Culinary Science & Hospitality Research 23(8), ; 2017 Information available at the Culinary Society of Korea ( Culin

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Culinary Science & Hospitality Research 23(8), 153-162; 2017 Information available at the Culinary Society of Korea

Journal & Article Management System: https://cshr.jams.or.kr/ http://dx.doi.org/10.2087

1 Culinary Science & Hospitality Research 23(8), ; 2017 Information available at the Culinary Society of Korea ( Culinary Science & Hospitality Research Journal & Article Management System: Probiotics 와미세분쇄원두커피가첨가된인스턴트커피의품질적특성 남양유업 중앙연구소, 2 고려대학교 BK21Plus 보건과학사업단 Quality Characteristics of Instant Coffee with Probiotics and Microground Coffee Bong Soo Ko 1 Sang Ho Lim 1 Sung Hee Han 2 1 Research and Development Center, Namyang Dairy Products Corporation 2 BK21Plus, College of Health Science, Korea University KEYWORDS Bacillus coagulans, Microground coffee, Instant coffee, Quality, Flavor, Probiotics. ABSTRACT Bacillus coagulans was added as probiotics in instant coffee with microground roasted coffee which is recently rising premium coffee and the instant coffee (ProBio coffee) was compared with 3 commercial instant coffees with microground roasted coffee in quality characteristics to understand the competitiveness of ProBio coffee. In sensory evaluation, ProBio coffee had inferior aroma intensity and overall acceptance compared with control group (brewed coffee) (p<0.05) but it had equal quality or more compared with 3 commercials. Total polyphenol content, chlorogenic acid content, DPPH free radical scavenging activity and caffeine content were ±1.99 mg/g, 2,700±20 mg/g, ±3.62 TEAC mg/g, 28.1±3.2 ppm respectively. In general quality characteristics, water content, solubility, particle size and particle strength of ProBio coffee were similar to general instant coffee but had darker color than it. In safety assessment, acrylamide content was 502±10 ppb, and residual pesticides and ochratoxin A were not detected. Over 70% of B. coagulans were maintained in 4 months of storage and 16 months of shelf life was predicted in ProBio coffee by Q 10 model. Therefore, ProBio coffee was confirmed to have a sufficient product competiveness compared with 3 commercials. 1. 서론 ( ), ( ),, (KFDA, 2016) , , %, 62.5%(4 ), 37.5%(2 4 ) %, * 1. ** 1. Corresponding author:, sungheeh3@gmail.com, 145, BK21Plus

2 154 고봉수 임상호 한성희 2 3, , % (Korea Agro-Fisheries & Food Trade Corporation, 2016) Starbucks VIA, Nescafé Azera Barista Style Instant Coffee, Kenco Millicano WholeBean Instant (Euromonitor International, 2015). Starbucks kore Via 2011,,,, ,000 (Korea Herald, 2017)., -196,. (probiotics), (FAO/WHO, 2002; Gorbach, 2002).,,,,,,, (Nagpal, Kumar, Kumar, Behare, Jain, & Yadav, 2012; Foligné, Daniel, & Pot, 2013).,,,, Bacillus coagulans (Ko, Lim, & Han, 2016)., (Schäffer, Keller, Szakály, & Lőrinczy, 2011; Benković, Srečec, Špoljarić, Mršić, & Bauman, 2015),.., (,, A). Bacillus coagulans (ProBio coffee), 3 ProBio coffee. 2. 연구방법 2.1. Probiotics 와미세분쇄원두커피가첨가된인스턴트 커피제조 ( L 42 45) 2. Ko, Lim Han(2016), 50%(w/w). Bacillus coagilans 0.2%(w/w), -45 B. coagulans. (L 30) μm. B. coagulans, ProBio( 8.2% w/w) 관능적특성 (optimization) 3 (scoring test), (Tanzania arabica, L 90). A 11,. 1%(w/ w). 5 (sweetness), (bitterness), (acidity), (body) (aroma) (overall acceptability). 7 (1 :,, 7 :, ) 향기성분분석 head space-solid phase micro

Probiotics 가첨가된인스턴트커피의품질적특성 155 extraction (HS-SPME) (Fig. 1). 1%(w/w), 95 (Waltham, Thermo scientific Co. Ltd, USA) 600 rpm 10, SPME fiber(carboxen/pdms, Supelco, Bellefonte, PA, USA).

