Suprasterol ProductionunderNovelUVBLED,SPERTILampandNaturalSunlight
NancyShaw1,TylerKalajian2,AliAldoukhi2,AngelineVeronikins2,KellyPersons2,Dr.MichaelFHolick2
PlanoWestSeniorHighSchool, 5601WParkerRd,Plano,TX75093
VitaminD,Skin,andBoneResearchLaboratory,DepartmentofMedicine,85ENewtonStreet,M-1013Boston,Ma02118
Abstract
Methods
Objective: ItisknownthatwhenvitaminD3 isexposedtohighintensity
UVradiationitisconvertedtoseveralcompoundsincludingsuprasterols.
ItishoweverunknownwhethervitaminDcanbephotoisomerized into
theseproductsduringexposuretonaturalsunlight.Sincesuprasterols
haveanti-proliferativeeffects,thismayhaveintegralanti-proliferative
andphotoprotective effects.Thegoaloftheprojectistodetermineif
vitaminDisabletoundergophotoisomerization toproducethese
productsduringexposuretosunlightinBoston.
Method:Twoborosilicateampoulescontaining150μg/mLofvitaminD3
dissolvedinethanolwereirradiatedatvaryingtimeintervalsunderLED
andtheSPERTIlampforatotalof25MEDandsunlightinBostonfor3
hours.TheLEDhasasolewavelengthof293nmwhilenaturalsunlight
emitsbroad-spectrumUVAandUVBradiation.Theorganicsolventinthe
ampouleswasrecovered,driedundernitrogenandredissolved ina
solventusedforseparatingphotoproductsfromvitaminDbyHigh
PerformanceLiquidChromatography.Thesampleswere
chromatographedandvitaminD3 anditsphotoproductsweredetected
withavariablewavelengthUVdetector.
Results: HPLCchromatographsindicatedthepresenceofmultiplephotobyproducts.
Conclusion: Therefore,duringtheexposuretosunlight,thevitaminD3
thatisproducedcaneitherenterthecirculationorabsorbsolarUV
radiationandbeconvertedtophotoproducts.Ifoneormoreofthese
photoproductsshouldhaveuniquebiologicalproperties,thenbeing
exposedtosunlightmayhaveadditionalhealthbenefitscomparedto
takingavitaminDsupplement.Thesefindingswouldbeabigstep
towardsunderstandingwhethertakingavitaminDsupplementhasthe
samebenefitasbeingexposedtosunlight.
Discussion
VitaminDdissolvedinethanolinborosilicateampoules(150μg/mL)
TheampouleswereexposedtoLED(30min.,50MED/hr)andSPERTIlamp(30min,50MED/hr)foratotalof
approximately25MEDs;anddirectsunlight(3hr.,2MED/hr,total6MED)atBostonduringsummer
Thepresenceofsuprasterols andotherphotobyproducts wasdeterminedusingnormalphasehigh
performanceliquidchromatographywithanisocraticmobilephaseconsistingof0.4%isobutanol inhexane
LED
BostonSunlight
SPERTILamp
Results
Absorbance
Introduction
TherearetwowaystomakeupforvitaminDdeficiency:exposureto
UVBfromthesunandthroughtakingsupplements
Time(min)
a) ChromatogramofControl(vitaminD3)using265nmdetector indicatingpresenceof
VitaminD3 only
b) UVSpectrumofVitamin D3
HumanshavenaturallydependedonsunlightfortheirvitaminD
requirement
VitaminD3 issynthesizedintheskinwhenUV-Bradiationfromthesun
converts7-dehydrocholesterol(7-DHC)intoprevitamin D3 (preD3),which
thermallyisomerizesintovitaminD3 .
Time(min)
a) ChromatogramofControl(vitaminD3)using210nmdetectorindicatingpresenceof
VitaminD3 only
b) UVSpectrum
Absorbance
Conclusions
PreD
S3
5,6trans
VitD
S1
VitaminD
S5 S6
S2
Time(min)
ChromatogramofvitaminD3 ampouleirradiatedunderLEDfor30minutesusing210
nmdetector
Pre-D
Theresultsofthestudiesconfirmsthepresenceofsuprasterols in
ampoulesirradiatedundervarioussourcesofUVlightincluding;LED,
SPERTIlamp,andnaturalsunlight
Thepresenceofsuprasterols indicatethatvitaminDnaturallybreaksdown
whenexposedtoUVlight
Suprasterols mayhavesignificantbiologicaleffects,beingcreatedwhen
exposingvitaminDunderUVlight,probablyactingasanti-proliferative,
photoprotective,feedbacklooptopreventskincancer
VitaminD
Suprasterols
Suprasterols mayplayanintegralroleinreducingtheriskofskincancerdue
topreliminaryexperimentsthatdemonstrateanti-proliferativeeffectsin
keratinocytes.SinceUVlightexposurecausestheproductionofthese
suprasterols fromvitaminDintheskin,thismaybeapartofafeedbackloop
thatpreventsskincancerundermoderatesunexposure.Itishighlylikelythat
providedwithmoderatesunexposure,thereislittlerisktoskincancerdueto
thenaturalcounterbalancingactthathasbeenbuiltin.
b
a
Thesephotobyproducts,specificallysuprasterol 5and6haveshownantiproliferativeactivityinkeratinocytes;theothersuprasterols mayhave
otherphotoprotective effectsandplayaroleinpreventingskincancer
Objective:
• TodetermineifvitaminDisabletoundergophotoisomerization to
producetheseproductsduringexposuretosunlightinBostondueto
UVlight
• NotequalitativedifferencesbetweenaspectrumofUVandasingle
UVBwavelengthonsuprasterol production
• Identifythesuprasterols producedbycomparisontopreviousHPLC
migrationpatterns
vs.
