• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.11
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• pp.1796-1800
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• 2014

Trienzyme digestion (AOAC Official Method 2004.05) procedure using protease, ${\alpha}$-amylase, and chicken pancreas conjugase was evaluated to determine its usefulness in the microbiological quantitation of total folate in foods. Folate values obtained by alkali hydrolysis (Korean Food Standards Codex) were compared to those obtained by the trienzyme method for four certified reference materials (CRM) representing diverse matrixes. Trienzyme treatment increased measurable folate from most CRM compared to levels found after alkali hydrolysis. The largest increases were observed with CRM 487 (pig liver, 5.8-fold) and CRM 121 (whole meal flour, 3.1-fold) after trienzyme digestion. Using trienzyme digestion method, total folate contents of raw and blanched edible plants were determined. Eleutherococcus senticosus ($146.9{\mu}g/100g$) showed the highest total folate content, followed by Aster glehni F. Schmidt ($142.8{\mu}g/100g$) and Ledebouriella seseloides H. Wolff ($140.4{\mu}g/100g$) on a wet weight basis. Blanching of samples resulted in an insignificant decrease in folate content for five samples and 11~63% reduction for nine samples. Our finding suggests that trienzyme digestion method is accurate for the determination of food folate in leafy vegetables.

• Korean Journal of Food Science and Technology
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• v.52 no.6
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• pp.580-586
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• 2020

Fishery resources have been widely consumed as protein- and vitamin-rich food sources in the Korean diet. However, information regarding their vitamin levels is extremely limited. In this study, trienzyme-Lactobacillus casei method was validated and used to determine the folate contents in fishery foods. The trienzyme-L. casei method for folate analysis showed excellent accuracy (85.2 to 95.3% recovery) and precision (repeatability 1.4% RSD and reproducibility 2.4% RSD). Folate contents of 20 fish foods (4 fish, 3 crustaceans, 3 sea algae, 3 cephalopods, 4 shellfish, and 3 others) ranged from 1.75 to 97.98 ㎍/100 g. Furthermore, we found that the folate content in seaweed fusiforme was the highest, followed by gulfweed (69.73 ㎍/100 g). Folate analysis using the trienzyme-L. casei method was determined excellent based on the z-score of -0.3 in the Food Analysis Performance Assessment Scheme test. Analytical and method validation data generated in this study could be used to update the national food composition table on vitamin B9 in Korean fishery resources.

• Food Science and Preservation
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• v.24 no.8
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• pp.1103-1112
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• 2017

This study was performed to investigate the effects of different cooking methods (boiling, roasting, stir-frying, and deep-frying) on folate retention in 6 kinds of mushrooms (Beech-, button-, Juda's ear-, oak-, oyster-, and winter-mushrooms) frequently consumed in Korea. In order to assure reliability of analytical data, trienzyme extraction-L casei method was verified and analytical quality control was also evaluated. Folate contents of mushrooms varied by 6.04-64.82 g/100 g depending on the type of mushrooms. and were significantly affected by cooking methods. Depending on cooking methods, folate contents of mushrooms decreased by 22-48%, 2-31%, and 17-56% for Juda's ear-, oak- and oyster-mushrooms, respectively, while 17-90% of folate was increased in Beech mushroom. Overall, the largest weight loss was found in boiled mushrooms, but the lowest one in deep-fried samples. True folate retention rates considering processing factor were less than 100% for all cooked mushrooms except for Beech samples. Overall, folate loss was the largest by boiling with water but the smallest by deep-frying. Both accuracy and precision of trienzyme extraction-L-casei method were excellent based on a recovery close to 100% and coefficient variations less than 3%. Quality control chart of folate analysis (n=26) obtained during the entire study and an international proficiency test (z-score=-0.5) showed that trienzyme extraction-L casei method is reliable enough for production of national folate database.

• Food Science and Biotechnology
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• v.18 no.1
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• pp.31-35
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• 2009

Total folate content was determined by microbiological assay using Lactobacillus casei spp. rhamnosis (ATCC 7469) with a 96-well microplate technique. Using roasted peanut and red kidney beans as representative legume samples, response surface methodology (RSM) was supplied to optimize the trienzyme procedures for the determination of folate in legumes. After response surface regression (RSREG), the second-order polynomial equation was fitted to the experimental data. Ridge analysis showed that the optimal digestion times were <2 hr for $Pronase^{(R)}$ and $\alpha$-amylase, and <5 hr for conjugase to obtain maximal folate values for legume samples. This study confirms that established digestion times for cereal products (AOAC Method 2004.05) of 3 for protease and 2 hr for $\alpha$-amylase are applicable to legumes. Conjugase treatment can be reduced to 5 from 16 hr and the conjugase level to 5 from 20 mg per sample, providing significant cost saving.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.5
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• pp.716-724
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• 2015

