European Journal of water quality, tome 37, fasc.

2, 2006, p 221 à 232

Action of pulsed low frequency electromagnetic fields

on physicochemical properties of water: incidence
on its biological activity
ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES
BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
PHYSICO-CHIMIQUES DE L’EAU :
INFLUENCE SUR SON ACTIVITÉ BIOLOGIQUE

Philippe VALLÉE*
24, rue Alphand
75013 Paris, France

* Correspondence should be addressed to: Email: phvallee@libertysurf.fr

a) Late of Laboratoire d’Optique des Solides (UMR CNRS 7601),
Université Pierre et Marie Curie, 140, rue de Lourmel, 75015 Paris, France

Abstract

The electromagnetic fields (EMF) effects on water is presently controversial mainly

because of variation in water composition and/or environmental conditions. In the
present study, we developed new experimental procedures in order to investigate the
physicochemical as well as the biological effects of EMF on water. We studied the action
of pulsed low frequency electromagnetic fields on the physicochemical properties of
water samples prepared under severely controlled conditions. The effect was
characterised by using elastic light scattering and photoluminescence: intensities
decreased by 25 and 70 % respectively. Our interpretation is (1) that low frequency
electromagnetic fields act on the gas/water interface by disturbing the ionic double-layer
which stabilises the gas nanobubbles in water (2) that photoluminescence results from
the excitation of these hydrated ionic compounds. These results open interesting
perspectives in biology. We present preliminary results of germination experiments.

Résumé :

L’eau étant le milieu majoritaire commun à tous les organismes vivants, nous avons
mis l’hypothèse que l’eau pouvait être la cible des champs électromagnétiques dans les
systèmes biologiques. Les études montrant des effets des champs électromagnétiques sur
l’eau sont actuellement controversées, car essentiellement dues à la variation de la
composition de l’eau et/ou des conditions environnementales. De ce fait afin de limiter
les artefacts, nous avons mis au point un protocole de préparation des échantillons
rigoureux permettant, d’étudier les effets des champs électromagnétiques sur les
propriétés physico-chimiques de l’eau et à travers son activité sur des systèmes
biologiques. Les efforts ont notamment porté sur : (i) la purification de l’eau par osmose

Article disponible sur le site http://www.water-quality-journal.org ou http://dx.doi.org/10.1051/wqual/2006006

222 ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
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inverse et par une étape finale appelée « polissage » (ii) le contrôle de l’environnement
atmosphérique (boîte à gants avec atmosphère contrôlée), électromagnétique et
acoustique (boîtes de traitement et de stockage du type cage de Faraday avec revêtement
de mu-métal et isolation acoustique), (iii) l’inertie chimique des récipients (utilisation
exclusive de silice fondue de haute pureté de qualité optique). Grâce à ce protocole
expérimental standardisé, l’action des champs électromagnétiques pulsés de basse
fréquence (< 1 kHz) sur les échantillons d’eau préparés a pu être caractérisé par deux
techniques : la diffusion élastique de la lumière et la photoluminescence. Ces expériences
de caractérisation ont permis d’observer des effets notables, à savoir une baisse
importante, de l’ordre de 25 %, pour l’intensité de diffusion et d’environ 70 % pour
l’intensité de photoluminescence après exposition au champ électromagnétique. Des
expériences complémentaires de diffusion dynamique de la lumière et de dégazage des
échantillons ont permis de conclure que l’action du champ électromagnétique avait porté
principalement sur les bulles de gaz présentes dans les échantillons d’eau, entraînant
notamment une forte baisse d’une population de nanobulles d’environ 300 nm de
diamètre. L’ensemble de ces résultats suggère que les champs électromagnétiques basse
fréquence pulsés agiraient sur l’interface gaz/eau, principalement en perturbant la
double-couche ionique qui stabilise les nanobubbles de gaz dans l’eau. La
photoluminescence des échantillons résulterait de l’excitation de ces composés ioniques
hydratés concentrés autour des bulles de gaz. Ces résultats ouvrent d’intéressantes
perspectives en biologie. Afin d’illustrer l’effet biologique de l’eau prétraitée par ces
champs électromagnétiques, nous avons effectué des expériences de germination. Des
études de faisabilité sur des graines de poireaux nous ont permis d’observer une
augmentation d’environ 25% du taux final de germination dans des conditions bien
précises. Une des hypothèses de cette action pourrait se traduire par une amélioration de
la bio-disponibilité et/ou par une meilleure hydratation des composés résiduels présents
dans l’eau.

