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Comparison of homeopathic globules prepared from high and ultra-high dilutions of various starting materials by ultraviolet light spectroscopy

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Abstract and Figures

Objective Homeopathic globules are commonly used in clinical practice, while research focuses on liquid potencies. Sequential dilution and succussion in their production process has been proposed to change the physico-chemical properties of the solvent(s). It has been reported that aqueous potencies of various starting materials showed significant differences in ultraviolet light transmission compared to controls and between different dilution levels. The aim of the present study was to repeat and expand these experiments to homeopathic globules. Methods Globules were specially produced for this study by Spagyros AG (Gümligen, Switzerland) from 6 starting materials (Aconitum napellus, Atropa belladonna, phosphorus, sulfur, Apis mellifica, quartz) and for 6 dilution levels (6x, 12x, 30c, 200c, 200CF (centesimal discontinuous fluxion), 10,000CF). Native globules and globules impregnated with solvents were used as controls. Globules were dissolved in ultrapure water, and absorbance in the ultraviolet range was measured. The average absorbance from 200 to 340 nm was calculated and corrected for differences between measurement days and instrumental drift. Results Statistically significant differences were found for A. napellus, sulfur, and A. mellifica when normalized average absorbance of the various dilution levels from the same starting material (including control and solvent control globules) was compared. Additionally, absorbance within dilution levels was compared among the various starting materials. Statistically significant differences were found among 30c, 200c and 200CF dilutions. Conclusion This study has expanded previous findings from aqueous potencies to globules and may indicate that characteristics of aqueous high dilutions may be preserved and detectable in dissolved globules.
Content may be subject to copyright.
Complementary
Therapies
in
Medicine
24
(2016)
111–117
Contents
lists
available
at
ScienceDirect
Complementary
Therapies
in
Medicine
jo
ur
nal
homep
age:
www.elsevierhealth.com/journals/ctim
Comparison
of
homeopathic
globules
prepared
from
high
and
ultra-high
dilutions
of
various
starting
materials
by
ultraviolet
light
spectroscopy
Sabine
D.
Klein
,
Ursula
Wolf
Institute
of
Complementary
Medicine,
University
of
Bern,
3012
Bern,
Switzerland
a
r
t
i
c
l
e
i
n
f
o
Article
history:
Received
19
May
2015
Received
in
revised
form
17
December
2015
Accepted
29
December
2015
Available
online
3
January
2016
Keywords:
UV
spectroscopy
High
dilutions
Globules
Homeopathy
Anthroposophically
extended
medicine
Complementary
medicine
a
b
s
t
r
a
c
t
Objective:
Homeopathic
globules
are
commonly
used
in
clinical
practice,
while
research
focuses
on
liquid
potencies.
Sequential
dilution
and
succussion
in
their
production
process
has
been
proposed
to
change
the
physico-chemical
properties
of
the
solvent(s).
It
has
been
reported
that
aqueous
potencies
of
various
starting
materials
showed
significant
differences
in
ultraviolet
light
transmission
compared
to
controls
and
between
different
dilution
levels.
The
aim
of
the
present
study
was
to
repeat
and
expand
these
experiments
to
homeopathic
globules.
Methods:
Globules
were
specially
produced
for
this
study
by
Spagyros
AG
(Gümligen,
Switzerland)
from
6
starting
materials
(Aconitum
napellus,
Atropa
belladonna,
phosphorus,
sulfur,
Apis
mellifica,
quartz)
and
for
6
dilution
levels
(6x,
12x,
30c,
200c,
200CF
(centesimal
discontinuous
fluxion),
10,000CF).
Native
globules
and
globules
impregnated
with
solvents
were
used
as
controls.
Globules
were
dissolved
in
ultrapure
water,
and
absorbance
in
the
ultraviolet
range
was
measured.
The
average
absorbance
from
200
to
340
nm
was
calculated
and
corrected
for
differences
between
measurement
days
and
instrumental
drift.
Results:
Statistically
significant
differences
were
found
for
A.
napellus,
sulfur,
and
A.
mellifica
when
nor-
malized
average
absorbance
of
the
various
dilution
levels
from
the
same
starting
material
(including
control
and
solvent
control
globules)
was
compared.
