{'fraction_non_alphanumeric': 0.034891835310537335, 'fraction_numerical': 0.016249626158907387, 'mean_word_length': 5.173538461538461, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 0, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
DISTRIBUTION AND COMPOSITION OF THE MAIN ACTIVE COMPONENTS FOUND IN STINGLESS BEE PROPOLIS FROM VARIOUS REGIONS IN INDONESIA

Objective: The aim of this study is to map out the distribution and composition of the main active components found in stingless bee propolis from various regions in Indonesia. Methods: The stingless bee propolis used was obtained from ten different provinces in Indonesia and the active components analysis using Gas Chromatography-Mass Spectrometer (GC-MS) pyrolyzer. Results: This study found 85 main types of active components with concentrations of ≥ 1%. The most frequently found active component w as alpha-d-glucopyranoside , which had an average concentration of 28.20%. Conclusion: There were differences between the main activ e components found in 14 samples of stingless bee propolis obtained from 10 provinces in Indonesia, which was due to the variety of bee species and plant origin.

Most stingless bee species have the potential to cultivate and produce high-quality propolis in large quantities, as much as 2.243 tons per 4 mo or 6.729 tons per year [3]. Propolis is a mixture of resin substances (plant sap), gum tree bark, and shoots of plants which are collected by bees, mixed with beeswax and bees saliva [4]. Propolis can strengthen the structural stability of bees hive to prevent decomposing of the inside. Currently, propolis is mainly used in the health industry as an anti-inflammatory and antibacterial treatment and also as antioxidant serum [5]. Stingless bee propolis can provide health benefits including the prevention and treatments of diseases and consumed in prescribed dosage [6,7]. Propolis has more than 300 different active components [8], with polyphenols (flavonoid, phenolic acid, and ester) as the main active components found in propolis, which are known to have antibacterial and antioxidant activities [9].
There are plenty of unidentified active components in stingless bee propolis due to various geographic locations, plant resins, and bees species [10]. Therefore, it is essential to discover the distribution and composition of the main active components found in stingless bee propolis from different regions in Indonesia.

Materials
The stingless bee propolis used were obtained from 10 different provinces in Indonesia, namely Tetragonula minangkabau and Sundatrigona moorei from North Sumatra, Tetragonula laeviceps from Banten, Tetragonula laeviceps from West Java, Tetragonula laeviceps from Central Java, Heterotrigona itama from West Borneo, Heterotrigona itama from East Borneo, Heterotrigona itama, Geniotrigona thorasica, Tetragonula laeviceps from South Borneo, Wallacetrigona incisa and Tetragonula biroi from South Sulawesi, Tetragonula fuscobalteata from West Nusa Tenggara, and Tetragonula fuscobalteata from North Maluku. These bees were harvested by bee farmers and delivered to Jatinangor, Sumedang. Materials for extraction and GC-MS Analysis, such as alcohol, ethanol, paraffin were obtained from Sigma-Aldrich, USA. Propylene glycol was obtained from Merck, USA.

Propolis extraction
The first step was mixing 1 kg of raw propolis (still in the process of glass transition) with ethanol 70% at a ratio of 1:2,5 (propolis: ethanol). Then, propolis was mashed into propolis pulp and filtered using a 30-mesh filter before being left for 12 h. The filtrate was separated while the rest of the propolis pulp was mixed with ethanol 70% at a ratio of 1:1,5 (propolis: ethanol), which was repeated 3 times. Afterward, the filtrate was condensed using a rotary evaporator at a maximum temperature of 50 °C, which was proceeded until the color of propolis extract turned dark brown, then it was mixed with propylene glycol and filtered using Whatmann 50 filter paper.

GC-MS analysis
This study used the GC-MS QP equipped with pyrolyzer, with the oven temperature set at 50 °C for 5 min, then raised up to 280 °C, with a pressure of 101 kPa, column flow 0,85 ml/min. MS detector was set at ion source temperature (200 °C), interface temperature 280 °C, detector temperature 280 °C, pyrolyzer temperature 300 °C. When stable,±1 µg/1 drop liquid propolis was injected into the pyrolyzer, and GC-MS started to operate for 50 min.

