Article(id=1153433750634483817, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250116002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1736956800000, receivedDateStr=2025-01-16, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929635914, onlineDateStr=2025-07-19, pubDate=1745510400000, pubDateStr=2025-04-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929635914, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929635914, creator=13701087609, updateTime=1752929635914, updator=13701087609, issue=Issue{id=1153433737141412332, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='8', pageStart='1', pageEnd='316', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929632696, creator=13701087609, updateTime=1757293087150, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1171735391666225233, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1171735391666225234, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=107, endPage=114, ext={EN=ArticleExt(id=1153433752018604173, articleId=1153433750634483817, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Rapid screening of 302 kinds of illegal additives in traditional Chinese veterinary medicine by ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry, columnId=1151895322692776479, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Analysis and Monitoring of Toxic and Harmful Substances in Food, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method based on ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry (UPLC-Q-TOF-MS) use as a rapid screening method for the illegal addition of 302 kinds of drugs in Chinese veterinary medicine. Methods A high-resolution mass spectrometry database containing information such as the names, molecular formulas, and exact mass numbers of 302 kinds of drugs was established. Samples were extracted with methanol and then analyzed by UPLC-Q-TOF-MS. A sample was considered as a suspected positive if the retention time deviation was less than 0.2 min, the mass error was less than 5 ppm, the isotopic ratio deviation was less than 5%, and the library matching score of the main characteristic fragment ions was greater than 80. Results The method exhibited good selectivity, with a minimum detection concentration as low as 50 ng/mL for the target substances (100 ng/mL for tetracyclines, penicillins, estrogens, and nonsteroidal anti-inflammatory drugs; 200 ng/mL for aminoglycosides). This enabled the efficient screening of 302 kinds of illegally added drugs. In the screening of actual samples, 7 batches of suspected positive samples were detected. Conclusion This approach is applicable for the screening of illegal additives in various veterinary Chinese medicines, providing effective technical support for the quality control of veterinary Chinese medicines.

, correspAuthors=Yan-Ning YANG, 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=Jing-Yuan GUAN, Yan-Ning YANG, Da-Ke ZHOU, Xue-Ming LONG, Can ZHOU, Gang ZHANG, Rui-Qi ZHANG, Ju-Fang HU, Shi-Qi LI), CN=ArticleExt(id=1153433773237588772, articleId=1153433750634483817, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=基于超高效液相色谱-四极杆-飞行时间高分辨质谱法快速筛查中兽药中302种药物非法添加, columnId=1151895322898297380, journalTitle=食品安全质量检测学报, columnName=本期专题:食品中有毒有害物质分析与监测, runingTitle=null, highlight=null, articleAbstract=

目的 建立一种基于超高效液相色谱-四极杆-飞行时间高分辨质谱法(ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry, UPLC-Q-TOF-MS)用于中兽药中的302种药物非法添加的快速筛查方法。方法 建立了包含302种药物名称、分子式、精确质量数等信息的高分辨质谱数据库。样品经甲醇提取后, 经过UPLC-Q-TOF-MS进行数据采集并分析, 当保留时间与数据库绝对偏差<0.2 min, 质量误差<5 ppm, 同位素比值偏差<5%, 且主要特征碎片离子的库比对分数>80时, 认为检出疑似阳性样品。结果 该方法选择性较好, 目标物最低检出质量浓度低至50 ng/mL(四环素类、青霉素类、雌激素类、及非甾体抗炎药检出质量浓度100 ng/mL; 氨基糖苷类检出质量浓度200 ng/mL), 实现了302种非法添加药物的高效筛查。实际样品筛查时, 检出了7批疑似阳性样品。结论 本研究建立的筛查手段具有快速、广谱的特点, 适用于多种中兽药中的非法添加药物的初步筛查, 为中兽药质量控制提供了有效技术支持。

, correspAuthors=杨彦宁, authorNote=null, correspAuthorsNote=
* 杨彦宁(1986—), 女, 硕士, 农艺师, 主要研究方向为兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:
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关静渊(1996—), 女, 硕士, 兽医师, 主要研究方向为兽医药理学与兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:

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关静渊(1996—), 女, 硕士, 兽医师, 主要研究方向为兽医药理学与兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:

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关静渊(1996—), 女, 硕士, 兽医师, 主要研究方向为兽医药理学与兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:

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Hunan Agricultural University, College of Veterinary Medicine, Changsha 410128, China), AuthorCompanyExt(id=1171733879787397920, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, companyId=1171733879774815006, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.湖南农业大学动物医学院, 长沙 410128)]), AuthorCompany(id=1171733879917421346, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, xref=null, ext=[AuthorCompanyExt(id=1171733879921615651, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, companyId=1171733879917421346, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3. Hunan Agricultural University, School of Food Science and Technology, Changsha 410128, China), AuthorCompanyExt(id=1171733879930004260, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, companyId=1171733879917421346, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.湖南农业大学食品科学技术学院, 长沙 410128)])], figs=[ArticleFig(id=1171733883637769089, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Fig.1, caption=Chromatogram of spectinomycin hydrochloride pentahydrate, figureFileSmall=6tU+wBJPHb+1uvsHr+bVsQ==, figureFileBig=utj7295unamq3f4+mWFE5Q==, tableContent=null), ArticleFig(id=1171733883767792514, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=图1, caption=盐酸大观霉素五水合物色谱图, figureFileSmall=6tU+wBJPHb+1uvsHr+bVsQ==, figureFileBig=utj7295unamq3f4+mWFE5Q==, tableContent=null), ArticleFig(id=1171733883843289987, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Fig.2, caption=Chromatographic profile of budesonide, figureFileSmall=GA3krk7Ew9Iwf8jg8BVunA==, figureFileBig=KqC/tiHtJtwca7Rtd+6w2A==, tableContent=null), ArticleFig(id=1171733883990090628, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=图2, caption=布地奈德色谱图, figureFileSmall=GA3krk7Ew9Iwf8jg8BVunA==, figureFileBig=KqC/tiHtJtwca7Rtd+6w2A==, tableContent=null), ArticleFig(id=1171733884136891269, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Fig.3, caption=Structures and chromatograms of formoterol fumarate dihydrate and phenylethanolamine A, figureFileSmall=xgzB31EN8tExyB75t93HQw==, figureFileBig=WrWfgzErcSUhh/WJ6Ss9QQ==, tableContent=null), ArticleFig(id=1171733884212388743, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=图3, caption=富马酸福莫特罗二水合物与苯乙醇胺A的结构图与色谱图

注: A. 富马酸福莫特罗二水合物; B. 苯乙醇胺A; C. 色谱图。

, figureFileSmall=xgzB31EN8tExyB75t93HQw==, figureFileBig=WrWfgzErcSUhh/WJ6Ss9QQ==, tableContent=null), ArticleFig(id=1171733884283691913, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Fig.4, caption=Comparison of characteristic fragment ions of atropine and scopolamine in the fertility-promoting intrauterine infusion liquid with reference standards, figureFileSmall=MtYMcj/faP/CckpAr8t20w==, figureFileBig=vu8tr1USr6WdvV91+02ufQ==, tableContent=null), ArticleFig(id=1171733884350800778, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=图4, caption=促孕灌注液中阿托品和东莨菪碱与对照品特征碎片离子比对图

注: A. 阿托品; B. 东莨菪碱。

, figureFileSmall=MtYMcj/faP/CckpAr8t20w==, figureFileBig=vu8tr1USr6WdvV91+02ufQ==, tableContent=null), ArticleFig(id=1171733884409521036, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Table 1, caption=

Gradient elution of mobile phase

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 流动相A/% 流动相B/%
0 95 5
3 70 30
8 50 50
17 5 95
19 5 95
19.5 95 5
), ArticleFig(id=1171733884510184334, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=表1, caption=

流动相的梯度洗脱

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 流动相A/% 流动相B/%
0 95 5
3 70 30
8 50 50
17 5 95
19 5 95
19.5 95 5
), ArticleFig(id=1171733884644402064, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Table 2, caption=

Database information of selected compounds (positive ion mode)

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文名称 提取离子质荷比 保留时间/min 中文名称 提取离子质荷比 保留时间/min
喹噁啉类 喹烯酮 307.1083 12.18 喹乙醇 264.0984 5.9
林可酰胺类 盐酸林可霉素 407.2220 6.34 盐酸吡利霉素 411.1728 8.39
大环内酯类 白霉素 泰乐菌素
白霉素A1 786.4669 13.59 泰乐菌素A 916.5291 11.29
白霉素A5* 772.4516 12.74 泰乐菌素B* 772.4515 10.22
白霉素A7 758.4364 11.88 泰乐菌素C 902.5124 10.77
白霉素A8 786.4319 11.76 泰乐菌素D 918.5473 10.7
白霉素A9 744.4201 11.11 螺旋霉素
白霉素A13 800.4823 14.46 螺旋霉素Ⅰ 843.5253 8.94
麦迪霉素A1 814.4622 13.39 螺旋霉素III 899.5509 10.01
维吉尼霉素 红霉素 734.4720 10.99
维吉尼霉素M1 526.2576 12.07 罗红霉素 837.5347 12.83
维吉尼霉素S1 824.3652 14.23 泰妙菌素 494.3311 12.73
乙酰氨基阿维菌素B1a 914.5300 16.65 伊维菌素B1a# 892.5459 18.09
β2-受体激动剂 盐酸利托君 288.1606 6.87 盐酸莱克多巴胺* 302.1766 7.68
富马酸福莫特罗二水合物 345.1824 8.47 盐酸苯氧丙酚胺* 302.1759 9.49
盐酸克仑特罗 277.0877 8.93 苯乙醇胺A 345.1814 11.9
硝基呋喃类 呋喃他酮 325.1158 7.02 呋喃唑酮* 226.0463 8.81
M胆碱受体阻断药 东莨菪碱 304.1551 6.91 阿托品 290.1754 7.63
氟喹诺酮类 环丙沙星 332.1420 7.86 诺氟沙星 320.1421 7.67
磺胺类 磺胺氯哒嗪* 285.0221 8.84 磺胺氯吡嗪* 285.0223 10.1
磺胺邻二甲氧嘧啶* 311.0816 9.13 磺胺间二甲氧嘧啶* 311.0815 10.37
磺胺对甲氧嘧啶* 281.0716 8.16 磺胺间甲氧嘧啶* 281.0715 8.53
磺胺二甲嘧啶* 279.0919 7.83 磺胺二甲异嘧啶* 279.0918 6.59
磺胺甲氧哒嗪* 281.0708 7.97 磺胺异噁唑* 268.0765 9.56
磺胺二甲唑* 268.0753 7.55 磺胺嘧啶 251.0609 6.95
非甾体抗炎药 双氯芬酸* 296.0256 14.34 依托度酸 288.1608 14
吡罗昔康* 332.0707 11.65 甲氯芬那酸* 296.0258 15.24
氟尼辛葡甲胺 297.0849 13.25 异丙氨基比林 246.1603 6.83
硝基咪唑类 甲硝唑 172.0722 6.25 洛硝哒唑 201.0632 6.76
氨基糖苷类 硫酸新霉素 硫酸庆大霉素
新霉素B 615.3246 7.62 硫酸庆大霉素C1 478.3256 2.65
新霉胺 323.1928 13.69 硫酸庆大霉素C2 464.3100 2.65
盐酸大观霉素五水合物 351.1719 3.18 硫酸庆大霉素C3 450.2942 2.64
糖皮质激素类 布地奈德 可的松* 361.2027 10.2
布地奈德 22R* 431.2450 13.01 倍他米松* 393.2090 10.88
布地奈德 22S* 431.2449 13.2 地塞米松* 393.2090 10.94
醋酸双氟拉松* 495.2202 14.73 泼尼松龙* 361.2029 9.85
甲基泼尼松龙 375.2184 10.67 氟轻松* 495.2198 14.58
性激素类 雌酮* 271.1703 13.38 双烯雌酚 267.1396 13.61
α-群勃龙* 271.1695 11.8 甲睾酮 303.2324 13.35
玉米赤霉酮类 玉米烯酮 319.1556 14.19
伏马菌素类 伏马菌素B1 722.3982 10.13 伏马菌素B2 706.4037 11.5
抗病毒类 盐酸金刚乙胺* 180.1753 9.16 盐酸美金刚* 180.1753 9.51
离子载体类抗球虫药 莫能菌素钠盐 693.4215 18.46 盐霉素钠 773.4838 17.01
苯并咪唑类 阿苯达唑 266.0963 11.05 阿苯达唑亚砜 282.0918 7.73
阿苯达唑砜 298.0869 9.2 芬苯达唑砜* 332.0715 10.85
头孢菌素类 头孢拉定 350.1191 6.9 头孢克洛 368.0487 6.14
头孢他啶 547.1094 6.06 头孢匹罗 515.1196 6.29
青霉素类 阿莫西林三水物 366.1135 5.78 氨苄青霉素三水合物 350.1182 6.71
四环素类 盐酸金霉素* 479.1248 8.72 盐酸四环素 445.1635 7.61
多西环素盐酸盐半乙醇半水合物 445.1634 7.99 差向金霉素盐酸盐* 479.1244 8.38
抗菌增效剂 甲氧苄啶 291.1452 7.29
), ArticleFig(id=1171733884791202706, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=表2, caption=