Keywords: Water, atmospheric environment, electromagnetic fields, light

scattering, nanobubbles, photoluminescence, gas/water interface, biological activity

INTRODUCTION

Water being the major component common to all living organisms, we

hypothesised that water could be the target of electromagnetic fields (EMF) in living
systems. Some studies showed that the physicochemical properties of water may be
modified by the magnetic and electromagnetic fields (Oshitani, et al., 1999 ; Ozeki, et
al., 1996). Nevertheless, the mechanisms by which such electromagnetic fields act on
water are still a controversial issue. The topic is complex by the fact that most of the
effects studied depends on water composition and/or environmental conditions.
Among the different assumptions put forward to explain the EMF effects, the water
interface appears to be the most probable, in particular colloid/water interface
(Chibowski, et al., 1995 ; Knez, et al., 2005) and hydrophobic/water interface (Colic, et
al., 1998) (Ozeki, et al., 2000 ; Ozeki, et al., 1996). In this regard, Colic and Morse
working with oscillating radio-frequencies EMF on colloids in water proposed that the
electromagnetic treatment (EMT) effect results from the perturbation of the
gas/liquid interface (Colic, et al., 1998). This interface has been shown to play a key role
in emulsion stability, cavitation, and biological reactivity (Karaman, et al., 1996).
Moreover, Ozeki et al. working on water-solid interactions under steady magnetic
 Ph. VALLÉE 223

fields showed that the magnetic fields induced adsorption or desorption of water
depending on the intensity fields. Water in multilayer, condensed water in pores and
clusterlike water around functional groups on hydrophobic surfaces will be altered
by magnetic fields (Ozeki, et al., 1996). Other studies (Chowdhury, et al., 1993 ;
Higashitani, et al., 1996) showed the effect of magnetic field on fluorescent probes
which is affected by the solvent nature and their environment.
In a previous work (Vallée, et al., 2003), we have shown that the optical properties
(Raman scattering) of purified water can be changed by the photoluminescence of
traces impurities arising from container/content interactions. Aware of these problems,
we have developed new experimental procedures to investigate the effect of pulsed
low-frequency EMF on water by performing elastic light scattering (Vallée, et al.,
2005a) and photoluminescence experiments (Vallée, et al., 2005b). Water samples were
prepared under particular severely controlled environmental conditions (purification,
atmospheric, electromagnetic and acoustic isolation, chemically inert containers). In
the experiments reported here, we show that a specific electromagnetic treatment
induces a modification in the relative elastic light scattering and photoluminescence
intensities of the water samples. We discuss the possible mechanisms at the origin of
these effects. The next step of this research is to check if the effects observed on the
physicochemical properties of water also occur in biological systems. With this aim, we
began germination experiments on Leek seeds (Allium Porrum L). The first results
concerning the feasibility of these experiments are presented here.

EXPERIMENTAL DESIGN
We describe briefly our experimental design which has been presented in details
in our previous papers (Vallée, et al., 2005a ; Vallée, et al., 2005b).

Water samples preparation

In order to avoid contamination, water is prepared inside a glove box under a
reconstituted atmospheric air. Relative humidity (60 ± 2%) and carbon dioxide
concentration (310 ± 20 ppm) were measured.
The water purification is carry out in two steps: inverse osmosis and “ final
polishing”. One gets a water resistivity of 18.2 M⍀.cm at 25°C, exempt from
compounds ≥ 5-10 nm (ultra-filtering). In order to minimise the fluctuations of residual
concentration of organic compounds [Total Organic Compounds (TOC) ≈ 0.05 mg/l],
a small amount of the spectroscopy grade ethanol (purity 0.999, UVASOL, Merck)
was added (TOC ≈ 0.95 mg/l) during water preparation. In order to minimise
container/content interaction (Vallée, et al., 2003), all glassware were made of pure
fused optical silica (Suprasil, Heraeus).

