Table Of ContentDerivative Spectrophotometry and Electron
Spin Resonance (ESR) Spectroscopy
for Ecological and Biological Questions
.
Vladimir S. Saakov Valery Z. Drapkin
l l
Alexander I. Krivchenko Eugene V. Rozengart
l l
Yuri V. Bogachev Mikhail N. Knyazev
l
Derivative
Spectrophotometry and
Electron Spin Resonance
(ESR) Spectroscopy for
Ecological and Biological
Questions
VladimirS.Saakov
SechenovInstituteofEvolutionary ValeryZ.Drapkin
Physiol.andBiochem. YuriV.Bogachev
RussianAcademyofScience MikhailN.Knyazev
ThorezAv.44 DepartmentofPhysics
Saint-Petersburg StateElectrotechnicalUniversity
Russia Saint-Petersburg
Russia
AlexanderI.Krivchenko
EugeneV.Rozengart
SechenovInst.ofEvolutionaryPhysiol.
andBiochem.
RussianAcademyofScience
Saint-Petersburg
Russia
ISBN978-3-7091-1006-5 ISBN978-3-7091-1007-2(eBook)
DOI10.1007/978-3-7091-1007-2
SpringerWienHeidelbergNewYorkDordrechtLondon
LibraryofCongressControlNumber:2012940214
#Springer-VerlagWien2013
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Preface
Thisbookiswrittenbyagroupofauthorswhohavealreadycooperatedformany
decades. The successful combination of the scientific approaches of biologists
interested in modern methods of physics and physical chemistry with those of
physicists and experts in the electronics fields, who have developed new methods
of derivative and EPR spectroscopy, has enabled the technical realization – with
Russian equipment – of Prof. Ernest Rutherford’s ideas about derivative spectra
registration.
Yearsagosomemembersofthisgroupofauthors–forthefirsttimeinRussia
(USSR)–usedactivationwithprotons andalphaparticlesforthedetectionofthe
stable oxygen isotope O18 incorporated in biologically active substances from
labelled water molecules. These results are unique to date. The authors of the
book have worked with great thoroughness in these fields for many years. One
stage of the above-mentioned work was the development of radiochemical purifi-
cationbasesforuseinbiochemicalpreparations.Theseexperimentsrequiredgreat
application throughout decades by these authors, when their interests extended
beyondradiationandspectroscopicresearch,ascoveredinoneofbook’schapters.
Thebookbasicallydescribesprinciplesandpracticeofworkingwithderivative
spectrophotometry and EPR spectroscopy. The abundance of graphic material
provides good visual perception of derivative spectra features and will help the
readertointerprethisownexperimentaldatafromvariousscientificorigins.
The authors freely cite Russian and European literature, and also publications
fromadjacentareasofthenaturalscienceslesswellknowntotheEuropeanreader.
Englishliteratureiswellcovered,attentionbeingpaidtosomedebatablequestions
ofbiochemistry,thesolutionofwhichbecamepossibleduetotheapplicationofthe
researchmethodsconsideredinthebook.
The width of approach of the book’s authors in various research directions
concerned with the usage of diverse genetic material, with features of pigment
metabolism of evolutionary forms of Eucaryota and Procaryota, and also with
demonstration of practical application of derivative spectrophotometry for the
assessmentofstatesofbiologicalobjectsundervariousanthropogenicandnatural
influencesandinmedicalbiochemistry,isimpressive.
Materialofferedbyauthorsandtheirinterpretationofitisofdoubtlessinterestto
the wide audience of biologists, physicians, biophysicists, postgraduate students,
v
vi Preface
and students of biological, physical, chemical, and medical research profiles, and
alsototheordinaryinquisitivereaderwishingtobecomefamiliarwithfeaturesof
theexperimentalworkofbiologistsofphysicalandchemicalareasandtowidenhis
viewofthisinterestingareaofknowledge.
AcademicianV.L.Svidersky
AdviseroftheRussianAcademyofSciences
Acknowledgements
To make all the information presented in this book clear for the reader and a
scientific audience, the publishing Senior Editor Dr. C. Panuschka had to limit
manyauthors’thoughtsandtocontroltheirwisheswithintherequirementsoflogic
andofthepublishinghouse.Weexpressourthankstoherfortheprofessionaland
kindatmosphereshecreatedwhileworkingonthebook,andforoperativesolution
ofallarisingproblems.Also,wearegratefultotheproductioneditorW.Dollha¨ubl
andtootherstaffmembersofthepublishinghouse responsiblefor illustrations as
well as to Martina Himberger, Production Coordinator in Heidelberg. Authors’
thanks go to I. Danilova who, as a biophysicist, found and corrected some inac-
curaciesandalsopreparedthefirstEnglishtranslationofthebookwithgreatmeti-
culousness.Additionally,wewouldliketoexpressourappreciationtothetranslator
of the Springer publishing house, A. V. Buldakov, to the publisher of Russian
editionTechnolitLtdPubl.House,SPbandthecollectiveeffortsofSpringerWien
New York. They worked with the authors in a friendly environment, considering
theopinionsofthespecialists,andnotstrayingfromtheirusualinsistenceonhigh
standards.
TheheadauthorofthisbookrepeatedlyworkedintheZentralinstitutfu¨rGenetik
und Kulturpflanzenforschung AdW, Gatersleben, GDR (renamed into Institut fu¨r
Pflanzengenetik und Kulturpflanzen-forschung, IPK, after German reunification).
The Institute has been organized by Professors H. Stubbe and K. Rosenstiel in
Vienna Tuttenhof in 1943. It has suffered in the Second World War and Prof.
