Analysis of biopesticide active compounds in Barringtonia asiatica L. Kurz using the GC-MS method

The objective of this study is to use gas chromatography mass spectrometry (GC-MS) to discover the biopesticide active components in bitung seeds (Barringtonia asiatica L. Kurz). research being carried out at Gadjah Mada University Integrated Research and Testing Laboratory in Yogyakarta (LPPT-UGM). The research was conducted from March 1, 2023, until April 2, 2023. The Rf number, color test, fragmentation pattern, and solubility of B. asiatica L. Kurz seed extract were the assessment standards that were used. Based on the area’s chromatogram results, quantitative analysis was carried out to ascertain the quantity of organic molecules that comprise secondary metabolites. After being gas chromatographically separated, the number of organic components that comprise secondary metabolites was enumerated in order to perform a qualitative study. The findings of tests using extracts from seeds of B. asiatica by Gas Chromatography-Mass Spectrometry (GC-MS) yield substances that are secondary metabolites, namely: Octasiloxane, hexadecamethyl-1,1,3,3,5,7,7,9,9,11,11,13,13,15,15-, has the chemical formula C16H50O7Si8 and has a heavy molecular mass of 578 g/mol. The formula chemistry of hexasiloxane, 1,1,3,3,5,7,7,9,9,11,11-dodecamethyl -, is C12H38O5Si6 heavy molecule 430 g/mol, while the formula chemistry of heptasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13-tetradecamethyl, is C14H44O6Si7 heavy molecule 504 g/mol.


Introduction
Fruit bitung is an endemic fruit found in the Indonesian Province of North Sulawesi.It is powerful enough to be used as an insecticide, but neither the public nor researchers have adequate knowledge of its uses to conduct innovative research.Barringtonia asiatica L. Kurz type plants are suitable as substitutes for pesticides used to control Crocidolomia pavonana.Because secondary metabolites of own content compounds [1,4].According to Umaru et al (2018), the cytotoxicity test findings showed that the leaf of B. asiatica was poisonous to brine shrimp larvae at a threshold lethal concentration (LC50) of 208.091 µg/ml, as opposed to 7,455 µg/ml for the control group.
According to cholinesterase test results, farmer sprayer pesticide synthetic plantations in Tomohon City experience poisoning in 18% of the sample blood.According to study findings by Mubushar et al [2], farmers engage in unhealthy practices and disregard advice to apply synthetic pesticides that are safe.About 48.2% of farmers do not follow directions, and more than half (54.4%) use practice spraying pesticide synthetic without safe onsite agriculture; nonetheless, spraying insecticide synthetic is done if population caterpillars on plants exceed threshold control on size four plant invested per 25 plants [3].Examine The goal is to use the GC-MS method to analyze the biopesticide-active chemicals in Barringtonia asiatica L. Kurz.
More than 80% of farmers use synthetic insecticides to control pests, and 90% of them are aware of the harm that these pesticides cause to human health and the environment.However, many farmers do not follow counseling recommendations or the instructions that are printed on insecticide bottles or receptacles [5].

Sample origin
The materials utilized are extracts of the local origin of the B. asiatica L. Kurz variety from the Malalayang City district of Manado, North Sulawesi Province.

Material preparation 1)
. Solution vortex for 5 minutes of sample powder with two milliliters of MeOH in a microtube 2) Centrifuge the Microtube for five minutes at 9500 rpm. 3) After transferring the supernatant to a GC container, instruments for research This is a description of gas chromatography mass spectrometry (GC-MS): kind GC-MS type Thermo Scientific Trace 1310 GC Thermo Scientific ISQLT Single Quadropole Mass Spectrometry with an automatic injector of type TriPlus RSH, a centrifuge, a soft Chromeleon device, UHP helium gas, type column, HP -5MS UITG-5MS and TG-WAXMS, and a votex mixer.Grate, knife, colander, plastic tray, and filter are needed for the preparation sample [3].

GCMS analysis and method
Utilizing labaoratorium based research apparatus, such as the Gas Chromatography Mass Spectrometer (GC-MS) with the following specifications: Thermo Scientific Trace 1310 GC, Type GC: Type MS Thermo Scientific ISQLT Single Quadropole Mass Spectrometer; HP-5MS UI, TG-5MS, and TG-WAXMS; TriPlus RSH type autoinjector; helium gas uhp; chromeleon software; column type.Condition of the column: Maximum temperature: 325/350 °C, HP-5MS UI, length: 30 m, film: 0.25µm, I. D.: 0.25 mm Temperature of the injector: 275 o C; split flow: 100 ml/min; split ratio: 200; front inlet flow: 0.50 ml/min; temperature of the MS transfer line: 260 o C; temperature of the ion source: 250 o C, Tool : Transporter Gas: UHP (He) helium; mass list range: 10-400 (amu); Column temperature: 1, Purge Flow: 3 ml/min, Gas Saver Flow: 5 ml/min, Gas Saver Time: 5 min.Hold time (min) 10, Hold time (min) 2, Rate ( o C/min) 0, Target value ( o C) 100.Target value ( o C) 300, Retention Time (min) 60.0, Rate ( o C/min) 5, Hold duration (min) 10 [4].Quantitative analysis set based on organic compound content and secondary metabolites based on chromatogram area results.Gas chromatography was used to separate the various organic composer metabolites that were determined through qualitative analysis [6].Fragmentation pattern, color test, solubility, and Rf number of seed extract are some of the measurement parameters B. asiatica L. Kurz.Analysis results form with technique and chart point peak for each compound Using a GC-MS technique, matching weight molecules and fragmentation patterns from compound findings isolation with compounds in the library [7,8].

