Food Sci. Technol. Res., +, (+), -2ῌ.,, ,**0

Deodorizing E#ect of on the O#ensive Odor of the Porcine Large Intestine

+ + , - +ῌ Kaori KOHARA , Reiko KADOMOTO , Hiroshi KOZUKA , Koji SAKAMOTO and Yasuyoshi HAYATA

+ Graduate School of Applied Biosciences Program, Hiroshima Prefectural University, /0, Nanatsuka-cho, Shobara, 1,1ῌ**,-, Japan , Hiroshima Bunkyo Women’s University, +ῌ,ῌ+ Kabehigashi, Asakita-ku, Hiroshima 1-+ῌ*,3/, Japan - Hiroshima Prefectural Food Technology Research Center, +,ῌ1* Hijiyamahon-machi, Minami-ku, Hiroshima 1-,ῌ*2+0, Japan

Received September +/, ,**/; Accepted February -, ,**0

The deodorizing e#ect of coriander (Coriandrum sativum L.) on the o#ensive odor caused by porcine large intestine was studied. Both *./ and ,.* g of coriander were found to deodorize the stench of ,.* g porcine large intestine almost completely, and the deodorant e#ect of coriander was maintained for 0 hrs or more even after the peculiar odor of the coriander disappeared. We detected four main compounds that contributed to the porcine large intestine odor: .-Methylphenol (a sludge-like substance), unknown com- pound I (porcine large intestine-like), unknown compound II (a sludge-like substance) and Indole (excrement- like). Eleven main compounds of coriander odor were detected: Decanal, ,-Decenal, Undecanal, ,-Un- decenal, +-Decanol, (E)-,-Undecen-+-ol, ,-Dodecenal, (E)-,-Tetradecenal, Hexadecanal, Octadecenal and 3- Octadecenal. Although the four main compounds were not significantly decomposed by the coriander treatment, the coriander had a remarkable deodorant e#ectontheo#ensive odor emitted by the porcine large intestine.

Keywords: coriander, deodorizing e#ect, o#ensive odor, porcine large intestine, sensuous deodorization, volatile compound

Introduction reported to have a deodorization e#ect until now. Coriander (Coriandrum sativum L.) is an herbaceous The porcine large intestine is a popular food material in annual of the cicely family. The , sometimes Southeast Asia and China, and as such is a precious food called “cilantro,” are widely used in Southeast Asia to add resource. This food material was used empirically with flavor to foods or to hide the unpleasant smell of certain coriander to remove its strong o#ensive odor. Up to cooking materials. The fresh has a unique odor that now, there have been no reports on deodorization of the is similar to the smell of Kamemushi (Halyomorpha picus), o#ensive odor of the porcine large intestine by the corian- and the seeds have a refreshing aroma and sweet , der leaf and stem. and are frequently used as a . The aim of this study was to examine the deodorizing One of the important attributes of coriander is aroma e#ect of coriander on the o#ensive odor of the porcine quality, and this quality as paramount to the culinary large intestine, to compare its e#ect with the reported value of the fresh , usually decreasing before the e#ect of , wild grasses and green tea, and to identify visual quality decreases (Loaiza and Cantwell, +331). The the characteristic odor compound of the porcine large aroma and volatile compounds in coriander have been intestine and coriander. extensively described in the literature (Fan and Sokorai, ,**,). A lot of information is available on the identific- Materials and Methods ation of coriander essential oils (Taskiness and Materials Coriander [Coriandrum sativum L. (Sakata Nykanen, +31/; Bandoni et al., +332; Smallfield et al., ,**+). Seed Co., Ltd., Kanagawa, Japan)] was cultured in a green- Recently, the volatile compounds of coriander leaves house at Hiroshima Prefectural University from +* March have been reported with regards to their functionality, through +* July ,**+. The stems and leaves were sam- such as their bactericidal e#ect against the microorgan- pled at the seedling stage (.* days after sowing). Wild isms of Salmonella choleraesuis (Kubo et al., ,**.). To grasses such as dandelion, spiny sowthistle (Taraxacum date, the deodorizing e#ects of various such as albidum Dahlst. and Sonchus asper) and two herbs, thyme, sage and hydrangea against o#ensive components and (Thymus vulgaris L. and Rosmarinus o$- have been recognized (Tokita et al., +32.; Nakatani et al., cinalis L.) were collected at the university farm, and green +323; Harasawa and Tagashira, +33.), whereas the corian- tea (Camellia sinensis) was purchased at a supermarket in der plant or coriander essential oil (seed oil) have not been Shobara City. The fresh leaves and stems of the plant samples, except for the green tea, were clipped from the * To whom correspondence should be presently addressed. plants using solvent-rinsed stainless steel scissors and E-mail: [email protected] (School of Agriculture, Meiji Uni- tweezers, and the surface moisture was lightly wiped versity) from the samples with a paper towel after the samples Deodorizing E#ect of Coriander on the O#ensive Odor of the Porcine Large Intestine 39