Additionally,
absorbance
within
dilution
levels
was
compared
among
the
various
starting
materials.
Statistically
significant
differences
were
found
among
30c,
200c
and
200CF
dilutions.
Conclusion:
This
study
has
expanded
previous
findings
from
aqueous
potencies
to
globules
and
may
indicate
that
characteristics
of
aqueous
high
dilutions
may
be
preserved
and
detectable
in
dissolved
globules.
©
2016
The
Authors.
Published
by
Elsevier
Ltd.
This
is
an
open
access
article
under
the
CC
BY-NC-ND
license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
1.
Introduction
Highly
diluted
remedies
are
applied
in
homeopathy
and
anthro-
posophically
extended
medicine.
Several
modes
of
action
and
models
have
been
discussed
on
how
the
production
of
these
aque-
ous
solutions
by
sequential
dilution
and
succussion,
also
termed
potentization,
might
feature
their
structural
properties.
1
It
has
been
proposed
that
a
network
of
hydrogen
bonds
develops
around
non-
polar
solutes
that
remains
with
successive
dilution,
even
when
the
molecules
of
the
starting
material
have
disappeared,
2,3
thus
giving
Corresponding
author
at:
Institute
of
Complementary
Medicine,
University
of
Bern,
Fabrikstrasse
8,
3012
Bern,
Switzerland.
Fax:
+41
31
6324262.
E-mail
addresses:
sabine.klein@ikom.unibe.ch,
kontakt@sabineklein.ch
(S.D.
Klein).
the
water
a
more
organized
state.
4
Additionally,
nanobubbles
have
been
discussed
to
contribute
to
these
supramolecular
structures,
5
which
are
assumed
to
disappear
upon
heating.
4
Differences
in
the
structure
of
the
water
in
these
homeopathic
preparations
may
be
reflected
in
small
but
measurable
changes
in
physico-chemical
properties.
Accordingly,
methods
such
as
ultraviolet
(UV)
spectroscopy,
2,6–12
nuclear
magnetic
resonance
spectroscopy,
4,13–17
calorimetry
18
or
thermoluminescence
19,20
have
been
employed
to
investigate
possible
differences
between
homeopathic
preparations
and
respective
controls.
It
has
been
found
that
dilution
and
succussion
can
lead
to
the
introduction
of
contaminants,
e.g.,
trace
elements
such
as
Si,
Li,
Na,
Mg.
9,21
This
can
be
avoided
when
dilutions
are
prepared
very
care-
fully
(e.g.,
by
washing
all
flasks
and
pipettes
with
the
same
solution
used
to
prepare
the
dilutions)
or
under
clean
room
conditions.
16,22
http://dx.doi.org/10.1016/j.ctim.2015.12.017
0965-2299/©
2016
The
Authors.
Published
by
Elsevier
Ltd.
This
is
an
open
access
article
under
the
CC
BY-NC-ND
license
(http://creativecommons.org/licenses/by-nc-nd/4.
0/
).
112
S.D.
Klein,
U.
Wolf
/
Complementary
Therapies
in
Medicine
24
(2016)
111–117
Fig.
1.
Obtaining
normalized
absorbance
by
correcting
for
instrumental
drift.
A
typical
example
from
one
measurement
day
is
shown.
Diamonds
represent
native
globules,
and
squares
represent
solvent
control
globules.
Open
symbols
show
the
average
absorbance
from
200
nm
to
340
nm.
Samples
were
measured
in
a
randomized
order,
and
plotting
absorbance
versus
sample
number
revealed
an
instrumental
drift
over
time.
The
intercept
of
the
linear
regression
(measure
for
the
differences
between
measurement
days)
and
the
slope
(measure
for
the
instrumental
drift)
were
used
to
correct
the
values,
yielding
the
normalized
absorbance
(filled
symbols).
The
same
equation
was
then
used
to
normalize
the
absorbance
of
the
various
dilution
levels
measured
on
the
same
day
(not
shown).
A
normalized
absorbance
can
thus
have
positive
or
negative
values.
This
is
crucial
for
experiments
testing
for
differences
between
high
dilutions
and
controls.
