Aquaphotomics for weak signals quantification in water

Roumiana Tsenkova Kobe University, Japan rtsen@kobe-u.ac.jp

 Bio Measurement Technology Laboratory http://nirslab.org/ Kobe University

Bio measurement Technology Laboratory, Kobe University, Japan

Dr. Jacques Benveniste

Understanding that the smallest amount of a substance affects the organism A person can enter a room two days after a cat has left it and still suffer an allergic response.
Yolene Thomas, Larbi Kahhak and Jamal Aissa (2006) The physical nature of the biological signal, a puzzling phenomenon: the critical contribution of Jacques Benveniste, In Water and the Cell, 325-340, Ed. Gerald Polack, Springer.

electromagnetic transmission of biochemical information can be stored in the electric dipole moments of water in close analogy to the manner in which magnetic moments store information on a computer disk.
A. Widom, Y.N. Srivastava, V. Valenzi The Biophysical Basis of Water Memory , http://jacquesbenveniste.org/bio_conf_widom.pdf

Vibrational Spectroscopy IR, NIR IR Infrared spectroscopy NIR Near infrared spectroscopy (overtone IR) IR active transition electric dipole moment of molecule changes NIR overtone spectroscopy can measure water in-vivo

WATER SPECTRUM
IR range
fundamental frequencies

NIR range
overtone region

Bio measurement Technology Laboratory, Kobe University, Japan

AQUAPHOTOMICS: THE CONCEPT

AQUAPHOTOMICS:

WATER as a MOLECULAR MIRROR

Perturbations

water

AQUA PHOTOME Spectral Data Base

WAMACS The LETTERS

Activated Water Absorbance Bands

Bio Aqueous System

NIR spectrum

Water Spectral Pattern THE WORD

WAMACS = Water Matrix Coordinates, i.e. water absorbance bands in VIS-NIR range
Bio measurement Technology Laboratory, Kobe University, Japan

Prof. Rustum Roy

Water structures should be the LETTERS and water functionalities will be described as WORDS

AQUAPHOTOMICS
Perturbations Experiments Data Analysis
Water

Prof. Rustum Roy

Sun
NIRS
UVVISIR

Bio Aqueous System

Medical Science, Pharmacy

.

Spectral Data Base

Multivariate Analysis
WAMACS
Activated Water Absorbance Bands
Database

Engineering

GOAL

Biotechnology

bio-system
HOLISTIC MODELS

Building

Basic Science
Analyzing of aqueous system Interaction of water with DNA Interaction of water with organic and non-organic molecules Aqueous systems in cells Aqueous systems in tissue Aqueous systems in organs Aqueous systems in whole body

AQUAPHOTOME

Database of water absorbance bands and patterns WAPS Database of water absorbance patterns according to change perturbation

analyzing spectral changes of WATER MOLECULAR SYSTEM

through

Applying in various fields

Bio measurement Technology Laboratory, Kobe University, Japan

SPECTRA and AQUGRAMS of WATER AND VAPOR

1518nm (V1, V2) 1344nm (V3) 3.5
3 2

1364nm (water shell)

1362 1382
1492nm (S4)

2.5 1.5 1 0.5 0

1372nm (V1+V3)

1452 1372

Absorbance

MPA

-0.5 1474nm (S3) -1 -1.5 1382nm (water shell)

VAPOR WATER

1464nm (S2)

1398nm (free OH)

1300

1410

1490 Wavelength (nm)

1580

1680

1444nm (S1)

1410nm (S0, free water)

moisture

1438nm (H5O2)

water

A'

A : Absorbance after EMSC (1300-1600 nm) Mean of Averaged spectra SD of absorbance each wave length

Bio measurement Technology Laboratory, Kobe University, Japan

APPROACHING WATER with MONITORING Following the idea of QUANTUM SPECTROSCOPY where
matter is excited and probed with a sequence of light pulses at defined frequencies IN ORDER TO control and characterize quantum dynamics of many-body states1

1. http://en.wikipedia.org/wiki/Quantum-optical_spectroscopy

Measurement of NIR Spectra Under Perturbations

NIR spectra of water and protein solutions measured every minute, for the first 5 minutes and every 15 minutes, for 690 min, at 37oC
Instrument: (NIRSystem 6500, wavelength range from 400 to 2500 nm)
Cuvette cell is inserted in to the instrument holder and kept at 37oC
ILLUMINATION as a perturbation 5 6 7 45 [I, times]

VIS NIR Light

1,2,3,4,5

20

35

690

[T, min]

TIME as a perturbation MODELS DEVELOPED: Y = F [ I ], Y = F [ T ]

