Technical Bulletin

Glutamine-S Stability
SAFC Biosciences provides a stabilized dipeptide form of
Figure 2
L-alanyl-L-glutamine, Glutamine-S 200 mM, which
postpones the degradation of L-glutamine, thereby Stability of 8 mM Glutamine-S in DMEM/High
prolonging supplemental degradation that could Stored at 2 to 8 C, 15 to 30 C and 37 C
compromise cell culture longevity. This unique dipeptide
structure not only resists degradation in medium, but 10

prevents the accumulation of cytotoxic ammonia because of 9

its enhanced stability.
8

An investigation using Glutamine-S 200 mM (Catalog No.

Concentration (mM)
6
59212) and L-Glutamine Solution 200 mM (Catalog No. 5
59202) supplemented at 8 mM in Dulbecco’s Modified
4
Eagle’s Medium/High Modified (DMEM/High) (Catalog No.
3
51440) demonstrated the consistent stability of the
2
dipeptide under various storage conditions as compared to
that of L-glutamine (see Figures 1 and 2). 1

0
0 1 2 3 4 5 6 7
Time (days)
Figure 1 2 to 8 C
Glutamine-S 2-8?C 15 to 30 C
Glutamine-S 15-30?C 37 C
Glutamine-S 37?C

Stability of 8 mM L-Glutamine in DMEM/High

Stored at 2 to 8 C, 15 to 30 C and 37 C The stability of Glutamine-S in medium over time is
9 important in demonstrating this product's advantages over
8 normal L-glutamine supplementation. The use of
7 Glutamine-S, aimed at maintaining consistent levels of
6 glutamine during 2 to 8 C storage of supplemented
Concentration (mM)

5
medium, can be applied further to the product's stability
4
under culturing conditions. An investigation looked at
L-glutamine and Glutamine-S at 8 mM concentrations in
3
DMEM/High and demonstrated the degradation trends
2
when stored at 37 C. Samples were taken daily for seven
1
days and analyzed for the remaining levels of their respective
0
0 1 2 3 4 5 6 7
glutamine formulations in order to observe the stability of
Time (days) each solution when simulating cell culture incubation (see
2 to 8 C
L-Glutamine 2-8?C L-Glutamine 15 to 30 C
15-30?C 37 C
L-Glutamine 37?C Figure 3).

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SAFC Biosciences, Inc. SAFC Biosciences Ltd. SAFC Biosciences Pty. Ltd.
13804 W. 107th Street Smeaton Road, West Portway 18-20 Export Drive
Lenexa, Kansas 66215 Andover, Hampshire SP10 3LF Brooklyn, Victoria 3025
USA UNITED KINGDOM AUSTRALIA
Phone
Toll free-USA
+1 913-469-5580
1 800-255-6032
Phone
Fax
+44 (0)1264-333311
+44 (0)1264-332412
Phone
Toll free-AUS
+61 (0)3-9362-4500
1 800-200-404
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 Figure 3 Figure 4
Stability of L-Glutamine and Glutamine-S in Equivalence of L-Glutamine and Glutamine-S in Culture
DMEM/High Stored at 37 C Cell Density and Viability vs. Time
10
4 120
9
3.5
8 100

3
7
80

Cell Density (x10 6)

Concentration (mM)

6 2.5

Viability (%)
5 2 60

4
1.5
40
3
1
2 20
0.5
1

0 0
0
0 1 2 3 4 5 6 7
0 1 2 3 4 5 6 7
Time (days) Time (days)

37 C
L-Glutamine 37?C 37 C
Glutamine-S 37?C Density, L-Glutamine Density, Glutamine-S Viability, L-Glutatmine Viability, Glutamine-S

Both L-glutamine and Glutamine-S in medium resisted
degradation at 2 to 8 C in DMEM/High for seven days
(Figure 5). These solutions remained at consistent
concentrations and demonstrated greater stability compared
to solutions stored under higher temperatures. In an
extended evaluation through six weeks, L-glutamine and
Glutamine-S in medium remained within 2 mM of their
original concentrations.
In an equivalence evaluation, Chinese Hamster Ovary (CHO)
cells supplemented with Glutamine-S were evaluated in
comparison to those supplemented with L-glutamine (see
Figure 4). CHO cells supplemented with the dipeptide
performed similarly to those supplemented with
L-glutamine. Further investigations have been conducted
which verify equivalent performance, and in some instances
enhanced performance, when using the dipeptide
Stability of L-Glutamine Solution 200 mM
In addition to the use of glutamine in working
concentrations with cell culture medium, the storage of
L-Glutamine Solution 200 mM at 2 to 8 C is of increasing
importance when determining the stability of product prior
to supplementation. The degradation of 200 mM
supplements could cause similar cytotoxicity issues once
supplemented into various applications. An investigation of
200 mM stock solutions of L-glutamine and Glutamine-S
stored at 2 to 8 C was carried out over eight weeks and
analyzed for amino acid concentration (see Figure 5).
Figure 5
Stability of L-Glutamine and Glutamine-S in
DMEM/High Stored at 2 to 8 C
210

supplement. This evidence suggests that similar 205

performance and even enhanced growth and viability could 200

be expected with the use of the Glutamine-S supplement 195

with several mammalian cell line applications.

Concentration (mM)

190

185

180

175

170

165

160
0 1 2 3 4 5 6 7 8
Time (w eeks)

L-Glutamine 2-8?C
2 to 8 C Glutamine-S 2-8?C
2 to 8 C

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Both L-glutamine and Glutamine-S remained stable
throughout eight weeks in 200 mM solutions stored at
2 to 8 C. By maintaining high levels of glutamine in
200 mM stock solutions, Glutamine-S further demonstrates
how it can serve as an ideal substitution for L-glutamine in
cell culture media.

Conclusions
The loss of glutamine concentration in media could
compromise cell growth and viability by decreasing the
amount of available nutrients to cells and increasing the
amount of cytotoxic ammonia present. For storage
purposes, cooler temperatures are required to prevent
compromising the stability of medium supplemented with
L-glutamine. The use of Glutamine-S as a substitute for
L-glutamine can serve to increase the stability of
supplemented medium, and can produce equivalent
performance when used in mammalian cell culture.

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 Warranty, Limitation of Remedies
SAFC Biosciences warrants to the purchaser for a period of one year from date of delivery that this product conforms to
its specifications. Other terms and conditions of this warranty are contained in SAFC Biosciences’ written warranty, a
copy of which is available upon request. ALL OTHER WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING THE IMPLIED
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ANY HUMAN OR THERAPEUTIC USE.
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© 2006 SAFC Biosciences

Issued March 2006 T074

0805 1005 0106

United States Europe Asia Pacific

SAFC Biosciences, Inc. SAFC Biosciences Ltd. SAFC Biosciences Pty. Ltd.
13804 W. 107th Street Smeaton Road, West Portway 18-20 Export Drive
Lenexa, Kansas 66215 Andover, Hampshire SP10 3LF Brooklyn, Victoria 3025
USA UNITED KINGDOM AUSTRALIA

www.safcbiosciences.com Phone
Toll free-USA
+1 913-469-5580
1 800-255-6032
Phone
Fax
+44 (0)1264-333311
+44 (0)1264-332412
Phone
Toll free-AUS
+61 (0)3-9362-4500
1 800-200-404
Fax +1 913-469-5584 E-mail info-eu@sial.com Fax +61 (0)3-9315-1656
E-mail info-na@sial.com E-mail info-ap@sial.com