Water electromagnetic treatment

Optical cells filled with purified water were placed in one of the two insulated
cages (thermally, acoustically and magnetic insulated) and exposed for 6 h to pulsed
electromagnetic fields generated by a solenoid coil (4367 turns/m). The signals
consisted of three successive sinusoidal signals in the frequency range from 10 to
1000 Hz. Optical cells were placed in a vertical position in the centre of the coil
(Bcalculated ≈ 1 mT and Einduced ≈ 4.1 mV/m). Of note, in the absence of current, the fields in
224 ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
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the solenoid coil were: Bo = 9.9 ␮T, BAC residual = 16 nT, EAC residual = 6.3 V/m. During the
electromagnetic treatment, the temperature variation was less than 0.01°C. Control
untreated (reference) optical cells were placed in the other insulated cage under the
same conditions as exposed optical cells.

Water samples degassing treatment (Figure 1)

In order to investigate a possible role of gasses during the EMF water treatment,
in some experiments the samples were degassed before exposure. The degassing
process was performed with a zeolithe pump for 20 min with a pressure of 1,6 ⳯ 103 Pa
(or 12 Torr).

Figure 1: Photograph of degassing process. The special silica cells used

for degassing water was composed of two compartments
(a round beaker connected to an optical cell) closed by a silica tap.
The pressure was measured by a differential manometer (on the left)
[Photographie du procédé de dégazage. Les cuves spéciales en silices utilisées
pour dégazer l’eau etaient composées de deux subdivisions (une partie sphérique
facilitant le dégazage reliée à une cuve optique) fermées par un robinet en silice.
La pression était mesurée par un manomètre différentiel (sur la gauche)]

Water samples analysis

Elastic light scattering analysis

Scattered light was collected at 90° to the incident beam and analysed using a
Jobin-Yvon HR460 spectrometer equipped with a multichannel CCD detector. The
light source was an argon laser (Coherent, Innova 300), with a wavelength ␭ = 488 nm,
a continuous wave power of 100 mW. All data were recorded at room temperature
(21.0 ± 0.5 °C).

Photoluminescence analysis
The selected excitation wavelength (Vallée, et al., 2005b) from the light source
a xenon lamp (450 W) was 310 nm (190 mW power). Light emitted from the sample
was recorded at right angle of the excitation beam, on a Fluorolog 3-2-Triax
spectrofluorometer (Jobin Yvon).
 Ph. VALLÉE 225

RESULTS
During the water samples preparation, an aliquot sample was taken and analysed
for physicochemical measurements. Water samples (R, “reference sample” ; T, “ pulsed
low frequency EMF treated sample ” ; DR, “ reference sample after degassing”; DT,
“pulsed low frequency EMF treated sample after degassing ”) have the following
characteristics: resistivity 4.0 M⍀.cm at 25 °C, TOC content: ≈ 1 mg/l (mainly ethanol),
pH: 5.9, redox potential: 280 mV, oxygen content: 2 mg/l.
Two physicochemical techniques (light scattering and photoluminescence) were
used to analyse the water samples before and after EMF exposure.

Elastic light scattering analysis

We first investigated the effect of a 6 h pulsed low frequency electromagnetic
treatment on water characterised by light scattering techniques. Typical results for
light scattering experiments before and after exposure to electromagnetic fields
are presented in Fig. 2, panel A. After electromagnetic exposure, a decrease of 22%
(SD 5%) was observed in non-degassed samples (R and T). After degassing, the EMF
effect between the maximum scattering peak intensity of DR and DT was abrogated
(Fig. 2, panel B). The maximum scattering peak intensity is a low limit value of light
scattering, that non-degassed treated samples reach this state after electromagnetic
exposure. The observed changes persist over several weeks (not shown).
Panel A Panel B
R
Scattering Intensity (A.U.)

