JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 524-532, E- ISSN: 2249 –1929 [DOI: 10.24214/jcbps.B.11.3.52432.]

Journal of Chemical, Biological and Physical Sciences

An International Peer Review E-3 Journal of Sciences Available online atwww.jcbsc.org Section B: Biological Sciences

CODEN (USA): JCBPAT Research Article

Proximate and microbial analysis of megono lactogenic canned packaging

Indar Widowati1, Asep Nurhikmat2, Afiyah Sri Harnany1 and Zaenal Amirudin1*

1Poltekkes Ministry of Health Semarang, Department of Nursing, Perintis Kemerdekaan Street and 51164 and Pekalongan, , , +62285421642 2Nature Materials Technology Research Institute of Science Indonesia - , Yogyakarta - Wonosari Street KM 31.5 Gading Village, Playen District, Gunung Kidul Regency, DI Yogyakarta,Indonesia, +62 274 392 570

Received: 13 June 2021; Revised: 24 June 2021; Accepted: 05 July 2021

Abstract: Research has been carried out on traditional megono lactogenic foods in cans as a solution for global food supply. The purpose of this study was to increase knowledge about the nutritional and microbial content of megono lactogenic. The Association of Official Analytical Chemists (AOAC) method was used for proximate analysis. While microbial analysis with Total Plate Count (TPC) to calculate the number of microbes in food. The basic proximate analysis revealed that the megono lactogenic contained protein (1.75%), fat (5.04%), water content (79.43%), ash content (3.11%), and carbohydrates (11.15%). Microbial analysis with the result 0 colony/gram anaerobic bacteria of the Clostridium parhingens, Salmonella sp, and Staphylococcus aureus groups. Megono lactogenic cans are suitable for consumption and developed for commercialization. Keywords: Proximate analysis, Microbial, Megono lactogen, Can packaging

1. INTRODUCTION

Food packaging and preservation technology continue to be developed by the Natural Material Technology Research Institute of the Indonesian Institute of Sciences (BPTBA-LIPI) Yogyakarta.

524 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 524-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al.

There have been 42 types of traditional archipelago culinary recipes packaged with this technology, namely Gudeg, , , Sayur Lombok Ijo, Mangut Lele, , and others[1]. This packaging technology has several advantages, including that food is more durable for more than 1 year, changes the taste, and is easy to carry anywhere including abroad and it is hoped that the traditional culinary of the archipelago can be worldwide[1]. The advantage of this packaging technology is that food is durable without preservatives for more than 1 year and does not change the taste. In addition, it is easy to carry outside the region and even abroad so that the traditional food of the archipelago can become global[1]. Megono is a typical Pekalongan food made from young mixed into small pieces, grated coconut, and spices, and shaped like a cone of rice. In the North Coast area of Java Island, this food used to be an offering to Dewi Sri, so that the harvest was abundant and the people lived prosperously[2,3]. Since Islamic teachings entered Pekalongan City, megono has become a treat in tahlil events at mosques around Pekalongan[2,3]. Megono lactogenic is a typical Pekalongan megono mixed with Sauropus androgynous leaf powder which is useful in increasing breast milk production and spurring baby growth[4]. This Megono has a short shelf life of only a few hours 8 to 12 hours. Several studies on the packaging process have been carried out, including: Effect of Temperature Bonding and Sterilization on Fo Value and Physical Condition of Gudeg Cans[5]; and empal cans are typical Cirebon food packaging[6]. Research on canned megono[4]: as well as lactogenics have been shown to increase breast milk production and stimulate infant growth[7]. In this research, an analysis of the megono-lactogenic quality in cans will be carried out as a basis for starting the production and marketing of megono-lactogenic products in cans more broadly. Processed vegetables suffer some loss of quality during processing and storage. The main quality attributes of vegetables undergoing thermal processing include color, aroma, taste, and texture. In addition, there are hidden quality attributes such as nutritional value and safety factors, namely chemical and microbial[8,9]. In this study, we will analyze the nutritional and microbial values.

