Sartorius Vivaspin 500 User manual

85030-511-58
85030-511-58
Technical data and operating instructions
Vivaspin®500 µl and 2 ml
Vivaspin®500 and 2 10K devices for in vitro diagnostic use
Vivaspin®500 and 2 3K, 5K, 30K, 50K, 100K, 300K, 1000K and 0.2 µm
devices for research use only; not for use in diagnostic procedures
For French, Italian, and Spanish manuals, please go to
www.sartorius.com/en/product-family/product-family-detail/m-centrisart-i/

2 |
Vivaspin®500 µl and 2 ml – Introduction
Storage conditions|shelf life
Vivaspin®ultrafiltration spin columns
should be stored at 15–30°C. The devices
should be used before the expiry date
printed on the box.
Introduction
Vivaspin®Concentrators are disposable
ultrafiltration devices for the concentration
of biological samples. Vivaspin®500 is suit-
able for sample volumes of 100–500 µl and
the Vivaspin®2 can handle samples up to
2 ml. Vivaspin®2 can effectively be used in
either swing bucket or fixed angle rotor
accepting 15 ml centrifuge tubes.
The patented vertical membrane design
and thin channel filtration chamber
(US 5,647,990) minimises membrane fouling
and provides high speed concentrations,
even with particle laden solutions.
Vivaspin®500 can be used in a benchtop
fixed angle rotor, accepting 2.2 ml
centrifuge tubes.
C
The Vivaspin®500 & 2 product line includes
9 different cutoffs (Molecular Weight Cutoff,
MWCO):
– Vivaspin®500 & 2 3K device: 3,000 MWCO
– Vivaspin®500 & 2 5K device: 5,000 MWCO
– Vivaspin®500 & 2 10K device:
10,000 MWCO
– Vivaspin®500 & 2 30K device:
30,000 MWCO
– Vivaspin®500 & 2 50K device:
50,000 MWCO
– Vivaspin®500 & 2 100K device:
100,000 MWCO
– Vivaspin®500 & 2 100K device:
300,000 MWCO
– Vivaspin®500 & 2 100K device:
1000,000 MWCO
– Vivaspin®500 & 2 100K device: 0.2 µm
Vivaspin®500 & 2 10K filter devices are for
in vitro diagnostic use and can be used to
concentrate serum, urine, cerebrospinal
fluid, and other body fluids prior to analysis.
Vivaspin®500 & 2 3K, 5K, 30K, 50K, 100K,
300K, 1.000K and 0.2 µm filter devices are
for research use only and not for use in
diagnostic procedures. The Vivaspin®500 &
2 devices are supplied non-sterile and are for
single use only.

| 3
Vivaspin®2
The Vivaspin®2 is specifically designed with
low internal surface and membrane area in
order to achieve superior recoveries from
very dilute solutions.
Another feature of the Vivaspin®2 is the
choice of directly pipetting the concentrate
from the dead stop pocket built into the
bottom of the concentrator, or alternatively
reverse spinning the concentrate into the
recovery cap.
Membrane Alternatives
In addition to the proven high flux
polyethersulfone (PES) membrane range
which is recommended with most solutions,
Vivaspin®2 is additionally offered with
cellulose triacetate (CTA) and Hydrosart®.
CTA is particularly recommended when
high recovery of the filtrate solution is of
primary importance. Hydrosart®is a stabilised
cellulose based membrane that has been
optimised for the biotechnological industry.
The Hydrosart®membrane is a stable polymer
that features a broad pH range. Hydrosart®
is also extremely hydrophilic, making it
non-protein binding, virtually non-foul,
and has extremely high flux. Hydrosart®is
available in 5k, 10k, and 30k molecular
weight cutoffs.
Please note that membrane behaviour largely
depends on the specific characteristics of the
solution being processed. Sartorius Stedim
Biotech recommends that users experiment
with alternative membranes in seeking to
optimise their process performance.
Equipment Required
1. Centrifuge with swing bucket of fixed
angle (minimum 25°) rotor.
Device Carrier Required
Vivaspin®500 2.2 ml/11 mm d
Vivaspin®2 15 ml/17 mm d
2. Pipettes for sample delivery and removal.
For maximum recovery a thin gel loader type
is recommended.