Results and discussion
These findings indicate that the extract seed bitung (Barringtonia asiatica L. Kurz) contains sixteen chemicals.The extract seed B. asiatica test results from gas chromatography mass spectrometry (GC-MS) reveal 16 peaks in the sample.Figure 1 displays the GC chromatograms of the organic chemical constituents and secondary metabolites in the fruit extract.Using chromatogram gas chromatography, the mass detected by the spectrometer yields the spectrum mass of each detected peak.The role of the mass spectrometer is to provide structural information on the compounds being analyzed as well as quantification of compounds that are not separated by chromatography.
A gas viscosity changes with temperature.If the pressure at the column entrance is constant during the chromatographic analysis, the carrier gas flow in the analytical column drops as the oven temperature rises.The analyte's elution speed decreases as a result, leading to peak expansion and a decline in the chromatographic profile's quality.For this reason, electronic flow regulators are a feature of contemporary injectors.This device makes consistent gas flow in the column by adjusting the inlet pressure of the carrier or column head based on the oven temperature, which significantly enhances chromatography performance.The pressure inside the mass spectrometer source is conditioned by the gas flow.Accordingly, this pressure is the outcome of a balance between molecules that are drawn to the source by chromatography and molecules that are expelled by aspiration, which is done continually by a pumping system.Constraints on gas flow differ depending on the mass spectrometer being used [6].
If the stationary phase of a mixture is polar, the compounds are essentially separated based on their polarity; if the phase is non-polar, the compounds are separated based on their volatility.Because the low polarity stationary phase is typically stronger and more thermally stable than its polar stationary phase, a low polarity column is typically chosen when working with mixed mixtures.GC-MS makes use of capillary columns with low bleed [9].
Using a mass spectrometer, the constituent organic components in the extract seed bitung were identified.The fractional mass spectra, peak base (base peak), and similarity index (SI) values will be compared to the spectral data from MainLib libraries.The findings of the analysis indicate that, as shown in Figure 2, there are 16 peaks and 48 possible component successful compounds that were separated using the solvent methanol.Peak capacity is the the most number of distinct peaks that can fit within a separation space defined as the earliest time at which a peak can elute from a column and the time at which the nth peak elute.The amount of time available for the last peak to come out of the column with elution under gradient or programmed conditions produces many more peaks than elution under constant conditions.In essence, programmed elution produces more peaks per unit time than in invariant elution conditions.All peaks should have the same resolution (R) otherwise some peaks will be too well separated thereby wasting space.Generally, the requisite resolution is taken as unity.Here, B and A denote the later and earlier of a pair of adjacent peaks, tR and σ are the retention times and standard deviations of these peaks R= t R,B-tR,A 2(σA+ σB) (1) If R is set to unity, and when the two peaks have essentially the same widths, then the two retention times will be separated by 4σ.It is clear that in a given chromatographic space the total number of 4σ intervals is finite and controls the number of peaks that can fit in this space.
Giddings argued that under conditions of constant elution we can take the 4σ width as The i + 1th peak must elute at a time relative to the ith peak equal to the mean of the two peak widths thus: Retention area allows for the determination of the level of abundance of separated chemicals within a sample using the GC-MS method [6] [8].Based on the total number of ions created of each component chemistry, area retention affects the production of chromatograms [8].There will be a large amount of ions created from the component molecules.A component in the mixture under analysis is at a certain percentage tall, thus the peak that shows up on the chromatogram also has a huge area.Table 1 displays the compounds' identification results from the peak with the highest area retention.
Table 1.Compound composition, metabolites, secondary extract from three peak highest identification results obtained using GC-MS for B. asiatica seed.

Figure 1 .
Figure 1.Chromatogram findings GC-MS examination of extracts from B. asiatica seeds.

Figure 3 .
Figure 3. Depiction/Illustration of Chemical Structure: Octasiloxane hexadecamethyl Formula get up C 16 H 50 O 7 Si 8 Source : Similarity Index (SI) of the National Center for Biotechnology Information.2023.

Figure 4 .
Figure 4. Displays the molecule Hexasiloxane dodecamethyl's Similarity Index (SI).Depiction/Illustration of Chemical Structure: Hexasiloxane dodecamethyl Formula get up C12H38O5Si6 Source: Similarity Index (SI) of the National Center for Biotechnology Information.2023.