Article(id=1217529307124126611, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250330001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1743264000000, receivedDateStr=2025-03-30, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768211207419, onlineDateStr=2026-01-12, pubDate=1752508800000, pubDateStr=2025-07-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768211207419, onlineIssueDateStr=2026-01-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768211207419, creator=13701087609, updateTime=1768211207419, updator=13701087609, issue=Issue{id=1217529305693864468, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='13', pageStart='1', pageEnd='320', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768211207077, creator=13701087609, updateTime=1768212057891, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217532874337730593, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217532874337730594, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=276, endPage=281, ext={EN=ArticleExt(id=1217529307921044377, articleId=1217529307124126611, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Determination of fat content in flavored fermented milk by papain-gravimetric method, columnId=1151923894560060071, journalTitle=Journal of Food Safety & Quality, columnName=Food Nutrition and Functional Foods, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method for determination the fat content in flavored fermented milk containing thickeners by papain-gravimetric method. Methods Papain was used to reduce the adsorption of fat by casein in dairy products. The amount of papain added was 0.5 g, the enzymatic hydrolysis temperature was 65 ℃, and the hydrolysis time was 20 min. A mixture of ether and petroleum ether with equal volumes [25:25 (V:V)] was used to extract fat twice from the flavored fermented milk hydrolysate containing thickening agents. The extraction solvent was evaporated in a water bath, and the fat content was quantified by gravimetric method after drying in an oven. Results The determination of fat content in flavored fermented milk with thickener flavor was consistent with the label value, with a relative standard deviation of 0.40% and a maximum deviation of 0.05 g/100 g, which met the precision requirements of GB 5009.6—2016 third method for determination of fat in foods. Conclusion This method is simple to operate and conform to the concept of green analytical chemistry, compared with GB 5009.6— 2016 third method, it reduces the total volume of mixed ethers used, thereby minimizing harm to human health and environmental pollution. With good reproducibility, it can effectively determine the fat content in flavored fermented milk containing thickeners, providing robust technical support for quality control of such products.

, correspAuthors=Chao HOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Chao HOU, Qing LV), CN=ArticleExt(id=1217529312387977291, articleId=1217529307124126611, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=木瓜蛋白酶-重量法测定风味发酵乳中脂肪含量, columnId=1151923894698472105, journalTitle=食品安全质量检测学报, columnName=食品营养及功能性食品, runingTitle=null, highlight=null, articleAbstract=

目的 建立木瓜蛋白酶-重量法测定含增稠剂风味发酵乳中脂肪含量的分析方法。方法 采用木瓜蛋白酶降低风味发酵乳中酪蛋白对脂肪的吸附, 木瓜蛋白酶添加量为0.5 g, 酶水解温度为65 ℃, 水解时间20 min, 使用乙醚和石油醚等体积[25:25 (V:V)]混合液对含增稠剂风味发酵乳水解液中脂肪进行2次提取, 水浴蒸干提取溶剂, 经过烘箱干燥后, 重量法定量脂肪含量。结果 含增稠剂风味发酵乳脂肪含量测定结果与标签值一致, 测定结果的相对标准偏差为0.40%, 测定结果最大偏差为0.05 g/100 g, 符合GB 5009.6—2016《食品安全国家标准 食品中脂肪的测定》第三法中精密度要求。结论 与GB 5009.6—2016第三法相比, 该方法符合绿色分析化学理念, 降低有机提取溶剂的使用量以及对人体危害和环境的污染, 该方法操作简单、重复性好, 可满足含增稠剂风味发酵乳中脂肪含量测定, 为含增稠剂风味发酵乳的质量控制提供有力的技术保障。

