Next Level - Nebulizing B12 & Glutathione 

                                              Final

Revision 1.5   Last updated:  03/30/09                            Page 1/6
 
Message #12913                                                        Author:  rr22rr44 (Rivka)
I'm simply forwarding this email below from Rich Van K. I asked Rich for the theory behind
nebulizing B12 and Glutathione. I need this to send to my doc and pharmacist, and I thought this CFS
Yasko list could use it, too, since I could not find the complete theory summarized in any of the
archives. This seems very helpful and maybe, Marti, should go in the basic info documents for
newbies?
 
 -- Rivka
_________
 
Date: Fri, 06 Mar 2009 18:09:13 -0500
From: richvank
Subject: Re: nebulizing: using glut to protect b12
 
Hi, Rivka.

Below are the abstracts of three papers that provide the basis for combining glutathione and
hydroxocobalamin.

The idea is that glutathione reacts with hydroxocobalamin to form glutathionylcobalamin.  This
protects the hydroxocobalamin so that it can be used to form its coenzyme forms, adenosylcobalamin
and methylcobalamin, rather than reacting with toxins.

This normally happens inside the cells of the body, but when glutathione is depleted, as in CFS, toxins
build up because the detoxication system is not working well (Glutathione is responsible for one of the
Phase II detox pathways, as well as handling hydrogen peroxide that is produced from superoxide,
which in turn is produced by the Phase I CYP450 enzymes. So when glutathione becomes depleted, the
detox system does not function well, and toxins build up.)  Then, because the concentration of
glutathione is low, while the concentrations of the toxins have been allowed to rise, the toxins are able
to compete successfully with glutathione to react with the hydroxocobalamin.  The hydroxocobalamin
is thus not available to be converted to its coenzyme forms.  That's why I say that vitamin B12 is
"hijacked" in CFS.

The ratio of 4 to 1 in mixing hydroxocobalamin with glutathione comes from the facts that these
substances react in a 1 to 1 mole ratio, but the molecular weight of hydroxocobalamin is 1346.37, while
the molecular weight of glutathione is 307.33.  The ratio of these two numbers is 4.38.  If you mix
these two substances at a ratio of 4 to 1, you will have excess glutathione with respect to
hydroxocobalamin, so that all the hydroxocobalamin will react with glutathione, and there will be some
glutathione left over.
I first heard about nebulizing glutathione together with hydroxocobalamin from Jim Seymour, a
pharmacist from Key Pharmacy in Kent, Washington, in a talk he gave to the Multiple Chemical
Sensitivity (MCS) workshop that was held at the DoubleTree Hotel in Burlingame, CA,  near the San
Francisco airport, on August 4-6, 2006.  He reported that Dr. Grace Ziem had been giving glutathione
and hydroxocobalamin separately by nebulizer as part of the protocol she and Prof. Martin Pall had
developed for MCS, and she decided to mix them together to speed up the process.  She found that
when she did this, her patients experienced more benefit, so she continued to do it. [RIVKA'S NOTE: I
CALLED KEY PHARMACY AND THEY SELL THE B12 AND GLUTATHIONE NEEDED TO
DO THIS NEBULIZING. THO THEIR GLUTATHIONE IS VERY PRICEY, AS THEY SEND IT
OVERNIGHT ON ICE. INSTEAD, SOME PEOPLE CHOOSE TO USE THE "REDUCED L-
GLUTATHOINE WITH SODIUM BICARBONATE" POWDER THAT COMES IN CAPSULES
FROM THE COMPANY CALLED THERANATURALS. NO PRESCRIPTION NEEDED. MUCH
CHEAPER. NOW BACK TO RICH'S EMAIL...]

I was curious about this, dug into the literature, and found these papers.  I concluded that mixing the
two together was protecting the hydroxocobalamin from toxins.
Please feel free to forward this email to your pharmacist, Dr., the CFS-Yasko group, and to whomever
else you think might be interested.
 
