Effects of Combined Microwaving/Atmospheric Steaming with Xylanase Enzymolysis on Endogenous Enzyme Activity and Antioxidant Activity in Wheat Bran-Germ

CHEN Peng; HAN Jing; SUN Binghua; ZHENG Xueling; LI Li; MA Sen

doi:10.13386/j.issn1002-0306.2024060397

Turn off MathJax

Article Contents

Abstract

References

CHEN Peng, HAN Jing, SUN Binghua, et al. Effects of Combined Microwaving/Atmospheric Steaming with Xylanase Enzymolysis on Endogenous Enzyme Activity and Antioxidant Activity in Wheat Bran-Germ[J]. Science and Technology of Food Industry, 2025, 46(9): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024060397.

Citation:

PDF (3145 KB)

Effects of Combined Microwaving/Atmospheric Steaming with Xylanase Enzymolysis on Endogenous Enzyme Activity and Antioxidant Activity in Wheat Bran-Germ

College of Grain, Oil and Food, Henan University of Technology, Zhengzhou 450001, China

More Information

Received Date: June 26, 2024
Available Online: February 28, 2025

Graphical Abstract

Abstract

Abstract

Oxidative rancidity plays a crucial role on the quality of wheat processing by-products—wheat bran-germ, and in order to attenuate its adverse effects on wheat bran-germ products. This study aimed to investigate the effect of six wheat bran-germ modification treatments (microwaving, atmospheric steaming, enzymolysis, atmospheric steaming combined microwaving, microwaving combined enzymolysis, atmospheric steaming combined enzymolysis) on endogenous enzyme activity, color, total phenolic contents, and antioxidant activity of wheat bran-germ. The results showed that all the single treatment techniques significantly reduced the activities of lipase, lipoxidase and peroxidase in wheat bran-germ under optimal conditions, especially atmospheric steam and enzymatic hydrolysis inactivated the three endogenous enzymes better than microwave. Atmospheric steaming treatment group achieved over 95% inactivation of lipase and completely inactivated peroxidase. Meanwhile, enzymolysis treatment group resulted in a lipase inactivation rate of over 97%. Compared with the single treatm, the combined treatment exhibited enhanced efficacy, resulting in inactivation rates exceeding 90% for the three endogenous enzymes in wheat bran-germ.In terms of total phenolic content, microwave combined enzymatic digestion resulted in a maximum increase of 1.40-fold in total phenolic content compared to untreated wheat bran-germ. This was followed by the atmospheric steam combined enzymatic hydrolysis group, showing a 1.05-fold elevation in total phenolic contents. Furthermore, microwaving combined enzymolysis resulted in the highest free-radical-scavenging rate towards ABTS⁺ and DPPH radicals in wheat bran-germ, reaching 91.13% and 91.79%, respectively. Given the eco-friendliness, high efficiency, mature equipment, standardized operation, and significant long-term economic benefits of microwaving combined enzymolysis technology, this approach could be a potential approach to deactivate endogenous enzyme activity and enhance the antioxidant properties of wheat bran-germ. These findings will aid future optimization of industrial wheat bran-germ modification.
- wheat bran-germ,
- pretreatment,
- endogenous enzyme activity,
- total phenols,
- antioxidant activity

FullText(HTML)

References (43)