, correspAuthors=侯超, authorNote=null, correspAuthorsNote=
*侯超(1988—), 男, 硕士, 工程师, 主要研究方向为食品安全检测。E-mail:
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articleId=1217529307124126611, language=EN, orderNo=1, keyword=papain), Keyword(id=1217901254592872774, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, orderNo=2, keyword=thickener), Keyword(id=1217901254697730385, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, orderNo=3, keyword=flavored fermented milk), Keyword(id=1217901254802588002, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, orderNo=4, keyword=fat content), Keyword(id=1217901254924222828, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, orderNo=1, keyword=木瓜蛋白酶), Keyword(id=1217901255087800699, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, orderNo=2, keyword=增稠剂), Keyword(id=1217901255209435526, tenantId=1146029695717560320, 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China Inspection Body & Laboratory, 2024, 32(4): 81-84, 59., articleTitle=Uncertainty analysis of fat content in milk by near infrared spectroscopy and alkaline hydrolysis method, refAbstract=null)], funds=[Fund(id=1217901256996209200, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, awardId=KY-2023-9-SP, language=CN, fundingSource=上海市质量监督检验技术研究院项目(KY-2023-9-SP), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1217901252118233289, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, xref=null, ext=[AuthorCompanyExt(id=1217901252126621897, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, companyId=1217901252118233289, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Shanghai Institute of Quality Inspection and Technical Research, Shanghai 200233, China), AuthorCompanyExt(id=1217901252135010509, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, companyId=1217901252118233289, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=上海市质量监督检验技术研究院, 上海 200233)])], figs=[ArticleFig(id=1217901255691780539, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, label=Fig.1, caption=Effects of amounts of papain added on the determination results of fat content, figureFileSmall=JStJHXVLjednhpjgRgcooA==, figureFileBig=ZRlV5E4lSg5kAMHVI1paDA==, tableContent=null), ArticleFig(id=1217901255800832455, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, label=图1, caption=木瓜蛋白酶添加量对脂肪含量测定结果的影响, figureFileSmall=JStJHXVLjednhpjgRgcooA==, figureFileBig=ZRlV5E4lSg5kAMHVI1paDA==, tableContent=null), ArticleFig(id=1217901255968604640, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, label=Table 1, caption=

Fat content measurement values corresponding to different sample weights

, figureFileSmall=null, figureFileBig=null, tableContent=
称样量/g 脂肪含量/(g/100 g) 平均值/(g/100 g)
1 3.30, 3.32, 3.34 3.32
3 3.29, 3.30, 3.31 3.30
5 3.30, 3.31, 3.30 3.30
7 3.23, 3.18, 3.21 3.21
10 3.11, 3.05, 3.07 3.08
), ArticleFig(id=1217901256111210986, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, label=表1, caption=

不同称样量下的脂肪含量测定值

, figureFileSmall=null, figureFileBig=null, tableContent=
称样量/g 脂肪含量/(g/100 g) 平均值/(g/100 g)
1 3.30, 3.32, 3.34 3.32
3 3.29, 3.30, 3.31 3.30
5 3.30, 3.31, 3.30 3.30
7 3.23, 3.18, 3.21 3.21
10 3.11, 3.05, 3.07 3.08
), ArticleFig(id=1217901256232845814, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, label=Table 2, caption=

Precision test results of fat content determination

, figureFileSmall=null, figureFileBig=null, tableContent=
本方法 GB 5009.6—2016第三法
脂肪含量/(g/100 g) 平均值/(g/100 g) 相对标准偏差/% 脂肪含量/(g/100 g) 平均值/(g/100 g) 相对标准偏差/%
3.30, 3.32, 3.34 3.31 0.40 2.81, 2.95, 2.75 2.79 2.90
3.30, 3.31, 3.30 2.85, 2.76, 2.65
3.30, 3.32, 3.31
3.29, 3.30, 3.31
2.75, 2.86, 2.77
2.85, 2.79, 2.69
), ArticleFig(id=1217901256375452158, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, label=表2, caption=

精密度试验脂肪含量测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
本方法 GB 5009.6—2016第三法
脂肪含量/(g/100 g) 平均值/(g/100 g) 相对标准偏差/% 脂肪含量/(g/100 g) 平均值/(g/100 g) 相对标准偏差/%
3.30, 3.32, 3.34 3.31 0.40 2.81, 2.95, 2.75 2.79 2.90
3.30, 3.31, 3.30 2.85, 2.76, 2.65
3.30, 3.32, 3.31
3.29, 3.30, 3.31
2.75, 2.86, 2.77
2.85, 2.79, 2.69
), ArticleFig(id=1217901256580973071, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=EN, label=Table 3, caption=