Best regards,
 
Rich
 
Chem Res Toxicol. 2004 Dec;17(12):1562-7.Links
 

A new role for glutathione: protection of vitamin B12 from depletion by xenobiotics.
 
Watson WP, Munter T, Golding BT.
Syngenta Central Toxicology Laboratory, Alderley Park, Cheshire, SK10 4TJ, United Kingdom.
william.watson@syngenta.com
NADPH in microsomes reduces the hydroxocob(III)alamin form of vitamin B12 to cob(II)alamin and
the supernucleophilic cob(I)alamin, which are both highly reactive toward xenobiotic epoxides formed
by mammalian metabolism of dienes such as the industrially important chemicals chloroprene and 1,3-
butadiene. With styrene, the metabolically formed styrene oxide is reactive toward cob(I)alamin but not
cob(II)alamin. Such reactions in humans could lead to vitamin B12 deficiency, which is implicated in
pernicious anemia, cancer, and degenerative diseases. However, glutathione inhibits the reduction of
hydroxocob(III)alamin by formation of the 1:1 complex glutathionylcobalamin. This blocks reactions
of the cobalamins with metabolically formed epoxides. The interaction between glutathione and
vitamin B12 could protect against diseases related to vitamin B12 depletion.

PMID: 15606130 [PubMed - indexed for MEDLINE]

 

 
Inorg Chem. 2004 Oct 18;43(21):6848-57.Links
 

Studies on the formation of glutathionylcobalamin: any free intracellular aquacobalamin is likely

to be rapidly and irreversibly converted to glutathionylcobalamin.
Xia L, Cregan AG, Berben LA, Brasch NE.
Research School of Chemistry, Australian National University, Canberra ACT 0200, Australia.
A decade ago Jacobsen and co-workers reported the first evidence for the presence of
glutathionylcobalamin (GSCbl) in mammalian cells and suggested that it could in fact be a precursor to
the formation of the two coenzyme forms of vitamin B(12), adenosylcobalamin and methylcobalamin
(Pezacka et al. Biochem. Biophys. Res. Commun. 1990, 169, 443). It has also recently been proposed
by McCaddon and co-workers that GSCbl may be useful for the treatment of Alzheimer's disease
(McCaddon et al. Neurology 2002, 58, 1395). Aquacobalamin is one of the major forms of vitamin
B(12) isolated from mammalian cells, and high concentrations of glutathione (1-10 mM) are also found
in cells. We have now determined observed equilibrium constants, K(obs)(GSCbl), for the formation of
GSCbl from aquacobalamin and glutathione in the pH range 4.50-6.00. K(obs)(GSCbl) increases with
increasing pH, and this increase is attributed to increasing amounts of the thiolate forms (RS(-)) of
glutathione. An estimate for the equilibrium constant for the formation of GSCbl from aquacobalamin
and the thiolate forms of glutathione of approximately 5 x 10(9) M(-1) is obtained from the data.
Hence, under biological conditions the formation of GSCbl from aquacobalamin and glutathione is
essentially irreversible. The rate of the reaction between aquacobalamin/hydroxycobalamin and
glutathione for 4.50 < pH < 11.0 has also been studied and the observed rate constant for the reaction
was found to decrease with increasing pH. The data were fitted to a mechanism in which each of the 3
macroscopic forms of glutathione present in this pH region react with aquacobalamin, giving k(1) =
18.5 M(-1) s(-1), k(2) = 28 +/- 10 M(-1) s(-1), and k(3) = 163 +/- 8 M(-1) s(-1). The temperature
dependence of the observed rate constant at pH 7.40 ( approximately k(1)) was also studied, and
activation parameters were obtained typical of a dissociative process (DeltaH++ = 81.0 +/- 0.5 kJ mol(-
1) and DeltaS++ = 48 +/- 2 J K(-1) mol(-1)). Formation of GSCbl from aquacobalamin is rapid; for
example, at approximately 5 mM concentrations of glutathione and at 37 degrees C, the half-life for
formation of GSCbl from aquacobalamin and glutathione is 2.8 s. On the basis of our equilibrium and
rate-constant data we conclude that, upon entering cells, any free (protein-unbound) aquacobalamin
could be rapidly and irreversibly converted to GSCbl. GSCbl may indeed play an important role in
vitamin B(12)-dependent processes.
PMID: 15476387 [PubMed - indexed for MEDLINE]
Biochem Biophys Res Commun. 1990 Jun 15;169(2):443-50.Links
 

Glutathionylcobalamin as an intermediate in the formation of cobalamin coenzymes.