Table +. Comparison of the deodorizing activity of various plant materials against porcine large intestine.

῍+῎ Deodorization intensity (*: perfectly deodorized, +: almost deodorized, ,: considerably deodorized, -: a little deodorized, .: completely not deodorized). ῍,῎ The deodorization activity was carried out in the porcine large intestine (,.* g) immediately after the addition of each plant sample (*.+, *./ or ,.* g). ῍-῎ Each plant sample (,.* g) was added to the porcine large intestine (,.* g), and is the deodorization intensity of the sample set for - or 0 hours after addition at .*῔. ῍.῎ Symbols are the odor intensity of each plant material (῏῏῏: very strong, ῏῏: strong, ῏: a little). ῍/῎ The green tea dry matter added was in a quantity converted into a fresh leaf quantity. a, b, c: Mean values with di#erent superscripts are significantly di#erent according to the LSD test (pΐ*.*/, nῒ-).

were washed with running water. All samples were performed with a Shimadzu GC-+1A with a flame ioniza- frozen with liquid nitrogen, and crushed and stored at tion detector and a Shimadzu Chromatopac C-R1A inte- ῐ2*῔ until the time of the deodorizing activity test or grator (Shimadzu Co., Ltd., Kyoto, Japan). The com- extraction of volatile compounds. pounds were separated on a -* mῑ*.,/ mm (i.d.) fused The porcine large intestine of a pig with its internal silica capillary column coated with a *.,/-mm film bonded organs was prepared at the slaughter house in Fukuyama polar DB-WAX (J&W Scientific Inc., Folsom, CA). The City, Hiroshima. The large intestine was washed by dis- capillary column was maintained at .*῔ for +* min after tilled water, and it was immediately transported to our injection and then programmed at -῔ min-+ to ,,*῔, laboratory under *῔, then rewashed and cut into minced which was maintained for -* min. Open split injection meat and stored at ῐ2*῔. Small samples were thawed was conducted with a split ratio of +: ,*; helium was used and used as the need arose during the experiment. as the carrier gas at ,/* Pa. For each column the injector Simultaneous Distillation Extraction method (SDEM) temperature was ,-*῔ and the detector temperature was We performed SDEM as described previously (Maneerat ,/*῔. Some individual components could be identified et al., ,**,). A sample flask with +*.* g coriander and ,.* by both the injection of pure compounds and by compar- g porcine large intestine was mixed with 0** ml distilled ison of their retention times (as a Kovats index). water in a +-L round-bottom flask. Ten mlof+ῌ cy- Gas chromatograph- mass spectrometer (GC-MS) analy- clohexanol solution was added as an internal standard. sis GC-MS was performed with a Shimadzu GC-+1Agas A ,**-ml V-shaped bottom flask containing +** ml of chromatograph coupled with a Shimadzu QP/*/* mass diethyl ether was attached to the solvent arm of the SDE spectrometer (Shimadzu Co., Ltd., Kyoto, Japan). The head. The separation of volatile compounds was carried column (DB-WAX, 0* mῑ*.,/ mm i.d.; J&W Scientific Inc., out under reduced pressure (1/ mmHg, 2*῔)for0* min. Folsom, USA) and temperature programs used were the The condenser of the SDE head was cooled with a mixture same as those used for the GC analysis. Split injection of water and ethylene glycol at ῐ/῔. The extract was was performed with helium as the carrier gas. The com- dried over anhydrous sodium sulfate and concentrated to pounds were identified by the use of NIST and our own ῌ+** ml under a nitrogen stream before + ml of the sample mass spectra libraries. was drawn for gas chromatograph (GC) analysis or gas Gas chromatograph-olfactometry (GC-O) analysis A chromatograph mass spectrometry (GC-MS) analysis. Shimadzu GC-+1A (Shimadzu Co., Ltd., Kyoto, Japan) was Gas chromatograph (GC) analysis GC analysis was equipped with a Y connector outside a capillary, allowing 40 K. KOHARA et al.