Fat content determination values of commercial samples (g/100 g)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号 脂肪含量 平均值 标签值
1 3.30, 3.32, 3.31 3.31 3.3
2 3.59, 3.60, 3.58 3.59 3.6
3 3.95, 4.00, 3.90 3.95 4.0
4 3.43, 3.38, 3.41 3.41 3.4
5 2.91, 2.88, 2.90 2.90 2.9
), ArticleFig(id=1217901256757133851, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529307124126611, language=CN, label=表3, caption=

市售样品脂肪含量测定值(g/100 g)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号 脂肪含量 平均值 标签值
1 3.30, 3.32, 3.31 3.31 3.3
2 3.59, 3.60, 3.58 3.59 3.6
3 3.95, 4.00, 3.90 3.95 4.0
4 3.43, 3.38, 3.41 3.41 3.4
5 2.91, 2.88, 2.90 2.90 2.9
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木瓜蛋白酶-重量法测定风味发酵乳中脂肪含量
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侯超 * , 吕庆
食品安全质量检测学报 | 食品营养及功能性食品 2025,16(13): 276-281
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食品安全质量检测学报 | 食品营养及功能性食品 2025, 16(13): 276-281
木瓜蛋白酶-重量法测定风味发酵乳中脂肪含量
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侯超* , 吕庆
作者信息
  • 上海市质量监督检验技术研究院, 上海 200233

通讯作者:

*侯超(1988—), 男, 硕士, 工程师, 主要研究方向为食品安全检测。E-mail:
Determination of fat content in flavored fermented milk by papain-gravimetric method
Chao HOU* , Qing LV
Affiliations
  • Shanghai Institute of Quality Inspection and Technical Research, Shanghai 200233, China
出版时间: 2025-07-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250330001
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目的 建立木瓜蛋白酶-重量法测定含增稠剂风味发酵乳中脂肪含量的分析方法。方法 采用木瓜蛋白酶降低风味发酵乳中酪蛋白对脂肪的吸附, 木瓜蛋白酶添加量为0.5 g, 酶水解温度为65 ℃, 水解时间20 min, 使用乙醚和石油醚等体积[25:25 (V:V)]混合液对含增稠剂风味发酵乳水解液中脂肪进行2次提取, 水浴蒸干提取溶剂, 经过烘箱干燥后, 重量法定量脂肪含量。结果 含增稠剂风味发酵乳脂肪含量测定结果与标签值一致, 测定结果的相对标准偏差为0.40%, 测定结果最大偏差为0.05 g/100 g, 符合GB 5009.6—2016《食品安全国家标准 食品中脂肪的测定》第三法中精密度要求。结论 与GB 5009.6—2016第三法相比, 该方法符合绿色分析化学理念, 降低有机提取溶剂的使用量以及对人体危害和环境的污染, 该方法操作简单、重复性好, 可满足含增稠剂风味发酵乳中脂肪含量测定, 为含增稠剂风味发酵乳的质量控制提供有力的技术保障。

木瓜蛋白酶  /  增稠剂  /  风味发酵乳  /  脂肪含量

Objective To establish a method for determination the fat content in flavored fermented milk containing thickeners by papain-gravimetric method. Methods Papain was used to reduce the adsorption of fat by casein in dairy products. The amount of papain added was 0.5 g, the enzymatic hydrolysis temperature was 65 ℃, and the hydrolysis time was 20 min. A mixture of ether and petroleum ether with equal volumes [25:25 (V:V)] was used to extract fat twice from the flavored fermented milk hydrolysate containing thickening agents. The extraction solvent was evaporated in a water bath, and the fat content was quantified by gravimetric method after drying in an oven. Results The determination of fat content in flavored fermented milk with thickener flavor was consistent with the label value, with a relative standard deviation of 0.40% and a maximum deviation of 0.05 g/100 g, which met the precision requirements of GB 5009.6—2016 third method for determination of fat in foods. Conclusion This method is simple to operate and conform to the concept of green analytical chemistry, compared with GB 5009.6— 2016 third method, it reduces the total volume of mixed ethers used, thereby minimizing harm to human health and environmental pollution. With good reproducibility, it can effectively determine the fat content in flavored fermented milk containing thickeners, providing robust technical support for quality control of such products.