Pezacka E, Green R, Jacobsen DW.
Department of Laboratory Hematology, Cleveland Clinic Foundation, Ohio 44195.
To evaluate the possible role of glutathionylcobalamin (GS-Cbl) in the intracellular metabolism of
cobalamin, the following reactions were analyzed using extracts of rabbit spleen: (i) decyanation of
cyanocobalamin; (ii) utilization of GS-Cbl by cobalamin reductase; (iii) participation of GS-Cbl in
methionine biosynthesis; and (iv) conversion of GS-Cbl to adenosylcobalamin. Decyanation of
cyanocobalamin required reduced glutathione which appeared to form a complex with the cobalamin.
This complex decomposed during the extraction steps to sulfitocobalamin which was identified by
high-performance liquid chromatography. Cobalamin reductase in spleen extract was more active with
GS-Cbl than with aquocobalamin or cyanocobalamin as substrates (specific activities: 10.4, 2.8 and
0.93 nmol/mg/min, respectively). Methionine synthase utilized GS-Cbl as cofactor more efficiently
than aquocobalamin or cyanocobalamin based on initial rates of enzyme activity. This suggests that
GS-Cbl is a more direct precursor of the coenzyme required for methionine synthase. Formation of
adenosylcobalaminm from GS-Cb1 was four times greater than from aquocobalamin alone. Based on
these results, we propose that GS-Cbl or a closely related thiol-cobalamin adduct is a proximal
precursor in cobalamin coenzyme biosynthesis.
PMID: 2357215 [PubMed - indexed for MEDLINE]

Here is info on a ratio of B12 and glutathione and some info re: nebulizer….This one has an additional
mouthpiece (purchased separately, that closes the valve when you are not breathing, saving medication.
 
 
THIS IS A STARTING POINT - Please search the archives for more recent posts but this is where it
stand at this point in time (March 2009)
 
 
Message #7735 of 7747
April 27, 2008 
 
Exact dose of GSH and HB12 to form glutathionylB12
 
Hi muralimanoharam,

I have made my numbers, and for maintaining the rate 1:1 moles in the
reaction: GSH+HB12=glutathionylB12, for each 10 mg of HydroxyB12 you
have to add 2.18 mg of GSH, so if you take 4 mg of HB12, you have only
to add 0.87 mg of GSH!!! this is a good news, since it is very cheaper!!

Now my concern is how magnesium sulfate can affect this recation...

That is the same said by Rich, but I wanted to understand by myself!!

Sergio

_____________________

--- In CFS_Yasko@yahoogroups.com, "muralimanoharam"

<muralimanoharam@...> wrote:
>
> Hi everyone,
>
> Rich has promptly answered my question about nebulizing hydroxy B12
> with reduced glutathione and the proportions necessary to form the
> compound hydroxocobalamin. I will repost the relevant section here:
>
> " Along with one Perque a day, I...have been nebulizing about 4000mcg
> of hydroxy B 12 with about 60 - 70 mg of reduced, buffered
> glutathione several times a week, hoping that combining them would
> form the compound hydroxocobalamin and protect the B12 for cellular
> use. Would this be true? I am keeping the reduced glutathione low so
> as not to provoke too much detox. My idea has been more to protect
> the B 12 since I am pretty sure I have a load of toxins to deal with.
>
> Rich***Your dosages would amount to about 3 micromoles of
> hydroxocobalamin and about 200 micromoles of glutathione. Since they
> react in a one-to-one mole ratio to form glutathionylcobalamin, you
> have more than enough glutathione to take care of all the cobalamin.
> Once inside the body, no doubt some will become unbound because of
> the usual dynamic equilibrium in chemistry and because there will be
> competing reactants. But this should give it a good start."
>
> Hope this helps shed light on the question. Would love to hear your
> comments.
>

