1 2 1 Digestibility of Ration was Added Nanocapsule of Turmeric Extract

2 Using Chitosan Cross-Linked with Sodium Tripolyphosphate on Broiler 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17ABSTRACT. The Use of turmeric is restricted by its low solubility in water, 18therefore it has low bioavailability. This obstacle can be solved by the 19development of nanoparticle technology to improve drug delivery profile. This 20study aimed to develop nanoparticle formulation using turmeric extract and 21industrial chitosan as the matrix and sodium-tripolyphosphate as cross linker, 22to study its ability to improve feed digestibility. Method used in the formulation 23of nanoparticle in this study was by ionic gelation followed by oven drying at 2450C. Method used to evaluate the digestibility
was total collection. One
3) 2) 1)

Chicken By Sundari1*, Zuprizal2, Tri-Yuwanta2, Ronny Martien3 Departement of Animal Husbandry , Faculty of Agroindustry, Mercu Buana University Yogyakarta. Jl. Wates km 10, Sedayu-Bantul 55753, Indonesia. Tilp 02746498212, Fax 02746498213. HP 081328746141. Faculty of Animal Science ,Gadjah Mada University. Jl. Fauna 3, Bulaksumur Yogyakarta 55281, Indonesia. Tilp. 0274-513363, Fax. 0274521578. Departement of Pharmaceutics, Faculty of Pharmacy ,Gadjah Mada University. Sekip Utara Yogyakarta 55281, Indonesia. Tilp.+62274 542739, Fax .+62274 543120. *Corresponding Author e-mail: sundari_umby@yahoo.com

25hundred and twenty broiler chickens with an average body weight of 900 g, 26were randomly divided into 20 treatments (one treatment was fasted and 19 27were treated with the ration and feed additive),
and six replicates were

2 1performed on each test. Chickens were fasted one day before and after they 2were treated with rations. Nutrient levels and the amount of feed consumed 3and excreta released were weighed to calculate the digestibility of the ration. 4It was found that the basal ration had dry matter digestibility of 70.48% 5significantly smaller (P<0.05) than 75.90% digestibility of the ration added 6with nanocapsule turmeric extracted by aquades 0.4% level. The result of this 7study concluded that formulation of nanocapsule using turmeric extract and 8industrial chitosan as the matrix and sodium tripolyphosphate as cross linker 9was potential to increase nutrient digestibility so can be an alternative for feed 10additive on broiler chicken. 11Key words: digestibility, nanocapsule, turmeric-extract, industrial-chitosan, 12broiler chicken. 13 14 Kecernaan Ransum yang Ditambah Nanokapsul dari Ekstrak Kunyit 15 16 17 18ABSTRAK. Penggunaan kunyit / kurkumin terbatas karena kelarutannya yang 19rendah dalam air sehingga bioavailabilitasnya juga rendah. Masalah ini dapat 20diatasi dengan teknologi nano yang dikembangkan sebagai alternatif 21penghantaran obat bagi bahan kimia yang mempunyai bioavailabilitas rendah. 22Penelitian
ini bertujuan untuk mengembangkan formulasi nanokapsul dengan Kitosan Berikatan Silang Sodium Tripolifosfat pada Ayam Broiler.

23menggunakan ekstrak kunyit dan kitosan industry (tehnis) sebagai matrik atau 24polimer serta sodium tripolifosfat sebagai cross linker, untuk dipelajari 25kemampuannya dalam meningkatkan kecernaan ransum. Metode yang 26digunakan dalam formulasi nanokapsul adalah gelasi ionik dilanjutkan

2 1pengeringan dengan oven pada suhu 50C. Sedangkan metode untuk 2mengevaluasi kecernaan menggunakan total koleksi. Seratus dua puluh ekor 3ayam broiler dengan bobot badan rata-rata 900 g, dibagi secara acak ke 4dalam 20 perlakuan (satu perlakuan dipuasakan dan 19 diberi ransum 5perlakuan dengan penambahan feed additive), masing-masing enam ulangan 6dan satu ekor untuk tiap ulangan. Ayam dipuasakan sehari pada sebelum dan 7sesudah ayam diperlakukan dengan ransum, kadar nutrien dan jumlah pakan 8yang dikonsumsi serta ekskreta yang dikeluarkan diketahui beratnya untuk 9menghitung kecernaan ransum. Telah ditemukan bahwa kecernaan bahan 10kering ransum basal (70,48%) nyata lebih kecil (P<0,05) dibanding ransum 11basal yang ditambah nanokapsul kunyit ekstrak air (EA) dengan level 0,4% 12sebesar
(75,90%). Dapat disimpulkan bahwa formulasi nanokapsul

