KOREAN J. FOOD SCI. TECHNOL. Vol. 44, No. 2, pp. 168~172 (2012) The Korean Society of Food Science and Technology 오메가 -3 연질캡슐의코팅조건에따른장용성코팅품질에미치는영향 고원화 홍준기 이성완 차자현 차재욱 백현호 박현진 1 * 삼성정밀화학, 1 고려대학교생명공학부 The Effects of Coating Treatments on Enteric Coating of the Soft Capsules Containing Omega-3 Fatty Acids Won Hwa Ko, Jun Kee Hong, Sung Wan Lee, Ja Hyun Cha, Jae Uk Cha, Hyon Ho Baek, and Hyun Jin Park 1 * Samsung Fine Chemicals Co., Ltd. 1 School of Life Sciences and Biotechnology, Korea University Abstract This article presents an evaluation of the effects of coating conditions on the enteric coating quality of soft gelatin capsules containing Omega-3 fatty acids. Three conditions were controlled: concentration of hydroxypropyl methylcellulose phthalate (6, 8, and 10 wt% in solution), temperature of the inlet air (32, 35, and 38 o C), and the coating solution feed rate (7.5, 11.25, and 15.0 g/min). The transparency of the enteric coated soft gelatin capsules was evaluated by measuring the degree of whiteness of the surface using a spectrophotometer. Results showed that the most important parameter in the enteric coating process was the coating solution feed rate. As the coating solution feed rate decreased and inlet air temperature increased, the degree of whiteness of coating surfaces decreased. We also evaluated the disintegration properties of the enteric coated capsules in accordance with the Korea Health Functional Food Code. Keywords: enteric-coated, omega-3, hydroxypropyl methylcellulose phthalate,, AnyCoat-P 서 2008 년 건강기능식품에관한법률 이개정되면서건강기능식품에서개발가능한제형이확대되었고, 이후국내산업의현실에맞는건강기능식품공전의개정이계속진행되고있다 (1). 특히건강기능식품에서장용성제제의개발은소재의기능성을최대화하기위해도입이필요함을인지하고있었으나, 코팅소재사용의제한과제품의원가상승등을이유로국내에서는개발이다소미흡했던것이사실이다. 또한이미개발된제품이라할지라도앞서언급한것과같이소재의사용제한, 즉, 코팅제로식품또는식품첨가제로인정되는일부소재를사용하여야하며용매는물과주정만을사용해야했기때문에개발된제품의장용성을입증하기위한붕해시험에부적합한사례가빈번히발생하고있으며해외에서수입되는제품에서는의약품에서사용되고있는소재가무분별하게사용되는사례도발견되고있다 (2). 다양한장용성제제중에서캡슐과정제를특수코팅을하여위에는그대로남아있고, 소장에서함유성분을노출하도록하는것을 장용코팅 (enteric coating) 이라고한다. 장용코팅은장의 ph 가높은환경에서붕해가된다는점에서 ph 의존적이고, 위장관을통과하면서시간이지남에따라수분에의해용해된다는점 *Corresponding author: Hyun Jin Park, School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea Tel: 82-2-3290-3450 Fax: 82-2-953-5892 E-mail: hjpark@korea.ac.kr Received October 18, 2011; revised January 18, 2012; accepted January 18, 2012 론 에서시간의존적이며, 또한장내효소의가수분해촉진작용에의해변화를받는다는점에서효소의존적이다 (3). 