Fig. +. Chromatogram of volatile compounds of porcine large intestine added to coriander or not. Abbreviations: I.S., Internal standard; BHT, ,,0-Bis (+,+-dimethylethyl)-.-methylphenol ῎ Porcine large intestine used ,.* g addition fresh weight coriander *./ g. ῎ The main o#ensive odors compounds (i: .-Methylphenol, ii: unknown compound I, iii: unknown compound II and iv: Indole) of the porcine large intestine were identified comparing their mass spectra with mass spectral databases and comparing their retention index. ῎ The coriander odors compounds (from Decanal; +, ,-Decenal; ,, Undecanal; -, ,-Undecenal; ., +-Decanol; /,(E)-,-Undecen- +-ol; 0, ,-Dodecenal; 1,(E)-,-Tetradecenal; 2, Hexadecanal; 3, Octadecenal; +* and 3-Octadecenal; ++) of coriander were identified comparing their mass spectra with mass spectral databases and comparing their retention index.

the e%uent to be split between a sni$ng port and a flame with diethyl ether in the ratio +: -n, where n was selected ionization detector (FID). The column was a DB-Wax from +, ,, - or n as the dilution factor. All odor qualities fused silica capillary column (0* mῌ*.,/ mm i.d.; J&W were defined by two assessors, whereas stepwise dilution Scientific Inc., Folsom, USA), and the oven temperature analysis was performed only one time. was maintained at .*῍ for +* min and then increased by The deodorizing activity test The o#ensive odor sam- -῍/min to ,,*῍, where it was maintained for -* min. ples (,.* g of the porcine internal organs) were placed in a The injector and detector temperatures were ,-* and +**-ml conical beaker and boiled for +* minutes, after ,/*῍, respectively. The flow rate of the helium carrier which *.+ g, *./ gand,.* g of the samples was added to the gas was -* cm/s. The split ratio of the injector was +: ,*. coriander or the wild grasses, herbs, and green tea, respec- The values of the relative amounts of volatile compounds tively. The odor of the porcine large intestine was were calculated by dividing the GC peak area by the sni#ed by the ten trained members (ten healthy adults, internal standard area. The calculations of these relative four men and six women, ranging in age from ,+ to .1 amount values were repeated four times. In the sni$ng years, who were recruited at Hiroshima Prefectural Uni- experiments, three panelists sni#ed directly from the versity; all subjects were non smokers), and the deodoriz- sni$ng port, and the odor attributes of each compound ing activity test was carried out. The deodorizing activ- were repeatedly assessed by di#erent panelists on sepa- ity against the o#ensive odor was measured using / rate runs. The expression of each aroma compound was grades (Perfectly deodorized: *, Almost deodorized: +, determined by agreement between at least two of the Considerably deodorized: ,, A little deodorized: -, Com- trained members. pletely not deodorized: .). Aroma extract dilution analysis (AEDA) The signifi- cance of each odor compound in the porcine large intes- Results and Discussion tine odor was evaluated by its potency using the AEDA Up to now, there have been many reports on the deo- method of Hayata et al (,**-). The volatile extract (/* ml) dorization e#ect on o#ensive odors of wild grasses and collected using the SDE method was used and diluted herbs, so we compared the deodorization e#ect of corian- Deodorizing E#ect of Coriander on the O#ensive Odor of the Porcine Large Intestine 41

Table ,. Comparison of main volatile compound of coriander and the porcine large intestine.

