• 대한본초학회지
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• 제33권3호
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• pp.25-35
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• 2018

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

• 대한본초학회지
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• 제34권5호
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• pp.13-20
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• 2019

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.

• 한국근적외분광분석학회:학술대회논문집
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• 한국근적외분광분석학회 2001년도 NIR-2001
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• pp.1246-1246
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• 2001

The quality of meat is highly variable in many properties. This variability originates from both animal production and meat processing. At the pre-slaughter stage, animal factors such as breed, sex, age contribute to this variability. Environmental factors include feeding, rearing, transport and conditions just before slaughter (Hildrum et al., 1995). Meat can be presented in a variety of forms, each offering different opportunities for adulteration and contamination. This has imposed great pressure on the food manufacturing industry to guarantee the safety of meat. Tissue and muscle speciation of flesh foods, as well as speciation of animal derived by-products fed to all classes of domestic animals, are now perhaps the most important uncertainty which the food industry must resolve to allay consumer concern. Recently, there is a demand for rapid and low cost methods of direct quality measurements in both food and food ingredients (including high performance liquid chromatography (HPLC), thin layer chromatography (TLC), enzymatic and inmunological tests (e.g. ELISA test) and physical tests) to establish their authenticity and hence guarantee the quality of products manufactured for consumers (Holland et al., 1998). The use of Near Infrared Reflectance Spectroscopy (NIRS) for the rapid, precise and non-destructive analysis of a wide range of organic materials has been comprehensively documented (Osborne et at., 1993). Most of the established methods have involved the development of NIRS calibrations for the quantitative prediction of composition in meat (Ben-Gera and Norris, 1968; Lanza, 1983; Clark and Short, 1994). This was a rational strategy to pursue during the initial stages of its application, given the type of equipment available, the state of development of the emerging discipline of chemometrics and the overwhelming commercial interest in solving such problems (Downey, 1994). One of the advantages of NIRS technology is not only to assess chemical structures through the analysis of the molecular bonds in the near infrared spectrum, but also to build an optical model characteristic of the sample which behaves like the “finger print” of the sample. This opens the possibility of using spectra to determine complex attributes of organic structures, which are related to molecular chromophores, organoleptic scores and sensory characteristics (Hildrum et al., 1994, 1995; Park et al., 1998). In addition, the application of statistical packages like principal component or discriminant analysis provides the possibility to understand the optical properties of the sample and make a classification without the chemical information. The objectives of this present work were: (1) to examine two methods of sample presentation to the instrument (intact and minced) and (2) to explore the use of principal component analysis (PCA) and Soft Independent Modelling of class Analogy (SIMCA) to classify muscles by quality attributes. Seventy-eight (n: 78) beef muscles (m. longissimus dorsi) from Hereford breed of cattle were used. The samples were scanned in a NIRS monochromator instrument (NIR Systems 6500, Silver Spring, MD, USA) in reflectance mode (log 1/R). Both intact and minced presentation to the instrument were explored. Qualitative analysis of optical information through PCA and SIMCA analysis showed differences in muscles resulting from two different feeding systems.

• 한국식품영양과학회지
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• 제39권12호
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• pp.1867-1872
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• 2010

신선편이 식품인 진공 포장된 상추를 보관하면서 미생물과 품질을 분석하고자 하였다. 염소수로 살균 소독되어 진공 포장된 상추를 $5^{\circ}C$, $15^{\circ}C$, $25^{\circ}C$에서 7일까지 보관하면서 일반세균, 대장균군, 대장균, 식중독세균, 젖산균 생균수 분석과 포장내의 가스 및 관능검사를 하였다. 보관 실험 전에 총 균수가 2 log CFU/g, 대장균군은 1 log CFU/g 오염되어 있었고 젖산균은 검출되지 않았다. $5^{\circ}C$에서의 7일까지 보관하는 동안에 일반세균수, 대장균군, 젖산균의 증식은 1 log CFU/g이었으며 다른 병원성 세균은 검출되지 않았다. 그러나 이취와 아삭아삭함 등의 관능적 특성을 고려했을 때 5일 보관 시에 최상의 절반정도의 관능도를 보여 주었다. $15^{\circ}C$에서의 보관 시 3일 보관할 때 일반세균수는 3 log CFU/g, 대장균군은 2 log CFU/g 증식하였으나 젖산균은 4 log CFU/g 증식하여 현저히 증식이 잘 일어났고 다른 병원성 세균 중 B. cereus만이 증량 배양 후에 검출되었다. 이취, 아삭아삭함 등은 3일 보관 시에 최상의 절반정도의 관능도를 보여주었다. $25^{\circ}C$에서의 저장 시 1일만의 일반세균수의 3 log CFU/g, 대장균군은 1 log CFU/g, 젖산균은 4 log CFU/g 증가하였다. 식중독 세균 중 B. cereus만이 증량 배양 후에 검출되었다. 관능검사평가도 1일 안에 모든 항목에서 최상의 절반 이하로 낮게 나왔다. 그러므로 신선편이 상추의 오염세균 증식을 저해하기 위하여 가능한 낮은 온도로 보관하는 것이 바람직하다. 흥미롭게도 검출 세균 중에 젖산균의 증식이 혐기조건에서 크게 일어났는데 이들 젖산균은 신선편이 식품의 품질 저하와 관련되고 있을 것으로 사료된다.