Trienzyme extraction coupled with microplate assay (Lactobacillus casei subsp. rhamnosus) was validated and applied for the determination of folate (vitamin B9) in Korean ancestral rite foods. Foods included five Guk (Tang), eleven Sookchaes, eight Jeoks, nine Jeons, six Jjims, and twenty desserts. Folate was detected in all samples: Guk (Tang) 4.62~18.84, Sookchae 6.13~48.40, Jeok 5.49~49.50, Jeon 6.96~30.77, Jjim 10.34~38.88, and desserts $3.33{\sim}49.55{\mu}g/100g$. The lowest folate content was observed in Sikhye ($3.33{\mu}g/100g$), whereas the highest was observed in Songhwa-dasik ($49.55{\mu}g/100g$). Folate analyses of certified reference materials, BCR-121 (whole meal flour) and BCR-487 (pig liver), showed good recoveries of 90.0% (0.45 mg/kg) and 92.4% (12.3 mg/kg), respectively. The recoveries (96.0 to 106.2%) obtained by analyzing eight spiked samples with different matrices also showed good accuracy. Both repeatability and reproducibility were less than 5%, indicating good precision. The quality control chart (n>30) obtained by running commercial folate fortified-wheat flour once a week for about 10 months showed that all assays were under control. All validation method and analytical quality control results showed that folate contents in Korean ancestral rite foods produced by microplate assay were reliable enough to be used for the construction of a national folate database.

• Journal of Nutrition and Health
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• v.38 no.7
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• pp.586-604
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• 2005

A reliable nutrient database is a prerequisite for accurate calculation of dietary intakes. The folate database currently available in Korea, however, is not reliable because the values were obtained from published data in other countries using ineffective methods to extract folates from the food matrix. The purpose of this study was to complement the folate database by analyzing folate content in foods using a more effective method to extract food folates (trienzyme treatment). Folate content per unit weight was highest in laver, fermented soybeans, soybean, spinach, black soybeans, crown daisy, mung beans, and quail's egg in descending order. Legumes, leafy greens, eggs, and seaweeds were rich in folate, and meats, chicken, fish, and some fruits contained less folate. Some of the analyzed values were 10 times higher than those in the currently available database. Folate values of 423 foodcodes out of 2,932 foodcodes ($14.4\%$) in the database in the 7th revision in the Recommended Dietary Allowances for Koreans can be replaced by those analyzed in this study. Since folate values of rice and Kimchi, which are core dishes of Koreans, in the newly established database are higher than those in the current database, folate intake assessed using our data will be higher than that using the current available database. Folate content in more foods commonly consumed in Korea are needed to update the folate database. Meanwhile, folate values presented here can be used to assess dietary folate intake of the Korean population.

• Journal of Nutrition and Health
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• v.42 no.4
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• pp.397-405
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• 2009

A trienzyme extraction method (use of ${\alpha}$-amylase, protease and folate conjugase) for food folate assay has been used to release folate from the food matrix. In order to reduce the incubation time with three enzymes, folate values were compared between two incubation protocols; separate incubation (SI, incubated with ${\alpha}$-amylase and conjugase separately for 2 hours after protease treatment) and combined incubation (CI, incubated with ${\alpha}$-amylase and conjugase together for 2 hours after protease treatment) using 88 food items from 12 kinds of fast foods and processed foods. We found that folate values by CI were comparable to or higher than those by SI, indicating that CI might be a better extraction procedure to shorten the entire incubation time. We measured folate contents in 49 fast foods and 26 processed foods by microbiological assay after CI. Mean folate contents of one serving of various burgers ranged from 43.1 to 62.0 ${\mu}g$. One serving of French fries, pizza, sandwich and triangled kimbab contained a mean of 53.3, 28.4, 47.4, and 25.7 ${\mu}g$ of folate, respectively. Folate contents of non-alcoholic beverages were very low, ranging from 1.0 to 5.2 ${\mu}g$/100 g. Some of our values were comparable to the values in the folate database published in Korean Nutrition Society, however, some of the published values were 140 times higher than the measured values in this study. Folate values measured by the more recent modifications here can be used to update Korean folate database to accurately estimate dietary folate intake.