13menggunakan ekstrak kunyit dan kitosan industry sebagai matrik dan 14sodiumtripolifosfat sebagai cross-linker berpotensi sebagai feed additive 15alternatif untuk meningkatkan kecernaan nutrien pada ayam broiler. 16Kata kunci: kecernaan, nanokapsul, ekstrak-kunyit, kitosan-industri, ayam 17broiler. 18Introduction 19
Antibiotics are widely used to gain higher income in broiler farming as

20growth promotors and high-fat or high-energy ration. Antibiotics has facilitated 21the efficient production of poultry, allowing the consumer to purchase at a 22reasonable price on high quality meat and eggs (Donoghue, 2003). A wide 23range of antibiotics are used in poultry not only to treat disease but also to 24maintain health, promote growth and enhance feed efficiency (Gaudin et al., 252004). In particular, broiler chicken are often grown actively with antibiotics to 26attain maximum weight within a short period of time (Nonga et al., 2009). The

2 1uncontrolled and unlimited use of these antibiotics may however lead to the 2accumulation of undesirable residues in the animals treated and their products 3(Wachira et al., 2011). 4
Coronary heart disease and arteriosclerosis are strongly related to the

5dietary intake of cholesterol and saturated fatty acids and are among the most 6important causes of human mortality (Sacks, 2002 cit. Omojola et al., 2009). 7The use of high fat or high energy on ration causes cholesterol content of thigh 8meat 87.6 mg/100 g on broiler chicken (Daneshyar et al., 2011) and at local 9poultry in Indonesia include Nature chicken (177.47 mg/100 g), Tegal duck 10(166.91 mg/100 g) and Muscovy duck (171.94 mg/100g)(Ismoyowati and 11Widiyastuti, 2003). Controversy over the use of aforementioned antibiotics and 12high-energy rations call for an effort to find a feed additive from natural 13ingredients that have the potential to substitute the function of antibiotics as 14well as to lower cholesterol. One potential herbal medicine in Indonesia is 15curcumin which is the main active ingredient of turmeric rhizoma. Several 16studies in vitro and in vivo studies have shown that turmeric activities as: 17antibacterial, anti-inflammatory, antitoxic, antihiperlipidemia, antioxidant and 18anticancer, but curcumin has low bioavailability (low solubility, poor 19absorption, fast pass, high rates of metabolism in the gut cells, rapid 20elimination) (Anand et al., 2007). One reason for the low bioavailability of 21curcumin is that it is water insoluble at acidic or neutral pH, and this causes 22absorb difficulty (Maiti
et al., 2007). This obstacle can be solved by the

23development of nanoparticle technology to improve drug delivery profile, 24especially for the less bio-available chemical. Therefore, the application of 25curcumin
needed polymers capable of carrying and being delivered to be

26absorbed properly, such as chitosan by nanotechnology (nanoencapsulation).

1 2 1

5 This study was aimed to develop nanoparticle formulation using and industrial chitosan as the matrix and sodium

2turmeric extract

3tripolyphosphate as cross linker, to study its ability to improve feed 4digestibility on broiler chickens. Furthermore, it is to see its potential as a feed 5additive for antibiotics substitute as well as lowering cholesterol meat. 6Materials and Methods 7
The nanocapsule with formula 221 was created by ionic gelation method

8using ratio turmeric extracted : chitosan : TPP (sodium tripolyphosphate) = 2% 9: 2% : 1% (w/v). Two kinds of nanocapsule (400-870 nm) were used from: (1). 10Turmeric extracted and dissolved by ethanol (EE) then added with chitosan 11dissolved in buffer acetic pH 4 by magnetic stirrer for 20 minutes, and (2). 12Turmeric extracted and dissolved by aquades (EA) and chitosan dissolved by 13citric acid pH 4 by magnetic stirrer for 20 minutes and then both added with 14TPP was dissolved in aquades by magnetic stirrer for 20 minutes. Furthermore, 15the dispersed mixture was precipitated, filtered and oven-dried at 50 C. 16
A total of 120 broiler chickens weighing 900 g were placed in individual