Our
group
reported
that
high
and
ultra-high
dilutions
of
various
starting
materials
showed
significant
differ-
ences
in
UV
light
transmission
to
controls
and
between
different
dilution
levels,
all
the
more
so
when
these
dilutions
were
produced
under
controlled
conditions.
10–12
In
clinical
practice,
globules
of
potentized
starting
materials
are
commonly
applied
besides
aqueous
dilutions.
In
a
prelimi-
nary
study,
we
found
differences
in
UV
absorbance
between
verum
and
placebo
globules
of
Aconitum
napellus
30c
or
calcium
carbon-
ate/quercus
e
cortice
6x
dissolved
in
water.
23
These
globules
had
been
produced
for
clinical
trials
and
not
for
laboratory
experiments.
It
was
unclear
whether
the
differences
in
UV
absorbance
originated
from
specific
characteristics
of
the
starting
materials,
from
differ-
ences
in
the
production
of
verum
and
placebo
globules,
and/or
other
unknown
interference
factors.
Therefore,
the
aim
of
the
present
study
was
to
repeat
and
expand
our
previous
experiments
with
globules
produced
under
controlled
conditions
accurate
for
comparison
of
UV
absorbance,
i.e.,
sucrose
globules
and
ethanol
from
the
same
batch
were
used
to
minimize
the
introduction
of
possible
artifacts.
2.
Methods
2.1.
Globules
The
globules
were
specially
produced
for
this
study
by
Spagyros
AG
(Gümligen,
Switzerland)
and
differed
only
in
the
starting
mate-
rials
of
the
potentized
dilutions.
Sucrose
globules
and
ethanol
from
the
same
batch
were
used
to
minimize
the
introduction
of
possi-
ble
artifacts.
The
following
starting
materials
were
investigated:
A.
napellus,
Atropa
belladonna,
phosphorus,
sulfur,
Apis
mellifica,
and
quartz
(2
plants,
2
non-metal
elements,
1
animal,
and
1
mineral).
Dilution
was
performed
in
a
43%
ethanol/57%
water
mixture
until
the
last
step,
for
which
73%
ethanol/27%
water
was
used.
Man-
ual
succussion
was
performed
vertically
with
30
strokes
in
each
step.
200CF
and
10,000CF
dilutions
were
produced
by
discontinuous
fluxion
in
a
machine
with
water
as
dilution
medium.
In
this
sin-
gle
glass
technique,
the
potentisation
vessel
is
alternately
filled
with
the
dilution
medium
and
emptied
again
in
each
step,
while
a
defined
amount
of
liquid
remains
in
the
vessel.
The
operating
mode
of
the
machine
has
been
described.
24
The
final
three
steps
were
manually
diluted
in
30%
ethanol/70%
water,
43%
ethanol/57%
water
and
73%
ethanol/27%
water
and
succussed
with
30
strokes.
Two
kinds
of
controls
were
generated
in
the
experiments:
native
globules
and
globules
impregnated
with
a
succussed
73%
ethanol/27%
water
mixture
(solvent
control
globules).
The
globules
were
produced
in
July
2012
and
the
measurements
were
carried
out
from
September
2012
to
March
2013.
Vials
were
coded
to
display
the
starting
material
but
the
dilution
level
and
the
controls
were
blinded.
Coding
was
unblinded
only
after
completion
of
all
measurements
and
preliminary
description
of
the
data
by
box
plots.
The
vials
were
stored
in
aluminum
boxes,
each
box
containing
the
same
dilution
level
of
the
6
starting
materials.
2.2.
UV
absorbance
measurements
Globules
were
gently
dissolved
in
ultrapure
water
(arium
®
pro
VF,
Sartorius
Stedim
AG,
Goettingen,
Germany)
at
10
mg/ml
in
Fiolax
®
test
tubes.
Samples
were
prepared
in
quadruplicates,
19–22
h
prior
to
the
measurements
to
allow
complete
dissolu-
tion,
wrapped
individually
in
aluminum
foil
and
stored
in
the
dark
at
room
temperature.
Absorbance
of
the
samples
in
the
UV
range
(from
190
to
340
nm)
was
measured
in
a
randomized
order
with
a
Shimadzu
UV-1800
double
beam
spectrophotome-
ter
(Reinach,
Switzerland)
equipped
with
an
auto
sampler
CETAC
ASX-260
(Omaha,
USA;
as
described
previously
in
Ref.