정제및캡슐의장용코팅에쓰는물질들로는지방, 지방산, 왁스, 쉘락, 초산프탈산셀룰로오스, 프탈산히드록시메틸셀룰로오스 (Hydroxypropyl methylcellulose phthalate, 이하 ) 가있다 (4). 앞에서언급된다양한장용성코팅소재중에서 는 1971 년부터의약품첨가제로, 주로정제및캡슐의장용코팅제로사용되기시작하였으며, 천연펄프를원료로하여화학적합성방법에의해만들어진 ph 의존적용해도를지닌고분자이다. 특히셀룰로오스고리에치환된카르복실기가낮은 ph 인위액에서는붕해및용출이일어나지않고, 높은 ph 인장액에서는붕해및용출이빠르게일어나는특성을가지고있다. (Fig. 1) 는해외에서는이미정제, 과립제, 캡슐의장용코팅제로의약및건강기능식품의첨가물로오랫동안널리이용되어왔다 (5-10). 특히 는기존에확립된코팅기술을이용하여다른가소제나필름의성형제첨가없이도정제나과립코팅에이용될수있다 (11). 트리아세틴 (triacetin), 트리에틸시트레이트 (triethyl citrate) 혹은디에틸타르트레이트 (diethyl tartrate) 와같은첨가제가함께함유된 의미세입자분산액이정제코팅액으로사용되기도한다 (12). 국내에서는의약품장용코팅으로 가 30 여년전부터소염진통제, 유산균, 효소제및 PPI(Proton Pump Inhibitor) 계열의항궤양제등에주로사용되어왔으며, 다른장용코팅기제 (Acryl resin, Shellac 등 ) 에비해안전성, 코팅의용이성등의장점이많아그수요가늘어나는추세이다. 특히해외에서는북미, 유럽등에서약전수재부형제를건강기능성식품에이용가능하게되어있어 를건강기능식품용장용코팅제로널리이용하고있다 (Table 1). 168
오메가 -3 연질캡슐의코팅조건에따른장용성코팅품질에미치는영향 169 Fig. 1. The structure of hydroxypropyl methylcellulose phthalate. characteristics, particularly at the ph where dissolution occurs, are determined by the degree of substitution of the three substituent groups (i.e. methoxy, hydroxypropoxy, and carboxybenzoyl). Basically, this polymer is prepared from phthalic acid-treated HPMC (20). Table 1. The states of enteric coated dietary supplements in the North America No. Product Manufacture Active Ingredient Effects of enteric coating 1 Vitalzyme Energeticnutrition Bromelain, Protease, Lipase, Lactobacillus sporogenes etc. 2 GNC Arginmax GNC Multi Vitamin 3 GARLIQUE Sunsource Garlique GARLIC (Allium sativum L.) preventing gastroenteric trouble and oil reflux 4 Fish Oil 1200mg Enteric Coated Softgel Kirkland Signature Omega-3 fatty acid preventing oil reflux 5 SAMe (S-adenosyl-methionine) Life extension S-adenosyl-methionine 6 Nattokinase Plus TM 60 caps (NAT59) 7 Wakeupontime Wakeupontime 8 Fe Tabs, 325mg, Enteric Coated Tablets 9 Equilon Equilon Herbal Equilon Davinci Labs Nattokinase Guarana seed extract, multi vitamin etc. Windmill Ferrus sulfate preventing gastroenteric trouble Herbal Actives Red Yeast Rice Omega 3/6/9 with CoQ1 10 GINKOGIN BLAIREX LABORATORIES Ginkgo Biloba, Ginseng, Garlic and oil reflux, oil reflux and gastroenteric trouble 오메가 -3 지방산이나 DHA/EPA 함유제품과같은 fish oil 이충진된캡슐의경우오일역류, 속쓰림, 구취, 부글거림, 설사와같은부작용을자주일으킨다 (13). 특히임신여성은위와같은부작용으로복용에불편을겪을수있다. 따라서이러한캡슐을내용물의방출을지연시키기위하여장용코팅을함으로써이러한부작용을감소시키고환자의복용성을향상시킨다. Belluzzi 등 (14,15) 의문헌에서장용코팅한오메가 -3 제품을복용할경우크론씨병의재발률및부작용을현격히낮추고복약편의성을높인다는것이임상시험결과를통해제시되어있다. 이러한사실은제형개발의필요성에도불구하고소재의제한으로개발이제한적이었던장용성제제에대한개발이가능하다는점을시사한다. 