• 한국식품위생안전성학회지
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• 제31권1호
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• pp.28-35
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• 2016

본 연구에서는 국내 대표 식육인 소, 돼지, 닭, 오리의 4종 식육과 염소, 양, 말, 칠면조의 4종 식육을 동시에 신속하게 감별할 수 있는 2 set의 multiplex PCR법을 개발하고자 미토콘드리아 16S RNA에서 종 특이부위를 선발하고 각 종에 대한 특이도를 높이기 위하여 인위적인 미스매치를 주어 프라이머를 제작한 후 8종 식육의 274개 시료를 대상으로 특이도와 민감도를 조사하였다. 그 결과 소, 돼지, 닭, 오리 모든 시료에서 각각 279, 94, 192, 477 bp의 증폭산물이, 말, 양, 염소, 칠면조의 모든 시료에서 각각 152 bp, 271 bp, 670 bp, 469 bp에서 뚜렷한 PCR 유전자 산물이 확인되어 모든 축종에서 100%의 특이도를 나타내어 축종별 감별력이 우수한 것으로 나타났다. 8종의 축종별로 DNA를 $10ng/{\mu}l$으로 정량한 후 혼합물을 10배씩 단계 희석하여 반응여부를 조사한 결과, 소, 돼지, 오리에서는 100 fg까지, 닭에서는 1 pg까지 검출됨을 확인할 수 있었다. 소, 돼지, 닭, 오리고기를 99.9%, 99%, 90%, 70%, 50%, 30%, 10%, 1%, 0.1%의 비율로 혼합한 식육과 $83^{\circ}C$ 20분, $100^{\circ}C$ 30분, $121^{\circ}C$ 10분에서 각각 열처리한 가열 혼합육에 대하여 검출한계를 조사한 결과 마지막 단계의 희석 비율인 모든 혼합육의 0.1%에서 검출이 가능하였으며, 열처리 혼합육에서는 닭에서는 1% 농도에서 소와 돼지의 혼합육에서 0.1% 농도에서 검출되어 민감도가 높음을 확인할 수 있었다. 본 연구에서 개발된 multiplex PCR법은 특이도 및 민감도에 있어서 국내 대표 식육을 감별하는데 있어서 유용한 것으로 평가된다.

• 한국식품위생안전성학회지
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• 제27권1호
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• pp.68-73
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• 2012

본 연구에서는 축산물가공품 중 식육원료의 진위여부를 판별하기 위하여 분자생물학적 기법을 이용한 시험법을 개발하였다. 식육원료의 종 판별을 위한 유전자로는 미토콘드리아 DNA에 존재하는 12S 또는 16S 유전자를 대상으로 하였으며, 가공식품에 적용하기 위하여 PCR 산물의 크기는 200bp 내외가 되도록 프라이머(species-specific primer)를 설계하였다. 대상 식품원료로는 축산물 10종을 대상으로 하였으며 프라이머를 사용하여 유전자증폭(PCR) 후 전기영동 하여 예상되는 PCR 산물의 생성유무를 확인하였다. PCR을 실시한 결과 가축인 소고기, 돼지고기, 염소고기, 양고기, 사슴고기, 말고기에 대하여는 각각 131, 138, 168, 144, 191, 142bp에서, 가금류인 닭고기, 오리고기, 칠면조 고기, 타조고기에 대하여는 각각 281, 186, 174, 238bp에서 예상크기의 PCR 산물을 확인하였다. 그리고 프라이머 별로 유사 종에서는 비 특이적 PCR 산물(non-specific PCR product)은 생성되지 않았다. 본 연구에서 개발된 프라이머를 이용한 유전자분석법을 돼지고기 및 닭고기를 함유하는 축산물가공품, 소고기를 함유하는 복합조미식품을 대상으로 시험한 결과 적용 가능함이 확인되어 향후 축산물가공품 중 식육원료의 진위여부 판별에 활용이 가능할 것으로 판단된다.