References

[1]	BHAT N A, WANI I A, HAMDANI A M, et al. Effect of extrusion on the physicochemical and antioxidant properties of value added snacks from whole wheat (Triticum aestivum L.) flour[J]. Food Chemistry,2019,276:22−32. doi: 10.1016/j.foodchem.2018.09.170
[2]	HU Q, CHEN Y Y, JIAO Q Y, et al. Polyphenolic compounds from Malus hupehensis and their free radical scavenging effects[J]. Natural Product Research,2018,32(18):2152−2158. doi: 10.1080/14786419.2017.1367784
[3]	贾婉婷. 全麦挂面脂质酸败的抑制研究[D]. 无锡:江南大学, 2021. [JIA W T. Study on inhibition of lipid rancidity in dried whole wheat noodles[D]. Wuxi:Jiangnan University, 2021.] JIA W T. Study on inhibition of lipid rancidity in dried whole wheat noodles[D]. Wuxi: Jiangnan University, 2021.
[4]	刘远晓, 李萌萌, 卞科, 等. 热处理在小麦储藏与加工中的应用研究进展[J]. 食品科学,2019,40(13):326−333. [LIU Y X, LI M M, BIAN K, et al. Recent progress in the application of heat treatment in wheat storage and processing[J]. Food Science,2019,40(13):326−333.] doi: 10.7506/spkx1002-6630-20180623-444 LIU Y X, LI M M, BIAN K, et al. Recent progress in the application of heat treatment in wheat storage and processing[J]. Food Science, 2019, 40（13）: 326−333. doi: 10.7506/spkx1002-6630-20180623-444
[5]	李合, 蔡红燕, 沈汪洋, 等. 挤压膨化技术在麦麸加工中应用及研究进展[J]. 食品工业,2020,41(11):271−274. [LI H, CAI H Y, SHEN W Y, et al. Application and research progress of extruding technology in wheat bran processing[J]. Food Industry,2020,41(11):271−274.] LI H, CAI H Y, SHEN W Y, et al. Application and research progress of extruding technology in wheat bran processing[J]. Food Industry, 2020, 41（11）: 271−274.
[6]	XUE Y, CUI X, ZHANG Z, et al. Effect of β-endoxylanase and α-arabinofuranosidase enzymatic hydrolysis on nutritional and technological properties of wheat brans[J]. Food Chemistry,2020,302:125332. doi: 10.1016/j.foodchem.2019.125332
[7]	叶文俊, 汪丽萍, 叶国栋, 等. 不同改性麸皮回填全麦粉馒头品质特性的比较[J]. 食品工业科技,2023,44(10):20−26. [YE W J, WANG L P, YE G D, et al. Comparative study on the quality of whole wheat flour steamed bread backfilled with different modified bran[J]. Science and Technology of Food Industry,2023,44(10):20−26.] YE W J, WANG L P, YE G D, et al. Comparative study on the quality of whole wheat flour steamed bread backfilled with different modified bran[J]. Science and Technology of Food Industry, 2023, 44（10）: 20−26.
[8]	张岩岩, 李雪杰, 张剑, 等. 微波辐照对直接研磨法全麦粉的储藏稳定性与品质的影响[J]. 河南农业大学学报,2020,54(3):506−512. [ZHANG Y Y, LI X J, ZHANG J, et al. The effect of microwave irradiation on the storage stability and quality of direct grinding whole wheat flour[J]. Journal of Henan Agricultural University,2020,54(3):506−512.] ZHANG Y Y, LI X J, ZHANG J, et al. The effect of microwave irradiation on the storage stability and quality of direct grinding whole wheat flour[J]. Journal of Henan Agricultural University, 2020, 54（3）: 506−512.
[9]	赵梅, 韩传武, 宋俊男, 等. 微波处理对全麦粉理化性质及全麦鲜湿面品质的影响[J]. 中国粮油学报,2019,34(1):18−23. [ZHAO M, HAN C W, SONG J N, et al. Effect of microwave treatment on the physicochemical properties of whole wheat flour and qualities of fresh noodles[J]. Journal of the Chinese Cereals and Oils Association,2019,34(1):18−23.] doi: 10.3969/j.issn.1003-0174.2019.01.005 ZHAO M, HAN C W, SONG J N, et al. Effect of microwave treatment on the physicochemical properties of whole wheat flour and qualities of fresh noodles[J]. Journal of the Chinese Cereals and Oils Association, 2019, 34（1）: 18−23. doi: 10.3969/j.issn.1003-0174.2019.01.005