papain  /  thickener  /  flavored fermented milk  /  fat content
侯超, 吕庆. 木瓜蛋白酶-重量法测定风味发酵乳中脂肪含量. 食品安全质量检测学报, 2025 , 16 (13) : 276 -281 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250330001
Chao HOU, Qing LV. Determination of fat content in flavored fermented milk by papain-gravimetric method[J]. Journal of Food Safety & Quality, 2025 , 16 (13) : 276 -281 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250330001
风味发酵乳一般是以各种乳粉或生乳为原料, 经过杀菌和发酵后制成的酸性产品。在发酵过程中可以选择性添加果蔬、谷物以及食品添加剂和营养强化剂。风味发酵乳可提供人类日常生活中所需求的益生菌、乳酸菌、维生素、钙和蛋白质[1], 食用后可增强免疫力[2-4]
然而在风味发酵乳生产过程中会出现品质缺陷, 如乳清析出、黏稠度偏低等。为解决上述问题, 生产企业通常会通过添加一些增稠剂以改善风味发酵乳的品质。单一的增稠稳定剂往往不能完全改善酸乳品质的缺陷, 因此常将亲水胶体[5-6]、酪蛋白[7-8]、变性淀粉[9]等增稠稳定剂复配后用于风味发酵乳生产中, 通过协同发挥增稠作用既提高了风味发酵乳的品质又降低了生产成本[10]
随着风味发酵乳中增稠剂添加比例和添加量不断变化, 其脂肪含量按照GB 5009.6—2016《食品安全国家标准 食品中脂肪的测定》第三法(以下称GB 5009.6—2016第三法)测定越来越困难。主要是由于增稠剂使得风味发酵乳中脂肪被酪蛋白钙盐包裹无法正常全部游离出来[11], 不能完全被提取溶剂萃取出, 导致脂肪含量测定结果偏低, 无法得到真实脂肪含量结果。刘芳芳等[12]研究了羧甲基纤维素钠对风味发酵乳中脂肪测定的影响, 也证实羧甲基纤维素钠等添加剂的添加会导致发酵乳中脂肪检测结果偏低, 此外, 一些胶体增稠剂也导致脂肪含量检测结果偏低[13]。这不仅增加了风味发酵乳生产企业内部质量控制难度, 同时对于监管机构在风味发酵乳脂肪质量判定上造成很大困扰。
刘芳芳等[14]使用2 mL盐酸代替氨水进行水解, 测定风味发酵乳中脂肪含量, 这需要对盐酸的用量和水解时间有严格的要求, 可能会导致水解产物焦糖化, 会导致一部分糖被混合醚萃取, 导致结果偏大。宋艳梅等[15]使用凝乳酶对风味发酵乳进行预处理后, 采用GB 5009.6—2016第三法检测风味发酵乳脂肪含量, 其结果与标签值一致, 但该方法增加了检测流程, 延长检测时间。
木瓜蛋白酶是一种含巯基(-SH)肽链内切酶, 具有蛋白酶和酯酶的活性, 有较广泛的特异性, 对动植物蛋白、多肽、酯、酰胺等有较强的水解能力, 在酸性、中性、碱性环境下均能分解蛋白质[16-17]。本研究基于木瓜蛋白酶活性中心与酪蛋白结合紧密, 利用木瓜蛋白酶的催化作用, 水解蛋白, 降低酪蛋白对脂肪的束缚, 促进脂肪被有机溶剂提取[18-22], 建立了一种能准确测定含增稠剂风味发酵乳中脂肪含量的木瓜蛋白酶-重量法, 以期为改善风味发酵乳品质和维护消费者合法权益提供科学依据。
风味发酵乳样品购于超市。
无水乙醚、95%乙醇、石油醚(沸程30~60 ℃)(分析纯, 国药集团化学试剂有限公司); 木瓜蛋白酶(活力800 U/mg, 上海普誉科贸有限公司)。
MS204电子天平(精度0.1 mg, 瑞士梅特勒-托利多仪器有限公司); FED720烘箱(德国Binder公司); TW20-A恒温水浴锅(德国JULABO公司); Relab-19309脂肪抽提瓶(国药集团化学试剂有限公司)。