3 Probiotics 가첨가된인스턴트커피의품질적특성 155 extraction (HS-SPME) (Fig. 1). 1%(w/w), 95 (Waltham, Thermo scientific Co. Ltd, USA) 600 rpm 10, SPME fiber(carboxen/pdms, Supelco, Bellefonte, PA, USA). GC(7890A, Agilent technologies, Palo Alto, USA). mass/mass spectrometry(7000, Agilent technologies, Palo Alto, USA), column HP-INNOWA X(60 m 0.25 mm 0.5μm, J&W, Folsom, CA, USA), Table 1., signal to noise 10 height peak GC-MS/MS Library (Wiley/ NBS) mass spectrum library matching quality 70% peak. peak % 일반품질특성및 B. coagulans 생균수 (HX 204, Mettler toledo), (Colortest II, NEO TECH Co., Ltd., Germany). 3 g (Retsch, sieve shakers AS200)., 100 g 8 sieve (3.35 mm, 2.8 mm, 2.0 mm, 1.4 mm, 1.0 mm, 0.6 mm, 0.2 mm, plate) (MPS). rheometer(cr-3000ex, Sun Rheometer). Bacillus coagulans Bacillus coagulans counting (Ko, Lim, & Han, 2016) 항산화특성및카페인분석 총폴리페놀함량 Velioglu (Velioglu, Mazza, Gao, & Oomah, 1998). 0.5 g 50 ml 100 μl 2% Na 2 CO 3 2 ml 3 50% Folin-Ciocalteu's phenol (Wako Co., Japan) 100 μl nm. 0.1% gallic acid, 3 polyphenol Total chlorogenic acid 분석 Fig. 1. Extraction process by HS-SPME fiber and desorption systems for GC analyses. Table 1. Operation conditions of GC-MS/MS for the analyses of volatile flavor compounds Instrument Column Injector temperature Oven temperature Detector temperature Gas flow rate Split ratio MS ionization GC-MS/MS Agiilent 7890A GC/7000 Triple Quard MS/MS HF-INNOWAX(60 m 0.25 mm 0.5μm) (Hold 4 mins) 2 /min 210 (Hold 11 mins) ml/min Splitless 70 ev chlorogenic acid 3-CQA, 4-CQA, 5-CQA, 3,5-diCQA, 3,4-diCQA, 4,5- dicqa, chlorogenic acid %area. 1 g 100 ml, 5 ml. shaker 5. Chlorogenic acid HPLC, Table DPPH 자유라디칼소거능 DPPH Kim, Lee, Lee Lee(2002). DPPH(0.2 mm) 1 ml 50 μl nm, Trolox (Trolox equivalent antioxidant capacity, TEAC).

4 156 고봉수 임상호 한성희 Table 2. Operation conditions of HPLC for the analyses of chlorogenic acid, calibration 13 C3-acrylamide. Instrument Column Mobile phase Flow rate HPLC Agilence HPLC(waters Co., MA, USA) Capecell pak AC(4.5 mm 250 mm, 5μm) or equivalent Time(min) A(%) B(%) A: 0.5% orthphosphoric acid buffer, B : acetronitrile 1.0 ml/min 잔류농약 (KFDA, 2013). Dr. Ehrenstorfer (Germany) Wako (Japan), HPLC (ECD: electron capture detector, Agilent 6890N, USA) (NPD: nitrogen phosphorus detector, Agilent 6890N, USA) (Agilent 5973, USA),, (Acuity Ultra performance LC, USA) (Thermo triple quadruple, USA). Detector Column temperature Time of analysis Injection amounts 카페인 Diod array detector 325 nm min 10 μl Bradbury Weers(1995), Agilent 1200 series(agilent Technologies Inc., Philadelphia, USA). C18 Capcell Pak(Shiseido Co., Tokyo, Japan) 40, 10 mm HCl:Methanol = 90:10, 0.5 ml/min. Diode array detector(uv nm). (Sigma, St. Louis, MO, USA) 안전성평가 아크릴아마이드 13 C3-labeled acrylamide(200 ng/ml in 0.1% Formic acid) OASIS HLB SPE(200 mg packing, Waters Corporation, USA) Varian SPE cartridge(200 mg packing, Varian Inc., USA) PE(Solid Phase Extraction) cartridge LC-MS/MS. 5, 25, 125, 250, 500 ppb working solution 오크라톡신 A 12.5 g (methanol: 3% sodium bicarbonate, 1:1 v/v) 100 ml. PBS immunoaffinity column(ochratest, Vicam Co., Watertown, MA, USA), PBS 10 ml 10 ml 2 ml. HPLC(JP/Nanospace SI-2, Shiseido, Tokyo, Japan), CAPCELL PAK C18 UG120( mm, 5 μm), : : (99:99:2) (Ex: 333 nm, Em: 460 nm). A LC/MS/MS (US/API4000, Applied Biosystem, Foster City, CA, USA) 대장균수 standard method(richardson, 1985) 50 μl violet red bile agar colony 보존중품질변화및유통기한예측 ProBio coffee (N 2 packing; 97%) 25, 35,