________________________

Message #7740 of 7748                             Author:  normaharman (Susan)

 
About lowering the Glutathione in the combo to be nebulized, having
this info from Rich, I intend to do that. I am using a buffered form
of reduced glutathione sold by Theranaturals without a prescription.
I think they have been able to do it without a prescription because
they put it in a capsule, though the bottle says something like not
for oral use. This is one of the companies Rich mentioned.

I called them and liked their answers. It is a group of biochemists

who have developed this product mainly to treat those who wish to
nebulize it for cystic fibrosis. The person I spoke to said to simply
open the capsule (which contains 200mg reduced, buffered glutathione)
and add it to 3 to 5 ml of distilled water. So the dosage is a bit of
guesswork, though he said they were planning to supply it simply as a
powder that could be more easily measured.

The reason I chose it is because it is buffered and Rich said that

could be advisable if one had sensitive lungs or any tendency to
asthma. I think it is the hydroxy that sometimes (not often) makes me
cough a bit on the first inhalations.

I bought the hydroxy from Key Pharmacy in Washington state as they

are the ones who supply people on the Pall-Ziem protocol and they
know all the ins and outs. It requires a prescription. They said to
be sure to add the B12 to the glutathione, and not the other way
around--don't know why.

As far as nebulizers, there are a confusing array of technologies and

prices to choose from. I did research it quite a bit and finally
found studies in the Journal of Respiratory Therapy that compared
their efficiency, particle size delivered, percentage of medicine
wasted etc. My choice was the Pari ProNeb Ultra II compressor with
the Sprint nebulizer. It comes with the Plus nebulizer but I asked to
get the Sprint as well because it is newer and faster. These
nebulizers only deliver medication when you inhale--a valve closes
when you exhale, so you don't waste medication.

They sell for a big range of prices for the same thing but after
searching through MANY pages on Google found this one for $42.95 at
http://www.bpimedicalsupply.com/mm5/merchant.mvc?
Screen=PROD&Store_Code=BP&Product_Code=PRP86F83LCPLUS&Category_Code=Ne
bulizers . The extra sprint nebulizer was $10. Some places want a
prescription for nebulizers, most don't. This one didn't. It you want
to get the Pari, be sure to order the II as it is newer and much
lighter. They also offer a smaller and lighte one for travelling but
it is much more expensive. Hope this is useful.

__________________
 
Message #11001                                          Author:  normaharman (Susan)
P.S. This was about the best rated nebulizer in the Journal of
Respiratory Therapy. The price you pay for it varies hugely. I hunted
down to about page 7 on the google listings and found a pharmacy that
sells it for much less, as they have volume sales. Here is a link:
http://www.bpimedicalsupply.com/mm5/merchant.mvc?
Screen=PROD&Store_Code=BP&Product_Code=PRP86F83LCPLUS&Category_Code=Ne
bulizers It came with a different nebulizer (the thing you actually
put in your mouth) but they let me add the Sprint for just $10. I
think the price has gone up a bit since I ordered, though this is
still a good price.

Susan
 
________________________

This message is from Jason after verifying with Rich regarding the ratio….

Message #13428                            Author:  jprutty

 
Re: [CFS_Yasko] Re: Nebulizing B12 Amounts
 
Rich was kind enough to respond already and explained what everyone else has been trying to convince
me of.  You need 4 times the mass of hydroxocobalamin to 1 part of glutathione. 
 
It still doesn't make sense to me, but neither does God and I believe in Him.
 
Thanks for letting me try to confuse everyone!  I gave it my best without wanting to.
 
Take Care,
Jason R
 
for questions, please contact marti_zavala….