특히산에서의안정성이떨어지는유산균 (16-18) 마늘제품류, 특이의냄새가있거나위장장애를일으킬수있는오메가 -3 지방산, 마늘, 박하, 철분제품류에적용이기대된다. 본연구에서는최근건강기능식품의장용성코팅소재로가장부각되고있는 를사용하여오메가 -3 지방산이충진된투명한연질캡슐을장용코팅시코팅조건에따라연질캡슐의표면에미치는영향을살펴보고장용성제제의다양한활용을위한기초자료로제시하고자실험을수행하였다. 재료및방법 캡슐및코팅조성오메가 -3 지방산이충진된연질캡슐은시판되는오메가 -3( 정제어유 100%, oblong type) 를 2011 년 5 월에구입하여변질되지않도록냉장보관하면서사용하였다. 연질캡슐의피막은젤라틴 40%, 글리세린 40%, 정제수 20% 로구성되었다. 코팅제로는장용성기제인프탈산히드록시메틸셀룰로오스 (AnyCoat-P, Samsung Fine Chemicals Co., Ltd., Incheon, Korea) 와필름코팅의유연성을높이기위해사용되는가소제로는 acethylated monoglyceride ( 이하 AMG, Esterol Sdn.Bhd., Selangor Darul Ehsan, Malaysia) 가쓰여졌다. 코팅용매로는발효주정 (95%, Daejung Chemicals & Metals Co., Ltd., Siheung, Korea) 과정제수를혼합하여사용하였다. 코팅액조성및제조방법코팅액은총중량이 500 g 이되도록제조하였으며, 그중고형분인 의사용량을변화시켜 농도가전체용액의중량대비 6, 8, 10% 세가지처방으로진행하였으며, AMG 은모두고형분대비 10% 를첨가하였다. 용매로는발효주정과정제수
170 한국식품과학회지제 44 권제 2 호 (2012) 를 85:15 중량비로구성하였다. 코팅용액제조방법으로는발효주정과정제수혼합용매를 homogenizer 로교반하면서 와 AMG 를투입하여 2500 rpm 에서한시간정도교반하여균질한코팅용액을제조하였으며, 교반조건하에서모두용해되었다. 코팅운전방법필름코팅기 (Hi-Coater, Freund Corporate, Tokyo, Japan) 를이용하여 농도, 흡기온도, 코팅분사액량이연질캡슐의코팅표면에미치는영향을알아보기위하여흡기온도는 32, 35, 38 o C 로각각조절하였으며, 코팅분사액량은수동으로조절가능한 peristaltic pump(watson Marlow 101U/R, Tokyo Rikakikai Co., Ltd., Tokyo, Japan) 의 7.5, 11.25, 15.0 g/min 으로각각조절하였으며, 농도는 6, 8, 10% 로각각조절하면서코팅후코팅피막의품질을평가하였다. 본필름코팅실험에서는연질캡슐을약 200 g 정도사용하였으며, 특히코팅량은전체캡슐무게대비 3% 로일정하게조절하였다. 붕해시험건강기능식품공전에기술되어있는붕해시험법으로측정하였다. 장용코팅캡슐을인공위액과인공장액에 6 개씩넣은다음, 인공위액은 120 분, 인공장액은 60 분간상하운동을한다음관찰하여캡슐의용해되었는지관찰하였다. 이때사용한인공위액 (ph 1.2) 은염화나트륨 2.0g 에염산 7.0 ml 및물을넣어녹여 1L 로제조하였고, 인공장액 (ph 6.8) 은 0.2 mol/l 인산이수소칼륨시액 250 ml 에 0.2 mol/l 및물을넣어 1L 로하여제조하였다. 백색도장용코팅후연질캡슐의코팅품질을확인하기위하여코팅된캡슐표면의백색도를측정하였다. 백색도를측정하기위하여 20 mm rectangular cell에캡슐을적층하여 spectrophotometer (ColorQuest XE, Hunter Associates Laboratory, Inc., Reston, VA, USA) 으로캡슐피막의백색도를코팅전캡슐과비교하여측정하였다. 측정 mode는 RSIN이고, 기기의 port size는 1 inch로설정하였다. 주사전자현미경관찰코팅피막에균일하게코팅이되었는지판단하기위하여코팅캡슐의표면을 Ion Sputter Coater(MCM-100, SEC, Suwon, Korea) 로 gold-ion coating 한다음단면을주사전자현미경 (SNE-3000M, SEC, Suwon, Korea) 을이용하여 100 배율에서관찰하였다. 통계분석 JMP ver. 8.0 통계프로그램을사용하여 Screening analyze 와 DOE(Augmented design) 를실시하였으며, 유의성검정은 P-value 가 0.05 이하일경우를유의한것으로판정하였다. 결과및고찰 코팅조건변화에따른백색도에미치는영향 로장용코팅시코팅후캡슐의표면이투명하지않고백색현상이발생하는것이코팅품질을저하시키는주요인자로작용하고있다. 특히발효주정과정제수를혼합용매로사용하는반수계코팅의경우, 용매의휘발성이낮기때문에코팅공정조건의설정이까다롭다. 본실험에서는 의농도, 흡기온도및코팅분사액량, 이세가지인자를각각하나씩조절하면서백 Fig. 2. Surface profilers showing the effects of process parameters on the experimental results of the solid contents, inlet air temperature and the solution feed rate on the response, whiteness. 색현상을최대한억제할수있는장용코팅조건을설정하였다. 먼저코팅기의코팅분사액량을 7.5 g/min 으로, 흡기온도를 32 o C 로고정한후, 코팅용액내 의농도를 6, 8, 10% 로각각변화시키면서캡슐을코팅하였다. 이때의백색도는 Table 2 와같으