10
).
This
wavelength
range
has
previously
proven
to
be
better
suited
to
detect
differences
than
visible
light
or
near
infrared
light.
25
The
spectrophotometer
had
been
switched
on
2
h
prior
to
the
measure-
ments
for
sufficient
warming
up.
Globules
of
each
starting
material
and
corresponding
controls
were
freshly
dissolved
and
measured
on
5
independent
days.
2.3.
Data
analysis
Due
to
the
high
absorbance
of
sucrose
below
200
nm,
the
main
component
of
the
globules
used,
only
absorbance
values
at
200
nm
and
higher
were
included
in
the
analysis
and
one
value
for
each
nm
was
recorded.
For
each
sample
the
mean
of
the
absorbance
values
from
200
nm
to
340
nm
was
calculated.
Fig.
1
shows
how
S.D.
Klein,
U.
Wolf
/
Complementary
Therapies
in
Medicine
24
(2016)
111–117
113
Table
1
Comparison
a
of
normalized
absorbance
b
of
globules
of
various
dilution
levels
c
for
each
starting
material.
Including
native
and
solvent
control
globules
Including
solvent
control
globules
H
(7)
p
H
(6)
p
Aconitum
napellus
14.31
.046
11.68
.070
Atropa
belladonna
4.88
.675
3.73
.713
Phosphorus
2.52
.925
1.48
.961
Sulfur
15.24 .033 9.52
.147
Apis
mellifica
14.71
.040
13.59
.035
Quartz
4.89
.673
4.12
.660
a
By
Kruskal–Wallis
test,
statistically
significant
results
(p
0.05)
are
displayed
in
bold,
H
=
test
statistic
with
degrees
of
freedom
in
parentheses.
b
Average
between
200
nm
and
340
nm.
c
6x,
12x,
30c,
200c,
200CF,
10,000CF,
control(s)
as
indicated
(n
=
5
for
each
starting
material
and
dilution
level).
Fig.
2.
Boxplot
showing
normalized
absorbance
of
control
and
Aconitum
napellus
globules
of
various
dilution
levels.
Globules
were
dissolved
in
water,
their
absorbance
was
measured
and
corrected
for
the
daily
variations
and
drift
in
the
spectrophotometer,
yielding
a
normalized
average
absorbance
from
200
nm
to
340
nm
(n
=
5
for
each
dilution
level).
Circles
represent
outliers
(that
lie
more
than
one
and
a
half
box
lengths
above
or
below
from
the
upper
or
lower
quartile,
respectively).
*Indicate
statistically
significant
differences
(p
0.05,
Mann–Whitney-U
test)
between
the
respective
dilution
level
and
the
solvent
control
globules.
this
value
was
corrected
for
the
differences
between
measurement
days
and
the
instrumental
drift
during
the
measurements
by
lin-
ear
regression
(intercept
as
measure
for
the
differences
between
measurement
days
and
slope
as
measure
for
the
drift),
yielding
a
normalized
absorbance.
Native
and
solvent
control
globules
were
thereby
regarded
as
references,
since
they
were
independent
of
the
starting
materials.
Since
some
data
were
not
normally
distributed,
non-parametric
statistical
tests
were
applied.
The
normalized
average
absorbance
from
200
to
340
nm
was
compared
between
various
starting
mate-
rials
within
equal
dilution
levels
and
between
various
dilution
levels
within
equal
starting
materials
using
a
Kruskal–Wallis
test.
Within
equal
starting
materials,
each
dilution
level
was
com-
pared
to
the
solvent
control
globules
using
a
Mann–Whitney-U
test.
p
0.05
was
considered
statistically
significant.
Effect
sizes
(r)
were
calculated
and
results
were
reported
according
to
Ref.
26
no
adjustments
for
multiple
comparisons
were
performed.
While
such
adjustments
(e.g.,
Bonferroni
adjustments)
decrease
the
likelihood
for
type
I
errors
(mistakenly
rejecting
the
null
hypothesis),
they
increase
the
likelihood
for
type
II
errors,
i.e.,
important
differences
are
deemed
non-significant.