2. MATERIALS AND METHODS

2.1. Materials:

Proximate Analysis materials and tools: the tools used in this study include 50 mL Burette, Porcelain Cup, Desiccator, Erlenmeyer, Hot Plate, Kjeldahl flask, Electric Oven, One Set of Distillation Equipment, One Set of Soxhlet Equipment, Electric Furnace, Analytical Balance. The materials used include: Boric Acid (H3BO3), Hydrochloric Acid, Sulfuric Acid (H2SO4), Alcohol, Phenolphthalein Indicator, Filter Paper, Methyl Red, Petroleum ether, Mixed Bromocresol Green, Sodium Hydroxide (NaOH), and food ingredients (megono lactogenic in cans). Microbial Analysis Tools and Materials: Tools for microbial analysis, including petri dish, incubator, measuring cup, Erlenmeyer 100 ml, test tube, test tube rack, Bunsen. The materials used include physiological NaCl, canned megono lactogenic

2.2. Methods:

2.2.1. Proximate Analysis: Samples were prepared by crushing the sample with a food processor. The sample is processed into fine and small particles (± 30 – 60 seconds), then the sample is transferred using a spoon. Water content, Indonesian National Standard/SNI (01-2891-1992)[10]. Take

525 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al. an empty cup in an oven at a temperature of 105oC, cool it in a desiccator for 5 minutes, weigh the weight of an empty cup, add 3 grams of sample, in an oven for 3 hours at a temperature of 100 - 105oC, cooled in a desiccator for 15 minutes, weighed, in the oven for approximately one hour at a temperature of 100-105°C, until the weight remains weighed (weight difference ± 0.02 mg). Then calculated by the formula

W = sample weight before drying (g); W1 = sample weight after drying (g) The cup is flattened at 105oC for 3 hours, cooled in a desiccator for 15 minutes, the weight of the empty cloud is weighed, add about 3 grams of the sample while 2-3 drops of alcohol are dripped, the sample is lit on the stove until it becomes charcoal and does not smoke, put in a silencer until the sample is in the form of white ash (for 6 hours, at a temperature of 600°C, 7) is put in a desiccator for 115 minutes, weighed and the weight is recorded.

Then it is calculated by the formula

W = sample weight (g); W1 = fat weight before extraction (g); W2 = weight of fat flask after extraction (g)

Take a sample of 0.5 grams, add 1 gram of Kjeldahl tablet and 15 ml of H2SO4, digest for 2 hours, cool, put into a 100 ml volumetric flask and add distilled water to volume, pour into a Kjeldahl flask, add 3 drops of PP and 15 NaOH 50 ml of distillate, the distillate container was filled with 25 ml of 3% boric acid, 80 ml of distillate was accommodated, the distillate was titrated with HCL until the color changed to yellow.

Then calculated by the formula

W = sample weight (g); V1 = 0.01 N HCl volume for titrated sample (ml) V2 = 0.01 N HCl volume for sample titration (ml); N = normality of HCl; f.k = conversion factor (6.25); f.p = dilution factor Weigh the sample about 3 grams, weigh the empty fat flask, install soxhlet, add hexane solvent 2 times, heat for 4 hours, remove lead, evaporate solvent and collect, put the song in the oven, flask weighed after being in the oven. Then calculated by the formula

W = sample weight (g); W1 = fat weight before extraction (g); W2 = weight of fat flask after extraction (g)

Take one gram of sample, add 100 ml of H2SO4 (0.225 N) and bring to a boil, filter with filter paper (in the form of liquid), the residue on the filter paper is washed with 100 ml of hot distilled water, the residue that has been washed add 100 ml of NaOH (0.225 N), bring to a boil for 15 minutes, filter

526 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al. using what paperweight is known (liquid sample removed), add 5 ml of K2SO4 10%, 9) add 10 ml of boiling distilled water and 95% alcohol (15 ml), oven-dried paper (105oC ), for 1 hour. Then calculated by the formula:

