Hi,Back to the Top
I am analyzing the presence of an antibacterial drug in bone by HPLC.
I want to know that in bone tissue calibration standard sample
preparation, shall one use dried bone powder or bone freshly excised
from
animal and why? Which of these two samples will serve as true
calibration standards of bone?
Regards
Dear Jimmy,Back to the Top
I Suggest you to use the Fresh excised bone and you can dissolve the
bone in Conc HCL, But one should know the Physico chemical property of
the drug, if it is Susceptable for HCl, then we cant work out this
method.
You can use Dried powder of the Bone also, but The drug should get
extracted through proper Extraction Sovent.
Since we for some of the drugs use 1N HCl for extraction of some Drugs.
Regards
Venkatesh.P
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Jimmy:
Whenever there is an oppurtunity to prepare calibration curves, the
most appropriate matrix is the one closiest to the the actual matrix
where the real sample is made. So the answer to your question is if
your final samples is prepared from freshly excise or biopsy bone, then
it will be best to use freshly excised bone for you spike matrix.
Ade
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I would like to ask for some additional info on this topic.
Does anyone know of some good references to assess the penetration of
antibacterials "into bone" to treat osteomyelitis?
What techniques are used to measure the drug?/Besides that outlined
below by
Jimmy.
Is drug in blood/plasma or in bone or in marrow?
How do you distinguish location?
Thanks, Dave
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Whatever the procedure of preparing calibration standards in Bone we
have to
make sure that the drug is uniformly distributed. For that the best
strategy
is to make fine powered of bone, suspend this bone powder in a drug
solution
and evaporate the solvent. Preserve the dried powder in sub zero
temperatures so that when reconstituted the bone powered does not lose
its
integrity. Bottom line is to maintain the bone integrity as closely as
possible with clinical samples. How close is desired depends on you are
trying to do quantitative or qualitative inferences.
Hope this helps.
Prasad Tata
St. Louis, MO 63134
Dear David BenzigerBack to the Top
The answer for your ,
How do you distinguish the bone and marrow location?
The Bone marrow usualy in the Epiphysis region that is the ends of the
bone shaft.
It is better to take a Epiphysial and metaphysial region of the bone as
bone marrow is abundent in this region.
Regards
Venkatesh. P
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Dear Jimmy,
In early 1990s, antibiotic residue in farmed salmon
was the interest of public, aquaculture industry and
Health Canada so we looked at accumulation of
oxytetracycline in fish bone (should be similar to rat
or mice bone). The data was published in Aquaculture
144, 27-38. We used fresh bone and 1 N HCl with a
little bit of pepsin (please see the details in the
pub).
Rostam
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Dear Jimmy,
It may help for you to look for a reference from James Booth (one of the
discovers of tetracycline at Lederle Labs-American Cyanamid) that
appeared in the mid-1980's on the identification and isolation of
tetracycline residues from mammalian bones.
Hope this helps.
Ving J. Lee
VP Discovery Research
Anacor Pharmaceuticals, Inc.
Palo Alto, California 94303
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Copyright 1995-2010 David W. A. Bourne (david@boomer.org)