6
1.0x10 1.0x10 6
- 22%
8.0x10
5 DR
8.0x10 5 - 2%
T
6.0x10
5 DT
6.0x10 5

4.0x10 5
4.0x10 5
5
2.0x10
2.0x10 5

0.0
0.0
485 486 487 488 489 490 491
Wavelength (nm) 485 486 487 488 489 490 491
Wavelength (nm)

Figure 2 : Elastic light scattering spectra under laser illumination at 488 nm.
[Spectres de diffusion élastique de la lumière sous une illumination laser à 488 nm]
Panel A : Non-degassed water samples: reference (R) and treated (T)
[Échantillons d’eau non-dégazés: référence (R) et traité (T)]
Panel B : Degassed reference (DR) and degassed treated (DT) water samples
[Échantillons d’eau de référence dégazé (DR) et dégazé traité (DT)]
(Reprinted with permission from Langmuir 2005, 21(6), 2293-2299 ;
Copyright 2006 American Chemical Society)
Photoluminescence analysis
In order to further characterise the EMF action on the interfaces,
photoluminescence experiments were also performed.
 226 ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
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In order to compare results between experiments, each spectrum was normalised

to the maximal values of the Raman peak whose the intensity remains constant
for each couple (reference and treated samples). Figure 3 depicted the emission
spectra of water samples excited at the wavelength of 310 nm. Wide structureless bands
centred at 425 nm (2.90 eV) were observed. Interestingly (Fig. 3, panel A), after
electromagnetic treatment (T), we noted a striking decrease (around 70 %) of the
425 nm band. Other experiments (Fig. 3, panel B), performed with degassed-samples
(DR, DT), show that the emission bands (position, shape, intensity) under the same

Panel A Panel B
Energy (eV) Energy (eV)
4 3.5 3 2.5 2 4 3.5 3 2.5 2
Normalised PL Intensity

1.0 Raman peak 1.0 Raman peak

0.8 0.8

0.6 0.6

0.4 0.4
R
0.2
- 70% 0.2 DT
T
0.0
0.0
DR
300 350 400 450 500 550 600
300 350 400 450 500 550 600
Emission wavelength (nm)
Emission wavelength (nm)
Figure 3: Photoluminescence (PL) emission spectra under excitation wavelength
of 310 nm
[Spectres d’émission de photoluminescence (PL) excités à une longueur d’onde
de 310 nm]
Each spectrum is normalised to the maximal values of the Raman peak.
[Chaque spectre est normalisé sur la valeur maximale du pic Raman]
Panel A: Non-degassed water samples: reference (R) and treated (T)
[Échantillons d’eau non-dégazés: référence (R) et traité (T)]
Panel B: Degassed reference (DR) and degassed treated (DT) water samples
[Échantillons d’eau de référence dégazé (DR) et dégazé traité (DT)]
(Reprinted with permission from J. Chem. Phys., (2005), 122, 114513 ;
Copyright 2006 American Institute of Physics)

excitation were very similar and close to the corresponding bands of the treated non-
degassed samples (Fig. 3, panel A).

DISCUSSION

Elastic light scattering experiments

In our previous work (Vallée, et al., 2005a), we concluded that the extra scattering
observed in the reference samples (Fig. 2) could not be due to residual organic
compounds (as for instance, the ethanol being the main organic compounds ≈ 1 mg/l),
 Ph. VALLÉE 227