27,28
Statistical
analysis
was
performed
with
IBM
SPSS
Statistics
21.0
(Armonk,
NY,
USA).
3.
Results
Normalized
average
absorbances
of
the
various
dilution
levels
and
native
and
solvent
control
globules
from
the
same
starting
material
were
compared
by
a
Kruskal–Wallis
test.
Statistically
sig-
nificant
differences
were
found
for
A.
napellus,
sulfur,
and
A.
mellifica
(
Table
1).
The
example
of
A.
napellus
is
illustrated
in
Fig.
2.
Native
and
solvent
control
globules
showed
a
statistically
signif-
icant
difference
from
each
other
(p
=
0.035,
r
=
0.272).
Absorbance
of
dilution
levels
were
then
individually
compared
to
the
solvent
control
globules,
which
were
considered
a
more
accurate
control
than
native
globules
that
had
not
been
impregnated
with
ethanol
114
S.D.
Klein,
U.
Wolf
/
Complementary
Therapies
in
Medicine
24
(2016)
111–117
Table
2
Comparison
a
between
normalized
absorbance
b
of
solvent
control
globules
and
globules
of
various
starting
materials
and
dilution
levels
c
.
Aconitum
napellus
Atropa
belladonna
Phosphorus
Sulfur
Apis
mellifica
Quartz
Solvent
control
Median
.00007
.00003
.00016
.00001
.00038
.00003
MAD
.00011
.00016
.00004
.00008
.00027
.00015
6x
Median
.00029
.00012
.00011
.00002
.00017
.00005
MAD
.00041
.00031
.00023
.00023
.00013
.00003
p
.117
.465
.834
.602
.175
.917
r
.496
.231
.066
.165
.429
.033
12x
Median
.00029
.00020
.00016
.00014
.00039
.00011
MAD
.00033
.00028
.00035
.00007
.00072
.00033
p
.175
.754
.675
.602
.251
.754
r
.429
.099
.132
.165
.363
.099
30c
Median
.00057
.00009
.00012
.00011
.00020
.00006
MAD
.00021
.00044
.00006
.00007
.00022
.00011
p
.009
.834
.465
.344
.028
.754
r
.826
.066
.231
.299
.693
.099
200c
Median
.00040
.00047
.00011
.00022
.00036
.00007
MAD
.00022
.00039
.00018
.00008
.00017
.00080
p
.047
.249
.834
.465
.028
.917
r
.627 .364 .066 .231
.693
.033
200CF
Median
.00021
.00008
.00019
.00022
.00012
.00002
MAD
.00026
.00006
.00005
.00006
.00016
.00006
p
.209
.530
.754
.009
.076
.917
r
.397
.199
.099
.826
.562
.033
10,000CF
Median
.00009 .00042
.00020
.00009
.00032
.00048
MAD
.00028
.00017
.00011
.00024
.00010
.00013
p
.465
.249
.675
.917
.009
.117
r
.231
.364
.132
.033
.826
.496
a
By
Mann–Whitney-U
test,
median
normalized
absorbance
with
median
absolute
deviation
(MAD)
is
shown,
statistically
significant
results
(p
0.05)
are
displayed
in
bold,
r
=
effect
size.
b
Average
between
200
nm
and
340
nm.
c
n
=
5
for
each
starting
material
and
dilution
level.
Table
3
Comparison
a
of
normalized
absorbance
b
of
globules
of
various
starting
materials
c
for
each
dilution
level.
H
(5)
p
Native
13.59
.018
Solvent
control
4.93
.424
6x
4.66 .459
12x
3.82
.575
30c
11.36
.045
200c
11.94
.036
200CF
11.95
.036
10,000CF
9.97
.076
a
By
Kruskal–Wallis
test,
statistically
significant
results
(p
0.05)
are
displayed
in
bold,
H
=
test
statistic
with
degrees
of
freedom
in
parentheses.
Pairwise
comparisons
of
the
6
starting
materials
were
not
performed
due
to
the
high
probability
of
a
type
I
error
when
doing
15
tests.
b
Average
between
200
nm
and
340
nm.
c
Aconitum
napellus,
Atropa
belladonna,
phosphorus,
sulfur,
Apis
mellifica,
quartz
(n
=
5
for
each
starting
material
and
dilution
level).
and
water.
