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14227/DT020295P1
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'"I
125
via an HPLC TEFLON solvent inlet filter through a
vessel of water placed in a thermostatted water bath.
95%
Except for water dispen sed by a Hanson Media Mate,
all media were prepared in a 4 liter thi ck-walled glass c
.2
vessel. then poured into the I liter flasks in a di ssolu- 1!
~
75 -- - --
tion rate apparatus. No stirring or swirling of media ...
was employed during deaeration. Combinations of '0" - - - - --
the following conditions were tested.
.
OJ 50
25 - - -
Initial Water Temperature Both room temperature and
hot deionized tap water was available in hou se. Room
0
temperature deioni zed water varied in temperature
between about 21 and 25C. Hot deionized tap water '"" 0-
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temperature varied between about 45 and 50 C.
Figure I. Percent oxygen saturation for deaerationmethods.
Vacuum House vacuum pressure was measured by See text for desc riptions of methods. Abbreviations: AS :;;;: Air
manometer to fluctuate between 7 to 10 mm H. When Saturated; RT = Room Temperature Initially; H = Hot Initially; S =
used, vacuum was applied for 5 minutes. Sonicated; V = Vacuum ; C = Coarse Frit Filter; M = Medium Frit
Sonication Sonication was performed with a Branson Filter; F = 0.45 pm Membrane Filter; He = He lium Sparged; MM =
5210 ultrasonic bath with 185 W input power and 47 Media Mate. The recommended 95% saturation level for deaeration
kHz frequency. When used, sonication was applied is al so di splayed.
for 5 minutes.
Filtration Water was vacuum filtered through either a 350 below saturation, but greater than Hanson's recommended
1111 Kimax Buchner funnel with coarse fritted disk (40-60 95% saturation value. Sonication appears to enhance
11'" pore size), a 350 ml Kimax Buchner funnel with degassing efficiency when combined with vacuum, but
medium fritted disk (10-151J11l pore size), or through a has no measurable effect when perfom1ed alone.
300 ml glass funnel/support assembly fitted with a 0.45 Adequate deaeration techniques include vacuum
1JI11 membrane filter. Disklfrit diameter was 47 mm. filtration, he lium sparging, hot water placed under a
vacuum with or without sonication, and use of a Media
Helium Sparging Helium was bubbled through water via Mate. The manual method suggested by the USP includes
an HPLC Teflon solvent inlet filter. Helium flow rate was both heating the water to 45C then vacuum filtering it
about 900 ml/ min. When used, sparging occurred for 5 through a 0.45 micron pore size filter. OUf data suggests
minutes. (COf/filllled 011 page 8)
(continued/rom page 7) gently) and directly into the center of the vessels. Oxygen
vacuum filtration alone provides sufficient deaeration levels from both practi ces were not statistically different :
and that healing is an unnecessary extra step. 4.38 + 0.04 and 4.35 + 0.08 ppm for ge ntle and " turbu-
In addition to acceptability of the deaeration level , lent" transfer respectively. We conclude that if good
one must also consider the factors of time and conve- laboratory practice is followed, i.e., there is no loss of
nience. i.e., how quick and easy is the deaeralion media from spilling or splashing, no extra-special care
method? Vacuum filtration with either the medium frit or need be taken when transfening media.
the 0.45 pm pore disk took a significant amount of time, Media are often.stirred in order to reduce thennal
equilibration times. A