Temperature Changes
Room temperature 26.6 26.4 26.2 26.0 25.8 25.6 25.4 25.2 25.0 0 5 10 15 20 25 30 35 40 45 50 55 60 time(minute)

room temperature()

experiment

1 2 3

Humidity Changes
Humidity 50.0 40.0
experiment

humidity(%)

30.0 20.0 10.0 0.0 0 5 10 15 20 25 30 35 40 45 50 55 60 time(minute)

1 2 3

Weight Changes
experiment (g) before after difference 1st 10.9324 10.9088 0.0236 2nd 11.0326 10.9981 0.0345 3rd 10.9596 10.9302 0.0294

LOSS of Water
1st exp 0.22% 2nd exp 0.31% 3rd exp 0.27%

11.08 11.04 11.00 10.96 10.92 10.88 10.84 1 2 experiment 3

weight(g)

difference after

PCA SCORE PLOT

experiment

1st 2nd 3rd

Water Matrix Coordinates: WAMACSPCA, EZ WAMACS, LOADINGS

1412 1364 1490 1842 1872 Factor1 Factor2

1482
1474 1530

934

1368

1490 1842
wavelength(nm)

Water Spectra

Wavelengths, nm

WATER DYNAMIC SPECTRA 2D and 3D Synchronous Correlation Maps 600- 2500 nm

represent negative correlation represent positive correlation

WATER ABSORBANCE PATTERNS, WAPs

(dynamic study)
Power Spectra at 1530nm

Loading of the first PC over the 600 850nm region

1.15

1.1

670nm 684nm 694nm

a.u.

1.05

724nm 774nm 792nm 806nm

1.2
0.95

00 1 00 4 03 5 08 0 12 5 17 0 21 5 26 0 32 0

1.15
626nm

Time, min

1.1

644nm 654nm 722nm

Real time monitoring of

a.u.
1.05 1 0.95

772nm 810nm

NIR water absorbance

at various wavelengths

001

004

035

080

125

170

215

260

Time, min

320

Spectral Differences in Waters

1 2 3 4 5 6

1446nm Log1/T

972nm

1192nm

nm

Magnetic VivoWater miliQWater

Blue: miliQ; Green:magnetic; Red: Vivo water MSC, Baseline correction: miliQ second measurement subtracted+ smooth 13nm

Measurement of Very Low Concentration of Progesterone

Samples
1 2 3 4 5 6 7 8 9 10 PdG (ng/ml) 100 50 25 12.5 6.25 3.13 1.56 0.781 0.391 0.195

Pregnanediol glucuronide

Each of samples is measured 10 consecutive spectra.

Raw spectra

Absorbance
400

700

1100

1880

2500

Wavelength (nm)

2nd derivative spectra

Absorbance

25ng/ml 50ng/ml 100ng/ml

2262 nm
Wavelength (nm)

Correlation absorbance at 2262 nm with concentration of PdG standard

0.96

R = 0.7195
Absorbance at 2262 nm
0.955

Exclude
0.95

0.945

0.94

10 samples 100 spectra

0 20 40 60 80 100

0.935

concentration of PdG (ng/ml)

Correlation absorbance at 2262 nm with concentration of PdG standard

8 samples 80 spectra

PLSR for PdG

Predicted concentration (PdG ng/ml)

(the water region 1300 - 1600 nm)

Preprocessing : mean-center Transformation: smooth

R=0.999 SEC=1.42

10 samples 100 spectra

Actual concentration (PdG ng/ml)

Regression vector in the first overtone of water

1528

1366

1580

1334

1432 1410

1470

EMS from DNA Solutions Measurement

Data set for analysis

15 sample data sets (EMS from DNA, Imprinted EMS, and Water)
DNA Patient HIV+ HIV LTR 104bp HIV LTR 137bp (1st Day) HIV LTR 137bp (2nd Day) HIV LTR 194bp HIV LTR 300bp E. Coli (1st Day) E. Coli (2nd Day) Borrelia Water Distilled Water (DW) MilliQ Kobe Imprinted EMS HIV LTR 104bp Borrelia Distilled Water (DW)

Spectral data of each sample are divided into two groups depending on amplitudes of electromagnetic signal (EMS). 1) Higher EShigher or normal EMS, 2) Lower ESlower EMS
34

Prediction of EMS by PLS

(Higher ES Samples)
Factor: 16 R: 0.90 SEV: 53 dB / Hz
Sample
DNA-HIV-Patient DNA-HIV-104bp DNA-HIV-137bp (1st) DNA-HIV-137bp (2nd) DNA-HIV-194bp DNA-HIV-300bp DNA-E.Coli (1st) DNA-E.Coli (2nd) DNA-Borr Water-DW Water-MilliQ Water-Kobe Wave-HIV-104bp Wave-Borr Wave-DW Cross Check LB
35

