Vitamin Mixture

Stable vitamins and other nutrients, modified to exclude glutamate, aspartate and ß-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:

Clean magnetic stirring rod, about 3 - 5 cm long
Put the magnetic stirrer into the hood and plug it in.

Mixing containers:

Clean glass beaker, washed twice with de-ionized water and then twice with distilled water.
One 13 ml plastic tube.

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.