部分化合物数据库信息(正离子模式)

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文名称 提取离子质荷比 保留时间/min 中文名称 提取离子质荷比 保留时间/min
喹噁啉类 喹烯酮 307.1083 12.18 喹乙醇 264.0984 5.9
林可酰胺类 盐酸林可霉素 407.2220 6.34 盐酸吡利霉素 411.1728 8.39
大环内酯类 白霉素 泰乐菌素
白霉素A1 786.4669 13.59 泰乐菌素A 916.5291 11.29
白霉素A5* 772.4516 12.74 泰乐菌素B* 772.4515 10.22
白霉素A7 758.4364 11.88 泰乐菌素C 902.5124 10.77
白霉素A8 786.4319 11.76 泰乐菌素D 918.5473 10.7
白霉素A9 744.4201 11.11 螺旋霉素
白霉素A13 800.4823 14.46 螺旋霉素Ⅰ 843.5253 8.94
麦迪霉素A1 814.4622 13.39 螺旋霉素III 899.5509 10.01
维吉尼霉素 红霉素 734.4720 10.99
维吉尼霉素M1 526.2576 12.07 罗红霉素 837.5347 12.83
维吉尼霉素S1 824.3652 14.23 泰妙菌素 494.3311 12.73
乙酰氨基阿维菌素B1a 914.5300 16.65 伊维菌素B1a# 892.5459 18.09
β2-受体激动剂 盐酸利托君 288.1606 6.87 盐酸莱克多巴胺* 302.1766 7.68
富马酸福莫特罗二水合物 345.1824 8.47 盐酸苯氧丙酚胺* 302.1759 9.49
盐酸克仑特罗 277.0877 8.93 苯乙醇胺A 345.1814 11.9
硝基呋喃类 呋喃他酮 325.1158 7.02 呋喃唑酮* 226.0463 8.81
M胆碱受体阻断药 东莨菪碱 304.1551 6.91 阿托品 290.1754 7.63
氟喹诺酮类 环丙沙星 332.1420 7.86 诺氟沙星 320.1421 7.67
磺胺类 磺胺氯哒嗪* 285.0221 8.84 磺胺氯吡嗪* 285.0223 10.1
磺胺邻二甲氧嘧啶* 311.0816 9.13 磺胺间二甲氧嘧啶* 311.0815 10.37
磺胺对甲氧嘧啶* 281.0716 8.16 磺胺间甲氧嘧啶* 281.0715 8.53
磺胺二甲嘧啶* 279.0919 7.83 磺胺二甲异嘧啶* 279.0918 6.59
磺胺甲氧哒嗪* 281.0708 7.97 磺胺异噁唑* 268.0765 9.56
磺胺二甲唑* 268.0753 7.55 磺胺嘧啶 251.0609 6.95
非甾体抗炎药 双氯芬酸* 296.0256 14.34 依托度酸 288.1608 14
吡罗昔康* 332.0707 11.65 甲氯芬那酸* 296.0258 15.24
氟尼辛葡甲胺 297.0849 13.25 异丙氨基比林 246.1603 6.83
硝基咪唑类 甲硝唑 172.0722 6.25 洛硝哒唑 201.0632 6.76
氨基糖苷类 硫酸新霉素 硫酸庆大霉素
新霉素B 615.3246 7.62 硫酸庆大霉素C1 478.3256 2.65
新霉胺 323.1928 13.69 硫酸庆大霉素C2 464.3100 2.65
盐酸大观霉素五水合物 351.1719 3.18 硫酸庆大霉素C3 450.2942 2.64
糖皮质激素类 布地奈德 可的松* 361.2027 10.2
布地奈德 22R* 431.2450 13.01 倍他米松* 393.2090 10.88
布地奈德 22S* 431.2449 13.2 地塞米松* 393.2090 10.94
醋酸双氟拉松* 495.2202 14.73 泼尼松龙* 361.2029 9.85
甲基泼尼松龙 375.2184 10.67 氟轻松* 495.2198 14.58
性激素类 雌酮* 271.1703 13.38 双烯雌酚 267.1396 13.61
α-群勃龙* 271.1695 11.8 甲睾酮 303.2324 13.35
玉米赤霉酮类 玉米烯酮 319.1556 14.19
伏马菌素类 伏马菌素B1 722.3982 10.13 伏马菌素B2 706.4037 11.5
抗病毒类 盐酸金刚乙胺* 180.1753 9.16 盐酸美金刚* 180.1753 9.51
离子载体类抗球虫药 莫能菌素钠盐 693.4215 18.46 盐霉素钠 773.4838 17.01
苯并咪唑类 阿苯达唑 266.0963 11.05 阿苯达唑亚砜 282.0918 7.73
阿苯达唑砜 298.0869 9.2 芬苯达唑砜* 332.0715 10.85
头孢菌素类 头孢拉定 350.1191 6.9 头孢克洛 368.0487 6.14
头孢他啶 547.1094 6.06 头孢匹罗 515.1196 6.29
青霉素类 阿莫西林三水物 366.1135 5.78 氨苄青霉素三水合物 350.1182 6.71
四环素类 盐酸金霉素* 479.1248 8.72 盐酸四环素 445.1635 7.61