ES
2 2 2 2 2 1 2 2 2 1 1 1 2 1 1 1 1

Predicted EMS (dB / Hz)

Higher ES Lower ES

Measured EMS (dB / Hz)

Mean-center Smooth[25] 1st, 2nd, and 3rd Step 3

Prediction of EMS by PLS (Higher ES Samples)

Regression Vector, WAMACS
1870

Sample

ES
2 2 2 2 2 1 2 2 2 1 1 1 2 1 1 1 1
36

DNA-HIV-Patient DNA-HIV-104bp DNA-HIV-137bp (1st) DNA-HIV-137bp (2nd)

Regression coefficient (10^5)

964 618 728

658 674

776 790
900 838

1084
1030 1184 1124

1392 1458 1526

1248 1296 1588 1638

1814

DNA-HIV-194bp DNA-HIV-300bp DNA-E.Coli (1st) DNA-E.Coli (2nd) DNA-Borr

1274 1616 1326 636 1208 808 930 1560 1364 1056 1136 1376 1114 754 1422 872 998 1486 702

1760

Water-DW Water-MilliQ Water-Kobe Wave-HIV-104bp

1858

Wave-Borr Wave-DW Cross Check LB

Wavelength (nm)

Mean-center Smooth[25] 1st, 2nd, and 3rd Step 3

Prediction of EMS by PLS (Lower ES Samples)

Predicted EMS (dB / Hz) Factor: 9 R: 0.89 SEV: 40 dB / Hz

Sample

ES
2 2 2 2 2 1 2 2 2 1 1 1 2 1 1 1 1
37

DNA-HIV-Patient DNA-HIV-104bp DNA-HIV-137bp (1st) DNA-HIV-137bp (2nd) DNA-HIV-194bp DNA-HIV-300bp DNA-E.Coli (1st) DNA-E.Coli (2nd) DNA-Borr Water-DW Water-MilliQ Water-Kobe Wave-HIV-104bp Wave-Borr Wave-DW Cross Check LB

Higher ES Lower ES

Measured EMS (dB / Hz)

Mean-center Smooth[25] 1st, 2nd, and 3rd Step 3

Prediction of EMS by PLS (Lower ES Samples)

Regression Vector
1884

Sample

ES
2 2 2 2 2 1 2 2 2 1 1 1 2 1 1 1 1
38

DNA-HIV-Patient DNA-HIV-104bp DNA-HIV-137bp (1st) DNA-HIV-137bp (2nd) DNA-HIV-194bp DNA-HIV-300bp DNA-E.Coli (1st) DNA-E.Coli (2nd) DNA-Borr Water-DW Water-MilliQ Water-Kobe Wave-HIV-104bp

Regression coefficient (10^4)

606

776 1004 1148 1052

1376

1442
1526 1646 1702 1744 1846 1788

814 884 928 1040 844 944 1084 900 792

1344 1482 1408

1668 1566

746

1872

Wave-Borr Wave-DW Cross Check LB

Wavelength (nm)

Mean-center Smooth[25] 1st, 2nd, and 3rd Step 3

E.Coli DNA 2012 Experiment in Kobe

Water for DNA dilution Filter 1 (450nm)

measurement

measurement

Filter 2 (100nm)
1/101/10 1/101/10 1/101/10 1/101/101/10

10 stage
40

NIR Spectroscopy
Sample : E.coli DNA Spectrometer : NIRSystems 6500 Temperature : room temperature Consective spectra : 3 times Date : 2011/06/23 (Exp. 1) 2011/06/25 (Exp. 2)

41

Results
D2NF D2F450NM D2F450+100 or 20nm D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13 D14 D15 Exp. 1 1042 1042 1060 1125 1107 1004 990 760 765 709 671 1188 1170 1596 1589 1480 Exp. 2 801 801 832 855 829 798 768 809 575 635 610 584 865 801 807 809

Exp. 1 ES : D7D10 Exp. 2 ES : D8D11

ES : Electromagnatic signal
42

Raw spectra
Exp. 1 Exp. 2

Absorbance (-)

Absorbance (-)

Wavelength (nm)

Wavelength (nm)

EMSignal (ES) No Signal (NS)

43

Raw spectra
EMS NS
Absorbance (-)

Exp. 1
Absorbance (-)

Exp. 2

Wavelength (nm)