Distribution and composition of main active components of stingless bee propolis
The study found 85 types of main active components with concentrations of ≥ 1% in 14 propolis samples obtained from 10 provinces in Indonesia. Table 1 shows alpha d-glucopyranoside as the most frequently found substance, which was observed in 8 different propolis samples: H. itama from East Borneo, H. itama, T. laeviceps, and G. thorasica from South Borneo, W. incisa and T. biroi from South Sulawesi, T. fuscobalteata from West Nusa Tenggara, and T. fuscobalteata from North Maluku. This finding aligns with a previous study which found alpha d-glucopyranoside as the sugar component in propolis [11]. Details: • Percentage of a Compound The second most commonly found active component was the 1,6 anhydro beta d-glucopyranose which were observed in 7 different propolis samples: Banten, East Borneo, South Borneo, West Nusa Tenggara Barat, and North Maluku. This active component was also found in Algeria [12]. The hexadecanoic acid/palmitic acid came in third after being found in 7 different propolis samples obtained from West Lampung and South Lampung [13], Bursa-Orhangazi, Bartin, and Ankara-Mamak regions [14]. The fourth most frequently found component was the dodecanoic acid (lauric acid), found in 6 different propolis samples, and this active component was also found in Maribaya Bandung region [15]. The fifth most commonly found component was the tetradecanoic acid (myristic acid), found in 6 different propolis samples; this active component can also be found in Turkey (North-West Anatolia) [14].

Concentration of main active components of stingless bee propolis
Out of the 85 active components, the ten most commonly found were analyzed.

Plant origin of stingless bee propolis
Every bee species has its own plant source based on its region; thus the active propolis component varies. The variety is caused by the difference in tree type, temperature, region, and harvest time [17].   [20]. The difference of plants origin aligns with a study which proved the active propolis components were affected by the plants origin [21]. The higher resins in the plant, the stronger biological activity in the propolis.

Biological activities of the main active components in stingless bee propolis
Active components in stingless bee propolis have the potential to be further developed in Indonesia's healthcare sector. Based on the GC-MS analysis and library, there were 16 main active components with different biological activities (table 3).  [37], inhibit receptor signals, neutralize micro-toxin, and inhibit virulent factor secretion [38]. Another advantage of alpha-D-glucopyranoside is its ability to act as an antiemetic substance by reducing gastrointestinal hyperactivity. In a previous study, which was conducted by giving vomiting agents to chicks, flavonoid compounds showed an effect of reduction of the stomach's excessive movement [39].
One of the components which act as an immunostimulator is 1,6-Anhydro-Beta-D-Glucopyranose, which functions as an immune system inducer to increase T cells, which will release granules to hydrolyze the Mycobacterium tuberculosis (M. tbc) cell wall [27]. This component is also hepatoprotective, which means it can protect the liver from the toxic effect of antituberculosis drugs and maintain the liver's function, which in turn will result in the maintenance of appetite. The two main active components, alpha-D-glucopyranoside and 1,6-Anhydro-Beta-D-Glucopyranose, act as antioxidants which can reduce the radical compound 2,2-diphenyl-1-picrylhydrazyl (DPPH) [24].
Tetradecanoic acid and hexadecanoic acid are long-chain fatty acid compounds, an essential oil that works by damaging bacterial cell membranes [37]. Both of these components can also reduce the radical compound 2,2-diphenyl-1-picrylhydrazyl (DPPH) [40]. The component of dodecanoic acid/lauric acid acts as an antibacterial, which has more antibacterial effects on gram-positive bacteria compared to gram-negative bacteria [29].
2-methyl-furancarboxaldehyde has the ability to be antidiabetic, which was proven in a previous study by screening 2.4 derivatives of substitution of furan for its antidiabetic activity and compared it to standard Acarbose drugs (diabetes medications). The result showed that most of the active components were equal to those of Acarbose drugs [13]. Cycloartenol is found in Brazilian red propolis, which has been identified as having antibacterial, antimycotic, and antiradical activities, independently of its plant origin and chemical composition. Plenty of studies have shown that propolis has antimicrobial and antioxidant activities due to the role of stingless bee in the hive, which uses these active components to protect themselves against pathogenic microorganisms and weather elements [34].
The component octadecatrienoic acid, the main component of Bauhinia purpurea leaf extract, has shown the presence of antibacterial activity against two gram-positive bacteria (S. aureus and B. subtilis) and also has the potential to be used in the treatment of infectious diseases caused by microorganisms resistant to commercial antibiotic drugs [35].
The biological activities of some active components identified, namely the Hexanoic acid butyl ester and Acetoin, has yet to be known.

CONCLUSION
In this study, each of the 14 stingless bee propolis samples from 10 provinces in Indonesia had different main active components. The differences of the propolis LTS samples were caused by the variety of bee species and plant resins. The most frequently found active component was alpha-d-glucopyranoside, with an average concentration of 28,20%. The component can be utilized for its antimicrobial, antibacterial, hepatoprotective, and antifungal activities.