17cages equipped with feed and water. Provision of feed additive has been done 18on 20 groups of chickens ( 1 group fasted and 19 groups feeding treatments 19shown in table 2) with 6 replications, each replication using 1 tail in each 20replication. Total collection method was appliedin which, the first 10-day 21adaptation period followed the total collection. All chickens were fasted but 22drink water ad-libitum on the first day of total collection. On the second day, 2319 groups of chickens were given fed basal rations (table 1) treated plus feed 24additive and 1 group remained fasting to test endogenous N. On the third day 25all chickens were fasted again. Excreta were collected from the second to the 26third day (completed). On the fourth day all chickens were slaughtered take

2 1the contents of intestinal ileum. Nutrient content was analyzed from the 2samples of feed and excreta or ileal (AOAC, 1995) to determine the effect of 3addition of nanocapsule feed additives on the digestibility of the ration. 4
Nutrient digestibility. Samples of feed given (consumed) and

5excreta were removed, weighed, dried and further ground to pass 1 mm 6sieve, then swabbed 40 g, included in labeled plastic bags and stored 7at 4 C for nutrient analysis use. 8Nutrient digestibility was calculated by the formula of as follows : 9 Nutrient digestibil ity =

( Nutrient

int ake ) ( Nutrien excreta Nutrient int ake

) x 100 %

(1)

10Description: 11 nutrients intake is feed intake multiplied by the feed nutrient levels 12 nutrients excreta is out excreta multiplied by excreta nutrient levels 13 14
Protein digestibility particularly used ileal digestibility of protein

15(Lee et al., 2004 modified Julendra, 2010). To determine the amount of 16crude protein in the excreta, ileal digestas protein value was multiplied 17by the dry matter excreta. Difference in crude protein intake with 18protein wasted in excreta (undigested) is called crude protein digestible 19(Lee et al., 2004). The formula referring 20Julendra (2010) are as follow : 211. 22 23 24 252. 26
True protein digestibility:
CP
True

to protein digestibility in

CP fi ( DM E xCP ile DM em xCP ilem CP fi

x100 %

..(2) Apparent protein digestibility

CP
Apparent

CP fi ( DM E xCPile ) CP fi

x100 % .

27..(3) 28 29 Specification formula:

1 2 1 2 3 4 5 6
CPTrue = true crude protein digested (%), CPApparent = apparent crude protein digested (%),CPfi =crude protein feed intake (% DM), DME = dry mattter excreta (% DM), CPile = crude protein ileum (% DM), DMem = dry matter excreta metabolic /chicken fasted (% DM), CPilem = crude Protein ileum metabolic /chicken fasted (% DM).

7 Insert table 1 about here 8

The data nutrient digestibility was analysis by analysis of variance

9and further significant differences were analyzed post hoc by List 10Significant Difference used SPSS-16 computer program . 11RESULT AND DISCUSSION 12
Nutrient digestibility of ration given a feed additive nanocapsule

13of turmeric extract using chitosan cross linked sodium tripolyphosphate 14shown in table 2. Ration was added with chitosan 0.1% (T2) indicated 15that it can lower fat digestibility compared to Basal ration (T1), because 16chitosan can bound fat feed to excreta. This is in line with an opinion 17that Chitosan from shrimp shell can bind fats (FBC) averaging 416.5% 18for soybean oil, 503% for corn oil, and 400.8% for the sesame oil, FBC 19value of shrimp shell chitosan origin is higher than the original 20commercial FBC crab shell chitosan (Sigma Co.)(Sofia et al., 2010). 21Ration added with
0.1% turmeric extract (T3) indicated to improve

22digestibility of dry matter, organic matter, and protein compared basal 23ration (T1), because the turmeric extract contains curcumin that lowers 24peristaltic intestine that lengthens digestive enzyme stimulation. This is 25in line with an opinion that curcumin may increase the secretion of bile 26and
pancreatic lipase activity increased, amylase, trypsin and

27chemotrypsin (Chattopadhyay et al., 2004). In mice and rabbits, 28curcumin can stimulate an increase in relaxation intestine resulting 29longer food in the small intestine and stimulates the secretion of 30hormones from the brunner glands of the small intestine (Martini (1998)