Wavelength (nm)

Absorbance (-)

Absorbance (-)

Wavelength (nm)

Wavelength (nm)
44

Different spectra (Non-Filtered sample)

EMS NS Exp. 1
936

Exp. 2 934 1068 750 770 778 832 916 1046

Absorbance (-)

770

910

736 748
1416 1384 1460 1362 Absorbance (-)

1382 1406 1362 Absorbance (-) 1488

1490 Wavelength (nm)

1410

Wavelength (nm)

45

Signal calibration with PLS

Exp. 1
Short range
Factor = 7 R2 = 0.68 SEV = 175.7 Factor = 5 R2 = 0.53 SEV = 66.2

Exp. 2

Middle range

Factor = 8 R2 = 0.61 SEV = 198.2

Factor = 8 R2 = 0.67 SEV = 59.4

Transform : Smooth (9) averaged spectra (1st 3rd )

46

Signal prediction with PLS: WAMACS

Model : Exp. 1
Factor = 5 R2 = 0.69 SEV = 188.4
regression vector (-)
60000

1436
1452

40000

1382

20000

1360 1560

1622 1652

-20000

1552
-40000

Test : Exp. 2
R2 = 0.61 SEP = 250.3

1404
-60000

1410

1200

1300

1400

1500

1600

1700

1800

wavelength (nm)

Model : averaged spectra (1st 3rd ) Test : averaged spectra (1st 3rd ) Transform : Smooth (9)
47

E.Coli All Signal DATA Assignment (2012)

D2NF D2F450NM D2F450+100 or 20nm D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13 D14 D15 Exp. 1 1042 1042 1060 1125 1107 1004 990 760 765 709 671 1188 1170 1596 1589 1480 Exp. 2 801 801 832 855 829 798 768 809 575 635 610 584 865 801 807 809

1336nm (aqueous proton [H+(H2O)6] - H2O asymmetric stretch, 1st overt.), 1358nm (H17O8+ free OH stech, 1st overt.), 1374-78nm(aqueous proton [H+(H2O)3] - H3O+ symmetric stretch, ), 1398nm 1421-28nm, 1460nm, 1482nm, 1678nm, 1742nm(1st overtone Superoxide Tetrahydrate O2.(H2O)2 ), 1792nm (1st overtone Superoxide Tetrahydrate O2-.(H2O)3)

E.Coli All Signal DATA Assignment (2013)

E.Coli EMS Aquagram Exp. 1

A= A -
Exp. 1 1042 1042 1060 1125 1107 1004 990 760 765 709 671 1188 1170 1596 1589 1480

A : Absorbance after MSC (1110 nm 1800 nm) : Mean of all the spectra (with NF, F1) : SD of absorbance each wavelength
1344
2.5

D2NF D2F450 nm D2F450+100 or 20 nm D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13 D14 D15

1518
1.5 0.5 -0.5
-1.5

1364

1492

1372

1474

-2.5

1382

1464

1398

1444

1410

1438
50

Lowest Signal
1344
2.5

1518
1.5 0.5 -0.5
-1.5

1364

1492

1372

1474

-2.5

1382

1464

1398

1444

1410

1438

51

1344
2.5

1518
1.5 0.5 -0.5
-1.5

1364

1492

1372

1474

-2.5

1382

1464

1398

1444

1410

1438

52

1344
2.5

1518
1.5 0.5 -0.5
-1.5

1364

1492

1372

1474

-2.5

1382

1464

1398

1444

1410

1438

53

Highest Signal
1344
2.5

1518
1.5 0.5 -0.5
-1.5

1364

1492

1372

1474

-2.5

1382

1464

1398

1444

1410

1438

54

2013 Experiment in Paris

Sample : E.coli DNA Spectrometer : XDS Methrom Temperature : room temperature Consective spectra : 3 times Date : 2013/06

55

E.Coli All Signal DATA Assignment (2013)

Less hydrogen bonded structures activated with high EMS

Contactless Influence
Samples
1) LB (LB medium) 2) LB+A (LB medium + Ampicillin)

How to prepare samples

We prepared 6 sample sets (see next slide) as the following. 1) Put LB or LB+A into small or big tubes 2) After closing the lids of a small tube, set a small tube in a big one. (polypropylene, BD Falcon) Then, close the lids of half big LB LB+A tubes and keep others open. e.g. Open-top Closed LB LB+A