W = sample weight (g); W1 = ash weight (g); W2 = weight of sediment (g) The chemical analysis includes determination of moisture content by thermogravimetric method, ash content by direct washing method, total protein content by micro-Kjeldahl, total fat content by Soxhlet extraction method, and carbohydrate content by difference method[11]. 2.2.2 Microbial Analysis: Total Plate Count (TPC) is a method used to calculate the number of microbes in foodstuffs that can be seen directly with the eye. Nutrient (NA) (used to calculate the total bacteria using the cup method[13]. Weigh 1 gram of the ingredients then enter the Erlenmeyer, add 9 ml of phosphate buffer, shake and leave for 2-3 minutes. Make a dilution of 10-1, 10-2, 10-3, 10-4, 10-5 then inoculate 1 ml of the 10dilution-5 into a petri dish, add warm PCA medium (40°C), and shake until evenly distributed. Incubate at 30°C for 72 hours, the cup upside down. Count colonies by the standard plate count method.

3. RESULTS AND DISCUSSION

The results of the proximate analysis of samples A and B according to the Indonesian National Standard (SNI) 01.2343-1991: Canned Food Inspection [14], are shown in Table 1.

Table 1: Proximate analysis test result

Parameter Unit Deuteronomy Amount Average

I II III

Water Content % 78.95 79.28 80.05 238.29 79.43

Ash Content % 3.11 3.09 3.09 9.30 3.10

Protein Content % 1.74 1.57 1.97 5.29 1.76

Fat Content % 5.04 3.92 3.12 12.09 4.03

Carbohydrat tes % 11.14 12.11 11.75 35.01 11.67

3.1. Analysis Proximate: Table 1. it can be seen that each sample (I, II, and III), which is included in the category of food high bulk density, has a fairly high water content with an average of 79.43%. Ash content (mineral) 3.10%, protein 1.76%, fat content 4.03% and carbohydrates 11.67%. 3.2 Analisis Mikroba: Microbial analysis was carried out quantitatively, with the results of 0 colonies/gram with a dilution factor of 10 x. This means that canned megono lactogenic does not contain anaerobic bacteria belonging to the Clostridium parhingens, Salmonella sp, and Staphylococcus aureus groups (Table 2). 527 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al.

Table 2: Microbial analysis test result

Parameters Unit Result Microbial Contamination: Total plate number 30°C, 72 hours colony/gram < 10 Thermophilic anaerobic bacteria colony/gram < 10 Spore-forming Thermophilic Anaerobic colony/gram < 10 Bacteria (BATPS) Coliform APM/gram < 3 Escherichia coli APM /gram < 3 Clostridium parhingens* /gram negative Salmonella sp /25 gram negative Staphylococcus aureus* /gram negative