Statistically
significant
differences
were
found
between
several
of
the
ultra-high
dilutions
of
A.
napellus,
sulfur,
and
A.
mel-
lifica
and
the
respective
solvent
control
globules
(Table
2).
Finally,
absorbance
within
dilution
levels
were
compared
between
the
various
starting
materials.
Statistically
significant
dif-
ferences
were
found
between
30c,
200c
and
200CF
dilutions
and
between
control
globules
(Table
3).
The
example
of
200CF
globules
is
illustrated
in
Fig.
3.
No
differences
were
found
between
6x,
12x
and
10,000CF
dilutions
and
among
solvent
control
globules.
4.
Discussion
To
the
best
of
our
knowledge,
this
is
a
logical
development
of
our
previous
study,
23
the
first
report
on
differences
in
UV
absorbance
between
globules
prepared
from
various
starting
materials
and
various
dilution
levels.
Apart
from
differences
between
control
globules
and
globules
impregnated
with
defined
dilution
levels,
we
found
significant
differences
for
30c,
200c
and
200CF
from
var-
ious
starting
materials,
which
indicates
that
the
starting
material
is
important
in
the
potentization
process,
although
at
these
high
dilutions
a
presence
of
molecules
of
the
starting
material
is
not
expected.
This
is
in
agreement
with
Rey,
who
was
able
to
distin-
guish
between
lithium
chloride
15c
and
sodium
chloride
15c
by
thermoluminescence.
19
Limitations
of
UV
absorbance
measurements
of
high
dilutions
in
general
include
e.g.,
the
detection
limit
of
the
spectrophotome-
ter
or
differences
in
absolute
absorbance
between
measurement
days,
and
they
have
been
discussed
previously.
12
In
the
present
study,
we
corrected
for
the
daily
variations
of
the
instrument
as
well
as
the
drift
during
the
measurements.
Additionally,
the
glob-
ules
were
produced
under
controlled
state-of-the-art
conditions
and
in
accordance
with
legal
regulations
by
a
pharmaceutical
com-
pany.
The
results
were
generally
consistent
with
former
studies
of
aqueous
dilutions,
although
by
freshly
weighing
in
and
dissolving
the
globules
for
each
measurement,
an
additional
preparatory
step
was
introduced,
which
may
have
added
to
the
variations
between
measurements.
Each
series
of
dilutions
from
each
starting
material
was
mea-
sured
on
5
different
days.
Thus,
we
were
limited
in
statistical
testing,
i.e.,
pairwise
comparisons
between
starting
materials
were
not
performed
and
Bonferroni
corrections
were
not
applied.
In
future
studies,
one
could
focus
on
fewer
starting
materials
and
dilution
levels
and
perform
more
measurements.
We
used
high
dilutions
to
impregnate
globules,
which
were
then
dissolved
in
water.
In
a
future
study
it
would
be
worthwhile
to
not
only
measure
absorbance
of
the
dissolved
globules
but
also
of
the
original
high
dilutions,
i.e.,
the
solutions
with
which
the
globules
were
impregnated,
in
order
to
investigate
whether
comparable
dif-
S.D.
Klein,
U.
Wolf
/
Complementary
Therapies
in
Medicine
24
(2016)
111–117
115
Fig.
3.
Boxplot
showing
normalized
absorbance
of
200CF
globules
from
various
starting
materials.
Globules
were
dissolved
in
water,
their
absorbance
was
measured
and
corrected
for
the
daily
variations
and
drift
in
the
spectrophotometer,
yielding
a
normalized
average
absorbance
from
200
nm
to
340
nm
(n
=
5
for
each
starting
material).
Circles
represent
outliers
(that
lie
more
than
one
and
a
half
box
lengths
above
or
below
from
the
upper
or
lower
quartile,
respectively).
Comparison
between
the
starting
materials
by
Kruskal–Wallis
test
revealed
a
statistically
significant
difference
(p
=
0.036).
Pairwise
comparisons
of
the
6
starting
materials
were
not
performed
due
to
the
high
probability
of
a
type
I
error
when
executing
15
tests.
ferences
are
apparent
in
both.