Small tube (15 mL) Input: 10 mL

Big tube (50 mL) Input: 35 mL

Sample Preparation - 2
Open-top

LB LB+A

Sample Name

In: A_IL_OA_I Out: A_IL_OA_O

In: A_IA_OA_I Out: A_IA_OA_O

In: A_IL_OL_I Out: A_IL_OL_O

Control

Closed

Sample In: C_IL_OA_I Name Out: C_IL_OA_O

Cont_LB Cont_LBA

In: C_IA_OA_I Out: C_IA_OA_O

In: C_IL_OL_I Out: C_IL_OL_O

Measurement of Near-infrared spectrum

Near-infrared spectrum of each sample was measured using NIRSystems 6500
Spectroscopy: NIRSystems 6500 (FOSS NIRSystems) Wavelength: 400-2500 nm Methods: Transmittance Path length: 1 mm Temperature: Room temperature Consecutive measurement times: 3 times per a sample 1) Moving a sample into a cuvette cell

2) Measuring a spectrum

NIRSystems 6500

Green: with lid Red: no lid

LB: inside and outside

LB LB + A

LB + A and LB in LB+A

LB: inside LBA: outside LB: measured LB: inside LBA: outside LBA: measured

LBA influence is stronger

LB: inside LBA: outside LB: measured

LB: inside and outside

Difference in WAMACS
Loadings of PC Factors 1-red,2-green,3-blue
Close Top C WAMACS: 1410nm, So

1462nm, S2

1362nm 1398nm solvation solvation shell shell 1420nm hydration Open Top A WAMACS: 1362, 1398, 1410, 1420, 1462, 1490, 1538nm

1412nm, So
1486nm 1366nm solvation 1368, shell 1386nm

1428nm hydration

Investigation of water structural changes induced by cellular phone irradiation

63

Sample preparation
Location of samples
Distilled water
10 mL at each sample

1
Call Milli-Q water
Duration of call 0 minute 5 minutes 10 minutes 15 minutes 20 minutes Receive

12/16/2013

Biomeasurement technology laboratory

64

Spectral measurement
Spectrometer: NIRSystems6500(Foss NIRSystems) Mode: Transmittance Path length: 1mm Temperature: Room temperature Consecutive times: 3 times

Materials Cellular phone (PT002, KDDI, 800MHz) Milli-Q water Distilled water

12/16/2013

Biomeasurement technology laboratory

65

PCA score plot

Wavelength range : 680-1090 nm + 1110-1800 nm Transform : SNV

66

SIMCA

Interclass distance CS1@8 CS2@6 CS1 0.000000 6.163845 CS2 6.163845 0.000000
67

Middle NIR wavelength range

1110-1800nm
Discrimination of irradiated water and non irradiated with PCA

1452nm

Absorbance (-)

1.2

Score plots: discrimination

0 1110nm 1800nm

Loading spectra: investigation of

Wavelength (nm)
wavelengths which contribute the discrimination

12/16/2013

Biomeasurement technology laboratory

68

PCA Score Plots Factor2 (-) [33.00%]

20 minutes 15 minutes 10 minutes 5 minutes 0 minute

Optimal factors: 10 Preprocessing: mean center Transforms: smooth(17) Validation: cross(1)

Factor1(-) [66.60%]

: 0 minute : 5 minutes : 10 minutes : 15 minutes : 20 minutes

Factor 2 showed the duration of call.

Factor 3(-) [0.37%]

Factor1(-) [66.60%]
12/16/2013 Biomeasurement technology laboratory

Factor3 (-) [0.37%]

Factor2(-) [33.00%]
69

Loading spectra: WAMACS

0.1

Optimal factors: 10 Preprocessing: mean center Transforms: smooth(17) Validation: cross(1) : Factor1 : Factor2 : Factor3 : Factor4

Loadings (-)

-0.1

Number

Wavelength 1364nm 1386nm 1412nm 1414nm 1426nm 1430nm 1490nm 1492nm 1508nm 70

1110nm

1800nm

Wavelength (nm)

12/16/2013

Biomeasurement technology laboratory

Classification Model: SIMCA

Misclassifications
Model Actual 1 Actual 2 Actual 3 Actual 4 Actual 5 Test Actual 1 Pred. 1 16 1 0 0 0 Pred. 1 8 Pred. 2 0 15 0 0 0 Pred. 2 0 Pred. 3 0 0 16 0 0 Pred. 3 0 Pred.4 0 0 0 16 0 Pred.4 0 Pred.5 0 0 0 0 16 Pred.5 0 No match 0 0 0 0 0

Preprocessing: mean center Transforms: smooth(17) Class Variable: duration Optimal factors (duration): 5 (0 minute) 4 (5 minutes) 5 (10 minutes) 5 (15 minutes) 4 (20 minutes)