Analysis was Microbial Carried out quantitatively, with the result 0 colonies/gram with a dilution factor of 10x. This means that megono lactogenic canned does not contain anaerobic bacteria Clostridium groups belonging to theparhingens, Salmonella sp, and Staphylococcus aureus. The results of laboratory tests prove that the largest content is water, as 79.4% is relatively high. This happens because during the canning process a product will experience an exhausting stage. Exhausting is the process of emptying air on lactogenic megono that has been put in a can and has not been closed. This process is to reduce or eliminate air gas contained in the packaged material because the air is a source of contamination. Exhausting was carried out at 121.10C, for 10 minutes, to destroy microorganisms. This is evidenced by the results of standardized analysis of canned megono that the product does not contain anaerobic bacteria Clostridiumgroups belonging to theparhingens, Salmonella sp, and Staphylococcus aureus. Thus, canned megono products are safe for consumption by the public. In general, canning a food product will affect the nutritional value as follows. The quality of a food depends on the nutritional content and organoleptic properties of the food. Nutrient content relates to protein, fat, carbohydrates, vitamins while organoleptic properties relate to color, texture and taste. Food quality will decline when the food is processed and will continue as long as the food is stored. A measure of the quality of a food is sensitive to acidity, oxygen, heat and light. By knowing the decline in quality, optimal processing and storage conditions can be developed [15,16,17]. Proximate analysis includes: water content, ash, protein, fat, fiber and extracts without nitrogen. Water content is the main component of food products, because it can affect the color, texture and taste. The water content in foodstuffs determines the freshness and durability of foodstuffs, so that water becomes very important in food ingredients or products[18]. Water content analysis is a fundamental and important analysis of food products[19,20]. The dry matter content in foodstuffs is inversely proportional to the water content, therefore the water content becomes very important economically for food producers[21]. Reduction of water in food products aims to preserve, reduce the size and weight of food ingredients so as to simplify and save packing[18]. Determination of water content in food or food products is very important, because it determines the processing and distribution process so that it gets proper handling[22,23]. Water content in food products, can affect chemical changes and 528 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al. determine microbial content kandungan[24]. Ash content is a residue produced in the combustion process of organic matter, in the form of inorganic compounds in the form of oxides, salts and minerals. High ash content in food products is one indicator that the product has the potential to be dangerous when consumed. High ash content indicates the presence of metal elements in food products[25]. Determination of ash content is intended to determine the content of non-volatile components (inorganic components or mineral salts) that remain on burning and igniting organic compounds[26]. The lower the ash content of a material, the higher its purity. The high and low ash content of a material is caused, among other things, by the different mineral content in the source of the raw material and can also be influenced by the demineralization process at the time of manufacture[25]. Total minerals are relatively stable, unaffected by heat conditions during the canning process, but are susceptible to changes in bioavailability due to interactions with other food components[27). Vitamins are unstable in hot conditions during the canning process. Fat-soluble vitamins are more stable than water-soluble vitamins. However, vitamin loss can occur during canning due to oxidation[27). Ash content can be used in determining whether or not a processing is good, knowing the type of food ingredients used, and determining nutritional value parameters[28). One component of macronutrients is protein, which plays an important role in the formation of biomolecules, apart from being a source of energy[18]. In addition, protein also acts as a regulatory and building agent. Proteins contain the elements carbon (C), hydrogen (H), and oxygen (O), nitrogen (N) as well as phosphorus, sulfur and other types of protein containing metal elements, including iron and copper[18]. The protein content in canned megono lactogen comes from the basic ingredients, namely young jackfruit and added with protein derived from kitchen spices. The protein content of structured foodstuffs depends on the additional amount of ingredients which is also influenced by the water content[28]. In addition, the vegetable canning process will cause a decrease in protein quality, due to the high temperature resulting in the Maillard reaction. This reaction will cause a lysine loss of up to 40%[27]. Fat is an important food substance to maintain the health of the human body which is composed of the elements carbon (C), hydrogen (H), and oxygen (O). Fat is insoluble in water but soluble in solvents of hexane, ether, benzene, and chloroform. Fats are esters of glycerol and fatty acids[18]. Fats and oils are found in almost all types of food with different amounts of content[29]. For this reason, it is very important to analyze fat content in foodstuffs to determine the calculation of calorie needs. Most foods will undergo lipid oxidation during the canning process. Saturated fatty acids are relatively more stable at certain temperatures in the canning process, but saturated fatty acids will be degraded in the heating process due to oxygen and temperature, this can affect the taste of a food product[17]. Explained that carbohydrates are divided into two types, namely simple carbohydrates and complex carbohydrates[29]. Simple carbohydrates can be found in food products, including honey, fruits and milk. Complex carbohydrates are found in food products, including rice, potatoes, corn, bread and other complex carbohydrates, namely starch, glycogen, cellulose, and fiber. Carbohydrates play an important role in determining the characteristics of food products, such as taste, color and texture. Carbohydrates in the human body can be formed from several amino acids and partly from fat glycerol[18]. Carbohydrates are useful for preventing the onset of ketosis, protein breakdown, and helping protein and fat metabolism. Carbohydrates have decomposition products, namely glucose, sugar phosphate, pyruvic acid and lactic acid[31]. During the vegetable canning process, carbohydrate content is not affected, but