• Journal of Nutrition and Health
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• v.54 no.2
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• pp.211-223
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• 2021

Purpose: A total of 39 seafood menus were prepared according to the Korean standard recipe, and analyzed for vitamin B₉ (folate) and B₁₂ (cobalamins) contents, using validated applied analytical methods. The menus included Guk/Tang/Jjigae (boiled or stewed dishes, n = 10), Bokkeum (stir-fried dishes, n = 10), Jjim/Jorim (braised or steamed dishes, n = 7), Gui (baked or grilled dishes, n = 7), Twigim (deep-fried dishes, n = 2) and Muchim (dried or blanched-seasoned dishes, n = 3). Methods: The contents of vitamin B₉ and B₁₂ in all food samples were determined by the trienzyme extraction-Lactobacillus casei and immunoaffinity-high-performance liquid chromatography/photodiode array detection methods. Analytical quality control was performed in order to assure reliability of the analysis. Results: Accuracy (97.4-100.6% recoveries) and precision (< 6% relative standard deviations for repeatability and reproducibility) of vitamin B₉ and B₁₂ analyses were determined to be excellent. The vitamin B₉ and B₁₂ contents of the 39 seafood menus evaluated, varied in the range of 1.83-523.08 ㎍/100 g and 0.11-38.30 ㎍/100 g, respectively, depending on the ingredients and cooking methods. The vitamin B₉ content was highest in Jomi-gim (523.08 ㎍/100 g), followed by Geonsaeu-bokkeum (128.34 ㎍/100 g) and Janmyeolchi-bokkeum (121.53 ㎍/100 g). Vitamin B₁₂ was detected in all seafood menus, with highest level obtained in Kkomack-jjim (41.58 ㎍/100 g). The seaweed dish was found to have high levels of both vitamin B₉ and B₁₂. All assays were performed under strict quality control. Conclusion: Guk and Tang menus, which contain a large amount of water, were relatively lower in the vitamin B₉ and B₁₂ contents than the other menus. Bokkeum menus containing various vegetables were high in the vitamin B₉ content, but the vitamin B₁₂ content was dependent on the type of seafood used in the menu.

• Journal of Nutrition and Health
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• v.48 no.1
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• pp.94-104
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• 2015

Purpose: The purpose of this study was to estimate dietary folate intake and food sources of children and adolescents using a nutrient database revised based on measured folate in selected foods. Methods: Folate content in 51 foods known as folate sources was measured by microbiological assay after trienzyme extraction. Folate intake was estimated from a part of the data of 'Dietary Intake Survey of Infants, Children and Adolescents in 2007~2008' conducted by the Korea Food and Drug Administration (KFDA) and the Korea Health Industry Development Institute (KHIDI). The study subjects were 567 children aged 1~19 years living in the Chungcheong area who completed two 24-hour recalls. Results: Folate values were revised by replacing the values in the current database with the analyzed values except when the value in the current database was between the analytical values or was not different from the mean analytical value by more than 10%. Among the revised values of 40 food items, folate values of 36 foods were lower than the current values. Mean folate intakes estimated with the revised database were approximately 70~80% lower than those estimated with the current database. Mean folate intakes of males aged 12~14 and females aged 12~19 were lower than the 2010 Recommended Nutrient Intakes (RNIs). Chicken's eggs, Kimchi, rice, mandarin, and laver were found to be main food sources of folate. Conclusion: In this study, mean dietary folate intakes were lower than those estimated with the current database. Further analyses for folate content especially in cooked foods commonly consumed in Korea are needed using a reliable assay in order to accurately assess folate intake of the Korean population. In addition, nutrition education should be provided for adolescent females in order to increase consumption of folate-rich foods.

• Korean Journal of Food Science and Technology
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• v.51 no.5
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• pp.425-431
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• 2019

Selected leafy vegetables, widely used for Korean Namul dishes, were heat-treated in different ways and their folate retention was investigated. The Lactobacillus casei method was applied for folate estimation and validated to ensure reliability of analytical data. The folate content in Namul highly varied, from 29.7 to $293.4{\mu}g/100g$, depending on the heating methods and the types of vegetables. Most of the Namul variants showed increased folate content on heat treatment. Frying yielded higher folate retention than the other cooking methods (blanching, steaming, baking, and panfrying), and pig weed showed the highest folate retention (3.3 times, $293.4{\mu}g/100g$). L. casei assay for folate estimation showed 95.7% recovery and relative standard deviations less than 2% for both reproducibility and repeatability, indicating good accuracy and precision. Quality of the folate assay was assured by monitoring a quality control chart and a proficiency test (z-score= -0.1) during the entire of study.