Waters Oasis Glass Cartridges User Manual

[ Care and Use ManUal ]

oasIs hlB glass CartrIdges

Contents

I. IntroduCtIon

II. QuICk start sPe ProCedure

III. adjustments to oPtImIze reCoverIes

Iv. PreParatIon of PhosPhate-Buffered salIne (PBs)

a. Sample Comminution

b. Extraction/Partitioning

c. Dispersive Solid-Phase Extraction

v. orderIng InformatIon

a. Extraction Solution Preparation

I. IntroduCtIon

Waters Oasis® HLB glass cartridges are available in 5 cc (200 mg)

®

configuration with Teflon

for trace analysis at parts per trillion level including monitoring

endocrine disruptors, such as phenols and phthalates. The Oasis

glass cartridge contains a unique (patented

designed to have a hydrophilic-lipophilic balance (HLB), that gives

high and reproducible recoveries for acidic, basic, and neutral com-

pounds - even if the cartridge runs dry.

E a c h l o t o f gl a s s c a r t r i d g e s a n d T e f l o n f r i t s a r e t e s t e d f o r t h e

presence of bisphenol A and other phenols and phthalates before

packing. These tests assure that endocrine disruptors, in water

samples, can be analyzed to part per trillion levels.

This Instruction Sheet outlines in Section II the Quick Start Solid-

Phase Extraction (SPE) Procedure for extracting endocrine disruptors

and many other types of compounds. Section III gives a simple

protocol to determine if any adjustments are necessary to optimize

recovery. Section IV describes the preparation of phosphate-buffered

saline (PBS) solution (required only for serum, plasma or urine

matrix).

frits. T he clean glass cartridge is designed

1

) sorbent, a copolymer

Oasis HLB Glass Cartridges 1

The Certificate of Analysis (COA) reports recoveries, with RSDs, for

three polar pharmaceutical compounds. The COA displays results

from stringent quality control tests on the batch of polymer sorbent

and the lot of packed cartridges.

1

US Patent # 5,882,521

[ Care and Use ManUal ]

II. QuICk start sPe ProCedure

I. If desired, add 10 to 50 μL of internal standard to the sample

(soil, food and other solid samples require pretreatment before

SPE).

2. Adjust the sample to pH 3.

3. Place (Oasis HLB extraction cartridges on vacuum manifoldand

set vacuum to approximately 5” Hg. T he extraction procedure

can also be done by positive pressure using the 5 cc Teflon

adaptors (part number 405000934).

No individual stopcocks are necessary

4. Solid-Phase Extraction Procedure: The following simple protocol

s h o u l d b e u s e d i n p r e p a r i n g a n d u s i n g t h e c a r t r i d g e s f o r t h e

isolation of a wide spectrum of acidic, basic, and neutral

analytes especially many classes of endocrine disruptors.

No step should be omitted.

Procedure optimization is discussed in Section III.

Note: Once t he H LB sorbent has bee n cond itioned and

equilibrated, there is no need to keep the cartridges wet prior to

sample loading. Maintain a continuous vacuum on all cartridges

throughout steps 4a-4d This convenience will save you time.

Note: For the load and elute steps, the recommended flow rate

is 10 mL/min for 5 cc cartridges. You may need to momentarily

increasethe vacuum to start the flow of aqueous solutions.

4a. Condition: Add to and draw through each cartridge 5-10 mL

10% methanol in methyl tertbutyl ether (MtBE) and then 3 mL

methanol.

4b. Equilibrate: 3 mL water

4c. Load: Draw sample through the cartridge. The maximum recom-

mended sample volume is 1 L for 5 cc cartridges.

4d. Wash: Add to and draw through each cartridge 3 mL of 5%

methanol in water (v/v). Release vacuum, remove manifold

cover, and discard waste fluids. Insert rack containing collection

vessels, replace cover, and turn on vacuum.

4e. Elute: Add to and draw through each cartridge 6 mL 10%

methanol in MtBE, If desired, evaporate eluates to dryness.

5. Reconstitute in acetonitrile and adjust to the mobile phase

concentration for LC analysis.

6. For GC analysis dry extract over sodium sulfate and reconstitute

to 1 mL.

III. adjustments to oPtImIze reCoverIes

Spike an appropriate volume of reagent water (for general analysis)

or PBS (for biological fluids analysis) with all analytes and internal/

s u r r o g a t e s t a n d a r d s . F o r p r e p a r a t i o n o f P B S s o l u t i o n s e e S e c t i o n I V .

F o l l o w s t e p s 4 a - 4 e i n S e c t i o n I I , b u t u s e a r a c k t o c o l l e c t t h e e l u a t e s

in the Load (4c), Wash (4d), and Elute (4e) steps in separate collec-

tion vessels. In addition, repeat step 4e with a second portion of

elution solvent and collect the eluate. Analyze all four collected

fractions. Use the table to determine adjustments, if necessary, to

optimize sample recovery.

If the fraction
from this step
contains the
analyte:

Load (4c) The Oasis HLB sorbent has been found to retain ionized

Wash (4d) Recoveries of very polar analytes can be increased by

First Elution
(4e)

Second
E l u t io n
(4e repeated)

Make this adjustment for optimum sample recovery:

analytes more strongly than silica-based reversed-phase
sorbents. However, recoveries may be enhanced when
analyte ionization is suppressed. For acidic analytes,
adjust the sample pH to at least two pH units below the
pKa of the acid. For basic analytes, adjust the pH to at
least two pH units above the pKa of the conjugate acid.

using only 1 mL of water (not 5% methanol in water) as
the wash solution.

If an acceptable recovery of analyte(s) is obtained in this
fraction (usually > 90%), no adjustments are necessary.

For very nonpolar analytes, stronger solvents such as
acetonitrile, methylene chloride or ethyl acetate may be
substituted, or used in sequence. In addition, for ionizable
analytes, methanol may needed to be modified with the
a d d i t i o n o f 2 % a c i d o r 2 % b a s e , a s a p p r o p r i a t e . I f s o l v e n t s
stronger than methanol or acetonitrile are used for the
elution, then a preliminary conditioning step (see step 4a)
should be performed prior to the methanol conditioning
step. For example, if ethyl acetate is to be used as an
eluent, condition the cartridge with 1 mL of ethyl acetate,
followed by 1 mL of methanol and 1 mL of water.

Oasis HLB Glass Cartridges 2