称取混合均匀样品5 g(精确至0.0001 g)置于脂肪抽提瓶中, 加入0.5 g木瓜蛋白酶和5 mL水, 振摇, 将样品溶液和木瓜蛋白酶混合均匀, 放入65 ℃水浴中加热, 期间不时取出振荡, 20 min后取出, 冷却至室温。
缓慢加入10 mL 95%乙醇于脂肪抽提瓶中, 混匀, 加入25 mL无水乙醚, 缓慢振摇, 加入25 mL石油醚, 轻轻振摇, 放置, 待上层溶液澄清, 水相与有机相分层比较明显时, 打开瓶塞, 将有机层溶液倒入脂肪收集瓶中。向脂肪抽提瓶中继续加入5 mL 95%乙醇, 用25 mL无水乙醚和25 mL石油醚进行第2次抽提, 步骤同第1次。同时用10 mL水和0.5 g木瓜蛋白酶代替试样进行空白实验。
将有机层溶液于脂肪收集瓶中, 沸水浴上蒸干溶剂后, 放入105 ℃烘箱中干燥1.5 h, 取出后置于干燥器内冷却0.5 h后称重, 重复以上操作至恒重, 两次称量差不超过2 mg, 恒重后脂肪收集瓶和抽提物的质量选择最小一次的称量结果。脂肪含量计算方法见公式(1):
$X=\frac{\left( {{m}_{1}}-{{m}_{2}} \right)-\left( {{m}_{3}}-{{m}_{4}} \right)}{m}\times 100$
式中: X为试样中脂肪的含量, g/100 g; m1为恒重后脂肪收集瓶和脂肪的质量, g; m2为脂肪收集瓶的质量, g; m3为空白实验中, 恒重后脂肪收集瓶和抽提物的质量, g; m4为空白实验中脂肪收集瓶的质量, g; m为样品的质量, g; 100为换算系数。
所有实验重复测定3次, 数据采用Microsoft Excel 2016进行分析; 采用OriginPro 2017软件进行绘制图表。
称取不同质量的风味发酵乳样品, 加入木瓜蛋白酶进行预处理, 然后进行水解, 测定脂肪含量结果列于表1。从表1中可以看出, 随着称样量的增大, 脂肪含量平均值测定结果逐渐变小。这是由于随着称样量的增大, 酶与底物比例偏小, 其次, 在水解过程中, 间歇性振摇会导致部分水解物吸附在脂肪提取瓶壁上, 导致水解、提取不完全, 脂肪含量测定值偏低。当称样量过小时, 不仅增加了称量相对偏差, 还会导致取样不均匀, 综合考虑, 选取称样量为5 g。
分别使用不同添加量的木瓜蛋白酶, 对风味发酵乳样品按照1.3方法处理, 脂肪含量测定结果见图1a。由图1a可知, 随着木瓜蛋白酶添加量的提高, 样品脂肪含量测定值随之升高, 最后趋向于稳定, 这可能是随着木瓜蛋白酶添加量的增加, 木瓜蛋白酶与酪蛋白接触面积增加, 提高了酪蛋白的水解效率, 当木瓜蛋白酶的添加量大于0.5 g时, 脂肪含量测定值变小了, 可能是由于酶量超过酪蛋白的承载能力, 多余的酶无法结合底物, 导致单位酶活性下降。因此选取木瓜蛋白酶添加量为0.5 g。
在酶添加量为0.5 g时, 分别加热5、10、15、20、25、30 min, 图1b考查了加热时间对脂肪测定结果的影响, 从图1b中可以看出, 加热5~20 min时, 脂肪含量测定值逐渐变大, 这可能是由于随着加热时间的增加, 木瓜蛋白酶水解酪蛋白的量变大, 脂肪提取率变高。当加热时间超过20 min时, 木瓜蛋白酶的活性降低, 水解酪蛋白能力变小, 由此得出最佳加热时间为20 min。
由于木瓜蛋白酶的水解活性受温度影响较大, 图1c考查了加热温度对脂肪含量测定值的影响, 从图1c中可以看出, 当加热温度为60~65 ℃时, 脂肪含量测定值基本不变, 这与以往研究[23-24]基本一致, 考虑到酶水解效率, 参照GB 5009.6—2016第三法中使用水解温度为65 ℃, 故选取加热温度为65 ℃。
脂肪水解物提取过程中, 使用乙醚提取时, 提取效率高, 但容易导致提取过程中出现乳化现象, 有机相与水相分层不明显, 很难直接提取乙醚提取物。石油醚可降低乙醚的极性, 使乙醚与水不混溶, 页面分层明显, 因此需要对无水乙醚和石油醚的使用体积进行优化[25-28]
