We have injected STZ with a citrate buffer at the above left thigh through IP to induce diabetes in rat. But after five days, a wound formed in the IP dosed area. Can I use distilled water for dissolving the STZ instead of a citrate buffer?
Hi sarvanan I agree with Imran, the citrate buffer should be prepared fresh, and STZ should be dissolved just before the injection as it has very short life. The mode of injection you have given is IM (intra muscular), IP should be given in the abdominal cavity. The delivery should be right, other wise the effect of induction will not be 100%. When you give IM in the leg, you should be very careful not to inject near the blood vessel and nerve which runs in the thigh muscle. If you inject near or on it that will cause wound also difficulty in moving. Hope this helps.
Hi, Saravanan, have you checked your citrate buffer pH before administration? It should be 4.5. It is not recommended to dissolve STZ in water. If you are having problems with the IP administration, try preparing fresh STZ in citrate buffer (adjusted to pH 4.5), and administer through the tail vein.
Hi sarvanan I agree with Imran, the citrate buffer should be prepared fresh, and STZ should be dissolved just before the injection as it has very short life. The mode of injection you have given is IM (intra muscular), IP should be given in the abdominal cavity. The delivery should be right, other wise the effect of induction will not be 100%. When you give IM in the leg, you should be very careful not to inject near the blood vessel and nerve which runs in the thigh muscle. If you inject near or on it that will cause wound also difficulty in moving. Hope this helps.
Just to endorse the above comments. Must be citrate, pH matters, essential to make up fresh and ip is probably best although iv OK. Not im. If I am injecting large numbers of animals I make up a new fresh batch of STZ halfway. A little obsessional but the variable potency/short half life of STZ preps can be enough of a problem best to be absolutely sure! I'm not sure of the exact position of your injection from your description. I have never seen wounds at injection sites with ip STZ in citrate pH4.5 so do not think the use of citrate buffer is the reason? Good luck
STZ solution is light, pH and releases NO at room temperature. It is recommended to to use ice cold, freshly prepared STZ solution in citrate buffer (pH 4 where, maximum stability is seen). The solution should be also protected from light as photodegradation can take place. The NO release can only be delayed by ice cold temperatures hence it has to be freshly prepared just before use. Sodium buffers also speeds up the NO release. NO donor function is essential for many of our experimental application.
Animals will die if you dissolve in distilled water. For STZ you must use Citrate buffer (pH:4.5).
Even there is chance of animal death.
So, before giving STZ injection you feed the rats or mice with 2% glucose solution. (2ml/kg). there will be less chance of animal death after injection.
Have you ever tested the efficiency of intramuscular versus intraperitoneal administration?
Thank you for all your comments!
I have recently been testing out dissolving STZ in HBSS. We always used to dissolve in citric acid buffer (pH4.5) and I would protect from light and keep in ice. However injecting a neat acid is very painful for the animals (we try to avoid anaesthetic) and I found a paper using Hanks Buffered Saline solution (HBSS). If I give the animals the animals 160mg/kg i.p. freshly prepared, protected from light ad used within 15 mins. I find that the animals all become diabetic within 1-3 days and the process of injecting is much less stressful for the animals. I would be very interested to hear of peoples thoughts about this approach as I have been reading about the alpha and beta anomeric forms Richard Mark Smith and also about the route of administration. Has anyone given STZ s.c.?
the most important thing that STZ must be prepared freshly ..intraperitoneal injection is very save there is no wound appear in rats.
citric acid buffer must be 4 to dissolve STZ
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