The nanoparticle formula optimization from Temulawak extract ( Curcuma xanthorrhiza ) as adjuvant therapy

Temulawak extract was synthesized using temulawak's sodium alginate as a base material. It was characterized by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. The size of Alg-Curcuma microbeads measured at SEM was 296 m. FTIR spectra showed that the size of the resulting beads was microparticle in size. The particle size distribution of the sample was also analyzed. The results show that the nanoparticle formula was obtained from temuwak extract material with parameters and characterization that have been analyzed and tested. This is an open access article under the CC BY-SA license.


INTRODUCTION
Currently, a nanomedicine method has been developed that can target drugs to tumor cells specifically, namely nanoparticle therapy (Aghebati-Maleki et al., 2020;Liu et al., 2007;Oroojalian et al., 2021;Tang et al., 2021;Thakkar et al., 2020).Nanoparticles are colloidal particles that can be amorphous or crystalline in shape that have a size smaller than 1 micron (Croissant et al., 2020;Dong et al., 2020;Ijaz et al., 2020;Rastogi & Asa-Awuku, 2022;Shrestha et al., 2020).The use of nanoparticles in cancer management aims to increase bioavailability and drug distribution (Chivere et al., 2020;Gavas et al., 2021;Hafeez et al., 2021;Raj et al., 2021;Wang et al., 2020), as well as improve drug targeting and release to tumor cells, so that it is expected to increase efficacy and reduce side effects (Ali et al., 2021;de la Torre et al., 2020;Gomes et al., 2021;Kashkooli et al., 2020;Zein et al., 2020).Therefore, it is necessary to develop a nanoparticle formula of temulawak extract which is expected to increase the effectiveness of temulawak extract as an anticancer.
The main ingredient used is standard temulawak powder.Other ingredients used were sodium alginate powder with a molecular weight of 216.12 g/mol purchased from Sigma-Aldrich® (Darmstadt, Germany), calcium chloride anhydrous powder (CaCl2.2H2O)purchased from Merck (Darmstadt, Germany) and distilled water.All materials have an analytical level.
The research would like to analyze the nanoparticle formula optimization from Temulawak extract (Curcuma xanthorrhiza) as adjuvant therapy.It is expected to give more insight into the formula optimization process of herbs in Indonesia, especially Temulawak since it is rarely explored.

METHOD
The equipment used in manufacturing microbeads is a syringe pump from KD Scientific® (Massachusetts, USA), an AC automatic voltage regulator from OET® (Australia), a high voltage gauge (Ormond Beach, Florida), a voltage regulating cable, an analytical scale, a magnetic stirrer, a hotplate (Gyeonggi-do, Republic of Korea), a homogenizer (Staufen, Germany), aluminum foil, a petri dish, a 23 needle, a 10cc/mL Terumo Syringe (Leuven, Belgium), parchment paper and other glassware.For grain analysis, the equipment used is Fourier Transform Infrared Spectroscopy (FTIR), UV-Vis Spectrophotometer, Zetasizer Light Scattering Instrument, SEM (Oregon, USA) and Oven.

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The nanoparticle formula optimization from Temulawak extract (Curcuma xanthorrhiza) as adjuvant therapy 51
2) Temulawak powder as much as 15% weighed and dissolved in 100 mL aquades then stirred using a glass rod.3) 3.5% alginate powder was weighed and dissolved in the remaining 150 mL of distilled water using a homogeniser at a speed of 4000 rpm.4) After the homogenized sodium alginate solution is produced, the temulawak solution is mixed.The mixture is then stirred with a homogeniser at a speed of 4000 rpm until thoroughly mixed for approximately 5 minutes.5) Next, the mixture is stirred continuously on a magnetic stirrer with a low temperature (the first temperature on the hotplate) for approximately 30 minutes.6) The prepared mixture is then set for the dripping process using a syringe pump.The syringe pump is equipped with a 10 cc/mL syringe, size 23 needle and collecting wire (black wire), and the distance between the tip and the collecting solution is set at a constant distance of 10 cm.7) A 2% w/v solution of CaCl2 is prepared by dissolving it in 250 mL of distilled water.It is used as a crosslinking solution for the formation of microbeads.8) Prepare a petri dish with aluminum foil for conductivity from the soil (green wire) and pour the poured CaCl2 into it along with a small magnetic stirrer.9) The mixture is sprayed into the CaCl2 solution with a certain flow speed and drop tension.10) The droplet flow rate is set first, then the voltage is set until the desired cone-jet is generated.Table 1 shows the droplet flow rate values and voltages used.11) The resulting microbeads are then stored in CaCl2 solution in falcon tubes.Alginat-Temulawak Analysis SEM Analysis 1) Some dry microbeads are taken for SEM analysis and taken to KOM for coating and analysis.

