Vitamin Mixture

Stable vitamins and other nutrients, modified to exclude glutamate, aspartate and -alanine.

Not sterile.

Identical to the "Stable Vitamin Mix" recipe from the Furshpan lab.

WARNING: Some of the ingredients in this mixture do not dissolve. This problem particularly applies to the biotin-thiotic acid mixture - when adding this mixture, it is important to have just shaken that suspension before pipetting several milliliters out from the middle of the tube. Also, when preparing 10 ml aliquots from the beaker to put into 13 ml test tubes, it is important to keep the mixture stirring with an electric stirrer so as to keep the solid material in suspension. 

WARNING: Choline chloride can absorb water if stored improperly. If it is wet or very lumpy, it should not be used.

Equipment and supplies:


Mixing containers:


Storage containers: 13 ml plastic tubes, labeled "mod SVM".

Vitamin additive mix :Add the following to the glass beaker:

INGREDIENT  SOURCE  For 200 ml  For 400 ml
       
Distilled water    198 ml  396 ml
L-proline  Sigma P-0380  600 mg  1200 mg
L-cystine  Sigma C-8755  600 mg  1200 mg
p-amino-benzoic acid
(GABA free acid, crystalline) 
Sigma A-9878  200 mg  400 mg
vitamin B-12  Sigma V-2876  80 mg  160 mg
i-Inositol (meso)  Sigma I-5125  400 mg  800 mg
choline chloride, crystalline  Sigma C-1879  400 mg  800 mg
fumaric acid  Sigma F-2752  1000 mg  2000 mg
coenzyme A P-L Biochemicals # 8300A  16 mg  32 mg
Solution from the biotin - thiotic acid suspension in the 13 ml tube just shaken to suspend the ingredients    2 ml  4 ml
Leave this stirring automatically about 2 hours before putting 10 ml into each 13 ml tube. 
     


Biotin - thiotic acid suspension: Add the following to the 13 ml tube:

INGREDIENT SOURCE  AMOUNT
     
Distilled water    10 ml
D-biotin  Sigma B-4501  0.4 mg
DL-6,8-thiotic acid, lyophilized  Sigma T-5625  100 mg


Notes:
P-L Biochemicals is 1037 West McKinlez, Milwaukee WI 53205, 800 558-7110.
D-biotin is sometimes called vitamin H.
Thiotic acid is sometimes called lipoic acid.

Copyright 2002 Dr. Michael M. Segal, Department of Neurosurgery, Brigham & Women's Hospital, Boston MA 02115, USA.  Many of the solutions are based on recipes from the Furshpan and Potter labs; the recipes given here are the versions we use now in the Segal lab.  Individual copies may be made of this page for use in educational institutions.