Accuracy
99.0%

No match 0
98.5%

98.75%

Actual 2
Actual 3 Actual 4 Actual 5

1
0 0 0

7
0 0 0

0
8 0 0

0
0 8 0

0
0 0 8

0
0

98.0%

Model Test

97.5%

97.50%

0 0
97.0%

12/16/2013

Biomeasurement technology laboratory

71

SIMCA Interclass distance

0 minute 0 minute 5 minutes 10 minutes 15 minutes 20 minutes 5 minutes 1.39 10 minutes 4.10 2.50 15 minutes 7.52 12.06 7.55

Preprocessing: mean center Transforms: smooth(17) Class Variable: duration Optimal factors (duration): 5 (0 minute) 20 minutes 4 (5 minutes) 9.79 5 (10 minutes) 5 (15 minutes) 10.94 4 (20 minutes)
7.24 2.61 : Over 3.00

SIMCA
Discriminating power: WAMACS
Discriminating power (-)

1800nm

1110nm Number
12/16/2013

Wavelength 1136nm 1174nm 1190nm 1200nm

Wavelength (nm) 1228nm 1270nm 1366nm 1424nm 1468nm 1482nm 1642nm 1674nm 1704nm 1732nm 1756nm

1544nm
1556, 1562, 1568nm 1598nm 1622nm

1796nm
73

Biomeasurement technology laboratory

Investigate whether duration of irradiation or measurement order influenced spectra.

Sample preparation
A cell phone called the opposite one near some kinds of waters in test tubes.
Cellular phone PT002, KDDI, 800MHz (another one is the same one)

5 kinds water (Milli-Q water, distilled water, gold water, silver water, and bronze water) 4 kinds of duration of call (5,10,15,20min) 2 (Irradiated or no) 2 locations =80 samples

call

receive

Two locations, 1 & 2.

Spectra measurement
Spectrometer : MPA Mode : Taransmittance Consecutive times : 3 times Path length : 1mm Temperature : room temperatrure Wavelength range : 800-2500nm

Raw spectra

Absorbance(-)

800

1100 Wavelength (nm)

1800

2500

PCA Score plots (irradiation)

Factor2 loadings factor2 Consecutive No 1 Consecutive No 2 Consecutive No 3 1412

1489
factor3 factor5 factor4

factor1

factor2

factor3

factor4

factor2 shows temperature influenced by consecutive measurement

PCA Score plots: waters

factor2 bronze Milli-Q distilled gold silver

factor5

factor4

factor3

factor1

factor2

factor3

factor4

PLS: Milli-Q water Model accuracy

Measurement order Duration of irradiation

Predicted duration (-)

Predicted order (-)

SEC=0.00080 R=1.00

SEC=0.013 R=1.00

Measured order (-)

Measured duration (-)

Preprocessing : Mean-center Smooth(21)

PLS: Milli-Q water Accuracy of prediction

Measurement order SEP=0.26 R=0.97 Duration of irradiation SEP=5.00 R=0.79

Predicted order (-)

Measured order (-)

Predicted order (-)

Measured order (-)

PLS: Distilled water Model accuracy

Measurement order Duration of irradiation

Predicted duration (-)

Predicted order (-)

SEC=0.0010 R=1.00

SEC=0.020 R=1.00

Measured order (-)

Measured duration (-)

Preprocessing : Mean-center Smooth(21)

PLS: Distilled water Accuracy of prediction

Measurement order SEP=0.34 R=0.96 Duration of irradiation SEP=5.67 R=0.69

Predicted order (-)

Measured order (-)

Predicted order (-)

Measured order (-)

PLS: bronze water Model accuracy

Measurement order Duration of irradiation

Predicted duration (-)

Predicted order (-)

SEC=0.0013 R=1.00

SEC=0.013 R=1.00

Measured order (-)

Measured duration (-)

Preprocessing : Mean-center Smooth(21)

PLS: Bronze water Accuracy of prediction

Measurement order SEP=0.42 R=0.93 Duration of irradiation

Predicted order (-)

Predicted order (-)

SEP=7.69 R=0.42 Measured order (-)

Measured order (-)

PLS: Silver water Model accuracy

Measurement order Duration of irradiation

Predicted duration (-)

Predicted order (-)

SEC=0.0007 R=1.00

SEC=0.10 R=1.00

Measured order (-)

Measured duration (-)

Preprocessing : Mean-center Smooth(21)

PLS: Silver water Accuracy of prediction

Measurement order SEP=0.25 R=0.98 Duration of irradiation SEP=6.10 R=0.67

Predicted order (-)

Measured order (-)

Predicted order (-)