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The nanoparticle formula optimization from Temulawak extract (Curcuma xanthorrhiza) as adjuvant therapy 53 2) Morphological analysis of the surface of microbeads at a specific magnification.

UV-Vis spectrophotometry analysis
1) UV-Vis spectrophotometry is first set with the desired wavelength of 350 nm to 800 nm.
2) CaCl2 solution is prepared and used as a blank.
3) 1 mL of CaCl2 solution and 1 mL of wet alginate-curcumin microbeads in CaCl2 solution for each sample were then analyzed.
2) The filtered microbeads are transferred to parchment paper and put in the oven to dry at 37°C for approximately 24 hours.3) After drying is complete, several dry samples are taken for FTIR analysis.4) Samples of sodium alginate powder and temulawak powder were also prepared.5) This sample is carried out for FTIR analysis using FTIR spectroscopy.

Particle size analysis
1) 1 ml of alginate-curcumin solution is introduced into the cuvette.
2) The particle size distribution of the sample is analyzed.
Zeta potential analysis 1) Using a syringe, 1 ml of alginate-curcumin solution is introduced into the zeta cells of the Malvern Panalytical DTS1070 fold capillaries.
2) The zeta potential of the sample is analyzed.

RESULTS AND DISCUSSION
A nanoparticle formula was obtained from Temulawak extract material with parameters and characterization that have been analyzed and tested.

Visual observation
Alg-Curcuma was first synthesized at flow rates of 10, 20, 30, 40, and μL/min (first batch synthesis).However, the particle size varies greatly and can be observed with the naked eye.All the results for particle size analysis did not meet quality criteria due to the presence of large sedimenting particles.

Zeta potential analysis
Flow rate: 0.1 ml/hr *Graphs cannot be generated by the computer.
The zeta potential for other flow rates (0.2, 0.3 and 0.4 ml/hr) was not performed because the zeta potential data was unreliable.

Limitation
1) During the synthesis process, the voltage exceeds 25 kV, giving rise to an electric spark.Sometimes, even at 20 kV it can produce sparks.Therefore, there is a limit to obtaining the desired voltage at a given droplet flow rate.Therefore, optimized parameters may be difficult to obtain.2) The synthesized beads are microparticle in size.CaCl2 solutions containing microbeads are not colloidal and tend to settle quickly.3) During voltage adjustment, the size of the resulting microbeads varies.4) Dry samples do not spread well after drying.They become lumpy and need to be separated manually.This can affect the surface morphology of microbeads.

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The nanoparticle formula optimization from Temulawak extract (Curcuma xanthorrhiza) as adjuvant therapy 63 CONCLUSION Temulawak's sodium alginate synthesis results in microparticle size, not nanoparticle size.Microparticle sizes also vary thus contributing to high PdI which indicates a very wide particle size distribution.In addition, the zeta potential of the particle is 4.41 mV which means the suspension is unstable and the particles tend to aggregate.Based on the results obtained, it can be seen that temulawak sodium alginate microbeads are quickly precipitated and not well dispersed.Therefore, there are several recommendations that can be done to optimize the synthesis of temulawak sodium alginate.First, although ginger powder is dissolved in distilled water, it is not so stable to be homogenized well over a long period of time in alginate.This may be the main reason why synthesized alginate microgranules can precipitate quickly.Therefore, the use of ethanol as a solvent needs to be considered to dissolve Temulawak powder because it is also known as the best solvent.Secondly, since the nanoparticle size of temulawak alginate granules was not obtained, it is recommended to add chitosan to the solution because it can help reduce and control the resulting sample size (Phan et al., 2021).

Figure 2 .
Figure 2. Real Setup of Electrospray Experiment

Table 1 .
The Values of Dripping Flow Rate and Voltages Used Interdisciplinary Social Studies, Vol. 3, No. 4, January 2024 52 Figure 1.Schematic Diagram of Electrospray Experimental Setup

Table 1 .
Peak and absorbance observed for each flow rate.