Proceedings of MatricesFor IITTEP – ICoMaNSEd 2015
ISBN: 978-602-74204-0-3
Biology Page 500
measuring 50x30x10 cm lined with blotting paper and fed pesticide-free leek. Second instar larvae used for bioassay and the remainder used further propagation. Larvae will pupasi
transferred to another plastic box containing sterile saws dregs. Pupa formed moved to the breeding of insects and allowed to imago.
2.4.2. Extraction and Partition
Extraction is done by soaking method and partition with Counter-current distribution method. Bitung seeds that have been shredded dried in an oven at a temperature of 400C for 1 x 24
hours and then pulverized with a blender. Powder is finely then soaked with methanol in the ratio 1: 5 w v in the Erlenmeyer flask and shaken with a magnetic strirrer for 2 x 24 hours.
Results marinade filtered in stages by using 2 Buchner funnel that had been covered with coarse filter paper for upper funnel and fine filter paper Whatman no.41 for lower funnel
and accommodated in another Erlenmeyer flask. The dregs of the filter is rinsed repeatedly until the filter is not colored clear. Liquid distillate united and included in the evaporator
flask that had been weighed, then metahnol evaporated by using a rotary evaporator at a temperature of 45 - 500C, a rotation speed of 50-60 rpm and low pressure 150-200 mmHg.
After evaporation was completed, the flask containing the extract weighed again. The difference between the two is the weight of the weighing of extract.
The crude extract obtained from the methanol fraction was then partitioned with hexane - methanol system 95 with a ratio of 1:10:10 w v v in a flask separator for ± 3 hours,
and the hexane phase was washed with 95 methanol. Hexane phase is removed, while 95 methanol phase was evaporated with a rotary evaporator. Methanol fraction obtained was then
partitioned back in ethyl acetate - water system, the water phase was discarded and the ethyl acetate phase was evaporated solvent. Ethyl acetate fraction obtained is then stored in a
refrigerator
≤ 4 0 C for later use in the bioassay.
2.4.3. Applied 2.4.3.1. Toxicity Test
Ethyl acetate fraction of the extract was diluted with a diluent solution containing methanol, emulsifiers Agriwet 100 S and distilled water until it reaches the desired concentration. The
final concentration of methanol and emulsifiers in a solution of each extract was 1 and 0.1 . Pesticide -free leek cut to a length of 3 cm. The leaves of each treatment was immersed in
a solution with a concentration of extract that has been specified for ± 5 seconds, then wind dried. Srbagai controls are used qauadest containing 1 methanol and 0.1 emulsifier
glycerol alkyl phthalates Agriwet 100 S without the extract. After wind dried leaves of treated and control each put in a petri dish ø 9 cm were paved with blotting paper, then put
15 instar larvae of
S. exigua
2. After 48 hours, the leek is required and control is replaced with fresh chives without treatment.
2.4.3.2. Tests on The Activity of Eating
Four leaves 2 leaves 2 leaves the treatment and control put in a petri dish ø 9 cm which had been covered with blotting paper are alternately. Prior to treatment, all four of these
leaves were weighed to determine the initial weight and drawn one example of each piece leaves are used and weighed to determine the water content of leaves. The leaf samples were
dried in an oven for 2 days at a temperature of 100 C and then weighed again to obtain a dry sample weight. Based on the values obtained water content, leaf dry weight of initial
treatment and control is taken then dried as means mentioned above and then weighed. The
Proceedings of MatricesFor IITTEP – ICoMaNSEd 2015
ISBN: 978-602-74204-0-3
Biology Page 501
difference between the dry weight of the initial and residual dry weight is the weight of the leaves are eaten
2.4.3.3. Testing the Effects of Extracts on Larval Development
In this test the surviving larvae continue to be maintained imago. It is intended to observe any changes that occur in larvae test. While that manages to be imago will be transferred to a jar
and fed with 10 honey solution to observe its development. The population at every level of treatment increased the amount to be used as a material in further treatment, especially in
determining the reproduction of imago
S. exigua.
2.4.3.4. Providing Plant Onion Leaves
Leek plants used in this study derived from the seeds from farmers. These seeds are then planted in polybags and fertilization with manure to obtain fertile plants. Maintenance is
carried out without pesticides to produce healthy plants. The number of plants adapted to the needs.
2.5. Parameter of Observation 2.5.1. Toxicity Testing