오메가 -3 연질캡슐의코팅조건에따른장용성코팅품질에미치는영향 171 Table 2. Effects of concentration on the whiteness of the coating surface of omega-3 fatty acids soft gelatin capsules at feed rate 4 and inlet air temperature 32 o C 6 7.5 32 11.1±0.622 8 7.5 32 10.1±0.205 10 7.5 32 9.14±0.650 며, 농도의변화에따른백색도는큰변화가없었다. 다음으로코팅분사액량을 15.0 g/min으로고정하고, 농도를 8% 로고정하였을때, 흡기온도를 32, 35, 38 o C로각각변화시키면서장용코팅하였으며, 이때측정한캡슐의백색도는 Table 3과같다. 그결과흡기온도가증가할수록백색도가약간감소하는경향을보였다. 이것은흡기온도가증가할수록코팅팬내부의온도가증가하여코팅액을분사하였을때쉽게용매가휘발할수있는환경이조성되어표면의투명도가증가하는것으로사료된다. 마지막으로 농도는 8%, 흡기온도는 35 o C로고정하였을때, 코팅분사액량을 pump gage 값으로 7.5, 11.25, 15.0 g/min 으로조절하였을때코팅후백색도를측정하였다. 이때측정한값은 Table 4에나타내었다. 코팅분사액량이증가할수록캡슐의백색도는큰폭으로증가하며, 코팅분사액량이 4일때백색도가가장낮은것으로보인다. 이는코팅분사액량이증가할수록코팅팬내부의온도가낮아지면서용매가쉽게휘발되지않아캡슐표면을하얗게만드는백색현상을일으키는것으로여겨진다 (Table 2-4). 코팅분사액량이백색현상을일으키는데상당한영향을주고있음을알수있으며, 장용코팅시표면의투명도를증가시키기위해서용매의건조조건을높여야하며따라서흡기온도를높이고코팅분사액량을줄여야하는것으로사료된다 (Fig. 3). 상기실험조건인코팅분사액량 7.5-15.0 g/min, 흡기온도 32-38 o C, 농도 6-8% 에서백색현상이발생된코팅조건에서는코팅용액이캡슐표면에부착되어건조되기전에 air spray로분사되는즉시가루형태로건조되어캡슐에부착되어결과적으로표면이매끄럽지않게된다. 필름코팅시 air spray에서분사된코팅 Table 3. Effects of inlet air temperature on the whiteness of the coating surface of omega-3 fatty acids soft gelatin capsules at feed rate 8 and concentration 8% 8 15.0 32 80.44±2.51 8 15.0 35 79.27±2.75 8 15.0 38 60.80±1.74 Table 4. Effects of feed rate on the whiteness of the coating surface of omega-3 fatty acids soft gelatin capsules at concentration 8% and inlet air temperature 35 o C 8 07.5 35 06.73±0.799 8 11.250 35 47.99±0.658 8 15.0 35 79.27±2.750 용액에캡슐의표면에부착되고건조되는과정을겪는데, 이때캡슐표면에균일하게코팅용액방울이균일하게붙어건조되면필름의표면이투명하게된다. 그러나백색현상이발생시에는필름형성을할때코팅용액이표면에불규칙하게배열되어서난반사가발생하여표면이하얗게보이는것으로판단된다 (19). 통계분석 농도, 흡기온도, 코팅분사액량, 이세가지인자가연질캡슐의장용성코팅의코팅표면에미치는영향을알아보기위하여 Screening analyze 와 DOE(Augmented design) 를실시하였다. Screening analyze 를통하여분석한결과, 주인자중코팅분사액량 (p-value: 0.0001), 흡기온도 (p-value: 0.0011) 가백색도에영향을미치는인자로통계적으로유의하다는결론을얻었다. DOE(Augmented design) 를실시하여코팅후연질캡슐의표면에인자들이미치는효과를분석한결과, 코팅분사액량이감소할수록코팅후캡슐표면의백색도는감소하였다. 또한흡기온도는증가할수록백색도가감소하였다. 