Additionally,
a
reproduction
of
these
experiments
by
another
and
independent
research
group
would
be
desirable.
In
accordance
with
our
preliminary
results,
23
we
found
a
statis-
tically
significant
difference
between
the
normalized
absorbance
of
solvent
control
and
A.
napellus
30c
globules.
The
differences
in
uncorrected
absorbance
in
the
present
study
were
less
pronounced
than
in
the
previous
study
(data
not
shown).
While
the
globules
in
the
previous
study
had
been
produced
for
a
clinical
trial,
the
ones
in
the
present
study
were
specifically
produced
for
our
labo-
ratory
study,
i.e.,
control
globules
and
globules
impregnated
with
high
dilutions
were
from
the
same
batch.
Thus,
if
minor
impuri-
ties
existed
they
would
be
equally
distributed
in
the
globules
of
all
groups
and
could
not
account
for
the
differences
in
absorbance.
Since
more
samples
were
analyzed,
an
auto
sampler
was
used
in
the
present
study.
In
previous
studies,
we
compared
UV
absorbance
of
high
dilu-
tions
of
sulfur
and
quartz
to
succussed
but
not
diluted
water
controls.
For
sulfur,
we
found
statistically
significant
differences
between
the
high
dilutions
(6c
to
30c
combined)
and
controls
on
2
out
of
5
measurement
days.
12
When
each
dilution
level
from
6c
to
30c
was
tested
individually
against
the
controls,
16c
and
29c
had
significantly
different
absorbance
even
after
Bonferroni
correction
for
multiple
testing.
In
the
present
study,
sulfur
200CF
differed
sig-
nificantly
from
the
solvent
control
globules.
For
quartz,
we
found
no
differences
between
any
of
the
dilution
levels
and
the
solvent
control.
Similarly,
in
a
previous
experiment
there
was
no
difference
between
quartz
and
the
respective
control.
11
Also
in
accordance
with
our
previous
studies,
10,11
we
found
no
significant
differences
between
UV
absorbance
of
decimal
dilutions
and
controls.
In
clinical
studies,
placebos
are
used
as
control
interventions
and
are
defined
as
substances
with
no
pharmacological
effect.
In
homeopathic
in
vitro,
plant
and
animal
studies
several
possible
controls
can
be
used:
the
same
solvents
with
which
the
high
dilu-
tions
were
prepared
(e.g.,
water
or
water/ethanol),
29–31
succussed
but
not
diluted
solvents,
10,30
or
diluted
and
succussed
solvents
can
serve
as
controls.
30,32,33
Particularly,
the
succussion
may
lead
to
several
effects
such
as
ion
leaching
and
gas
exchange
and
in
turn
altering
the
properties
of
the
dilutions.
Therefore,
pure,
i.e.,
unsuc-
cussed
solvent
is
not
regarded
as
adequate
when
used
as
single
control.
30,34
Diluted
and
succussed
solvents
also
show
effects
dif-
ferent
from
the
undiluted
solvents.
29,31
Two
different
controls
were
used
in
this
study,
native
globules
and
globules
impregnated
with
an
ethanol/water
mixture
that
had
been
succussed
(solvent
control
globules).
The
normalized
average
absorbance
of
these
controls
differed
between
the
two.
Solvent
con-
trol
globules
were
considered
as
the
more
appropriate
control
and
thus
used
for
comparisons
with
the
high
dilutions
(e.g.,
in
Table
2).
Physico-chemical
properties
of
globules
have
rarely
been
inves-
tigated.
Aabel
et
al.
compared
T
1
relaxation
times
of
globules
impregnated
with
Betula
alba
30c
and
dissolved
in
water
to
control
globules
and
found
no
differences
between
the
two.
35
Sukul
et
al.
prepared
potassium
bromide
pellets
from
several
high
dilutions
and
solvents
as
replacement
for
sucrose
globules
and
reported
significant
differences
in
Fourier
transform
infrared
spec-
tra
between
the
high
dilutions
and
between
the
high
dilutions
and
the
solvents.
