APPLICATION NOTE

HyPerforma Single-Use Mixer

Mixing efficiencies for the 100 L
HyPerforma Single-Use Mixer
Goal
The next-generation Thermo Scientific™ HyPerforma™
Single-Use Mixer (S.U.M.) provides enhanced functionality,
ease-of-use, and efficiency for critical up- and downstream
process liquid preparation. In this study, the 100 L
HyPerforma S.U.M. was tested to demonstrate its
mixing efficiency.
Materials and methods
To demonstrate mixing efficiency across the workflow,
four separate mixing materials representing buffers,
classical media, and specialty media (Table 1) were usedat both the nominal volume (100 L) and at the minimum
turndown ratio (20 L).
Table 1. Mixing studies performed on the 100 L S.U.M.
Mixing study

Solutionmaterial

concentration

Sodium chloride*

1M

RPMI medium*

1X

Gibco™ CD
OptiCHO™ AGT™

1X

mediumGlycerol10%* 100 L studies were done in duplicate

Analytics
Osmolality,
conductivity
Osmolality, glucose
concentration
Osmolality, glucose
concentration
Osmolality, specificgravity

A standard 100 L S.U.M. and custom bioprocess containers(BPCs) with sampling ports located at the top, middle,
and bottom of the unit were used. For the 20 L studies,
only two sample points were used at the top and bottomof the volume.
The agitation speed for 100 L studies was set to 330 rpm(approximately 95% of maximum agitation speed) and for
20 L studies at 150 rpm (recommended for this volume).
The powder or liquid for each study was added directly tothe top of the mixer. To represent a worst-case scenario,
agitation was not initiated until all of the powder or liquid
was added.

Each mixing study was done in an hour; where25 mL (+/- 5 mL) samples were taken from each levelwith the following procedure:• First 10 minutes, samples were collected at 1-minuteincrements including time 0.

200%

150%

Percent of final conductivity• Mixing study materials were added 5 to 10 seconds prior
to agitation and sample collection.

Sodium chloride (1 M)(Powder - liquid)
Conductivity-averaged results

100%50%• Next 20 minutes, samples were collected at 5-minuteincrements.

0%012345678910152025304050605060Time (minutes)• Last 30 minutes, samples were collected at 10-minuteincrements.

100 L

20 L

Figure 2. The average conductivity results of sodium
chloride (1 M) for both the 20 L and 100 L volumes.

• Each sample was analyzed at a later time.
Data was normalized based on the results from the last
10 minutes of each study.
200%

150%

Percent of final osmolality

Results
Sodium chloride (1 M)
Osmolality and conductivity studies were performed for the20 L and 100 L volumes. Samples taken from the respective
locations on the S.U.M. were averaged. Within 2 minutes,the osmolality (Figure 1) reached homogeneity for bothvolumes. The conductivity study (Figure 2), reachedhomogeneity at the 1-minute time point.

RPMI medium (1X)
(Powder-liquid)
Osmolality-averaged results

100%

50%

0%

RPMI medium (1X)
Osmolality and glucose concentration studies wereperformed for the 20 L and 100 L volumes. Samples
taken from the respective locations on the S.U.M. wereaveraged. Within 3 minutes, the osmolality (Figure 3)
and the glucose concentration (Figure 4) reachedhomogeneity for both volumes.

200%Percent of final osmolality

150%

123456789101520253040Time (minutes)
100 L20 LFigure 3. The average osmolality results of RPMI medium(1X) for both the 20 L and 100L volumes.

RPMI medium (1X)
(Powder-liquid)
Glucose concentration–averaged results
200%

Percent of final glucose concentration

Sodium chloride (1 M)(Powder - liquid)
Osmolality-averaged results

0150%

100%

50%

100%

0%

012345678

100 L0%91015202530405060

Time (minutes)

50%01234567891015202530Time (minutes)
100 L20 L

Figure 1. The average osmolality results of sodiumchloride (1 M) for both the 20 L and 100 L volumes.

40506020 L

Figure 4. The average glucose concentration results of RPMImedium (1X) for both the 20 L and 100 L volumes.

CD OptiCHO AGT medium (1X)
Osmolality and glucose concentration studies wereperformed for the 20 L and 100 L volumes. Samples takenfrom the respective locations on the S.U.M. were averaged.
For both osmolality (Figure 5) and glucose concentration(Figure 6) studies, the 20 L sample reached homogeneity
within 1 minute and the 100 L sample reached homogeneity
within 5 minutes.

Glycerol (10%)
Osmolality and specific gravity studies were performed
for the 20 L and 100 L volumes. Samples taken from therespective locations on the S.U.M. were averaged. Within
2 minutes, the osmolality (Figure 7) and the specific gravity(Figure 8) reached homogeneity for both volumes.

Glycerol (10%)(Liquid-liquid)
Osmolality-averaged results

CD OptiCHO AGT medium (1X)
(Powder-liquid)
Osmolality-averaged results
200%

300%

250%

Percent of final osmolality

Percent of final osmolality

150%
200%

150%

100%

100%

50%
50%0%0%0123456789101520253040506001234567100 L

89100 L

20 L

Figure 5. The average results of CD OptiCHO AGT medium(1X) for both 20 L and 100 L volumes.

1015202530

405060405060Time (minutes)

Time (minutes)

20 L

Figure 7. The average osmolality results of glycerol(10%) for both the 20 L and 100 L volumes.

CD OptiCHO AGT medium (1X)
(Powder-liquid)
Glucose concentration–averaged results

Glycerol (10%)(Liquid-liquid)
Specific gravity–averaged results
200%

3.0

Percent of final specific gravity

Percent of final glucose concentration

2.5

2.0

1.5

1.0

150%

100%

50%

0.5

0.0

0

123456789101520253040Time (minutes)
100 L20 L

Figure 6. The average glucose concentration resultsof CD OptiCHO AGT medium (1X) for both the 20 L
and 100 L volumes.

50600%

01234567891015202530Time (minutes)
100 L20 L

Figure 8. The average specific gravity results of glycerol(10%) for both the 20 L and 100 L volumes.

Conclusions and recommendations
The 100 L HyPerforma Single-Use Mixer performs
consistently within the specifications of our mixing systems
from 5:1 to nominal volumes. The 100 L S.U.M. is capableof homogeneous mixing of typical powder to liquid and liquid
to liquid materials within 5 minutes (Table 2).
Table 2. Average mixing time of 20 L and 100 L
volumes using the 100 L S.U.M.
Mixing studymaterialIn-solution time

Sodium chloride

1 to 2 minutes

RPMI medium

2 to 3 minutes

CD OptiCHO AGT
mediumGlycerol

4 to 5 minutes
1 to 2 minutes

These studies represented a worst-case scenario,
where the agitation was not started until all of the mixing
study materials had been added. For typical use, it isrecommended that the operator fill up to 80% of the desired
volume with process water, enable the mixer, and set theagitation speed. Then over 5 minutes, add either powders
or liquids to the top of the S.U.M. with a total mixing time of10 minutes.

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