따라서코팅분사액량의조절 Table 5. Screening analyze of the effects of process parameters on the experimental results of the solid contents, inlet air temperature, and the solution feed rate Term Contrast Length t-ratio Individual p-value Simultaneous p-value rate 26.9500 15.75 <.001* 0.0003* -11.4358-6.68 0.0006* 0.0060* concentration 0.1463 0.09 0.9380 1.0000 rate * rate -0.4170-0.24 0.8240 1.0000 rate * -7.3765-4.31 0.0044* 0.0425* * 5.1793 3.03 0.0173* 0.1728 rate * concentration -1.1976-0.70 0.4774 1.0000 * concentration -1.1405-0.67 0.5159 1.0000 concentration * concentration -0.1631-0.10 0.9304 1.0000 rate * rate * 7.5759 4.43 0.0039* 0.0384* rate * * 1.3516 0.79 0.4115 1.0000 * * 1.1012 0.64 0.5434 1.0000 rate * * concentration -3.3866-1.98 0.0617 0.5340 rate * concentration * concentration 1.3406 0.78 0.4173 1.0000
172 한국식품과학회지제 44 권제 2 호 (2012) Fig. 3. Scanning electron microscopic photographs of soft gelatin capsule and enteric-coated soft gelatin capsule. A, soft gelatin capsule; B, enteric-coated soft gelatin capsule. Each photo was taken at 100 magnification. The even coating layer is observed picture B compared to picture A. 을통하여연질캡슐의표면의투명도가우수한캡슐을얻을수있을것으로사료된다 (Table 5). 붕해시험표면의백색도가가장낮은시료를조건별로각각 6 개씩취하여건강기능식품공전기준에의거하여붕해시험을진행한결과, 로코팅을한연질캡슐의경우제 1 액 (ph=1.2) 에서 120 분동안 6 개의캡슐모두붕해가되지않았다. 제 2 액 (ph=6.8) 의경우 60 분간붕해시험한결과시료 6 개모두붕해되었다. 장용코팅을하지않은대조군을붕해시험을한결과제 1 액 (ph=1.2) 에서 30 분내에용해되었고, 제 2 액 (ph=6.8) 에서도또한 30 분내에용해되었다. 이와같은결과로붕해시험에는코팅조건에따른영향은없었으며, 로코팅을한연질캡슐은장용성제제로활용이가능할것으로판단된다. 주사전자현미경관찰장용코팅을하기전의시료와코팅후백색도가가장낮고, 투명도가높은시료를가지고미세구조를관찰하였다. 연질캡슐상부에균일하게피막이도포되어있는데이는필름코팅을통하여연질캡슐에장용코팅이캡슐의피막에균일하게코팅되었음을알수있다 (Fig. 3). 요 본연구는프탈산히드록시메틸셀룰로오스를장용성기제로이용하여오메가 -3 지방산이충진된연질캡슐을코팅조건에따른연질캡슐의표면에미치는영향을살펴보고자하였다. 농도, 코팅분사액량및흡기온도를변화시켜장용코팅시캡슐표면의백색도를측정비교한결과, 코팅분사액량이감소할수록흡기온도가증가할수록백색도가감소하는것을알수있었다. 그리고붕해시험으로코팅후캡슐이장용특성을가지는것을확인할수있었다. 주사전자현미경으로미세구조를관찰한결과, 피막에균일하게 가코팅이된것을알수있었다. 이와같이 를이용하여오메가 -3 연질캡슐의장용코팅효과를확인할수있었으며, 투명도가높은코팅표면을얻을수있는코팅조건을확인할수있었다. 문 1. KFDA. Health functional food code proclaim. 2011-68. Korea Food & Drug Administration. Seoul, Korea. pp.2-4 (2011) 2. KFDA. Health functional food code proclaim. 2011-20. Korea 약 헌 Food & Drug Administration. Seoul, Korea. pp.6-7 (2011) 3. Guo HX, Heinämäki J, Yliruusi J. Diffusion of a freely water-soluble drug in aqueous enteric-coated pellets. AAPS PharmSciTech 3: 97-104 (2002) 4. Allen Jr. LV, Popovich NG, Ansel HC. Ansel s pharmaceutical dosage forms and drug delivery systems. Lippincott Williams & Wilkins, Philadelphia, PA, USA. p.222 (2009) 5. Ehrhardt L, Patt L, Schindler E. Optimization of film coating systems. Pharma. Ind. 35: 