1 2 1cit. Sinaga, 2010). Curcumin can affect the tone and contraction of the 2intestine, given at the low-dose and repeated will accelerate contraction 3intestinal tone, at high doses it will slow down and even stop the 4contractions of the small intestine, while at the right dose it will cause 5spontaneous contraction, i.e. digestibility and absorption of food will 6increase (Gupta et al. 1980 cit. Sinaga, 2010). The same finding was 7reported by Rao et al. (2003) that curcumin increases spending 8stimulation of pancreatic enzymes and small intestine and decreased 9intestinal peristalsis, thereby providing nutrients absorption of products 10of digestion more. Digestibility of dry matter ration plus 0.4% (T 18) of 11the nanoparticles using extracted turmeric by aquades (EA) and 12chitosan dissolved by citric acid were significantly (P <0.05) better than 13P13 using extracted turmeric by ethanol (EE) and chitosan dissolved in 14acetic acid or the other rations. It is assumted that EA nanoparticles 15solubility in water makes it easier to interact with intestinal cells as well 16as in bacterial cells. The same finding was reported by Islam et al, 17(2008) that supplementation 0.5% citric acid (B) on drinking water of 18broiler chickens shown positive effect on live weight, feed intake and 19feed conversion efficiency with no detrimental effect on carcass 20characteristics when compared with 0 % citric or acetic acid (A) , 0.5% 21acetic acid (C) and their combinations 0.5% citric acid and 0.5% acetic 22acid (D). Ration added with TPP 0.1% (T4) indicated that it can improve 23digestibility of dry matter, organic matter, and fat but decrease protein 24digestibility compared to basal ration (T1), this is possible because the 25TPP has alkalin pH 9.5 to 9.9 (Bhumkar and Pokharkar, 2006) and is 26potential to inactivate protease in proventriculus and activate lipase in 27intestine. Ration added with nanoparticle (NP) using either ethanol or

1 2 1aquades extract turmeric (T9 T19) indicated that it can improve 2digestibility of dry matter, organic matter, protein and fat compared to 3basal ration (T1), ration combinations (T5-T8) of the building blocks of 4nanoparticle, both with and without the encapsulation process showed 5better digestibility than the basal ration. 6Insert table 2 about here 7CONCLUSIONS 8

The successfully created nanokapsule of turmeric extract with chitosan

9industry as matrix and sodium tripolyphosphate technical as cross linker, gave 10new hope as an alternative feed additive as replacement for antibiotic growth 11promoters as well as lowering cholesterol meat, it was evident that the 12addition of nanokapsule to ration could improve ration digestibility including : 13dry matter, organic matter, crude fat, and protein. 14 15 REFERENCES 16 17Anand, P, AB Kunnumakkara, RA Newman, and BB Aggarwal. 2007. 18 19 20 21AOAC. 1995. Official Methods of Analysis. 16th ed. Association of Analytical 22 23 24Bhumkar DR and VB Phokarkar. 2006. Studies on Effect of pH on Cross-linking 25 26 27
of Chitosan With Sodium Tripolyphosphate: A Technical Note. AAPS PharmSciTech: 7(2) Article 50. Communities. Washington DC. USA. Bioavailability of Curcumin: Problems and Promises. Reviews. Mol. Pharmaceutics, 4(6):807-818.

10

2 1Chattopadhyay I, K Biswas, U Bandyopadhyay and RK Banerjee. 2004. 2 3

Turmeric and curcumin: Biological actions and medicinal applications. Riview. Current Science, 87(1):44-53.

4Daneshyar M, MA Ghandkanlo, FS Bayeghra, F Farhangpajhoh and M Aghaei. 5 6 7

Effects of dietary turmeric supplementation on plasma lipoproteins, meat quality and fatty acid composition in broilers. 2011. S. Afr. J. Anim. Sci. vol. 41(4) : 420-428.

8 Donoghue DJ. 2003. Antibiotic residues in poultry tissues and eggs: Human 9 10 11 Gaudin, V., Maris, P., Fusetier, R., Ribouchon, C., Cadieu, N. and Rault, A. 12 13 14 15 16Islam MZ, ZH Khandaker, SD Chowdhury and KMS Islam. 2008. Effect of citric 17 18 19 20Ismoyowati and T Widiyastuti, 2003. Fat and Cholesterol Content of Breast and 21 22 23 24Julendra, H. 2010. Earthworm meal (Lumbricus rubellus) addition as feed 25
additives for growth promoters in broiler chickens.Tesis, studiprogram Thigh Meat of Local Fowls. Animal Production 5(2):79-82. (In Indonesia with abstract in English). acid and acetic acid on the performance of broilers. J. Bangladesh Agril. Univ. 6(2): 315320. 2004. Validation of a microbiological method: The Star protocol, a five plate test for screening of antibiotic residues in milk. Food Additives and Contaminants 21(5): 422-433. health concerns? Poultry Science, 82: 618-621.