Abbreviations: PN, Peak number; KI, Kovats index; nd, not detected. ῌ+῍ Values are GC peak area of compound/GC peak area of internal standard. ῌ,῍ Peak number (see in Figure +.). ῌ-῍ Kovats index on DB-WAX ῌ.῍ Symbols are the odor intensity of each volatile compound in the odor character of a porcine large intestine with coriander *./ g(῎῎῎: very strong, ῎῎: strong, ῎: a little strong). ῌ/῍ The odor character of the volatile compounds by GCO. ῌ0῍ Values presented as means and standard errors for - times. a, b, c: Mean values with di#erent superscripts are significantly di#erent according to the LSD test (pῐ*.*/). der to several of these plants. A comparison of the deo- *./ g coriander by SDEM revealed that a total of +23 and 21 dorization e#ects against the o#ensive odor of porcine volatile compounds in both treatments were contained, large intestine is shown in Table +. The deodorization respectively (Fig. +). Among these compounds, 23 and -0 e#ect of coriander was significantly elevated when the of the volatile compounds in the porcine large intestine treatment amount increased from *.+ gto,.* g. Wild with and without coriander were clarified to have odors grasses such as dandelion and spiny sowthistle had a by GC-O. The four main compounds were detected as strong deodorant e#ect against the o#ensive odor in the porcine large intestine odor, and the eleven main Methyl mercaptan (Urabe et al., +333), but not against the compounds were sni#ed as coriander odor, as shown in porcine large intestine. Whereas the deodorization e#ect Table ,. By GC-MS and the Kovats index (KI), the four of coriander was higher than thyme, and that rosemary volatile compounds were .-Methylphenol (i: sludge-like and green tea had the deodorization e#ect against the odor), unknown compound I (ii: KI῏,,0/, sludge-like o#ensive odor in raw meat or fish. In this study, the odor), unknown compound II (iii: KI῏,-,3, porcine large e#ect of coriander was maintained longer than 0 hrs, even intestine-like odor) and Indole (iv: excrement-like odor). though the smell of coriander had almost disappeared In the test of odor intensity by GCO, .-Methylphenol and within - hrs after the treatment. On the other hand, the Indole were very strong (῎῎῎), and the unknown com- deodorization e#ect of green tea declined rapidly as time pound II and the unknown compound I were strong (῎῎) passed concomitant with the decrease of its green tea and a little strong (῎), respectively. The FD values of odor. In both thyme and rosemary, the strong odor and Indole and unknown compound II were the highest deodorant e#ects lasted for 0 hrs. These results sug- among all the main compounds (FD῏1,3), followed by .- gested that the deodorization e#ect of coriander is not Methylphenol and unknown compound I (FD῏,.-) (data dependent on its odor as a masking e#ect, whereas the not shown). .-Methylphenol is the main end product of odor of green tea plays a role in its deodorant e#ect on the fermentation of the amino acid tyrosine, and Indole o#ensive odors. results from the microbial degradation of tryptophan The chromatogram of the volatile compounds in the (Mackie et al., +332). It has been reported that .-Meth- porcine large intestine with and without the treatment of ylphenol or Indole are o#ensive odors from feces in pigs 42 K. KOHARA et al.