H.StubbehasbeenengagingintheorganizationofthisInstituteinGaterslebenin
1946–1949 and the representative of the Soviet military administration botanist
Prof. S.G. Saakov (senior)has been assistinghim. After 1969 the management of
theInsistutehasbeenheadedbyProf.H.Bo¨hme. Prof.SaakovV.S(junior)would
like to especially thank both Professors, H. Stubbe and H.Bo¨hme. Great friendly
help was provided by Professors H. Sagromsky, K. Mothes (the president of
Leopoldina Society), A. Rieth and P. Metzner, and also by colleagues Doctors
A. Meister, E. Brecht, K. Adler and M. Pejßker and by technical assistants of the
physicalphysiologydepartment.
vii
.
Contents
1 Introduction ................................................................ 1
References ................................................................... 3
2 BasisofDerivativeSpectrophotometry ................................... 5
2.1 TheMainLawofLightAbsorptionbyaSubstance ................... 5
2.1.1 ReasonsforDeviationfromBouguer’sLaw ..................... 7
2.2 CorrectnessandAccuracyofSpectrophotometricData .............. 10
2.2.1 MainFactorsInfluencingtheAccuracyofSpectrophotometric
Measurements ................................................... 11
2.2.2 Difference(Differential)Spectrophotometry ................... 15
2.2.3 MeasurementErrorsofDifferenceSpectrophotometry ........ 17
2.3 DerivativeSpectrophotometry ......................................... 19
2.3.1 MethodsofDerivativeSignalRegistrationandDiagramsof
DifferentialAnalyzers .......................................... 38
2.3.2 ParameterOptimizationoftheDifferentiatingCircuit ......... 43
2.3.3 DerivativeSpectrophotometryofDifferenceSpectra .......... 53
2.3.4 MethodofthePulseAmplitude-ModulatedFluorescenceforthe
SolutionofEcological-BiochemicalProblems ................. 54
References .................................................................. 57
3 TheDerivativeSpectrophotometryMethodforAnalysisof
BiologicallyActiveSubstances ........................................... 71
3.1 DerivativeSpectrophotometryforAnalysisofaNumberof
GuanidinePreparations ................................................ 71
3.2 ChelatingAbilityof1,3-Bis-((p-Chlorobenzylidene)amino)
Guanidine:ComplexeswithCa2+andLa3+Ions ...................... 79
3.3 TheSpecialFeaturesoftheCa2+Bindingbymono-,bis-andtris-
SubstitutedGuanidineDerivatives .................................... 84
3.4 SpecialFeaturesofInteractionofBis-((Chlorobenzylidene)amino)
GuanidineDerivativeswithCa2+DependingontheChlorineAtom
PositionintheMolecule ............................................... 93
ix
x Contents
3.5 TheSpecificCharacterofCa2+Interactionwith((Benzylidene)amino)
GuanidineDerivativesContainingElectron-DonororElectron-Acceptor
Substituters ............................................................ 101
3.6 SpecialFeaturesofCalciumIonsInteractionwithBis-((4-hydroxy-3-
methoxybenzilidene)amino)GuanidineandBis-((4-cyanobenzilidene)
amino)Guanidine ..................................................... 107
3.7 TheProofofPolycomponentCompositionofthePromising
AntitumorDrug“Ukrain” ............................................ 113
3.8 DerivedSpectraApplicationfortheAnalysisofDerivedFormsof
NondepolarizingMuscleRelaxantTercuronium,ofVitaminsand
Hormones ............................................................. 125
3.8.1 ComparativeAnalysisofTercuroniumDerivatives ........... 125
3.8.2 TheReasonabilityofDerivedSpectraApplicationforthe
AnalysisofCommercialPreparationsofVitamins
andHormones .................................................. 128
3.9 TheImportanceofDerivativeSpectrophotometryin
ModernStudiesofAromaticAminoAcidsandProteins ............ 133
3.9.1 SpecialFeaturesofDerivedSpectraofPhenylalanine,
Tyrosine,andTryptophan ..................................... 135
3.9.2 SpecialFeaturesofPhenylalanineSpectraChangeCausedby
InfluenceofGammaRadiation ................................ 143
3.9.3 SpecificCharacterofTyrosineSpectraChanges
UnderInfluenceofGamma-Radiation ........................ 149
3.9.4 TheCharacterofTryptophanDerivedSpectraChangeUnder
InfluenceofGamma-Irradiation ............................... 154
3.9.5 TheComparativeCharacteristicofAlbuminDenaturation
SpectralChangesUnderThermaland
RadiationExposures ........................................... 160
3.9.6 ChangesoftheGammaGlobulinOpticalSpectraUnder
g-Irradiation .................................................... 167
3.9.7 SpecialFeaturesofg-GlobulinSpectraChangesDuring
g-GlobulinDenaturationCausedbyThermalandRadiation
Exposure ....................................................... 174
3.9.8 TheCouplingofAlbuminDerivedSpectraChangewiththe
DeterminationAccuracyofAlbumin/GlobulinCoefficientfor
RadiationInjuries .............................................. 182
References ................................................................. 189
4 ApplicabilityoftheDSHOMethodinWorkwithPigmentsof
PlantsandAnimals ....................................................... 197
4.1 DerivedSpectraofHighOrdersforSomeCarotenoids ............. 204
4.2 NeoxanthinasaProbableKeyProductofFormationofa-and
b-CarotenoidDerivatives ............................................. 213
4.3 MetabolicTransformationsofLabeled14C-or3H-Carotene
inAnimalTissues ..................................................... 219