多西环素盐酸盐半乙醇半水合物 445.1634 7.99 差向金霉素盐酸盐* 479.1244 8.38
抗菌增效剂 甲氧苄啶 291.1452 7.29
), ArticleFig(id=1171733884975752085, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=EN, label=Table 3, caption=

Database information of compounds (negative ion mode)

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文名称 提取离子质荷比 保留时间/min 中文名称 提取离子质荷比 保留时间/min
基因重组药物 多拉菌素 897.5017 17.44
磺胺类 磺胺硝苯 334.0505 11.92
氨基糖苷类 阿米卡星 584.2789 2.63
雌激素类 己烷雌酚* 269.1553 13.82 己烯雌酚 267.1398 14.48
袢利尿剂 呋塞米 329.0006 11.01
非甾体抗炎药 卡洛芬 272.0489 14.29 菲诺洛芬 241.0872 13.78
抗寄生虫类 碘醚柳胺 623.8128 19.62 地克珠利 404.9727 14.77
尼卡巴嗪 301.0583 14.6 硝米特 210.0157 8.95
二硝托胺* 224.0314 9.22
呋喃类 呋喃苯烯酸钠 258.0407 13.03
玉米赤霉酮类 α-玉米赤霉醇* 321.1707 12.77 β-玉米赤霉醇* 321.1707 12.77
α-玉米赤霉烯醇* 319.1550 12.11 β-玉米赤霉烯醇* 319.1550 13.12
玉米赤霉酮* 319.1550 14.06
β-内酰胺酶抑制剂 舒巴坦 232.0286 6.39
), ArticleFig(id=1171733885084803991, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433750634483817, language=CN, label=表3, caption=

化合物数据库信息(负离子模式)

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文名称 提取离子质荷比 保留时间/min 中文名称 提取离子质荷比 保留时间/min
基因重组药物 多拉菌素 897.5017 17.44
磺胺类 磺胺硝苯 334.0505 11.92
氨基糖苷类 阿米卡星 584.2789 2.63
雌激素类 己烷雌酚* 269.1553 13.82 己烯雌酚 267.1398 14.48
袢利尿剂 呋塞米 329.0006 11.01
非甾体抗炎药 卡洛芬 272.0489 14.29 菲诺洛芬 241.0872 13.78
抗寄生虫类 碘醚柳胺 623.8128 19.62 地克珠利 404.9727 14.77
尼卡巴嗪 301.0583 14.6 硝米特 210.0157 8.95
二硝托胺* 224.0314 9.22
呋喃类 呋喃苯烯酸钠 258.0407 13.03
玉米赤霉酮类 α-玉米赤霉醇* 321.1707 12.77 β-玉米赤霉醇* 321.1707 12.77
α-玉米赤霉烯醇* 319.1550 12.11 β-玉米赤霉烯醇* 319.1550 13.12
玉米赤霉酮* 319.1550 14.06
β-内酰胺酶抑制剂 舒巴坦 232.0286 6.39
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基于超高效液相色谱-四极杆-飞行时间高分辨质谱法快速筛查中兽药中302种药物非法添加
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关静渊 1 , 杨彦宁 1, * , 周大可 1, 2 , 隆雪明 1 , 周灿 1 , 张港 1 , 张瑞祺 1, 3 , 胡菊芳 1 , 李诗琪 1
食品安全质量检测学报 | 本期专题:食品中有毒有害物质分析与监测 2025,16(8): 107-114
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食品安全质量检测学报 | 本期专题:食品中有毒有害物质分析与监测 2025, 16(8): 107-114
基于超高效液相色谱-四极杆-飞行时间高分辨质谱法快速筛查中兽药中302种药物非法添加
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关静渊1 , 杨彦宁1, * , 周大可1, 2, 隆雪明1, 周灿1, 张港1, 张瑞祺1, 3, 胡菊芳1, 李诗琪1
作者信息
  • 1.湖南省兽药饲料监察所, 长沙 410006
  • 2.湖南农业大学动物医学院, 长沙 410128
  • 3.湖南农业大学食品科学技术学院, 长沙 410128
  • 关静渊(1996—), 女, 硕士, 兽医师, 主要研究方向为兽医药理学与兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:

通讯作者:

* 杨彦宁(1986—), 女, 硕士, 农艺师, 主要研究方向为兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:
Rapid screening of 302 kinds of illegal additives in traditional Chinese veterinary medicine by ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry
Jing-Yuan GUAN1 , Yan-Ning YANG1, * , Da-Ke ZHOU1, 2, Xue-Ming LONG1, Can ZHOU1, Gang ZHANG1, Rui-Qi ZHANG1, 3, Ju-Fang HU1, Shi-Qi LI1
Affiliations
  • 1. Hunan Province Institute of Veterinary Drug and Feed Control, Changsha 410006, China
  • 2. Hunan Agricultural University, College of Veterinary Medicine, Changsha 410128, China
  • 3. Hunan Agricultural University, School of Food Science and Technology, Changsha 410128, China
出版时间: 2025-04-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250116002
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目的 建立一种基于超高效液相色谱-四极杆-飞行时间高分辨质谱法(ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry, UPLC-Q-TOF-MS)用于中兽药中的302种药物非法添加的快速筛查方法。方法 建立了包含302种药物名称、分子式、精确质量数等信息的高分辨质谱数据库。样品经甲醇提取后, 经过UPLC-Q-TOF-MS进行数据采集并分析, 当保留时间与数据库绝对偏差<0.2 min, 质量误差<5 ppm, 同位素比值偏差<5%, 且主要特征碎片离子的库比对分数>80时, 认为检出疑似阳性样品。结果 该方法选择性较好, 目标物最低检出质量浓度低至50 ng/mL(四环素类、青霉素类、雌激素类、及非甾体抗炎药检出质量浓度100 ng/mL; 氨基糖苷类检出质量浓度200 ng/mL), 实现了302种非法添加药物的高效筛查。实际样品筛查时, 检出了7批疑似阳性样品。结论 本研究建立的筛查手段具有快速、广谱的特点, 适用于多种中兽药中的非法添加药物的初步筛查, 为中兽药质量控制提供了有效技术支持。

超高效液相色谱-四极杆-飞行时间高分辨质谱法  /  非法添加  /  化学药物  /  中兽药

Objective To establish a method based on ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry (UPLC-Q-TOF-MS) use as a rapid screening method for the illegal addition of 302 kinds of drugs in Chinese veterinary medicine. Methods A high-resolution mass spectrometry database containing information such as the names, molecular formulas, and exact mass numbers of 302 kinds of drugs was established. Samples were extracted with methanol and then analyzed by UPLC-Q-TOF-MS. A sample was considered as a suspected positive if the retention time deviation was less than 0.2 min, the mass error was less than 5 ppm, the isotopic ratio deviation was less than 5%, and the library matching score of the main characteristic fragment ions was greater than 80. Results The method exhibited good selectivity, with a minimum detection concentration as low as 50 ng/mL for the target substances (100 ng/mL for tetracyclines, penicillins, estrogens, and nonsteroidal anti-inflammatory drugs; 200 ng/mL for aminoglycosides). This enabled the efficient screening of 302 kinds of illegally added drugs. In the screening of actual samples, 7 batches of suspected positive samples were detected. Conclusion This approach is applicable for the screening of illegal additives in various veterinary Chinese medicines, providing effective technical support for the quality control of veterinary Chinese medicines.

ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry  /  illegal additives  /  chemical drugs  /  Chinese veterinary medicine
关静渊, 杨彦宁, 周大可, 隆雪明, 周灿, 张港, 张瑞祺, 胡菊芳, 李诗琪. 基于超高效液相色谱-四极杆-飞行时间高分辨质谱法快速筛查中兽药中302种药物非法添加. 食品安全质量检测学报, 2025 , 16 (8) : 107 -114 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250116002
Jing-Yuan GUAN, Yan-Ning YANG, Da-Ke ZHOU, Xue-Ming LONG, Can ZHOU, Gang ZHANG, Rui-Qi ZHANG, Ju-Fang HU, Shi-Qi LI. Rapid screening of 302 kinds of illegal additives in traditional Chinese veterinary medicine by ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry[J]. Journal of Food Safety & Quality, 2025 , 16 (8) : 107 -114 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250116002
中兽药, 作为传统兽医学的重要组成部分, 其在动物疾病预防和治疗中发挥着不可替代的作用。随着“禁抗”政策的实施, 中兽药产业迎来了广阔的发展前景, 据中国兽药协会数据, 2021年我国中兽药行业规模为61.26亿元, 预计到2028年市场规模将达到91亿元。然而, 市场上的中兽药产品良莠不齐, 一些不法商家为了追求短期效益, 生产或售卖掺假中兽药, 这种行为不仅破坏了中兽药的天然属性, 还可能对动物健康和食品安全造成严重威胁。
我国中药掺假现象主要可分为3大类: 一是通过添加未申报的药物以增强供临床效果; 二是添加工业染料, 冒充优质药材; 三是用非药物成分替代以增加重量或数量[1]。本研究聚焦于中兽药中非法添加药物的现象, 其中涉及的非法添加的化学药物种类繁多, 包括但不限于抗菌药[2]、抗病毒药物[3]、抗胆碱药物[4]、激素类[5]、解热镇痛药物[6]、中枢神经系统药物[7]β-受体激动剂[8-9]等。这些药物的添加虽然可能短期内能提高中兽药的疗效, 但长期使用会导致动物耐药性增强[10]、内分泌失调[11-12]等一系列健康问题, 更重要的是, 这些非法添加物可能残留在动物性食品(如肉、蛋、奶等)中, 最终进入人类食品链, 对消费者健康构成严重威胁。比如, 抗生素类药物的滥用可能导致药物在动物体内残留, 进而进入食品链, 影响消费者健康。抗生素类药物的过度使用还可能导致水体、土壤中抗药性菌株的滋生, 进而影响生态系统的稳定。此外, 由于中草药在收获、加工或储存过程中管理不善, 可能导致霉菌毒素污染; 同时, 一些不法商家为降低成本, 采购低价劣质原料, 或因生产工艺落后、存储条件不当等原因, 进一步加剧了中兽药安全风险[13]
食品安全是公众健康的重要组成部分, 而动物源食品的质量安全直接关系到消费者健康。因此, 建立高效、精准的检测手段, 不仅对保障中兽药的质量至关重要, 也对确保食品安全、防止非法药物进入食品链具有重要意义。超高效液相色谱-四极杆-飞行时间高分辨质谱法(ultra performance liquid chromatography-quadrupole time of flight high resolution-mass spectrometry, UPLC-Q-TOF-MS)因其高灵敏度、高选择性和高通量的特点, 成为快速筛查中兽药中非法添加化学药物的理想技术。与轨道阱质谱相比, 飞行时间质谱提供了更快速的扫描速度; 与离子阱质谱相比, 又提供了更高的质量分辨率, 有助于提高复杂样品中非法添加物的检测能力。此外, 飞行时间质谱具有优异的定性分析能力, 能够提供更精确的碎片离子图, 支持非法添加物的快速识别。该技术能够通过提供精确的质量测量和碎片离子图谱, 结合质谱数据库并通过串级质谱法谱图解析, 推断分子式和分子结构, 从而实现对非法添加物的快速识别。
本研究通过甲醇提取样品, 同时结合UPLC-Q-TOF-MS技术进行数据分析, 来构建包含302种非法添加药物的高分辨质谱数据库, 以期为中兽药的质量控制提供一种新的检测手段, 为中兽药行业的健康发展提供坚实的技术支撑。
SCIEX X500R QTOF液相色谱-四极杆/飞行时间质谱仪(上海SCIEX爱博才思分析仪器贸易有限公司); ME203/02万分之一电子分析天平[梅特勒托利多科技(中国)有限公司]; EFAA-HM-01多管涡旋震荡仪(上海安谱实验科技股份有限公司); SB-800DTD超声波清洗机(宁波新芝生物科技股份有限公司); 75005289高速离心机(美国赛默飞世尔科技公司); PGILSCXM2纯水仪[威立雅水处理技术(上海)有限公司]; 0.2 μm亲水性聚四氟乙烯(water-wettable polytetrafluoroethylene, wwPTFE)滤膜[颇尔(中国)有限公司]; Kinetex® F5 100 Å色谱柱(100 mm×4.6 mm, 2.6 µm)(天津博纳艾杰尔科技有限公司)。
甲醇、乙腈、乙酸铵(色谱纯, 德国默克有限公司); 甲酸(色谱纯, 美国ACS恩科化学公司); 302种兽药标准品溶液(质量浓度100 μg/mL, 天津阿尔塔科技有限公司); 采用空白样品基质液稀释样品。