5 Probiotics 가첨가된인스턴트커피의품질적특성 (, ). 5 3 (KFDA, 2015), Q 10 model 통계처리 SPSS(statistical package for social science, version 12.0) package, t (t-test), (One-way ANOVA) Tukey s test (p<0.05). 3. 결과 3.1. 관능적특성 11 Table 3. control (p< 0.05), ProBio coffee A B (p<0.05). control A, B, ProBio coffee C (p<0.05). ProBio coffee B (p<0.05), Body ProBio coffee C (p<0.05). C Body. ProBio coffee control A, B, C (p<0.05), control, ProBio coffee, C, B, A. control (p<0.05) ProBio coffee A, C (p<0.05). ProBio coffee control., 3., A, B, C Probio coffee 향기성분분석 3 ProBio coffee control (total ion chromatogram) RT(retention time) Fig. 2. BP 100 pyridine Table 3. Sensory score of commercial instant coffee and ProBio coffee Intensity score 1) Preference score 2) Control 3) ProBio Commercial coffee F-value coffee 4) A B C (p-value) Sweetness 5.02±0.41 a 4.48±0.42 b 3.77±0.37 c 3.94±0.29 c 4.58±0.35 b 5.58 (0.01) Bitterness 6.38±0.32 a 5.72±0.44 b 6.19±0.22 a 6.23±0.20 a 5.59±0.23 b 4.09 (0.03) Acidity 5.47±0.43 a 4.97±0.28 b 4.76±0.32 b 4.25±0.46 c 5.45±0.41 a 5.24 (0.02) Body 6.20±0.30 a 5.15±0.45 b 5.80±0.56 ab 5.45±0.61 b 4.35±0.55 c 5.76 (0.01) Aroma 6.48±0.26 a 5.90±0.24 b 4.45±0.65 d 5.15±0.30 c 5.33±0.45 c (0.00) Overall acceptability 6.56±0.20 a 6.18±0.18 b 5.90±0.30 c 6.16±0.17 b 5.55±0.35 d 6.69 (0.01) 1) 2) 3) 4) 1: extremely weak, 7: extremely strong. 1: extremely dislike, 7: extremely like. Drip coffee(brix : 1.6, bean origin: Tanzania arabica 100%, roasting degree(l): 90). ProBio coffee: instant coffee with probiotics and microground coffee. a,b Values with different superscripts in the same rows are significantly different (ANOVA and Duncan's multiple range test, p<0.05). All values are mean±s.d.

158 고봉수 임상호 한성희 Control tanal, 3-methyl butanal, 2,3-butandione, 2,3-pentadione 4-hydroxy-2,5-dimethyl-3(2H)-furanone(HD3F), 5-ethyl-4-hydro, sulfurous, roasty group 2-furfurylthiol,

6 158 고봉수 임상호 한성희 Control tanal, 3-methyl butanal, 2,3-butandione, 2,3-pentadione 4-hydroxy-2,5-dimethyl-3(2H)-furanone(HD3F), 5-ethyl-4-hydro, sulfurous, roasty group 2-furfurylthiol, 2-methyl-3-furanthiol methional, 3-mercapto-3- methyl butyl-formiate, 3-methyl-2-butene-1-thiol methanethiol, dimethyl trisulfide., smoke phenolic group guaiacol, 4-ethylguaiacol, 4-vinylguaiacol, fruity group acetaldehyde, propanal, (E)-β-damascenone. aroma grouping Table 4. Sweet, caramellike group control 1.54% (p<0.05), %, B C (p<0.05). Sulfurous, roasty group B 0.32%, %. ProBio coffee control, A, B, C (p<0.05)., smoke phenolic group %, fruity group. ProBio coffee ProBio coffee Fig. 2. Total ion chromatogram of commercial instant coffee by GC-MS/MS. light aroma, BP 150 furfural medium aroma, RT heavy aroma., control, light aroma 10.3%, C 7.58%, A 5.96%, ProBio coffee 3.91%, B 2.93%. Medium aroma %, heavy aroma % (Fig. 3). Wang (2012) sweet, caramel-like group methylpropanal, 2-methylbu- Fig. 3. Comparison of aroma peak % area in coffee samples. (1) Light aromas: peaks before pyridine(b.p, 100 ). (2) Medium aromas: peaks before furfural(b.p, 150 ). (3) Heavy aromas: peaks beginning with furfural(b.p, ).

