Waters Oasis HLB User Manual

[ CARE AND USE MANUAL ]

Oasis HLB Cartridges and 96-Well Plates

CONTENTS

I. INTRODUCTION

II. SAMPLE PRE-TREATMENT

a. Biological Samples

b. Solid Samples: Soil, Whole Foods, Tissue

c. Aqueous Samples: Water, Beverages

d. Non-Aqueous Liquid

III. SOLID PHASE EXTRACTION FOR ACIDIC,
NEUTRAL, AND BASIC COMPOUNDS USING
OASIS HLB

IV. OASIS HLB PROTOCOL CHARTS

a. Generic Method Oasis HLB SPE

b. Strategy for Optimizing the Generic SPE Method

V. OASIS HLB 20 BOTTLE
OPTIMIZATION APPROACH

VI. TROUBLESHOOTING

a. Adjustment to Optimize Recoveries

I. OASIS HLB

Universal Sorbent for Acidic, Neutral, and Basic Compounds

Oasis® HLB is a Hydrophilic-Lipophilic-Balanced, water-wettable,

reversed-phase sorbent for all your SPE needs. It is made from a

specific ratio of two monomers, the hydrophilic N-vinylpyrrolidone

and the lipophilic divinylbenzene. It provides superior reversed-

phase capacity with a neutral polar ‘hook’ for enhanced retention of

polar analytes.

Waters® has built a family of SPE sorbents which inherit some

key features of this unique substrate: stability at pH extremes

and in a wide range of solvents, extraordinary retention of polar

compounds, and a relative hydrophobic retention capacity 3X

higher than that of traditional silica-based SPE sorbents like C18.

Water-wettable Oasis Sorbents exhibit excellent retention capacity

for a wider polarity spectrum of analytes, even if the sorbent bed

runs dry during conditioning or sample loading. This means that

your SPE methods will be more rugged and robust, obviating the

need for repeat preparation.

The advantage of having higher retention capacity [k] is that

more analytes are retained with less breakthrough, improving the

recovery and overall reproducibility of your SPE method.

Available in five particle sizes [60 μm, 30 μm, 25 μm, 15 μm, and

5 μm], Oasis HLB sorbent, in cartridge, plate, or column format,

allows you to select the appropriate product based on the volume,

viscosity, and turbidity of your sample.

[ CARE AND USE MANUAL ]

III. SAMPLE PRE-TREATMENT

a. Biological Samples

This section contains recommendations for preparing your

biological samples (plasma, serum, urine, etc.) prior to solid-

phase extraction.

1. Prepare acidified or basified water diluents.

Note: (To prepare 4% phosphoric acid, Dilute 4.7 mL of 85%

phosphoric acid (the most common available formulation) to

100 mL final volume with water. Or 11.76 mL to 250 mL). To

prepare 5% concentrated ammonia, dilute 5 mL of concentrated

ammonia solution to 100 mL with water. Users will need to

prepare large volumes 100 -250 mL) of pretreatement buffer to

accommodate multiple samples and ensure consistency in the

buffer composition.)

2. Dilute plasma or urine, 1:1 with acidified or basified water. Add
10 to 50 μL of internal standard.

Note: Final concentration should be no more than 10% organic

otherwise protein precipitation can occur and polar compound

retention may be compromised.

3. If necessary, clarify samples by centrifugation at 8,000 x g
for 10-30 minutes.

b. Solid Samples: Soil, Whole Foods, Tissue

1. Homogenize the sample with an appropriate solvent to obtain
an aqueous based or an organic solvent based extract of the
sample. Initial extraction conditions are chosen to maximize
analyte recovery, while minimizing matrix interference. In
many cases, it may be beneficial to add buffers, dispersive
salts, or co-solvents to improve extraction efficiency.

2. Adjust the initial extract to optimize analyte retention onto
the SPE sorbent. This can include pH adjustment, solvent
adjustment, or solvent exchange through evaporation and
reconstitution (refer to Sections IV and V). It may be necessary
to centrifuge or filter the sample prior to loading.

If sufficient dilution has occurred, the sample may be treated in a

manner similar to an aqueous sample.

Note: If necessary, filter samples for suspended solids

(Eg; Environmental/Waste water/Water analysis/Food, etc.).

IV. SOLID PHASE EXTRACTION FOR ACIDIC,
NEUTRAL AND BASIC COMPOUNDS USING
OASIS HLB

1. Place Oasis HLB cartridge or plate on the vacuum manifold
and set the vacuum to 5” Hg.

2. Condition with Methanol.

3. Equilibrate with Water
a. In each case (conditioning and equilibration) add the
solvent before applying vacuum.

4. Switch off the vacuum pump or Stop vacuum by closing the
valve ( before switching off the vacuum pump, please reduce
the vacuum to the lowest possible setting).

5. Load your diluted sample.

6. Switch on or open valve at lowest possible vacuum and
gradually increase as needed in order to load the entire
sample onto the sorbent bed.

7. Switch off the vacuum pump or stop vacuum by closing the valve.

8. Apply 5% methanol in water wash solvent.

9. Switch on vacuum to 5” Hg (adjust/increase as needed).

10. Pull vacuum for another 30 sec to a minute to eliminate
residual wash solvent.

11. Switch off the vacuum pump or stop vacuum by closing the
valve (before switching off the vacuum pump, please reduce
the vacuum to the lowest possible).

12. Release vacuum and discard waste fluids, insert collection
device and replace the cover.

13. Apply 100% organic elution solvent and let it flow through
by gravity before switching on the vacuum pump.

c. Aqueous Samples: Water, Beverages

Adjust pH to maximize analyte retention on the SPE sorbent.

Buffer salts and dispersive agents may be used to increase

partitioning onto the SPE sorbent. Pretreatment to remove

suspended matter prior to SPE treatment may include filtration or

centrifugation.

d. Non-Aqueous Liquid

When appropriate the sample may be diluted with aqueous buffers

and organic co-solvents for reversed-phased or mix-mode SPE.

14. Switch on or open valve at lowest possible vacuum and
gradually increase as needed.

15. Pull vacuum for another 30 sec to a minute (to collect all
elution solvent).

16. Remove collection device.

17. Evaporate / Reconstitute or Dilute as needed.

18. Transfer to vial or plate for analysis.

19. If using plates, cover prior to analysis.

Oasis HLB Cartridges and 96-Well Plates

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