实验所用实际检测样品均来自市售。
柱温40 ℃; 流动相为2 mmol/L乙酸铵+0.2%甲酸水溶液(A)与乙腈(B), 线性梯度洗脱, 共22 min, 流速0.3 mL/min; 进样量10 μL(表1)。
AB SCIEX X500R QTOF高分辨四极杆串联飞行时间质谱仪的质谱数据采用信息依赖扫描(information- dependent acquisition, IDA)的模式在正负离子模式下采集。
正离子模式下, 离子源参数为: 雾化气gas1为50 psi; 干燥气gas2为50 psi; 气帘气 curtain gas为30 psi; 离子源温度为550 ℃。TOF一级全扫描模式参数: 扫描范围50~1000 Da; 离子源电压为5500 V; 去簇电压50 V; 累计时间0.15 s; IDA参数: 最大候选离子数12; 触发二级碎裂扫描的离子强度阈值100 cps; 动态背景扣除; 质荷比误差范围±5 ppm。TOF二级碎片扫描模式参数: 扫描范围: 35~1000 Da; 碰撞电压为35 V; 累计时间0.06 s。
负离子模式下: 离子源电压-4500 V; 去簇电压-80 V, 碰撞电压为-35 V, 其他参数与正离子模式相同。
液体制剂: 移液枪精确移取20 μL, 甲醇10 mL定容;
固体制剂: 片剂、颗粒剂药物研磨粉碎后称取50 mg, 粉剂直接称取50 mg, 甲醇溶解并超声后, 10 mL定容。
样品定容后经0.2 μm wwPTFE滤膜过滤, 待上机。
X500R QTOF-MS可采集得到化合物的一级高分辨质谱(≥3500)和二级数据, 质量精确度≤5 ppm, 得到的谱图既有母离子的精确质量数、又有二级质谱全扫描信息。
本研究通过IDA模式扫描, 建立了302种兽药质谱数据库, 对其进行一级质谱全扫描, 得到高分辨质谱信息, 包括化合物的名称、CAS号、分子式、加荷方式、提取离子质荷比、保留时间, 见表2表3。所有兽药标准品均采用甲醇稀释至200 ng/mL后进样扫描, 以获取相关数据。
根据欧盟《食品饲料中农残分析的质量控制和方法确认的指导文件》(SANTE/11312/2021 (V2)及农业农村部公告第312号《饲料中风险物质的筛查与确认导则 液相色谱-高分辨质谱法(LC-HRMS)》, 物质定性需满足母离子质量偏差<5 ppm, 特征碎片离子质量数偏差小于10 ppm, 在不同碰撞能量下进行测定, 采集叠加所有子离子信息, 得到二级谱库。
提取目标物质质荷比±10 ppm窗口的离子图, 进行一级谱库检索, 当目标母离子精确质荷比≥200, 相对偏差≤10 ppm; 当目标母离子精确质荷比<200, 相对偏差应≤1 mDa, 提取特征碎片离子±20 ppm窗口的离子图, 各主要特征碎片离子均有检出。当保留时间与对照品绝对偏差<0.2 min, 质量误差<5 ppm, 同位素比值偏差<5%, 且主要特征碎片离子的谱库比对分数>80时, 认为筛查到可疑化合物或者疑似阳性样品。
本研究采用SCIEX OS软件(SCIEX OS Software, 版本号1.7.0.36606)进行数据处理, 包括质谱数据采集、峰检测、谱库建立、成分匹配、定性筛查及分析。
大多数物质在质谱分析中会经历离子化过程, 在此过程中不稳定的盐、酸、水等分子会脱除, 比如盐酸克仑特罗(C12H18Cl2N2O·HCl)在离子化过程中会失去HCl分子, 转化为C12H18Cl2N2O, 同样, 阿莫西林三水物(C16H19N3O5 S· 3H2O)会脱去3个水分子, 转化为C16H19N3O5S等。然而, 盐酸大观霉素五水合物(C14H24N2O7·2HCl·5H2O)进入质谱仪后, HCl和部分H2O分子被去除, 形成C14H24N2O7·H2O。检测结果显示, C14H24N2O7· H2O的离子响应更高且更稳定, 如图1
其次, 一些共晶体药物在进入质谱仪后, 会经历解共晶化的过程。如尼卡巴嗪是4,4’-二硝基苯胺(4,4’-dinitroaniline, DNC)和2-羟基-4,6-二甲基嘧啶(2-hydroxy-4,6-dimethylpyrimidine, HDP)分子以1:1的比例组合形成的共晶体[14], 质谱仪中, 这种共晶体分解为单独的DNC和HDP分子, 本研究条件下, DNC响应更高, 因此尼卡巴嗪以DNC (C13H10N4O5)计。同理, 氟尼辛葡甲胺是由2-(2-甲基-3-三氟甲基)苯胺基-吡啶-3-羧酸与1-脱氧-1-甲胺基-D-山梨醇以1:1的比例组合形成的共晶体[15], 因此, 氟尼辛葡甲胺以氟尼辛(C14H11F3N2O2)计算。
本研究共分离了19对同分异构体, 共41种物质, 见表1。这些同分异构体或色谱分离度良好, 或二级特征离子碎片不相同, 均可进行有效区分。比如, 布地奈德通常以22R和22S这两种差向异构体的1:1混合物形式存在[16], 是一种具有手性中心的化合物, 如图2
但是, 某些化合物虽并非同分异构体, 但在质谱分析过程中, 由于它们在离子源中的特定碎裂行为, 导致产生了与同分异构体相似的质谱特征, 比如, 富马酸福莫特罗二水合物与苯乙醇胺A不是同分异构体, 富马酸福莫特罗二水合物是一种多晶体结构[17](图3A), 在质谱中, 会解离成福莫特罗, 福莫特罗和苯乙醇胺A(图3B)是同分异构体, 其色谱图见图3C, 因此需要按照分析同分异构体的流程去进行甄别。与此相似的还有盐酸利托君和依托度酸、氨苄青霉素三水合物和头孢拉定、多西环素盐酸盐半乙醇半水合物和盐酸四环素。