[10]	杜昱蒙, 任国宝, 任晨刚, 等. 常压蒸汽处理对全麦粉脂肪相关酶活性及微生物的影响[J]. 粮油食品科技,2021,29(6):156−162. [DU Y M, REN G B, REN C G, et al. Research on the effects of atmospheric steam treatment on lipase related enzyme activities and microorganisms of whole wheat flour[J]. Science and Technology of Cereals, Oils and Foods,2021,29(6):156−162.] DU Y M, REN G B, REN C G, et al. Research on the effects of atmospheric steam treatment on lipase related enzyme activities and microorganisms of whole wheat flour[J]. Science and Technology of Cereals, Oils and Foods, 2021, 29（6）: 156−162.
[11]	毛濛兰. 麦麸生物改性及其在全麦馒头中的应用[D]. 南京:南京农业大学, 2020. [MAO M L. Biological modification of wheat bran and itsapplication in whole wheat steamed bread[D]. Nanjing:Nanjing Agricultural University, 2020.] MAO M L. Biological modification of wheat bran and itsapplication in whole wheat steamed bread[D]. Nanjing: Nanjing Agricultural University, 2020.
[12]	QU C, WANG H, LIU S, et al. Effects of microwave heating of wheat on its functional properties and accelerated storage[J]. Journal of Food Science and Technology-Mysore,2017,54(11):3699−3706. doi: 10.1007/s13197-017-2834-y
[13]	孟续. 预处理方式对黑小麦麸皮及其对面团品质的影响研究[D]. 无锡:江南大学, 2023. [MENG X. Study on the effect of pretreatment on triticale bran and its effect on the quality of dough[D]. Wuxi:Jiangnan University, 2023.] MENG X. Study on the effect of pretreatment on triticale bran and its effect on the quality of dough[D]. Wuxi: Jiangnan University, 2023.
[14]	ZHUANG K, SUN Z, HUANG Y, et al. Influence of different pretreatments on the quality of wheat bran-germ powder, reconstituted whole wheat flour and Chinese steamed bread[J]. LWT,2022,161:113357. doi: 10.1016/j.lwt.2022.113357
[15]	汪文丽, 朱晓明, 刘小玲. 多糖生物活性影响因素研究进展[J]. 化学与生物工程,2024,41(8):9−17. [WANG W L, ZHU X M, LIU X L. Research progress in factors influencing biological activity of polysaccharides[J]. Chemistry Bioengineering,2024,41(8):9−17.] doi: 10.3969/j.issn.1672-5425.2024.08.002 WANG W L, ZHU X M, LIU X L. Research progress in factors influencing biological activity of polysaccharides[J]. Chemistry Bioengineering, 2024, 41（8）: 9−17. doi: 10.3969/j.issn.1672-5425.2024.08.002
[16]	管骁, 夏吉安, 张妤, 等. 抗性淀粉的性质、制备及其对慢性肾脏病的影响[J]. 食品与生物技术学报,2024,43(1):1−12. [GUAN X, XIA J A, ZHANG Y, et al. Properties, preparations and effects of resistant starch on chronic kidney disease[J]. Journal of Food Science and Biotechnology,2024,43(1):1−12.] doi: 10.12441/spyswjs.20211106003 GUAN X, XIA J A, ZHANG Y, et al. Properties, preparations and effects of resistant starch on chronic kidney disease[J]. Journal of Food Science and Biotechnology, 2024, 43（1）: 1−12. doi: 10.12441/spyswjs.20211106003
[17]	WU Y, YE G, LI X, et al. Comparison of quality characteristics of six reconstituted whole wheat flour with different modified bran[J]. LWT,2022,153:112543. doi: 10.1016/j.lwt.2021.112543
[18]	LI M, SUN Q J, ZHU K X. Delineating the quality and component changes of whole-wheat flour and storage stability of fresh noodles induced by microwave treatment[J]. LWT,2017,84:378−384. doi: 10.1016/j.lwt.2017.06.001
[19]	杨磊. 重组全麦面粉的稳定化研究[D]. 无锡:江南大学, 2013. [YANG L. Study on the stability of reconstituted whole wheat flour[D]. Wuxi:Jiangnan University, 2013.] YANG L. Study on the stability of reconstituted whole wheat flour[D]. Wuxi: Jiangnan University, 2013.
[20]	张书静. 改性麦麸对含麸馒头的影响及其机理探究[D]. 武汉:武汉轻工大学, 2022. [ZHANG S J. Effects of modified wheat bran on bran-containing steamed bread and its mechanism[D]. Wuhan:Wuhan Polytechnic University, 2022.] ZHANG S J. Effects of modified wheat bran on bran-containing steamed bread and its mechanism[D]. Wuhan: Wuhan Polytechnic University, 2022.
[21]	王旭. 菌酶协同发酵麦麸及其对鹅饲用价值的研究[D]. 扬州:扬州大学, 2023. [WANG X. Study on synergistic solid-state fermentation of wheatbran by bacteria and enzymes and its value for goose feeding[D]. Yangzhou:Yangzhou University, 2023.] WANG X. Study on synergistic solid-state fermentation of wheatbran by bacteria and enzymes and its value for goose feeding[D]. Yangzhou: Yangzhou University, 2023.