but most likely to the gas bubbles present in water. Indeed, addition of large amounts
of ethanol (800 times the concentration of 1ppm measured in our samples) slightly
affected (≤ 5%) the maximum intensity of light scattering. Dynamic light scattering
technique allowed us to identify the presence of nanobubbles (around 300 nm) in non-
degassed water samples, that disappeared under the action of EMT and degassing.
Furthermore, as described before in the literature, our experimental conditions for
preparing purified water are favourable to the nucleation and stabilisation of gas
bubbles. Indeed, a state of gas supersaturation is reached during the water purification,
which may lead to the nucleation of gas bubbles in our samples, stable over a long
period of time since our cells are filled to the brim and hermetically sealed. In support
of this notion, Bunkin and co-workers (Bunkin, et al., 1997a ; Bunkin, et al., 1996 ;
Bunkin, et al., 1997b ; Vinogradova, et al., 1995) working on cavitation of gas bubbles
in liquids, suggested that gas nanobubbles occur and are stabilised by a shell of
negative ions present in the water, even when ion concentration is as low as 0.1 ppm.
Indeed, such concentration can be found in our water samples, due to (i) water
autodissociation (OH– ions) ; (ii) carbon dioxide gas present in water, indeed,
resistivity (4.0 M⍀.cm) and pH (5.9) measurements before closing the optical cells
correspond to a HCO3- content of ≈ 0.1 mg/l ; (iii) a release of silica ions traces from
the optical cell during the storage.
In experiments close to ours, on an aqueous electrolyte solution, Beruto et al.
(Beruto, et al., 2003) showed that low frequency electromagnetic fields (250 ␮T, square
pulsed fields at 75 Hz), have the same effect (vaporisation of carbon dioxide) as
degassing. As a consequence, the decrease of ionic concentration modifies the pH and
the surface tension of the interfacial area. Similarly, our pulsed low frequency
electromagnetic treatment could act on the bubble/water interface leading to
destabilisation of bubbles, in particular by disturbing the ionic balance between the
shell of adsorbed negative ions and counter ions. Others authors (Gamayunov, 1994 ;
Lipus, et al., 2001) envision the action of EMF directly on the ionic double-layer.
All these results allow us to conclude that EMF act likely on the gas
nanobubbles/liquid interface via the ionic double-layer which contributes to gas
nanobubbles stabilisation in water samples.
To further characterise the EMF action on the gas nanobubbles/water interfaces,
photoluminescence experiments were also performed.

Photoluminescence experiments
Regarding the origin of photoluminescence (PL), we showed in the previous work
(Vallée, et al., 2005b) that the emission band intensity observed in reference water
samples could not be attributed to impurities alone coming either from the
container/content interaction or simply present in water since: (1) all glassware was
made of pure fused optical silica to minimise the container/content interaction (see
below). (2) the water purification process guarantees the lowest amount of impurities
under controlled environmental conditions. (3) the 1 mg/l spectroscopy grade ethanol
present in water did not interfere with the UV 310 nm transmittance. We develop a
little these arguments below.
Interestingly, our data (Fig. 3) also indicate that degassing decreased (by about
70 %) the photoluminescence intensity in the reference samples. The light scattering
228 ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
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experiments pointed out the role of gas bubbles in the observed effect. We would
emphasise, however, that it is difficult to formally rule out at present, the possibility
that trace amount of residual impurities in our samples may have contributed to the
5-9 % relative emission intensity, observed after either EMT or degassing (Fig. 3).
The precise origin of the “ extra-photoluminescence ” intensity remains an open
question. Nevertheless, since degassing water decreases the photoluminescence
intensity (by about 70 %) in the water samples, we tend to assign the origin of the
extra-photoluminescence intensity to trace amount of impurities adsorbed at the gas
nanobubble/water interface. Different hypotheses regarding the relationships between
the decrease in PL intensity and the disappearance of gas bubbles could be considered.
Most of them are related to the electric charge density of ionic compounds present at
the bubble/water interface which has been shown to play a crucial role in colloids.
Indeed, the molecular interactions occurring in the ionic shell present at the bubble
surface could produce collective electronic effects being thus possibly at the origin of
the observed photoluminescence.
To conclude, these results indicate that electromagnetic treatment or degassing
decrease in a similar manner the photoluminescence intensity of water. The effect is
most likely attributed to the hydrated ionic compounds (OH-, H3O+, HCO3-, silica ions
traces, …) concentrated around gas nanobubbles (which are not intrinsically
luminescent) present in water samples (Vallée, et al., 2005b).