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Proximate… Indar Widowati et al. nutritional value is affected by interactions with other food ingredients, constituents and overall food quality[27,31]. Megono lactogen cans still contain protein, fat, water, ash and carbohydrates even though they are degraded during the heating process. This proximate content is not only derived from its basic ingredients, namely young jackfruit, it also comes from the addition of other food ingredients, such as grated young coconut, sugar, salt and spices

4. CONCLUSION

Megono lactogenic cans contain nutrients: water content, ash content, protein, fat and crude fiber. This food product is suitable for consumption and development because it does not contain anaerobic bacteria belonging to the Clostridium parhingens, Salmonella sp, and Staphylococcus aureus groups.

ACKNOWLEDGEMENTS

We thank you very much for getting support from the Center for Research and Development of the Health Polytechnic of the Ministry of Health Semarang. Guidance from the Research Institute for Natural Materials Technology, Indonesian Institute of Sciences - Yogyakarta, as well as cooperation from CV. Uniform Tester Lab .

REFERENCES

1. H.Julendra, 42 Types of Traditional Foods Already Packaged with Packaging Technology, 2017.LIPI News.

2. M.A.Pribadi, Pekalongan, Formerly an Offering for Dewi Sri. 2017. wartakota. tribunnews.com

3. L.Nurhadi, Recipes and How to Make Megono Typical Pekalongan, 2019. Yogyakarta: wartakota. tribunnews.com.

4. I.Widowati, Hartati, N. Asep and Z.Amirudin, Development of Lactogenic Megono Local Foods to Increase Breast Milk Production and Promote Baby Growth. Pekalongan City Research and Development Journal,2019a, 16 (1): 65-80.

5. Nurhikmat, A., Suratmo, B., Bintoro, N., Suharwadji, the Effect of Temperature and Time of Sterilization on the F Value and the Physical Cans Conditions in Canned Gudeg. Agritech, 2016, 36(1), pp. 71-78.

6. A.Nurhikmat, A. Susanto, A. Kusumaningrum, A.F. Amri and E.Masruroh, Empal gentong and empal asem with packaged cans: traditional foods from Cirebon. IOP Conf. Series: Earth and Environmental Science, 2020b,462. Cirebon, Indonesia: ICONPROBIOS.

7. I .Widowati, Hartati, A. Zaenal, Traditional Culinary Food Packaging: Megono As An Effort To Extend The Shelf Time And Product Competitivenes. Pekalongan City Research and Development Journal, 2019b,15(1):17-25.

8. J.Ahmed and U. Shivhare, Thermal Kinetics of Color Change, Rheology, and Storage Characteristics of Puree/Paste. Journal of Food Science, 2001, 66: 754-757. https://doi.org/10.1111/j.1365-2621.2001.tb04633.x

530 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al.

9. K.A.Amania, The effect of the level of sterility on the canning process on the physical properties of the resulting gudeg. Bogor Agricultural University Indonesia: Thesis,2011, Faculty of Agricultural Engineering. http://repository.ipb.ac.id/handle/123456789/47301

10. National Standardization Agency, 1992, Document SNI-01-2891-1992-cara-test-food- beverage-55c8046400086: How to Test Food and Drinks. Indonesia: National Standardisation Agency.

11. Association of Official Analytical Chemists (AOAC), Official Method of Analysis. London: Arlington; AOAC International,2020

12. N.G.Asp, T.F. Schweizer, D.A.T.Southgate, O. Theander, Dietary Fiber Analysis. In: Schweizer T.F., Edwards C.A. (eds) Dietary Fiber — A Component of Food. ILSI Human Nutrition Reviews. Springer, London, 1992, 57-101. https://doi.org/10.1007/978-1-4471- 1928-9_4

13. M.J.Pelczar, University, industry, government partnerships for improvement of response to social needs. Mircen Journal,1986, 2: 51–59. https://doi.org/10.1007/BF00937183

14. Ministry of Agriculture, Decree of the Minister of Agriculture. Indonesia:1998, No. 481/Kits/OT.210/5/98: Application of the Indonesian National Standard (SNI) for Agricultural Commodities.