1 2 1 2 3

11 Animal Husbandry, Graduate school, Faculty of Animal Husbandry, UGM. Yogyakarta. (In Indonesia with abstract in English).

4Lee KW, H Evert, HJ Kappert, H Wouterse, M Frehner, and AC Beynen. 2004. 5 6 7 8 9Maiti K, K Mukherjee, 10 11 12 13Nonga HE, M Mariki, ED Karimuribo, and RH Mdegela. 2009. Assessment of 14 15 16 17 18NRC, 1994. Nutrien Requirements of Poultry: Ninth Revised Edition, 1994. 19 20 21Rao RR, K Platel, K Srinivan. 2003. In vitro influence of spices and spice-active 22 23 24 25Sauland S, DTH Sihombing, M Bintang, ad Kartiarso. 2010. The Effect of Ration 26
Containing Curcumin (Extrac Tumeric/Curcuma domestica) in Pigs Rations principles on digestive enzymes of rat pancreas and small intestine. Nahrung 47(6): 408412. National Academy Press, Washington, D.C. antimicrobial usage and antimicrobial residues in broiler chickens in Morogoro Municipality, Tanzania. Pakistan Journal of Nutrition 8(3): 203207. phospholipid A Gantait, BP Saha, PK Mukherjee. 2007. Curcumin Preparation, therapeutic, evaluation and Cinnamaldehyde but not thymol counteracts the carboxymetyl cellulose induced growth depretion in female broiler chickens. Int J. of Poultry Sci. 3(9):608-612.

complex:

pharmacokinetic study in rats. Int. J. Pharm. 330(1-2), 155-63.

1 2 1 2 3

12 to Replace Antibiotic Sintetic as Growth. Forum Pascasarjana 33(2) : 123131. (In Indonesia with abstract in English).

4Sofia I, Pirman, and Z Haris. 2010. Characterization and functional chitosan 5 6 7 8 Wachira WM, A Shitandi and R Ngure, 2011. Determination of the limit of 9 10 11
detection of penicillin G residues in poultry meat using a low cost microbiological method. International Food Research Journal 18(3): 12031208. physicochemical obtained from waste shrimp shell. Jurnal Teknik Kimia Indonesia 9 (1): 11-18. (In Indonesia with abstract in English).

12 13TABLE 1. Composition and nutrient contents of the basal ration (BR) * :

FEED MATERIALS PERCENTAGE (%) Milled yellow corn 52.00 Rice bran 12.50 Soybean meal 19.50 Fish meal 9.50 Palm oil 5.10 Limestone 0.30 NaCl salt 0.40 Topmix mineral-vitamin ** 0.40 L-Lysine HCl 0.20 DL Methionine 0.10 TOTAL 100.00 NUTRIENT CONTENTS Crude protein (%) 20.13 Metabolizable Energy (kcal/kg) 3201.77 Crude fat (%) 5.41 Crude fiber (%) 3.35 Calcium (%) 0.90 Available phosphorus (%) 0.43 Lysine (%) 1.29 Methionine (%) 0.50 Description: * Standard nutrient requirements of broiler chickens 3-6 weeks (NRC, 1994): 20% protein; 1.0% Lys; 0.38% Met; energy 3200 kcal/kg, 0.9% Ca, 0.35% P.av ** Composition per kilogram of topmix mineral-vitamin : Ca 32.5%; P 10.0%; Fe 6.0 g; Mn 4 g; Iod 0.075 g; Zn 3.75 g; vit B12 0.5 mg; vit D3 50000 IU.

