Copyright © 2001, 2002 by Galen Daryl Knight and VitaleTherapeutics,
Inc.
MASS SPECTRAL
STRUCTURAL PROOF? JUDGE FOR YOURSELF!
UNM and its licensee through
the testimony of Drs. Wallace and Murray have alleged that the authentic
Sulfenate-Linked Benzyl Carbamate Dimer as originally prepared by Drs. Knight
and Scallen are taurine derivatives, or essentially sulfonates. However,
there is little, if any, scientific evidence that the authentic compounds
are sulfonates. Drs. Murray’s and Wallace’s attempts to make the authentic
compounds when allegedly using their corruption of the authentic procedures,
failed to yield “taurine derivatives” in any appreciable amount. The true
identity of the authentic compounds is graphically illustrated by the following
careful analyses of MALDI mass spectra of authentic sulfenate-linked benzyl
carbamate dimer (SLBCD), as obtained from Los Alamos National Laboratories
and Bruker Instruments, respectively. Although these are not high resolution
spectra, the LANL data being ± 1, the fairly good resolution of the
Bruker instrument makes the true identity of the SLBCD obvious:
Fragmentation patterns of the authentic sulfenate-linked benzyl carbamate
dimer, especially the calcium salt that is resistant to loss of the counterions
in the high vacuum of the mass spectrometer, indicate several signature,
mass ion fragments, following, such as 548.144 (547+1) and 563.116, the former
being observed at Los Alamos (supra) and the latter being observed
at Bruker whether KI is present in the analyses or not:
Another key, definitive mass ion was observed
at Los Alamos National Laboratories. Despite allegations of UNM’s attorneys
that the dihydrate of the sulfenate-linked benzyl carbamate dimer cannot
exist, the structure of the authentic dihydrate is actually stabilized by
hydrogen bonds and is observed as its mass ion in in the LANL spectrum as
a finely split peak at about 726-728:
POTASSIUM
IODIDE AS SHIFT REAGENT
Potassium iodide is
an extremely valuable shift reagent for analyzing MALDI (Matrix Assisted
Laser Desorption and Ionization) mass spectra of sulfenates, sulfinates,
and sulfonates. Note that in KI's presence, another signature mass is observed
at 777.712 for an authentic preparation that was contaminated with HCl, oxidized
by ICl to the bis-sulfinate-linked artifact, and then reacted with iodide
to form a characteristic adduct in the MALDI mass spectrometer.
KI's presence in the MALDI analyses also causes reduction of sulfenates
and sulfenate-linked compounds to thiols. It has long been known that sulfenates
react with iodide to form sulfenyl iodides
, which are then reduced to thiol compounds by ultraviolet light, such
as that emitted by the UV laser of the MALDI mass spectrometer. (Thiols
and unreacted sulfenates or sulfenyl iodides form disulfides.) Thus, sulfenates
and sulfenate-linked compounds tend to be reduced by iodide to smaller
sulfhydryl products in the MALDI mass spectrometer, whereas sulfinates
and sulfinate-linked compounds tend to form large complexes with iodide
that apparently do NOT reduce into the smaller thiol compounds. A second
signature peak for the artifactually oxidized authentic compound is observed
at 739.622 when KI is present under these acidic conditions:
Presumably, sulfonates and sulfonate-linked compounds, due to their high
oxidation states, are even more resistant to reduction to thiols under
these conditions, and may even be resistant to adduct formation with iodide.
This last suspicion remains to be tested, since there is absolutely no
evidence for sulfonates or sulfonate-linked compounds in the authentic
preparations, even when the authentic synthetic process using iodine was
contaminated with HCl (ICl) to control a pH overshoot. It is highly unlikely
that iodide and UV light could completely reduce a sulfonate to a thiol
in the split second between its desorption with a UV laser and its analysis
by the detector of the mass spectrometer, especially when even the sulfinate-linked,
-S(O)-O-, compounds appear resistant to such reduction. Full reduction
of the sulfonic acid to sulfide or thiol would require the highly unlikely
chance meeting of the sulfonic acid with several iodide ions in the UV-illuminated
flight path of the mass spectrometer.
Having chloride ions from HCl
present in the iodine reaction constitutes a serious departure from the
authentic procedure, since the iodine monochloride (
ICl
) so generated is known to be extremely reactive, and capable of artifactually
oxidizing sulfur compounds to higher oxidation states. Similar polarity
problems exist for reactive mixtures of bromine and
chloride
.
Additional proof that the authentic
sulfenate-linked benzyl carbamate dimer, -S-O-, is not the bis-sulfinate-linked
structure, -S(O)-O-, is provided by mass ion adducts that are intermediary
between the two, especially in preparations known to be contaminated with
HCl. While a mechanism can be drawn for the sulfenate-linkage being reduced
by iodide, presumably the sulfinate-linkage is less likely to be reduced
to thiol and, therefore, more likely to form a stable adduct with iodide,
the doubly charged mass ion of which is observed in the MALDI mass spectrometer
when KI is present:
Some of the more compelling evidence that the authentic compound is sulfenate-linked,
and NOT the alleged sulfonates or taurine derivatives, stems from the
fragmentation of the HCl-contaminated preparation to the free sulfenates
of vitaletheine, presumably as derived from the substantial amount of
authentic sulfenate-linked compound remaining in this preparation that
was only partially artifactually oxidized:
Still, the most compelling evidence for the authentic sulfenate-linked
structure is the actual appearance, upon reduction, of mass ions for the
sulfide or thiol forms, vitaletheine. These reduced forms are among the
more abundant masses in the spectrum of the authentic, albeit somewhat artifactually
oxidized preparation, but only when analyzed in the presence of KI. The
prevalence of the mass ions observed at 231.25 and 193.19, and the somewhat
surprising lack of the smaller mass ions expected for beta-aletheine and
its adducts, indicate that the authentic structure is as assigned by the
true inventors, i.e., the sulfenate-linked benzyl carbamate dimer.