Measured order (-)

PLS: Gold water Model accuracy

Measurement order Duration of irradiation

Predicted duration (-)

Predicted order (-)

SEC=0.0010 R=1.00

SEC=0.015 R=1.00

Measured order (-)

Measured duration (-)

Preprocessing : Mean-center Smooth(21)

PLS: Gold water Accuracy of prediction

Measurement order SEP=0.40 R=0.95 Duration of irradiation SEP=7.56 R=0.61

Predicted order (-)

Measured order (-)

Predicted order (-)

Measured order (-)

PLS Regression Results

measurement order model test SEC R SEP R 0.00080 1.00 0.26 0.0010 1.00 0.34 0.0013 1.00 0.42 0.00070 1.00 0.25 0.001 1.00 0.4 duration of irrudiation model test SEC R SEP R 0.97 0.013 1.00 5 0.96 0.02 1.00 5.67 0.93 0.013 1.00 7.69 0.98 0.1 1.00 6.1 0.95 0.015 1.00 7.56

Milli-Q distilled bronze silver gold

0.79 0.69 0.42 0.67 0.61

All models showed high accuracy Measurement order tests showed high accuracy, but duration of irradiation time tests showed very low accuracy.

SIMCA : Model misclassification

Duration of irradiation
actual actual actual actual actual 5min 10min 15min 20min 0min predicted 5min predicted 10min predicted 15min predicted 20min predicted 0min No match 20 0 0 0 0 0 0 20 0 0 0 0 0 0 20 0 0 0 0 0 0 20 0 0 0 1 3 0 76 0

Measurement order
predicted 1(5min) predicted 2(15min) predicted 3(10min) predicted 4(20min) No match actual 1(5min) 40 0 0 0 actual 2(15min) 0 40 0 0 actual 3(10min) 0 0 40 0 actual 4(20min) 0 0 0 40 0 0 0 0

Preprocessing : Mean-center

SIMCA : Test misclassification

Duration of irradiation
actual 5min actual 10min actual 15min actual 20min actual 0min Unmodeled predicted 5min predicted 10min predicted 15min predicted 20min predicted 0min No match 3 0 0 0 7 0 0 6 0 0 4 0 0 0 4 0 6 0 0 0 0 3 7 0 1 2 0 1 36 0 0 0 0 0 0 0

Measurement order
PredCS1@5 1(5min) 2(15min) 3(10min) 4(20min) Unmodeled 20 0 0 0 0 PredCS2@5 0 20 0 0 0 PredCS3@5 0 0 20 0 0 PredCS4@6 0 0 0 20 0 No match 0 0 0 0 0

Classification: SIMCA
Both of two patterns of model sets showed high accuracy. Interclass distance of duration of irradiation model set showed high values, but they showed very low accuracy in prediction. Measurement order prediction showed 100% accuracy. The effect of cell phone could be controlled by the water

NAFION AND EZ

Naphion

WAMACS: 1378nm, 1388, 1394, 1404 1414nm(-), 1438-1440nm(+), 1454nm(+), 1464nm(+), 1480nm(+), 1496nm last sharp max Blue: Water : consecutive spectra (first subtracted) WAMACS: 1386nm (+) first max 1414nm (-) 1446 -1448nm second max 1464min 1474 1480nm max

Water

ONLY Naphion: 3 consecutive measurements, 1st one subtracted WAMACS: 1324nm (-), 1340nm (+), 1420nm (+), 1444nm (+)?, 1458nm(+), 1498nm (-), 1520nm, 1526nm (+), 1538 1558nm (+) These are water molecules in the air

Effect of magnetic force on the electrical conductivity and spectra of milliQ water

Materials and methods

mQ water used Electrical conductivity was measured in a 20 mm cuvette (by a portable conductometer) Magnets attached at the two sides of the cuvette during the experiment
Place of the magnets

NS

NS

Orientation of the magnets

Two neodymium magnets (rare-earth magnet) were used using the following orientations:

NS

NS

2mm sample cell

NS

2mm sample cell

NS

NS

3. S-S southsouth magnetic poles

4. N-N northnorth magnetic poles

2mm sample cell

NS

NS

NS

1. S-N southnorth magnetic poles

2mm sample cell

2. N-S northsouth magnetic poles

RESULTS

Red line are 9 consecutive spectra of mQ water (second consecutive subtracted). S-N blue, N-S dark green, S-S brown, NN light green
1432nm OHR,, ncII) and DDA OH-(H2O)4, 1448 1458nm, O-HCl, deionized water, 1stover tone of O-H stretch, DDA symmetric stretch of OH-(H2O)4,5, 1492nm,second overtone of bending and symmetric str vibr., S4. Very small increase in 1464nm, S2, working horses and 1518nm.; 1667nm 1682nm O-HR, H13O6+ H-bonded OH strech, 1st overt. Decrease in S1, 1441nm!!!Dicrease in H-bonded OH stretch1477nm; 1479.3nm, 1482nm, IHB/HOH bend (OH-H2O) and H5O2 (terminal water) at. Decrease of aqueous protons. Isosbestic point moves from 1438nm to 1420nm.