New investigation in biology: Germination experiments

The feasibility of germination experiments was demonstrated with the
collaboration of Laboratory of Applied Plant Physiology (university of Paris VI) on
leek seeds (Allium porrum L.). The whole seeds were from the same batch and
stored dry in darkness in paper bag under ambient conditions, i.e. at 20-25°C and 70-
75 % relative humidity (RH) of the air for at least 6 months after harvest. Leek seeds
were germinated in darkness, at 20°C, on a layer of cotton wool moistened with 10ml
of Reference(R)/EMF-preTreated(T) water (used for the physicochemical
experiments presented above) in plastic covered Petri dishes (100 grains each). The
Temperature was unfavourable for leek seed in order to maximise the EMF-
pretreated water effects. A seed was regarded as germinated when the radicle had
pierced the seed coat. Germination counts were made daily up to 10 days. The results
presented in Figure 4 are the cumulative germination percentages measured every
day. After ten days, germination of 78 (SD 5%) and 58 (SD 5%) were observed with
EMF-pretreated water samples and reference samples, respectively (Fig.4) ; i.e.
around a 25% relative increase when the EMF-pretreated water was used as
compared to reference water.
These preliminarily results need to be confirmed on a larger number of
experiments in order to conclude that the specific EMF-pretreated water has a
biological effect on the seeds germination. They merit to be put together with the
reports on the increase of germination rate of seeds (Reina et al., 2001 ; Smith et al.,
1995). According to Garcia Reina et al., the low-intensity magnetic fields (< 10 mT)
modify ionic flows in lettuce seeds and thus enhance the water transport mechanism
across the cellular membrane (Reina et al., 2001).
The precise origin of the biological effect of EMF-pretreated water, still remains
a puzzling and open question. We suggest that the action of EMF-pretreated water on
 Ph. VALLÉE 229

80

70

T
60
Germination rate (%)

50

R
40

30

20

10

0
1 2 3 4 5 6 7 8 9 10

Number days after sowing

Figure 4: Germination rate of leek seeds after moistened with Reference(R)/ EMF-
pretreated water (T)
[Taux de germination de graines de poireaux après imprégnation avec de l’eau
référence/prétraitée par les champs électromagnétiques]

the leek germination could be due to an improvement of the bio-availability (Vallée,

2005) and/or a better hydration of the released ionic compounds (OH-, H3O+, HCO3-,
silica ions, …) which adsorbed at the bubble/water interfaces. Furthermore, the
230 ACTION DE CHAMPS ÉLECTROMAGNÉTIQUES BASSE FRÉQUENCE PULSÉS SUR LES PROPRIÉTÉS
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nanobubble vanishing may also increase of the dissolved oxygen rate which play a key
role in the seed germination.

CONCLUSIONS AND APPLICATION FIELDS

A thorough process of water sample preparation and characterisation has been

developed to investigate the effect of pulsed low frequency electromagnetic fields on
purified water. (i) Light scattering experiments, together with the degassing of water
sample, allowed us to point out the action of electromagnetic fields on gas
nanobubbles. (ii) Photoluminescence experiments allowed us to interpret the EMF
effect on the hydrated ionic compounds concentrated at the bubble/water
interface(Vallée et al., 2005a), (Vallée et al., 2005b).
We have demonstrated the feasibility of germination experiments on leek seeds.
Further studies will be necessary to directly address the possible role of hydrated ions
released from the ionic double-layer in biological activity.
Important potential biological applications of EMF-pretreated water are also in
several domains: (1) in biotechnology processes, EMF can generate a biocide effect in
combination with antibiotics. They can stimulate or inhibit the multiplication of yeast
cells. Moreover, magnetic fields can also affect endogenous peptide secretion (2) in
colloids science by considering the EMF treatment has already been used with more
and less success to reduce the scale deposit in tap water ; one can hope that our process
may improve the efficiency of these treatments.

ACKNOWLEDGEMENTS
This work was supported by a grant from the Odier Fondation.
The assistance of R. Nectoux, and C. Naud in the design and realisation of the
electromagnetic device was highly appreciated. I also thank C. Bailly for help in
germination experiments and procedures. I am especially grateful to J. Lafait and Y.
Thomas for helpful advices and fruitful discussions.

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