This authentic sulfenate-linked structure is the only one that can account
for the many definitive mass ions observed in the authentic and partially
oxidized preparations. Similar analyses of pristine, pharmaceutical grade
material, prepared by Dr. Knight, is expected to confirm these conclusions,
the oxidative artifacts being expected to drop out of the MALDI mass spectrum
of this pristine material, as the sulfenate and thiol mass ions and adducts
of vitaletheine increase in relative abundance when analyzed in the presence
of KI. The following table summarizes the compelling mass spectral evidence
for the authentic sulfenate-linked benzyl carbamate dimer, and for oxidative
artifacts generated, therefrom, when the otherwise authentic preparation
was contaminated with chloride ions:
*Asterisk indicates
good to excellent agreement of observed values with calculated ones.
UNM’s licensee and Dr. Murray, and UNM, for some reason, seem doggedly
fixated upon the taurine derivatives when there is absolutely NO discriminating
scientific evidence that these were the compounds used by Drs. Knight and
Scallen to perform the enabling biological experiments with the authentic
vitaletheine modulators. Scientists at Los Alamos's mass spectra group have
stated “(sic., Some) Scientists lie. Mass spectra tells the truth.
” Furthermore, Dr. Knight has been presented with NO credible evidence
that decarboxylated taurine derivatives have any biological activity, whereas
the authentic compounds were phenomenally active. As illustrated in the following
table, even when Dr. Murray allegedly used the authentic procedure, he did
NOT make product having any of the above mass spectral signatures observed
by the true inventors for the authentic compounds. There is absolutely NO
mass spectral NOR any IR evidence for the alleged sulfonates in the authentic
preparations, even when artifactually oxidized by exposing them to contaminating
HCl, and there is virtually NO evidence for the sulfonate mass at 723 in
Dr. Murray’s alleged attempt to make authentic compound when using his corrupted
version of the authentic procedure:
**Attempts by Dr. Murray allegedly using UNM’s method produced an oxidized
artifact of the authentic vitaletheine modulator (755.89), but this product
from Hauser Chemical clearly lacks the mass spectra fingerprint of the authentic
compound. His attempt produced only tiny traces of the 723 mass expected
for the alleged sulfonates. The artifactually oxidized authentic preparation,
contaminated with HCl and discussed previously, apparently did not fully
reduce, and instead produced some large, artifactually oxidized adducts
with iodide. Similar resistance to KI/UV reduction is expected for the sulfonates
alleged by Drs. Wallace and Murray. In contrast, the unoxidized SLBCD remaining
in the partially oxidized preparation appears to be readily reduced by KI
and the MALDI's UV laser to vitaletheine and other small fragments and adducts.
One possible explanation for Dr. Murray’s artifact, produced through his
admitted departures from the authentic procedures, including much longer
reaction times with iodine, is the much higher oxidation state at 755.621
than observed for the authentic compound. The calculated value, following,
is in good agreement with Dr. Murray’s observed mass ion adduct at 755.89,
the 723 mass for the alleged sulfonate being much smaller and virtually
absent from the mass spectrum of Dr. Murray’s alleged attempt:
The striking fit of the many
chemically defined masses for the authentic compound and its oxidized artifacts
(previous analyses, supra, and when contaminated with HCl) is in
stark contrast to the products generated by the significant departures from
the authentic procedures in Dr. Murray's alleged attempts. Dr. Knight could
deduce few, if any, mass ions for the alleged CBZ-beta-alanyl-taurines (sulfonates),
or iodine adducts thereof, following, that could substitute for the fit
observed with the authentic assignments:
This failure of the alleged
sulfonates to fit the observed data for the authentic compounds is further
illustrated by calculations performed for the alleged sulfonates, the UV
laser of the MALDI instument conceivably capable of removing the benzyl
groups from the alleged sulfonates, but NOT capable of reducing the sulfonates
to thiols, especially in the absence of iodide. As before, the following
table provides few, if any, alternative "sulfonic" assignments that provide
better "fits" than those graphically illustrated, supra, for the authentic,
and for the somewhat artifactually oxidized preparations:
In conclusion, UNM, its licensee,
and their so-called "experts" appear to be simply wrong about the structure
of the authentic sulfenate-linked benzyl carbamate dimer. The masses assigned
in the first table, supra, for the authentic sulfenate-linked benzyl
carbamate dimer, and for oxidative artifacts thereof, appear to produce
an accurate "mass signature" for the authentic sulfenate-linked compound.
This hasn’t stopped UNM and its licensee from promoting the alleged sulfonate
or taurine derivatives for human use, as if they were the same as the authentic
compounds, and apparently without much testing of these alleged sulfonate
and taurine derivatives in animals for biological efficacy and safety.
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