[22]	YE G, WU Y, WANG L, et al. Comparison of six modification methods on the chemical composition, functional properties and antioxidant capacity of wheat bran[J]. LWT-Food Science and Technology,2021,149:111996. doi: 10.1016/j.lwt.2021.111996
[23]	周颖, 杨震, 郭晓娜, 等. 过热蒸汽对燕麦挂面储藏稳定性及风味的影响[J]. 中国粮油学报,2022,37(2):54−61. [ZHOU Y, YANG Z, GUO X N, et al. Effects of superheated steam treatment on storage stability and flavor of dried oat noodles[J]. Journal of the Chinese Cereals and Oils Association,2022,37(2):54−61.] doi: 10.3969/j.issn.1003-0174.2022.02.010 ZHOU Y, YANG Z, GUO X N, et al. Effects of superheated steam treatment on storage stability and flavor of dried oat noodles[J]. Journal of the Chinese Cereals and Oils Association, 2022, 37（2）: 54−61. doi: 10.3969/j.issn.1003-0174.2022.02.010
[24]	先兆君. 超声辅助冷冻对预制调理红糖馒头面团品质的影响及作用机制研究[D]. 合肥:合肥工业大学, 2022. [XIAN Z J. Effect of ultrasound-assisted freezing on quality of prepared brown sugar steamed bread dough and its mechanism investigation[D]. Hefei:Hefei University of Technology, 2022.] XIAN Z J. Effect of ultrasound-assisted freezing on quality of prepared brown sugar steamed bread dough and its mechanism investigation[D]. Hefei: Hefei University of Technology, 2022.
[25]	侯珺瑶. 功能性麸皮发酵剂的研制及其在馒头中的应用[D]. 泰安:山东农业大学, 2023. [HOU J Y. Development of functional bran starter and its application in steamed[D]. Taian:Shandong Agricultural University, 2023.] HOU J Y. Development of functional bran starter and its application in steamed[D]. Taian: Shandong Agricultural University, 2023.
[26]	SHARMA P, SHARMA N, PATHANIA S, et al. Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry[J]. Journal, Genetic Engineering & Biotechnology,2017,15(2):369−377.
[27]	CHEN Z, LI Y, WANG L, et al. Evaluation of the possible non-thermal effect of microwave radiation on the inactivation of wheat germ lipase[J]. Journal of Food Process Engineering,2017,40(4):e12506. doi: 10.1111/jfpe.12506
[28]	朱松, 宋善武, 李进伟, 等. 胚芽米微波灭酶工艺[J]. 食品与生物技术学报,2018,37(6):603−609. [ZHU S, SONG S W, LI J W, et al. Study on the technology of inactivation enzyme in embryo-remaining rice by microwave[J]. Journal of Food Science and Biotechnology,2018,37(6):603−609.] doi: 10.3969/j.issn.1673-1689.2018.06.006 ZHU S, SONG S W, LI J W, et al. Study on the technology of inactivation enzyme in embryo-remaining rice by microwave[J]. Journal of Food Science and Biotechnology, 2018, 37（6）: 603−609. doi: 10.3969/j.issn.1673-1689.2018.06.006
[29]	张楠, 石琳, 孟继坤, 等. 不同处理技术对小麦胚芽钝酶效果及贮藏稳定性的影响[J]. 食品与机械,2021,37(8):140−145. [ZHANG N, SHI L, MENG J K, et al. Effects of different technologies on enzyme deactivation and storage stability of wheat germs[J]. Food Machinery,2021,37(8):140−145.] ZHANG N, SHI L, MENG J K, et al. Effects of different technologies on enzyme deactivation and storage stability of wheat germs[J]. Food Machinery, 2021, 37（8）: 140−145.
[30]	CHAKRABORTY S, PULIVARTHI M K, RAJ A S, et al. Inactivation of lipase and lipoxygenase in whole wheat flour using atmospheric cold plasma and steam treatments:Kinetics, mechanism, and impact on its compositional properties[J]. Journal of Cereal Science,2024,117:103889. doi: 10.1016/j.jcs.2024.103889
[31]	卞科, 郑学玲. 谷物化学[M]. 科学出版社, 2017. [BIAN K, ZHENG X L. Cereal chemistry[M]. Science Press, 2017.] BIAN K, ZHENG X L. Cereal chemistry[M]. Science Press, 2017.
[32]	ROSE D J, OGDEN L V, DUNN M L, et al. Enhanced lipid stability in whole wheat flour by lipase inactivation and antioxidant retention[J]. Cereal Chemistry,2008,85(2):218−223. doi: 10.1094/CCHEM-85-2-0218