7 Probiotics 가첨가된인스턴트커피의품질적특성 159 Table 4. Comparison of aroma group in commercial instant coffee and ProBio coffee Characters of aroma 1) Control ProBio coffee Commercial coffee A B C F-value (p-value) Sweet/caramel-like group 2) 1.54±0.12 a 0.41±0.04 b 0.26±0.01 c 0.31±0.09 bc 0.30±0.09 bc (0.00) Sulfurous/roasty group 3) 0.10±0.02 b 0.21±0.02 a 0.11±0.03 ab 0.32±0.08 a 0.16±0.01 a (0.00) Smoky/phenolic group 4) 1.80±0.16 b 2.45±0.21 ab 2.87±0.16 a 1.68±0.11 b 1.95±0.64 ab 7.19 (0.01) Fruity group 5) ) % sum of area, values with different superscripts in the same columns are significantly different (ANOVA and Duncan's multiple range test, p<0.05). 2) Sweet/caramel-like group: methylpropanal, 2-methylbutanal, 3-methylbutanal, 2,3-butandione, 2,3-pentadione, 4-hydroxy-2,5-dimethyl-3 (2H)-furanone (HD3F), 5-ethyl-4-hydro. 3) Sulfurous /roasty group: 2-furfurylthiol, 2-methyl-3-furanthiol methional, 3-mmercapto-3-methylbutyl-formiate, 3-methyl-2-butene-1-thiol methanethiol, dimethyltrisulfide 4) Smoky/phenolic group: guaiacol, 4-ethylguaiacol, 4-vinylguaiacol. 5) Fruity group: acetaldehyde, propanal, (E)-β-damascenone. All values are mean±s.d..,, ProBio coffee,,, ProBio Coffee 일반품질특성및 B. coagulans 생균수 ProBio coffee,,, Table 5,. Table 5. Analysis of general specification and B. coagulnace Commercial freeze-dried coffee ProBio p- coffee 2) value Moisture (%) 4.5± ±0.20 >0.05 Color (L value) 130.0± ±5 <0.05 Solubility 1) Particle size (MPS) 1.8 mm± mm±0.20 >0.05 Particle strength (g) 12,864.3± ,2557.0±334.6 >0.05 B. coagulans (cfu/g) ) Insoluble particle numbers of hot water surface for about 10 sec. 2) ProBio coffee: instant coffee with probiotics and microground coffee. All values are mean±s.d. (p<0.05),. B. coagulans cfu/g 70% ProBio Coffee 항산화특성및카페인분석 ProBio coffee chlorogenic acid, DPPH Table 6.,,, (Farah & Donangelo, 2006)., (Kim & Han, 2009). ProBio coffee ±1.99 μg/g, chlorogenic acid 2.7±0.2 mg/g. chlorogenic acid 100%,. Kim Park(2006) chlorogenic acid.,

8 160 고봉수 임상호 한성희 Table 6. Analysis of antioxidant activity and caffeine content ProBio coffee 1) Total phenolic content(μg/g) ±1.99 Chlorogenic acid(mg/g) 2.7±2.0 DPPH (TEAC mg/g) 14.6±3.62 Caffeine(ppm) 28.1±3.2 1) ProBio coffee: instant coffee with probiotics and microground coffee. All values are mean±s.d. chlorogenic acid, chlorogenic acid., chlorogenic acid. ProBio coffee DPPH, ProBio coffee 14.6±3.62 TEAC mg/g., 28.1±3.2 ppm.,. ProBio coffee A,., ProBio coffee 보존중품질변화및유통기한예측 4, B. coagulans 70% (> cfu/g) , ProBio coffee 3, Kinetic plot Fig ProBio coffee 446 ( 16 ) ( 12 ) ProBio coffee 안전성평가 ProBio coffee,, A Table 7.. 1,000 ppb, (ESFA) 900 ppb. ProBio coffee ±9.23 ppb. A green bean Table 7. Safety analysis of ProBio coffee ProBio coffee 1) Acrylamide(ppb) ±9.23 Chemical Residual pesticides 0 Ochratoxin A(ppb) 0 Microbial E. coli(cfu/g) 0 1) ProBio coffee: instant coffee with probiotics and microground coffee. All values are mean±s.d. Fig. 4. Kinetic plot for shelf-life prediction during the days of storage at 45, 55.