4 |
Operation
1. Select the most appropriate membrane
for your sample. For maximum recovery
select a MWCO at least 50% smaller than
the molecular size of the species of interest.
2. Fill concentrator with up to maximum
volumes shown in table 1. (Ensure lid is fully
seated).
3. Insert assembled concentrator into
centrifuge (when fixed angle rotors are
used, angle concentrator so that the
printed window faces upwards|outwards).
4. Centrifuge at speeds recommended
in table 2, taking care not to exceed the
maximum g force indicated by membrane
type and MWCO.
5. Once the desired concentration is
achieved, (see tables 3a & 3b for guide
to concentration times), remove assembly
and recover sample from the bottom of
the concentrate pocket with a pipette.
Removing the Vivaspin®2 body from
the filtrate tube
The sleeve (seen from the end) is oval in
cross section. The tube is round in cross
section to give a tight fit to the sleeve. To
release the tube from the sleeve, you must
pinch the tube – to press it into an oval
shape – before removing it with a twisting
action.
Press
here Press here
Tube Sleeve

Reverse spin with Vivaspin®2
Depending on user preference and need
for sample storage, the concentrate can be
reverse spun into the concentrate recovery
cap (when fixed angle rotors are used, angle
concentrator so that the printed window
faces upwards|outwards). In this procedure
remove filtrate tube, invert the concentrator
body, insert concentrate recovery cap into
filtrate tube and then spin at up to 3,000 g
for 2 minutes.
Desalting|Buffer Exchange
1. Concentrate sample to desired level.
2. Empty filtrate container.
3. Refill concentrator with an appropriate
solvent.
4. Concentrate the sample again and repeat
the process until the concentration of
contaminating microsolute is sufficiently
reduced. Typically 3 wash cycles will remove
99% of initial salt content.
| 5
Vivaspin®2 Reverse Spin
Filtrate and
concentrate
can be sealed
for storage.
Concentrate
recovery cap Concentrate
Position with printed
window upwards|outwards
Concentrator body
Filtrate tube
Filtrate
Equipment required Vivaspin®500 Vivaspin®2
Centrifuge
Rotor type Fixed angle Swing bucket or Fixed angle
Minimum rotor angle 40° 25°
Rotor cavity To fit 2.2 ml (11 mm) To fit 15 ml (17 mm)
conical bottom tubes conical bottom tubes
Concentrate recovery
Pipette type Fixed or variable volume Fixed or variable volume
Recommended tip Thin gel loader type Thin gel loader type

6 |
Technical Specifications
Table 1: Technical specifications
Vivaspin®500 Vivaspin®2
Concentrator capacity
Swing bucket rotor do not use 3 ml
Fixed angle rotor 500 µl 2 ml
Dimensions
Total length 50 mm 126 mm
Width 11 mm 17 mm
Active membrane area 0.5 cm21.2 cm2
Hold-up volume, <5 µl <10 µl
membrane and support
Dead stop volume* 5 µl 8 µl
Materials of construction
Body Polycarbonate Polycarbonate
Filtrate vessel Polypropylene Polycarbonate
Membrane Polyethersulfone PES, CTA, HY
Table 2: Recommended Spin Speed (x g)
Device Vivaspin®500 Vivaspin®2
Membrane Fixed angle Fixed angle Swing bucket
3–50,000 PES 12,000 8,000 4,000
>100,000 PES 12,000 8,000 4,000
5–20,000 CTA – 8,000 4,000
Hydrosart®– 8,000 4,000
* Dead stop volume as designed in moulding tool. This volume may vary depending on
sample, sample concentration, operation temperature and centrifuge rotor.