为验证该筛查手段的特异性, 分别选取具有代表性的空白样品基质进行实验, 包括注射液、口服液、灌服液、预混剂、散剂、颗粒剂和片剂。各空白样品按照1.3下前处理方法处理后进样分析。结果表明, 在302种目标化合物的保留时间窗口内, 各空白样品基质均未出现干扰峰, 表明该方法具有良好的选择性, 不受常见制剂成分的干扰, 能够满足不同类型中兽药样品中非法添加药物的筛查要求。
在正离子模式下, 50 ng/mL质量浓度下的目标化合物检出率为73.1%, 其中四环素类、青霉素类、雌激素类、氨基糖苷类及非甾体抗炎药未检出; 100 ng/mL质量浓度下, 检出率提升至90.6%, 主要为氨基糖苷类化合物未被检出; 当质量浓度达到200 ng/mL时, 所有目标化合物均被检出, 检出率达到100%。在负离子模式50 ng/mL质量浓度下的检出率为90.5%, 100 ng/mL质量浓度下为95.2%, 且在200 ng/mL质量浓度下同样实现了100%的检出率。基于非法添加药物的作用主要是增强药效, 本研究建立的筛查手段能够有效覆盖常见非法添加药物的化学结构特征, 其灵敏度、选择性和筛查范围均能满足中兽药中非法添加药物的初步检出要求。该手段可为中兽药质量控制的初步筛查提供可靠的技术支持。
样品经试验方法处理净化, 上机检测, 读取总离子流图扫描数据, 对36批次中兽药中非法添加物进行了筛查, 对比302种化合物数据库信息, 发现了7批疑似阳性样品, 其中, 促孕灌注液中疑似添加了阿托品和东莨菪碱, 如图4, 阿托品和东莨菪碱属于抗胆碱药物, 通过抗M型胆碱受体的机制作用于子宫平滑肌, 能够显著降低子宫蠕动波、抑制子宫肌层收缩[18], 并维持血流灌注的充盈状态[19-20], 功效与促孕灌注液相似, 可提高母畜配种率, 促进恶露排出、缩短产后发情间期[21]。四黄止痢颗粒中疑似添加了泰妙菌素。四黄止痢颗粒在临床上广泛应用于细菌性、病毒性和湿热性下痢腹泻的防治[22], 而泰妙菌素作为截短侧耳素类动物专用抗生素, 对多种革兰氏阳性菌[23-24]以及支原体[25]和某些螺旋体[26]都有良好的抗菌活性, 同样多用于治疗猪的痢疾、肺炎[27]等病症, 还可以促进动物生长, 提高饲料利用率[28], 二者功效类似。其他筛查结果还有杨树花口服液中疑似添加碘醚柳胺, 藿香正气口服液中疑似添加氟尼辛葡甲胺、地克珠利, 盐酸甜菜碱预混剂中疑似添加林可霉素、甲氧苄啶、恩诺沙星和环丙沙星, 穿心莲注射液中疑似添加了林可霉素, 麻杏石甘散中疑似添加了甲氧苄啶、磺胺嘧啶、恩诺沙星。分析中兽药与非法添加药物的功效发现, 其添加行为主要是为了增强中兽药的“疗效”, 而非法添加者会利用监管盲区或检测技术的局限性, 规避检测, 因此大范围的非法添加筛查技术是遏制非法添加行为的重要措施。由于使用者无法知晓添加物的准确剂量, 且在养殖户缺乏药物使用安全意识的情况下, 盲目依赖中兽药的天然属性, 可能导致药物过量使用, 进而引发动物病情的恶化或程度不等的副反应[29-30], 严重威胁动物的生命安全。最终, 这会导致动物产品的兽药残留等问题, 并通过食物链传递至人类, 还会导致耐药菌风险的增加[31]。非法添加行为破坏了中兽药行业的信誉, 可能导致消费者对中兽药安全性的质疑, 进而影响整个行业的健康发展。
本研究利用UPLC-Q-TOF-MS, 成功构建了涵盖302种非法添加药物的高分辨质谱数据库, 为中兽药中的非法添加药物的快速筛查提供了一种高效、精准的技术手段, 并有助于降低食品中的药物残留风险。结果表明, 该手段能够快速、准确地识别多种非法添加物, 为中兽药质量控制提供了技术支持。
中兽药中非法添加药物现象可能由多个因素引起。一方面, 生产条件简陋和技术水平低的企业可能因疏忽引入额外药物; 另一方面, 一些商家可能为了提高疗效、追求经济利益而故意添加功效药物。此外, 市场上对快速见效药物的需求也可能促使非法添加行为。虽然这些药物能短期提升疗效, 但长期或不当使用可能引发剂量非依赖性、不良反应及健康风险。为了从源头上确保食品安全, 应加强对中兽药及动物性食品的质量监管, 推广高效的检测方法, 防止非法药物进入食品链, 保障公众健康。
  • 湖南省自然科学基金-科市联合项目(S2022JJKSLH0129)
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2025年第16卷第8期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250116002
  • 接收时间:2025-01-16
  • 首发时间:2025-07-19
  • 出版时间:2025-04-25
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  • 收稿日期:2025-01-16
基金
湖南省自然科学基金-科市联合项目(S2022JJKSLH0129)
作者信息
    1.湖南省兽药饲料监察所, 长沙 410006
    2.湖南农业大学动物医学院, 长沙 410128
    3.湖南农业大学食品科学技术学院, 长沙 410128

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* 杨彦宁(1986—), 女, 硕士, 农艺师, 主要研究方向为兽药饲料畜禽产品药物残留检测及相关技术研究。E-mail:
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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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