(Schaefer, +331). In this study, we could not identify of g-irradiated fresh cilantro leaves during cold storage. J. unknown compounds I and II because these compounds Agric. Food Chem., /*, 10,,ῌ10,0. +33. , 0 were at a very low level in the o#ensive odor for GC-MS Harasawa, A. and Tagashira, A. ( ). Isolation of , -Dimethoxy +,.-benzoquinone from Hydrangea (Hydrangea macrophylla analysis, and the results suggest that the compounds .- Seringe var. otaksa Makino) and its deodorant activity against # Methylphenol and Indole are responsible for the o ensive methyl mercaptan. Biosci. Biotech. Biochem., /2, ,*1-ῌ,*1.. odor of the porcine large intestine, regardless of feces. Hayata, Y., Sakamoto, T., Maneerat, C., Li, X., Kozuka, H. and Eleven main compounds having a coriander odor were Sakamoto, K. (,**-). Evaluation of aroma compounds con- identified: Decanal, ,-Decenal, Undecanal, ,-Undecenal, +- tributing to muskmelon flavor in Porapak Q extracts by aroma Decanol, (E)-,-Undecen-+-ol, ,-Dodecenal, (E)-,-Tetrade- extract dilution analysis. J. Agric. Food Chem., /+, -.+/ῌ-.+2. +330 cenal, Hexadecanal, Octadecenal and 3-Octadecenal. In Ikemoto, M. ( ). Deodorants of natural plant extract. J. Odor ,1 +-0ῌ+., , , , 1 Research and Eng., , (in Japanese). particular, -Decenal (No. ), -Dodecenal (No. )and ,**. +* Kubo, I., Fujita, K., Kubo, A., Nihei, K. and Ogura, T. ( ). Anti- Octadecenal (No. ) had the strong characteristic odor bacterial activity of volatile compounds against Salmonella +330 , of coriander. Potter ( ) previously reported that - choleraesuis. J. Agric. Food Chem., /,, --,3ῌ---,. Decenal, ,-Dodecenal and (E)-,-Tetradecenal are the main Loaiza, J. and Cantwell, M. (+331). Postharvest physiology and components in coriander leaf oil. Those compounds quality of cilantro. HortScience, -,, +*.ῌ+*1. were also detected, and Octadecenal was clarified to be an Mackie, R.I., Stroot, P.G. and Varel, V.H. (+332). Biochemical iden- important compound for the coriander odor in our re- tification and biological origin of key odor components in livestock waste. J. Anim. Sci., 10, +--+ῌ+-.,. search. The peak area ratios of eleven volatile com- Maneerat, C., Hayata, Y., Kozuka, H., Sakamoto, K. and Osajima, pounds were significantly increased by the treatment of Y. (,**,). Application of the Porapak Q column extraction * + , * coriander from . gto . g fresh weight. method for tomato flavor volatile analysis. J. Agric. Food As for the deodorization mechanism, four types of Chem., /*, -.*+ῌ-.*.. mechanisms have been reported: chemical, physical, bio- Nakatani, N., Miura, K. and Inagaki, T. (+323). Structure of deodor- logical and sensuous deodorant activities (Ikemoto, +330). ant biphenyl compounds from Thyme (Thymus vulgaris L.) and /- Those deodorant activities, except sensuous activities, are their activity against methyl mercaptan. Agric. Biol. Chem., . +-1/ῌ+-2+ # . due to the decomposition or absorption of o ensive odor +330 # Nishida, K. and Ishikawa, Y. ( ). The neutralize deodoriza- compounds. The sensuous deodorizing e ect is thought tionῌmasking and modification of deodorization e#ect on # to be a masking and modification e ect (Nishida and o#ensive odor. PPM., /, 1-ῌ2, (in Japanese). Ishikawa, +330). The former refers to hiding the o#en- Potter, T.L. and Fagerson, I.S. (+33*). Composition of coriander sive odor with another strong odor, and the latter refers leaf volatiles. J. Agric. Food Chem., -2, ,*/.ῌ,*/0. to modulation of the o#ensive odor compound to be a Schaefer, J. (+311). Sampling characterization and analysis of - +,+ῌ+,1 di#erent odor compound. malodors. Agric. Environ., , . Smallfield, B.M., Klink, J.W., Perry, N.B. and Dodds, K.G. (,**+). We found that the deodorization mechanism of corian- # # Coriander spice oil: E ects of fruit crushing and distillation der to the o ensive odor of the porcine large intestine was time on yield and composition. J. Agric. Food Chem., .3, ++2ῌ not due to chemical or physical activities, because the +,-. treatment of coriander did not decompose the main o#en- Taskiness, J. and Nykanen, L. (+31/). Volatile constituents ob- sive odors. Moreover, coriander’s deodorization e#ect is tained by the extraction with alcohol-water mixture and by not considered to be a masking deodorization by the steam distillation of coriander fruit. Acta. Chem. Scand., B,3, .,/ῌ.,3 coriander smell, because the remarkable deodorant e#ect . Tokita, F., Ishikawa, M., Shibuya, K., Koshimizu, M. and Abe, R. was maintained for 0 hrs, even though the smell of corian- (+32.). Deodorizing activity of some plant extracts against der rapidly disappeared. methyl mercaptan. Nippon Nogeikagaku kaishi., /2, /2/ῌ/23 (in Japanese). References Urabe, K., Nadamoto, T., Kawamura, M. and Yasumoto, K. (+333). Bandoni, A.L., Mizrahi, I. and Juares, M.A. (+332). Composition E#ects of Houttuyniae cordata and refinery final molasses on and quality of the essential oil of coriander (Coriandrum the development of o#ensive odor in porcine small intestine sativum L.) from Argentina. J. Essent. Oil Res., +*, /2+ῌ/2.. during storage. J. Nutr. Sci. Vitaminol., .*, 0-ῌ1+. Fan, X. and Sokorai, K.J.B. (,**,). Changes in volatile compounds