Usage Tips
1. Flow Rate
Filtration rate is affected by several parame-
ters, including MWCO, porosity, sample con-
centration, viscosity, centrifugal force and
temperature. Expect significantly longer spin
times for starting solutions with over 5% sol-
ids. When operating at 4°C, flow rates are
approximately 1.5 times slower than at 25°C.
Viscous solutions such as 50% glycerine will
take up to 5 times longer to concentrate
than samples in a predominantly buffer
solution.
2. Pre-rinsing
Membranes fitted to Vivaspin®concentrators
contain trace amounts of Glycerine and
Sodium azide. Should these interfere with
analysis they can be removed by rinsing fill
volume of buffer solution or deionised water
through the concentrator. Decant filtrate
and concentrate before processing sample
solution. If you do not want to use the
pre-rinsed device immediately, store it in the
refrigerator with buffer or water covering
the membrane surface. Please do not allow
the membrane to dry out.
3. Sanitization of Polyethersulfone
Membranes
Vivaspin®devices should not be autoclaved
as high temperatures will substantially
increase membrane MWCO. To sanitize, use
a 70% ethanol solution or sanitizing gas
mixture.
4. Chemical Compatibility
Vivaspin®concentrators are designed for use
with biological fluids and aqueous solutions.
For chemical compatibility details, refer to
table 4.
| 7

8 |
Performance Characteristics
Table 3a: Typical Performance Characteristics Vivaspin®500
Time to concentrate up Concentrate
to 30x [min.] at 20°C recovery %
Start volume 500 µl 500 µl
Aprotinin 0.25 mg/ml (6,500 MW)
3,000 MWCO PES 30 96%
BSA 1.0 mg/ml (66,000 MW)
5,000 MWCO PES 15 96%
10,000 MWCO PES 5 96%
30,000 MWCO PES 5 95%
IgG 0.25 mg/ml (160,000 MW)
30,000 MWCO PES 10 96%
50,000 MWCO PES 10 96%
100,000 MWCO PES 10 96%

Table 3b: Typical Performance Characteristics Vivaspin®2
Time to concentrate up Concentrate
to 30x [min.] at 20°C recovery %
Start volume 2 ml 2 ml
Insulin chain A 0.1 mg/ml (2,535 MW)
2,000 MWCO Hydrosart®35 95%
Aprotinin 0.25 mg/ml (6,500 MW)
3,000 MWCO PES 50 96%
BSA 1.0 mg/ml (66,000 MW)
5,000 MWCO PES 12 98%
5,000 MWCO CTA 50 96%
5,000 MWCO Hydrosart®22 98%
10,000 MWCO PES 8 98%
10,000 MWCO CTA 10 96%
10,000 MWCO Hydrosart®12 98%
20,000 MWCO CTA 5 96%
30,000 MWCO PES 8 97%
30,000 MWCO Hydrosart®5 97%
IgG 0.25 mg/ml (160,000 MW)
20,000 MWCO CTA 6 97%
30,000 MWCO PES 10 96%
50,000 MWCO PES 10 96%
100,000 MWCO PES 8 95%
| 9

10 |
Chemical Compatibility
Table 4: Chemical Compatibility (2hr contact time)
Solutions PES CTA HY
Compatible pH range pH 1–9 pH 4–8 pH 1–9
Acetic Acid (25.0%) OK NO OK
Acetone (10.0%) NO NO NO
Acetonitrile (10.0%) NO NO NO
Ammonium Hydroxide (5.0%) ? OK OK
Ammonium Sulphate (saturated) OK ? ?
Benzene (100%) NO NO NO
n-Butanol (70%) ? NO ?
Chloroform (1.0%) NO NO NO
Dimethyl Formamide (10.0%) ? NO NO
Dimethyl Sulfoxide (5.0%) OK NO NO
Ethanol (70.0%) OK OK OK
Ethyl Acetate (100%) NO NO NO
Formaldehyde (30%) OK OK OK
Formic Acid (5.0%) OK ? OK
Glycerine (70%) OK OK OK
Guanidine HCI (6 M) OK ? OK
Hydrocarbons, aromatic NO NO NO
Hydrocarbons, chlorinated NO NO NO
Hydrochloric Acid (1 M) OK NO OK
Imidazole (300 mM) OK NO ?
Isopropanol (70%) OK OK OK
Lactic Acid (5.0%) OK NO OK
Mercaptoethanol (1.0 M) NO NO OK
Methanol (60%) ? ? OK
Nitric Acid (10.0%) OK NO NO