14 15 16 17 18 19 20 21

1 2 1 2 3 4 5 6 7 8 9 10 11 12 13 15 16

13

1 2
Table 2. Nutrient digestibility of ration were given a feed additive in broiler chickens (%) True Apparent Organic Dry matter Crude Fat Protein Protein matter Digestibility Digestibility Digestibilit Digestibilit TREATMENTS Digestibility y y % 51.74b 51.05b T1. BASAL RATION (BR) 70.48b 0.42 73.04c 0.39 69.70d 0.76 1.12 1.12 52.69bc 51.98bc a a a T2. BR + CHITS. 0.1% 69.79 0.13 72.13 0.14 42.80 4.99 0.23 0.22 57.08de 56.39de c d d T3. BR + TURM. EXT. 0.1% 70.84 0.15 73.82 0.23 68.59 0.27 0.57 0.58 49.43a 48.79a e d f T4. BR + TPP 0.1% 71.81 0.14 73.88 0.20 80.96 2.38 0.55 0.56 ef h 79.01 66.35 65.69h T5. BR +(TPP+CHITS 0.1%) 72.28g 0.15 74.98f 0.20 2.04 0.35 0.35 49.07a 48.39a T6. BR +(TPP+ TURM. EXT. 0.1%) 72.44g 0.07 75.32g 0.19 84.84g 2.63 0.51 0.552 T7. BR +(CHITS+TURM. EXT. 53.24c 52.59c 73.31i 0.15 75.41g 0.22 77.80e 0.33 0,1%) 0.28 0.28 T8. BR + NON CAPSULATION 65.92cd 58.01e 57.31e 0.1% 71.29d 0.80 72.89c 0.18 3.82 0.58 0.59 (CHIT+TURM. EXT.+TPP) bc 52.49 51.79bc T9. BR + NP D 0.1% 70.81c 0.20 74.34e 0.16 76.49e 0.55 0.26 0.26 62.62g 61.91g T10. BR + NP EE 0.1% 70.47b 0.12 73.60d 0.16 63.46c 1.07 2.26 2.30 62.71g 62.02g T11. BR + NPEE 0.2% 71.96ef 0.28 74.37e 0.31 69.76d 0.46 0.89 0.91 61.05f 60.35f T12. BR + NP EE 0.3% 72.04f 0.05 73.88d 0.15 59.07b 3.95 1.52 1.55 78.14ef 68.55i 67.83i h h T13. BR + NP EE 0.4% 73.11 0.08 75.68 0.22 0.89 0.32 0.32 66.17cd 68.76i 68.12i a b T14. BR + NP EE 0.5% 69.88 0.03 72.43 0.06 0.94 1.01 1.03 82.79fg 65.43h 64.77h j j T15. BR + NP EA 0.1% 74.21 0.10 76.51 0.17 0.44 0.26 0.26 68.68i 67.99i hi f d T16. BR + NPEA 0.2% 73.23 0.16 74.87 0.14 66.83 2.49 0.19 0.19 56.38d 55.67d j i e T17. BR + NP EA 0.3% 74.19 0.17 76.09 0.11 77.37 5.48 0.82 0.83 68.48i 67.79i l l e T18. BR + NP EA 0.4% 75.90 0.24 78.52 0.24 76.05 2.30 1.65 1.68 63.31g 62.61g k k e T19. BR + NP EA 0.5% 74.77 0.25 77.57 0.28 77.53 2.01 0.31 0.31 Crude Fiber Digestibilit y 27.42def 2.02 26.24cde 1.64 27.24def 0.50 26.26de 0.20 24.98bc 0.53 23.82b 2.18 21.63a 0.08 29.04g 0.27 25.50cde 0.62 29.96g 2.15 26.61def 0.82 26.70def 0.24 24.96b 1.32 26.46def 0.82 27.60f 0.73 27.88fg 0.37 26.77def 0.79 26.20cde 0.60 27.43ef 0.92

14

1 2 1 2

15

3Description: 4The average value in the same column with different superscripts indicate significant differences (P <0.05). 5CHITS. is chitosan, TURM.EXT. is turmeric extracted with ethanol 96%, TPP is sodium tri poly phosphate, 6NON CAPSULATION is chitosan plus turmeric extract plus TPP added in ration without encapsulation process, 7NP D is powder of nanoparticle formula D have concentration turmeric extracted ethanol 0.2% w/v added in ration with encapsulation 8process by ionic gelation, 9NP EE is powder of nanoparticle formula E have concentration turmeric extracted ethanol 2% w/v added in ration with encapsulation 10process by ionic gelation. 11NP EA is powder of nanoparticle formula E have concentration turmeric extracted Aquades added in ration with encapsulation process by 12ionic gelation.

13