15. N. Cayot, Sensory quality of traditional foods. Food Chemistry, 2007, 101(1): 154–162.

16. D.Lund, Effects of Heat Processing on Nutrients. In: Karmas E., Harris R.S. (eds) Nutritional Evaluation of Food Processing. Springer, Dordrecht, 1988. https://doi.org/10.1007/978-94- 011-7030-7_12.

17. G.Awuah, H. Ramaswamy & A. Economides, Thermal processing and quality: Principles and overview. Chemical Engineering and Processing, 2007, 46, 584-602.

18. F.G.Winarno, Food Chemistry and Nutrition. P.T, Main Library Scholastic,2002.

19. G.K.Harris, M.R.Marshall, Ash Analysis. In: Nielsen S. (eds) Food Analysis. Food Science Text Series. Springer, Cham.2017, https://doi.org/10.1007/978-3-319-45776-5_16

20. R.L.Bradley, Moisture and Total Solids Analysis. In: Food Analysis. Springer, Boston, M.A.2010, https://doi.org/10.1007/978-1-4419-1478-1_6

21. Y.W.Park, Moisture and Water Activity. In: Handbook of Processed Meats and Poultry Analysis. L. Nollet and F. Toldra, Eds., CRC Press. Boca Raton, FL.2008, Pp. 35-67

22. E.S.E.Hafes, Proximate Analysis Method. Jakarta, Indonesia,2000, : Erlangga

23. M.S.Fedha, M. A. Mwasaru, C.K. Njoroge, N.O. Ojijo and G.O.Ouma, Effect of drying on selected proximate composition of fresh and processed fruits and seeds of two pumpkin species. Agriculture and Biology Journal of North America, 2010, 1 (6): 1299-1302

24. H.Herawati, Determination of shelf life in food products. Journal of Agricultural Research and Development,2008, 27(4):124–130.

25. S.Sudarmadji, B. Haryono, Suhardi, Analysis of food stuffs and agriculture, 2010, 2 ( 4): Liberty, Yogyakarta

531 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]

Proximate… Indar Widowati et al.

26. M. Nurilmala, M. Wahyuni, H. Wiratmaja, Improvement of the added value of tuna bone waste into gelatin and Physico-chemical analysis of bone waste improvement Tuna (Thunnus sp) Becomes Gelatin and Physics-Chemistry Analysis. Bulletin of Fishery Products Technology, 2006, 9(2): 22-33.

27. P.Jason, Climate tolerance and interspecific variation in geographic range size. Publisher: The Royal Society,2003, 270: 475-481

28. J.G.Sebranek, Basic Curing Ingredients. In: Tarté R. (eds) Ingredients in Meat Products. Springer, New York, NY.2009. https://doi.org/10.1007/978-0-387-71327-4_1

29. S.Hermanto, A. Muawanah, P. Wardhani, Analysis of the Level of Damage to Vegetable Fats and Animal Fats Due to the Heating Process. Journal of Valence Chemistry, 2010, 1 (6): 262– 268. DOI: 10.15408/jkv.v1i6.237

30. S.Almatsier, Basic Principles of Nutrition Science. Gramedia Pustaka Utama. Jakarta, Indonesia, 2009.

31. H.E.Irianto, S. Giyatmi, Fishery Products Processing. Open University Publisher, Jakarta, Indonesia, 2000.

Corresponding author: Zaenal Amirudin,

Poltekkes Kemenkes Semarang, Department of Nursing, Perintis Kemerdekaan Street and 51164 and Pekalongan, Central Java, Indonesia, +62285421642 [email protected]

Online publication Date: 07.07.2021

532 JCBPS; Section B; May 2021 –July 2021, Vol. 11, No. 3; 424-532. [DOI:10.24214/jcbps.B.11.3.52432.]