Absorbance
Suprasterols,photobyproducts ofVitaminD,havebeenpreviously
identifiedafterirradiatingVitaminD,withUVlight.Thesesuprasterols
havebeencategorizedbyspecificHPLCelutiontimes,UVabsorption
spectra,andtheirstructuresdeterminedbyH-NMR.
AcomparisonwasmadebetweenRayVio’s novelUVBLEDandthe
commerciallyavailableSPERTIlamp(producedbyKBDInc.)thatputsoutUVA
andUVBradiation
ExposuretoUVBradiationmaybepreferabletosupplementsbecauseofthe
emergenceofseveralphotoproducts,includingsuprasterols,which
supplementsdonotpossess..Thesephotoproductscouldpotentiallyhavea
widerangeofhealthbenefitsthatwouldbelostinsupplements.
a
b
ThechromatogramindicatethatvitaminD3 convertstoseveralsuprasterols
whenexposedto3differentUVlightsources- LED,SPERTIlamp,andnatural
sunlight.Theirelutiontimesappeartocomebeforethevtamin D3 peak.Itis
highlylikelythattheproductsdecreaseandchangethroughoutexposure
sincetheyarenotphotostable.
Therewasagreaterpresenceofsuprasterols underbroadspectrumUVlight
fromtheSPERTIlampthancomparedtotheLED’ssinglewavelengthUVB
light.Naturalsunlightalsoresultedinalargervarietyofphotobyproducts.
Absorbance
2
S3
PreD
5,6trans
VitD
S1
S5
S6
VitaminD
Time(min)
ChromatogramofvitaminD3 ampouleirradiatedunderSPERTILampfor30minutesusing210
nmdetector
Furtherstudyinthisregionisneededtoassesstheanti-proliferativeeffects
ofsuprasterols inkeratinocytesandsurgically-obtainedskin
References
Webb,A.R.;Decosta,B.R.;Holick,M.F.Sunlightregulatesthe
cutaneousproductionofvitaminD3bycausingitsphotodegradation.
TheJournalofClinicalEndocrinology&Metabolism1989,68(5),
882-887
Absorbance
1
Acknowledgements
Time(min)
ChromatogramofvitaminD3 ampouleirradiatedunderBostonsunlightfor3hoursusing
using 210nmdetector
Thankyouformakingthisresearchinternshipanamazingexperience:
Dr.Holick,KellyPersons,TylerKalajian,AliAldoukhi,AngelineVeronikins,
ClaudiaDurbin,HaileyClark,Teja Mogasala,andNahee Kim.
Suprasterol Production under Novel UVB LED and Natural Sunlight
Nancy Shaw1,2, Tyler Kalajian2, Ali Aldoukhi2, Angeline Veronkins2, Kelly Persons2, Dr.
Michael F Holick2
1
Plano West Senior High School, 5601 W Parker Rd, Plano, TX 75093; 2Vitamin D, Skin, and
Bone Research Laboratory, Department of Medicine, 85 E Newton Street, M-1013 Boston, Ma
02118
It is known that when vitamin D3 is exposed to high intensity UV radiation it is converted to several
compounds including suprasterols. It is however unknown whether vitamin D can be
photoisomerized into these products during exposure to natural sunlight. Since suprasterols have
anti-proliferative effects, this may have integral anti-proliferative and photoprotective effects. The
goal of the project is to determine if vitamin D is able to undergo photoisomerization to produce
these products during exposure to sunlight in Boston. Two borosilicate ampoules containing 50
ng/mL of standard vitamin D3 dissolved in ethanol were irradiated at varying time intervals under
LED and sunlight in Boston during August for a total of 25 MED. The LED has a sole wavelength
of 290 nm while natural sunlight emits broad-spectrum UVA and UVB radiation. The organic
solvent in the ampoules was recovered, dried under nitrogen and redissolved in a solvent used for
separating photoproducts from vitamin D by High Performance Liquid Chromatography. The
samples were chromatographed and vitamin D3 and its photoproducts were detected with a variable
wavelength UV detector. Therefore, during the exposure to sunlight, the vitamin D3 that is
produced can either enter the circulation or absorb solar UV radiation and be converted to
photoproducts. If one or more of these photoproducts should have unique biological properties,
then being exposed to sunlight may have additional health benefits compared to taking a vitamin
D supplement. Findings would be a big step towards understanding whether taking a vitamin D
supplement has the same benefit as being exposed to sunlight.