted1:1 in H 0) were run through the MUP-PPT procedure. The
same samples were also run through a conventional spinmethodology which utilizes a 1.5 mL microfuge tube in place
O Compatible with all major autosamplers
of the Mini-UniPrep, and a 5 minute centrifuge @ 10,000 xg to bring the proteins down. Supernatant was aspirated into a separate tube for analysis. Micro-BCA was used as previously
Whatman Mini-UniPrep Syringeless Filters offer a highly
described to quantitate proteins. All results are reported in %reduction in protein concentration.
efficient, simple alternative for removing unwanted proteinprior to HPLC/MS analysis. Ideal for performing analyticalcharacterization in drug research, the method utilizesacetonitrile precipitation and filtration by compression to
Ordering Information
remove protein from plasma, serum, whole blood, and
Catalog No. Pore Size (µm) Membrane
other biological fluids. It is a single tube method that
saves time and eliminates the manual transfer steps
which make spin clarification problematic. Mini-UniPrep Compressor Catalog No. Description Quantity
The MUP Compressor enables up to six Mini-UniPrep
filter devices to be processed simultaneously. Thishandy tool will speed sample prep processing andreduce the risk of carpal tunnel syndrome.
For more information on the NEW Protein Precipitation (PPT) Protocol for Mini-UniPrep™
Syringeless Filters, or any other Whatman products call 1.800.WHATMAN
or visit our web site at www.whatman.com
Mini-UniPrep™ Protein Precipitation Protocol
Add 400 µL Insert MUP Add 100 µL acetonitrile to MUP filter plunger until plasma to MUP sample chamber first locking ring is sample chamber Vortex sample compressor unit and mixture for compress until second Analyze sample lock ring is engaged 3-5 seconds Reproducibility of Protein Removal HPLC Quantitation of Caffeine in Plasma Observed (ng/mL) % Reduction in [pr Replicate # Expected (ng/mL)
The reproducibility of protein removal from plasma samples was
Plasma samples containing varying amounts of caffeine were prepared
measured by applying the MUP-PPT procedure to 40 replicates of
in blinded fashion and processed using the Mini-UniPrep along with
normal human plasma. Filtrate was removed from the Mini-UniPrep
known plasma standards containing caffeine. After compression
and dried down in a microfuge tube prior to saline resuspension
filtration, the standards and “unknowns” were analyzed by HPLC
and micro-BCA protein quantitation. Percent protein removal was
using 10% Acn/90% H 0 mobile phase and a Partisil® 5-ODS-3 reverse
calculated by comparing the protein concentrations of pre- and
phase column (4.6 x 100 mm) at a flow rate of 2 mL/min. The OD275
nm caffeine peaks as measured used to determine the caffeine concen-tration of the unknown samples by comparison to the standard set. Productivity Advantages Versus Spin Method SPIN METHOD
Single sample processing time (plasma tube ➤HPLC) ~30 seconds
• close vial screw cap• place in autosampler
~45 minutes(>45 minutes if centrifugecapacity is <48 samples)
1.800.WHATMAN • 973.773.5800FAX: 973.773.0168www.whatman.com • [email protected]
Copyright, Whatman Inc., 2002 Printed in USAWhatman is a trademark of Whatman International Ltd.
Mini-UniPrep™ is a trademark of Whatman Inc. Partisil® is a registered trademark of Whatman Inc.
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