4 replies to this topic

[megashock]

does anyone have a method on how to prepare a Methyl Cellulose solution ?

i have to use 1% methyl cellulose as a vehicle for Prednisone injections.
the vehicle is made of water+ 0.9% NaCl+ 1% Methyl Cellulose.

when i've tried to just add the methyl cellulose to a saline solution it didn't dissolve good and there were a lot of clumps.

right now i've tried to prepare the solution by:
1) warm a Saline solution to 65C and then to add the Methyl Cellulose.
2) then to cool it down by attach the tube to a Vortex and shake it in a 8C room for 20Min, or until it cooled down.

there were no clumps, but the problem is that there was a lot of foam in the tube.

Iv'e found another protocols but they require to shake it, but the shaking makes a lot foam, or that they require to freeze the solution for a long time before the use.
http://www.protocol-online.org/biology-for...osts/31408.html
http://76.162.25.80/.../1/100/m100.htm

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Edited by megashock, 12 July 2009 - 05:52 AM.

[klinmed]

megashock, on Jul 12 2009, 01:41 PM, said:

does anyone have a method on how to prepare a Methyl Cellulose solution ?

i have to use 1% methyl cellulose as a vehicle for Prednisone injections.
the vehicle is made of water+ 0.9% NaCl+ 1% Methyl Cellulose.

when i've tried to just add the methyl cellulose to a saline solution it didn't dissolve good and there were a lot of clumps.

right now i've tried to prepare the solution by:
1) warm a Saline solution to 65C and then to add the Methyl Cellulose.
2) then to cool it down by attach the tube to a Vortex and shake it in a 8C room for 20Min, or until it cooled down.

there were no clumps, but the problem is that there was a lot of foam in the tube.

Iv'e found another protocols but they require to shake it, but the shaking makes a lot foam, or that they require to freeze the solution for a long time before the use.
http://www.protocol-online.org/biology-for...osts/31408.html
http://76.162.25.80/.../1/100/m100.htm

Methocel is easy to handle, if you know how!

In a 500 ml conical flask bring 100 ml water to the boil. Remove from heat and SLOWLY add 2 g of methylcellulose. Add over about 2-3 mins with swirling (be careful as it has a tendency to boil over) . Add a large magnetic stirbar and bring to the boil briefly. Allow to cool to ca 30 - 40 oC with stirring. Then add 100ml 1.8% saline. Stir overnight at 4 oC. The methocel will slowly dissolve at 4 oC and the solution become very viscous.

Dispense aliquots with a syringe and add the concentrated solution of drug. Mix by pumping up and down with the syringe. If you are careful there will not be too many bubbles. Allow the mix to settle for 10 mins, draw up the bubble-free solution from the bottom of the tube.

Hope this helps

[megashock]

Thank's, klinmed.

Here is my method for preparing a methyl cellulose solution.
i've tried it with 1%, and 2% methyl cellulose solutions.

First, all the methods for preparing methyl cellulose are based on the fact that methyl cellulose dissolve better if he added to hot water first, and than cool down while being vigorous shaking.


If you don't need to do an autoclave you can use that protocol :

1) Add the methyl-cellulose powder to a warm water 65oC or more.
2) Here is the trick, you have to shake it like crazy until it cool's down to room temperature or lower.
you can do it by hand, or that you can use a vortex machine and attach the methylcellulose tube to the vortex machine with a tape and vortex it in a 4oC room until it cools down. use a small tube so it will be easier to attach it the vortex. stirring with a magnet will not be good enough. you have to do it by hand, or with a vortex machine.
Don't worry about the foam, leave the tube in 4oC for a few hours and the foam will disappear.
In the end you will get a clear viscous solution.


If you want to use an autoclave, use that protocol:

1) Do an autoclave for the methyl-cellulose solution.
2) Shake the tube with a vortex machine inside a 4oC room until it cools down.

As for my original post on how to prepare a saline solution with methylcellulose.
i took klinmed advice and did the autoclave separately in different tubes for the saline and the methyl cellulose solutions. i used double concentrations and then mixed them together. and stir them with a magnet over night at 4oC.

P.S

I've found another protocol that is similar to the one that I've did.
http://openwetware.o...Methylcellulose

Edited by megashock, 15 July 2009 - 01:57 AM.

[megashock]

Methyl Cellulose solution Storage :
According to protocols 1,2 the methyl cellulose solution can be stored for at least 1 year at -20oC, and thaw at 4oC.

1) http://76.162.25.80/...ts/1/29/m29.htm
2) http://76.162.25.80/.../1/100/m100.htm

[megashock]

Here is my preparation method of Methyl Cellulose with Prednisone .

the Vehicle for Prednisone is: 0.9% sodium chloride, 0.5% methyl cellulose, 0.5% benzyl alcohol, 0.1% Tween 80, DDW.

1) mix 10mg Prednisone with 1.5ml Methanol.
Warm in a 37oC bath and vortex until the solution is clear and there are no floating particles in the tube.
store at 4oC, and check before every use if there are floating particles in the tube, if the are floating particles, then prepare a fresh solution.

This will be the prednisone stock solution.

2) prepare 0.5% methyl cellulose + 0.9% sodium chloride solution as described in the previous post.

3) take the 0.5% methyl cellulose + 0.9% Nacl solution and mix with 0.5% benzyl alcohol.
shack very gently by hand, at first you will see a lot of white pieces shake until they will disappear and you will see a lot of bubbles, leave the tube at 4oC until the bubbles will disapear.

4) add 0.1% Tween 80, the tween 80 will not dissolve at first, so shack the tube very gently by hand and try not to make any foam until there is a clear solution and all the Tween 80 chunk is gone .
store at 4oC until use.

5) before each treatment mix the desire concentration of the methyl-cellulose vehicle and prednisone.

the recommended dosage for prednisone is 1mg/kg daily.

mix 940microL from (4) with 60microL from (1) and inject S.C 25microL for each 10gr of body weight.

Reference:
Hinkle RT, Lefever FR, Dolan ET, Reichart DL, Dietrich JA, Gropp KE, Thacker RI, Demuth JP, Stevens PJ, Qu XA, Varbanov AR, Wang F, Isfort RJ. Corticortophin releasing factor 2 receptor agonist treatment significantly slows disease progression in mdx mice. BMC Med. 2007 Jul 12;5:18.


Another easier option is to use a slow release pellets of prednisone that can save a lot of time.
the pellets are coming at a defined dosage, and the Pellet dosage reflect the total amount of net active product released over the indicated period of time.

for example the the 2.5 mg pellet, will release 41microgr each day, for 60 days.

1) you can order pellets from here:

and you will also need the Placebo pellets.

2) the 2.5mg pellet is equal to a 1.3mg/kg dosage for a 60 day treatment for a 30gr mouse.

References:
1) Baltgalvis KA, Call JA, Nikas JB, Lowe DA. Effects of prednisolone on skeletal muscle contractility in mdx mice. Muscle Nerve. 2009 Jul 17.

2) Herrmann M, Henneicke H, Street J, Modzelewski J, Kalak R, Buttgereit F, Dunstan CR, Zhou H, Seibel MJ. The challenge of continuous exogenous glucocorticoid administration in mice. Steroids. 2009 Feb;74(2):245-9.

Edited by megashock, 11 August 2009 - 06:15 AM.