1st experiment (in every 40s)

N-N 4. S-S 5. conductivity, S/cm (2nd measurement)

Measurement order
conductivity, S/cm (1st measurement)

No magnet

S-N

N-S 3.

2. 1. 3. 5. 1. 2. 2nd experiment 1st part (continuous) 4.

number of data

Legend S-S pink N-N blue S-N red N-S green mQ brown

AQUAPHOTOME DATA BASE

Systems
Waters: mQ, distilled, deionised, mineral, treated etc. Pressure Water solutions: second overtone disease

Perturbations

Wavelengths, LETTERS Spectral Patterns, WORDS

Functiona lities
stress tolerance

Temperature

first overtone

DNA
biomolecules: proteins, sugars, lipids, alcohols etc.

Illumination Concentration

third overtone combination bands

probiotics dehydration

minerals, salts, metals etc.

Bio fluids blood, urine, serum, rumen juice etc. Cells CHO, bacteria Tissue: (in vitro and in vivo) organs: liver, kidney, stomack etc. mammary gland, mussels, plants tissue, etc.

Magnetic field

solvation shells

hydration

Dillution

oxidation

Water Bands (WAMACS) ASSIGNMENT SOURCE

1368.7 1368.7 1369.5 1369.5 1369.5 1369.9 1370.5 1370.6 1371.0 1371 1371.4 1372.1 1374.0 1375.9 1377.4 1379 1379.3 1380 1380.85 1383 1383.15 1388.7 1388.9 1388.9 1389 1394.7 1394.75 1396.6 1398.141 1399 1407.3 1st overtone OH-str (OH-H2O) H15O7+, 1st overt. H17O8+, 1st overt. H15O7+ free OH stech, 1st overt. H17O8+ free OH stech, 1st overt. aqueous proton [H+(H2O)6] - H2O symmetric stretch, 1st overt. 1st overtone (OH-(H2O)5) H13O6+ free OH stech, 1st overt. aqueous proton [H+(H2O)5] - H2O symmetric stretch, 1st overt. 1st overtone Superoxide Tetrahydrate O2-.(H2O)4 H11O5+ free OH stech, 1st overt. aqueous proton [H+(H2O)4] - H2O symmetric stretch, 1st overt. aqueous proton [H+(H2O)3] - H2O symmetric stretch, 1st overt. H9O4+ free OH stech, 1st overt. aqueous proton [H+(H2O)3] - H3O+ symmetric stretch, 2nd overt. n1+n3 H2O - n1+n3 1st overtone OH-str (OH-H2O) 1st overtone (OH-(H2O)5) 1st overtone Superoxide Tetrahydrate O2-.(H2O)4 1st overtone interwater / DD stretch (OH-(H2O)4) H2O - 2*n1 H+(H2O)10, 1st overt. H bond 18=<n=<24, 1st overt. O-H str 1st overtone H2O (1%) - n1 1st overtone H2O (1%) - n2 aqueous proton [H+(H2O)3] - H3O+ free-OH stretch, 1st overt. free water H5O2 asymmetric H15O7+ H-bonded OH strech, 1st overt. Science28 Wei and Salahub 1997: The Journal of Chemical Physics, 106: 6086. Wei and Salahub 1997: The Journal of Chemical Physics, 106: 6086. Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868. Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868. Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Science28 Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868. Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Weber, Science 2000 Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868. Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868. Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Tsenkova Ozaki 1982 Xantheas, 1995 Science28 Weber, Science 2000 Science28 Ozaki 1982 Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Shin et al. (Mark Johnson) 2004: Science, 304: 1137. Tsenkova Ozaki 1982 Ozaki 1982 Headrick et al. (Mark Johnson) 2005: Science, 308: 1765. Tsenkova Tsenkova Mizuse and Fijii 2012: The Journal of Physical Chemistry, 116: 4868.

Conclusion
Water Matrix Coordinates are the Letters or the Characters related to water structures Water Spectral Patterns are the words that speak about functionalities Huge data base, i.e. Aquaphotome has to be organized in order to learn the language of water and to measure and understand weak signals