• 대한예방한의학회지
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• 제6권2호
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• pp.66-85
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• 2002

Traditional herbal medicine is used extensively among the Korean populations, and other Asian countries employ similar therapies as well In recent years, extensive focus was laid on adulteration of the herbal medicine with heavy metals. This may be mainly due to a soil contamination by an environmental pollution. The objective of this study is to identify the contents of various heavy metals in the blood from OhJeokSan-Decoction (OD) treated-rats. For this study, 13 kinds of metals including essential and heavy metals, i.e. Al, As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn were analyzed by a slight modification of EPA methods and the following results are obtained. ; 1. There is no significant difference between the OD-treated groups and control group in liver, kidney, bone, brain, weight, food intake. 2. The amount of each metal analyzed in the liver are as follows; Al ; $0.235{\sim}4.215mg／kg$, As ; $0.103{\sim}0.461mg／kg$, Cd ; $0.005{\sim}0.010mg／kg$, Co ; $0.017{\sim}0.046mg／kg$, Cr ; $0.137{\sim}0.403mg／kg$, Cu ; $1.736{\sim}4.827mg／kg$, Fe ; $54.472{\sim}381.447mg／kg$, Hg ; not detected, Mn ; $1.159{\sim}2.803mg／kg$, Ni ; $0.007{\sim}0.095mg／kg$, Pb ; not detected, Se ; $0.682{\sim}1.887mg／kg$, Zn ; $10.213{\sim}26.119mg／kg$, by groups, respectively. In control and other experimental group, several metal (Co, Cu, Mn, Zn, As, Cr) has a significant difference, but in experimental I and other experimental II, III, IV, V groups, there are no significant difference. 3. The amount of each metal analyzed in the kidney are as follows; Al ; $1.712{\sim}31.230mg／kg$, As ; $0.062{\sim}0.439mg／kg$, Cd ; $0.010{\sim}0.062mg／kg$, Co ; $0.000{\sim}0.101mg／kg$, Cr ; $0.125{\sim}0.636mg／kg$, Cu ; $3.385{\sim}12.502mg／kg$, Fe ; $41.148{\sim}99.709mg／kg$, Hg ; $0.000{\sim}0.270mg／kg$, Mn ; $0.433{\sim}2.301mg／kg$, Ni ; $0.000{\sim}0.221mg／kg$, Pb ; $0.000{\sim}0.584mg／kg$, Se ; $0.540{\sim}1.600mg／kg$, Zn ; $8.775{\sim}17.060mg／kg$, by groups, respectively. The concentration of Cu, Se, Cr, and Hg are variated significantly in control and other experimental group, and Cu, Se, Cd, Cr are variated significantly in experimental I and other experimental II, III, IV, V groups. 4. The amount of each metal analyzed in the bone(tibia and fibula) are as follows; Al ; $9.557{\sim}119.464mg／kg$, As ; $0.139{\sim}12.250mg／kg$, Cd ; $0.000{\sim}0.295mg／kg$, Co ; $0.022{\sim}0.243mg／kg$, Cr ; $0.239{\sim}1.999mg／kg$, Cu ; $0.000{\sim}2.291mg／kg$, Fe ; $240.249{\sim}841.956mg／kg$, Hg ; $0.000{\sim}0.983mg／kg$, Mn ; $0.214{\sim}7.353mg／kg$, Ni ; $5.473{\sim}11.453mg／kg$, Pb ; $0.000{\sim}8.502mg／kg$, Se ; $0.000{\sim}3.005mg／kg$, Zn ; $61.158{\sim}195.038mg／kg$, by groups, respectively. The concentration of Se, Cd are variated significantly in control and other experimental groups, and Se is variated significantly in experimental I and other experimental II, III, IV, V groups. 5. Exceptionally several metal concentration is increased or decreased. but there is no significant harmful difference of metal concentration in the liver, kidney and bone, from the OD-treated-rats compared to those of the control group even if higher dosage($1{\sim}8$ times dosage of person) of OD was administered. Thus, it is expected that the herbal decoction in the traditional herbal medicine would not lay any burden on the body and the heavy metal toxins would not affect our physiological system. However, other kinds of herbal treatment, such as i.v. and i.p. should be considered in terms of metal toxicity in the body since the level of certain metal.