ConstantIndependentDependent AOS Species of larvae Environmental conditions Extraction Method (after determination) Extraction Method (before determination) S. Oleracea sub- species. Concentration of extracts Mortality Rate of T. molitor Observable deformities of T. Molitor HCI

Buy grocery spinach and perform Fractionated precipitation Solvent- Solvent DistributionChromatography Purification of ecdysteroids Bio- Assay

Buy organic spinach and seeds determine best extraction method Plant Spinach and freeze-dry when mature HPLC Bio- Assay

HCI Timeline Proposal Oct Discussion with AOS29 Oct – 7 Nov Finalisation of Proposal8 Nov Preliminary Experimentation – to determine the best method of extraction 9 Nov – 7 Dec & 16 Jan - Prelims Preliminary Round of Projects Day CompetitionMarch Experimentation on MealwormsBetween Prelims and Semi finals Semi-FinalsJuly Final Experimentation on mealwormsBetween semifinals – 2 weeks before final judging Compilation of Results with AOSBefore Finals, after final experimentation Research paperAfter finalizing results Finals JudgingAug

AOS Timeline Background research Oct Discussion with HCI29 Oct – 7 Nov Finalisation of ProposalNovember Testing Extraction methods to determine the best extraction method Growing and harvesting of spinachNovember Extracting Phytoecdysteroids and determining the best extraction method November - January Experimentation on MealwormsFebruary to April Travel to Singapore for finals presentation and finalization August

Number of Mosquito Larvae used Solvent ControlPetri Dish 1Petri Dish 2Petri Dish 3 Concentration 110 Concentration 210 Concentration 310 Negative Control10