36
Detecting
delayed
luminescence,
Lenger
et
al.
showed
differences
between
homeopathic
argentum
metallicum
globules
and
controls
37
and
found
that
the
stability
of
ethano-
lic
argentum
metallicum
declined
within
a
month,
while
globules
were
stable
over
1
year.
38
While
we
found
significant
differences
between
potencies
of
3
starting
materials
and
controls
in
this
study,
3
other
starting
materials
did
not
produce
such
differences.
This
might
not
seem
116
S.D.
Klein,
U.
Wolf
/
Complementary
Therapies
in
Medicine
24
(2016)
111–117
to
be
in
accordance
with
the
hypothesis
of
a
change
in
the
solvent’s
structure;
however,
we
would
not
expect
the
same
resulting
struc-
tures
for
all
starting
materials.
Therefore,
it
may
well
be
that
some
changes
are
not
detectable
with
our
measurement
system.
Unexpectedly,
we
found
differences
in
absorbance
between
native
globules
used
as
controls
for
the
various
starting
materials,
while
no
difference
was
detected
between
solvent
control
globules
(
Table
3).
This
can
presently
not
be
explained
and
may
be
a
result
obtained
by
chance.
It
may
also
raise
questions
about
the
possible
transfer
of
a
therapeutically
active
ingredient
from
high
dilutions
to
a
control,
a
phenomenon
that
has
been
observed
but
not
yet
investigated
systematically.
For
example,
Endler
et
al.
39
observed
an
effect
of
thyroxine
30c
on
the
development
of
frogs,
even
when
the
animals
were
in
the
water
and
had
no
direct
contact
to
the
thyroxine
dilution.
Variations
in
controls
have
been
observed
before
in
one
of
our
previous
studies
with
liquid
high
dilutions.
12
However,
the
variations
in
the
present
and
previous
studies
occurred
irregularly
and
did
not
indicate
a
systematic
influence
of
high
dilutions
on
controls.
As
described
above,
several
modes
of
action
were
proposed
that
lead
to
a
change
in
water
structure
during
potentization,
but
it
is
yet
unclear
how
a
changed
structure
may
be
preserved
on
globules.
As
an
explanation
the
importance
of
lattice
defects
in
lactose
monohy-
drate
or
small
pores
on
sucrose
globules
to
transfer
therapeutically
active
ingredients
from
high
dilutions
has
been
pointed
out.
40
It
has
also
been
suggested
that
in
the
solid
phase,
nanostructures
may
retain
their
properties
without
dissipating
energy
from
the
environment
and
could
return
to
their
preceding
state
when
water
became
available,
41
but
precise
models
of
this
process
are
lacking.
Therefore,
it
is
important
to
gain
insights
in
physical
properties
of
globules
as
these
may
facilitate
to
developing
models
to
better
understanding
their
effects
and
modes
of
action.
Finally,
one
of
the
next
achievements
will
be
to
transfer
findings
from
such
physico-
chemical
measurements
on
effects
observed
in
plants,
animals
and
humans.
5.
Conclusions
Globules
prepared
from
high
dilutions
of
A.
napellus,
sulfur
and
A.
mellifica
showed
significantly
different
UV
absorbance
compared
to
solvent
control
globules
when
dissolved
in
water.
This
study
has
expanded
our
findings
from
aqueous
high
dilutions
to
globules,
and
it
suggests
that
characteristics
of
aqueous
high
dilutions
may
be
preserved
and
detectable
in
dissolved
globules.
Conflict
of
interest
None.
Funding
The
study
was
financially
supported
by
Spagyros
AG
(Gümli-
gen,
Switzerland)
and
the
Swiss
Medical
Society
for
Homeopathy
smsh.
The
sponsors
had
no
influence
whatsoever
on
the
design,
collection,
analysis
and
interpretation
of
the
data,
or
in
the
writing
of
the
manuscript
or
in
the
decision
to
submit
the
manuscript
for
publication.
Acknowledgements
Globules
were
specially
produced
and
provided
by
Spagyros
AG
(Gümligen,
Switzerland).
We
gratefully
acknowledge
the
sponsors
of
the
study.
We
also
thank
the
Department
of
Clinical
Research,
University
of
Bern,
for
providing
laboratory
space
and
Stephan
Baumgartner
for
helpful
discussion.
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