[33]	ZHANG X, TAO N, WANG X, et al. The colorants, antioxidants, and toxicants from nonenzymatic browning reactions and the impacts of dietary polyphenols on their thermal formation[J]. Food Funct,2014,6(2):345−355.
[34]	吴佳, 王燕, 陈星. 麦胚稳定化处理的研究[J]. 中国农学通报,2019,35(29):134−142. [WU J, WANG Y, CHEN X. Stabilization treatment of wheat germ[J]. Chinese Agricultural Science Bulletin,2019,35(29):134−142.] doi: 10.11924/j.issn.1000-6850.casb18050165 WU J, WANG Y, CHEN X. Stabilization treatment of wheat germ[J]. Chinese Agricultural Science Bulletin, 2019, 35（29）: 134−142. doi: 10.11924/j.issn.1000-6850.casb18050165
[35]	白宇皓, 李超, 杨志国, 等. 梨多酚氧化酶特性与酶促褐变抑制研究进展[J]. 食品科技,2022,47(2):75−81. [BAI Y H, LI C, YANG Z G, et al. Advances in polyphenol oxidase properties and enzymatic browning inhibition of pear[J]. Food Science and Technology,2022,47(2):75−81.] doi: 10.3969/j.issn.1005-9989.2022.2.spkj202202012 BAI Y H, LI C, YANG Z G, et al. Advances in polyphenol oxidase properties and enzymatic browning inhibition of pear[J]. Food Science and Technology, 2022, 47（2）: 75−81. doi: 10.3969/j.issn.1005-9989.2022.2.spkj202202012
[36]	HU Y, WANG L, LI Z. Superheated steam treatment on wheat bran:Enzymes inactivation and nutritional attributes retention[J]. LWT,2018,91:446−452. doi: 10.1016/j.lwt.2018.01.086
[37]	冯健超, 许倍铭, 江薛丽, 等. 小麦籽粒不同层次酚类物质与抗氧化活性差异及氮肥调控效应[J]. 作物学报,2022,48(3):704−715. [FENG J C, XU B M, JIANG X L, et al. Distribution of phenolic compounds and antioxidant activities in layered grinding wheat flour and the regulation effect of nitrogen fertilizer application[J]. Acta Agronomica Sinica,2022,48(3):704−715.] doi: 10.3724/SP.J.1006.2022.11007 FENG J C, XU B M, JIANG X L, et al. Distribution of phenolic compounds and antioxidant activities in layered grinding wheat flour and the regulation effect of nitrogen fertilizer application[J]. Acta Agronomica Sinica, 2022, 48（3）: 704−715. doi: 10.3724/SP.J.1006.2022.11007
[38]	叶国栋, 汪丽萍, 沈汪洋, 等. 麸皮稳定化处理方式对全麦粉品质的影响[J]. 食品工业科技,2021,42(6):15−21. [YE G D, WANG L P, SHEN W Y, et al. Effect of bran stabilization on the quality of whole wheat flour[J]. Science and Technology of Food Industry,2021,42(6):15−21.] YE G D, WANG L P, SHEN W Y, et al. Effect of bran stabilization on the quality of whole wheat flour[J]. Science and Technology of Food Industry, 2021, 42（6）: 15−21.
[39]	何若男, 刘思师, 陈雅欣, 等. 嗜热木聚糖酶的研究与优化进展[J]. 精细与专用化学品,2023,31(8):16−24. [HE R N, LIU S S, CHEN Y X, et al. Progress in the research and optimization of thermophilic xylanase[J]. Fine and Specialty Chemicals,2023,31(8):16−24.] HE R N, LIU S S, CHEN Y X, et al. Progress in the research and optimization of thermophilic xylanase[J]. Fine and Specialty Chemicals, 2023, 31（8）: 16−24.
[40]	连彩霞. 脱脂麦胚提取物抗氧化、抗肿瘤活性的研究[D]. 无锡:江南大学, 2012. [LIAN C X. Study on antioxidant and anticancer activity of extracts from defatted wheat germ[D]. Wuxi:Jiangnan University, 2012.] LIAN C X. Study on antioxidant and anticancer activity of extracts from defatted wheat germ[D]. Wuxi: Jiangnan University, 2012.
[41]	BETA T, NAM S, DEXTER J E, et al. Phenolic content and antioxidant activity of pearled wheat and roller-milled fractions[J]. Cereal Chemistry,2005,82(4):390−393. doi: 10.1094/CC-82-0390
[42]	VERMA B, HUCL P, CHIBBAR R N. Phenolic acid composition and antioxidant capacity of acid and alkali hydrolysed wheat bran fractions[J]. Food Chemistry,2009,116(4):947−954. doi: 10.1016/j.foodchem.2009.03.060
[43]	LIU L, ZHAO M, LIU X, et al. Effect of steam explosion-assisted extraction on phenolic acid profiles and antioxidant properties of wheat bran[J]. Journal of the Science of Food and Agriculture,2016,96(10):3484−3491. doi: 10.1002/jsfa.7532