719-722 (1973) 6. Delporte JP, Jaminet F. Influence of formulation of enteric-coated tablets on the bioavailability of the drug. J. Pharm. Belg. 31: 263-276 (1976) 7. Patt L, Hartmann V. Solvent residues film forming agents. Pharma. Ind. 38: 902-906 (1976) 8. Stafford JW. Enteric film coating using completely aqueous dissolved hydroxypropyl methylcellulose phthalate spray solutions. Drug Dev. Ind. Pharm. 8: 513-530 (1982) 9. Thoma K, Heckenmuller H, Oschmann R. Resistance and disintegration behavior of gastric juice resistant drugs. Pharmazie 42: 832-836 (1987) 10. Thoma K, Heckenmuller H, Oschmann R. Impact of film formers and plasticizers on stability of resistance and disintegration behavior. Pharmazie 42: 837-841 (1987) 11. Rowe RC. Molecular weight studies on hydroxypropyl methylcellulose phthalate (HP-55). Acta Pharm. Technol. 28:127-130 (1982) 12. Muhammad NA, Boisvert W, Harris MR, Weiss J. Evaluation of hydroxypropyl methylcellulose phthalate 50 as film forming polymer from aqueous dispersion systems. Drug Dev. Ind. Pharm. 18: 1787-1797 (1992) 13. Fetterman F, Hussain MM. Therapeutic potential of n-3 polyunsaturated fatty acids in disease. Am. J. Health- Syst. Ph. 66: 1169-1179 (2009) 14. Belluzzi A, Brignola C, Campieri M, Camporesi EP, Gionchetti P, Rizzello F, Belloi C, De Simone G, Boschi S, Miglioli M. Effect of new fish oil derivative on fatty acid phospholipid-membrane pattern in a group of Crohn s disease patients. Digest. Dis. Sci. 39: 2589-2594 (1994) 15. Belluzzi A, Brignola C, Campieri M, Pera A, Boschi S, Miglioli M. Effect of an enteric-coated fish-oil preparation on relapses in Crohn s disease. New Engl. J. Med. 334: 1557-1560 (1996) 16. Haghes DB, Hoover DG. Viability and enzymatic activity of bifidobacteria in milk. J. Dairy Sci. 78: 268-276 (1995) 17. Kailaspathy K, Rybka S. Lactobacillus acidophilus and Bifidobacteria spp.- their therapeutic potential and survival in yogurt. Aust. J. Dairy Technol. 52: 28-35 (1997) 18. Klaver FAM, Kingma F, Weekamp AH. Growth and survival of bifidobacteria in milk. Neth. Milk Dairy J. 47: 151-164 (1993) 19. McGinity JW. Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms. Vol. 79. Mercel Dekker, Inc., Piehurst, NC, USA. pp. 315-323 (1997) 20. Lai X, Sun C, Tian H, Zhao W, Gao L. Evaluation of poly(styrene-alt-maleic anhydride) ethanol as enteric coating material. Int. J. Pharm. 352: 66-73 (2008)