图1d所示, 当乙醚:石油醚的体积比例为25:25 (V:V)时, 脂肪含量测定值最高, 该比例提取溶剂能有效提取脂肪。因此最终确定混合醚比例为25:25 (V:V)。
按照1.3方法对同一样品酶水解物分别提取1~5次, 其脂肪含量测定值见图1e。结果表明, 当提取2次以上时, 样品中脂肪含量测定值保持稳定, 表明2次提取后, 脂肪基本被提出, 这主要是由于实验称取样品量少和单次提取溶剂体积比GB 5009.6—2016第三法多, 在保证实验结果准确前提下, 尽量节省时间, 最终选择提取次数为2次。
按照所建立的脂肪测定方法平行测定12次, 计算其平均值和相对标准偏差, 测定结果列于表2。由表2可知, 本方法测定结果的相对标准偏差为0.40%, 小于5.0%, 满足GB 5009.295—2023《食品安全国家标准 化学分析方法验证通则》对重复性相对标准偏差的要求。王瑜等[29]研究过发酵乳脂肪测定的扩展不确定度为0.2 g/100 g。本方法的最大偏差为0.05 g/100 g, 与杨爱君等[30]、孙婷婷等[31]研究基本一致, 均满足GB 5009.6—2016第三法中两次独立测定结果之差不大于0.1 g/100 g的精密度要求, 从表2可以看出GB 5009.6— 2016第三法脂肪检测结果分散, 最大偏差为0.30 g/ 100 g, 脂肪测定平均值也小于本方法, 相对标准偏差大于本方法, 表明本方法精密度良好。
采购市售风味发酵乳样品, 按照本方法进行脂肪含量的检测, 如表3所示, 结果表明以上各种样品脂肪含量均符合GB 19302—2010《食品安全国家标准 发酵乳》要求, 且和标签明示值一致。
本研究通过优化条件建立了含增稠剂风味发酵乳中脂肪含量检测方法, 其优化后的前处理方法为样品中加入0.5 g木瓜蛋白酶, 65 ℃水解20 min, 采用等比例乙醚/石油醚混合液提取酶水解物2次。经过验证, 本方法测定结果的相对标准偏差为0.40%, 最大偏差为0.05 g/100 g, 重复性相对偏差符合GB 5009.295—2023的要求, 通过对市售样品的检测, 满足实际应用需求, 弥补了相关检测方法的不足, 为相应标准的修订提供了技术支持。
  • 上海市质量监督检验技术研究院项目(KY-2023-9-SP)
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2025年第16卷第13期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250330001
  • 接收时间:2025-03-30
  • 首发时间:2026-01-12
  • 出版时间:2025-07-15
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  • 收稿日期:2025-03-30
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上海市质量监督检验技术研究院项目(KY-2023-9-SP)
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    上海市质量监督检验技术研究院, 上海 200233

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*侯超(1988—), 男, 硕士, 工程师, 主要研究方向为食品安全检测。E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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