9 Probiotics 가첨가된인스턴트커피의품질적특성 결론및시사점 Bacillus coagulans (Pro- Bio coffee), 3 ProBio coffee., control( ) (p<0.05),., chlorogenic acid, DPPH caffeine ±1.99 μg/g, 2.7±2.0 mg/g, 14.6±3.62 TEAC mg/g, 28.1±3.2 ppm.,,,. ProBio coffee 4 B. coagulans 70%, Q 10 model 16. ProBio coffee,.,, prebiotics synbiotics. REFERENCES Benković, M., Srečec, S., Špoljarić, I., Mršić, G., & Bauman, I. (2015). Fortification of instant coffee beverages-influence of functional ingredients, packaging material and storage time on physical properties of newly formulated, enriched instant coffee powders. Journal of the Science of Food & Agriculture, 95(13), Bradbury, A., & Weers, M. (1995). Non-volatile compounds analysis in coffee, Kraft Eur. Anal Method, Euromonitor International. (2015). Micro-ground instant coffee takes off. Retrieved from 09/premium-instant-coffee-takes-share-in-uk-reflecting-stron g-potential-in-tea-drinking-markets.html [cited 15 August 2017]. FAO/WHO. (2002). Guidelines for the Evaluation of Probiotics in Foods. Food and Agriculture Organization of the United Nations and World Health Organization Expert Consultation Report (online). Farah, A., & Donangelo, C. M. (2006). Phenolic compounds in coffee. Brazilian Journal of Plant Physiology, 18(1), Foligné, B., Daniel, C., & Pot, B. (2013). Probiotics from research to market: the possibilities, risks and challenges. Current Opinion in Microbiology, 16(3), Gorbach, S. L. (2002). Probiotics in the third millennium. Digestive & Liver Disease, 34, S2-S7. KFDA. (2009). Food 4ode, 9-2. Seoul, Korea: Korean Food and Drug Administration. KFDA. (2013). Food code practical affair manual for residual pesticide analysis method. Seoul, Korea: Korean Food and Drug Administration. KFDA. (2015). Experiment Guideline for Prediction of Shelf Life of Food & Animal Products. Seoul, Korea: Korean Food and Drug Administration. Kim, D. O., Lee, K. W., Lee, H. J., & Lee, C. Y. (2002). Vitamin C equivalent antioxidant capacity (VCEAC) of phenolic phytochemicals. Journal of Agricultural & Food Chemistry, 50(13), Kim, J. Y., & Han, Y. S. (2009). Influence of roasting time on antibacterial and antioxidative effects of coffee extract. Korean Journal of Food & Cookery Science, 25(4), Kim, K. J., & Park, S. K. (2006). Changes in major chemical constituents of green coffee beans during the roasting. Korean Journal of Food Science & Technology, 38(2), Ko, B. S., Lim, S. H., & Han, S. H. (2016). Selection of lactic acid bacteria suitable for manufacture of freeze-dried coffee. Korean Journal of Food & Nutrition, 29(6), Ko, B. S., Lim, S. H., & Han, S. H. (2017). Optimization of coffee extract condition for the manufacture of instant coffee by RSM. Korean Journal of Food & Nutrition 30(2), Korea Agro-Fisheries & Food Trade Corporation (2016). Processed food market status -Coffee market. Seoul, Korea: Agro- Fisheries & Food Trade Corporation. Korea Herald. (2017). Instant regular coffee threatening coffee mix. Retrieved from [cited 6 April 2017]. Nagpal, R., Kumar, A., Kumar, M., Behare, P. V., Jain, S., & Yadav, H. (2012). Probiotics, their health benefits and applications for developing healthier foods: a review. FEMS Microbiology Letters, 334(1), Richardson, G. H. (1985). Page 351 in standard methods for the examination of dairy products. Washington, DC, USA:

10 162 고봉수 임상호 한성희 Publ. Health Assoc. Schäffer, B., Keller, B., Szakály, Z., & Lőrinczy, D. (2011). Examination of a coffee product enriched with calcium by the isoperibolic method. Journal of Thermal Analysis & Calorimetry, 108(1), Velioglu, Y. S., Mazza, G., Gao, L., & Oomah, B. D. (1998). Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products. Journal of Agricultural & Food Chemistry, 46(10), Wang, N. (2012). Physicochemical changes of coffee beans during roasting (Doctoral dissertation). The University of Guelph

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