Supplements (0)

Cited By

Cited by

Periodical cited type(12)

1.	王琳，王耀，胡骁飞，王成宾，吴佳蓓. 免疫分析技术在金刚烷胺残留检测中的应用. 食品与机械. 2023(04): 205-210 .
2.	孙玲玲，尹营，耿旭浩，霍思宇，高家政，陈冬东，彭涛，薛占永. 动物源性食品中金刚烷胺类药物残留检测技术研究进展. 食品科技. 2023(08): 294-301 .
3.	任宏彬，张鑫鑫，贾晓婷，杨晓伟. 高效液相色谱-质谱联用法同时测定鸡肉中2种金刚烷胺类药物和12种喹诺酮类药物残留. 中国家禽. 2022(01): 77-82 .
4.	张艳，王全胜，吴银良. 分散固相萃取-液相色谱-串联质谱法同时测定鸡蛋中7种抗病毒类药物和利巴韦林代谢物残留量. 食品安全质量检测学报. 2022(06): 1872-1879 .
5.	郭东光，陈明艳，冯春花，王丰，卞爽丽，李文明，王凯露，吕薇薇，郑文珺，陈子怡，孙国鹏，李雪华，田金河，李鹏，朱艳平，岳锋，王选年. 齐帕特罗人工抗原合成及其多克隆抗体制备. 动物医学进展. 2022(05): 6-12 .
6.	林瑶. 畜禽肉食品中金刚烷胺金刚乙胺残留量的超高效液相色谱串联质谱法测定. 预防医学论坛. 2022(06): 438-441 .
7.	赵军强，韩典峰，田秀慧，王景，刘鸽，丁玉竹，彭中校，薛敬林，刘永春，乔瑞光，徐英江. 食品中金刚烷胺的危害、检测方法和残留风险研究进展. 中国渔业质量与标准. 2022(03): 64-71 .
8.	姜英杰，葛子欣，翟玮玮，苏晶. EMR-Lipid固相萃取结合液相色谱串联质谱法快速测定鸡蛋中金刚烷胺和金刚乙胺残留量. 中国食品添加剂. 2022(11): 218-224 .
9.	徐少华，聂婵，谭磊. 内标法测定鸡肉中金刚烷胺含量方法的优化——高效液相色谱-串联质谱法. 养禽与禽病防治. 2022(07): 20-23 .
10.	王丰存，端礼钦，王静，郑云天. 超声提取-固相萃取-超高效液相色谱串联质谱法同时测定畜禽产品中金刚烷胺及8种氟喹诺酮类药物残留. 中国检验检测. 2022(06): 52-55+79 .
11.	曾海英，王家磊，刘志敏，任召珍. 肉鸡中金刚烷胺残留测定及其残留代谢规律研究. 食品安全质量检测学报. 2021(08): 3118-3122 .
12.	彭娟，赖科洋. 食品中金刚烷胺残留检测方法新进展. 食品科学. 2021(19): 325-333 .