Article(id=1228268591108256432, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, articleNumber=null, orderNo=null, doi=10.12048/j.issn.1674-229X.2025.08.008, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1726761600000, receivedDateStr=2024-09-20, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1770771652246, onlineDateStr=2026-02-11, pubDate=1756051200000, pubDateStr=2025-08-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770771652246, onlineIssueDateStr=2026-02-11, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770771652246, creator=13701087609, updateTime=1770771652246, updator=13701087609, issue=Issue{id=1228268587459216114, tenantId=1146029695717560320, journalId=1227999776411738115, year='2025', volume='35', issue='8', pageStart='561', pageEnd='640', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1770771651377, creator=13701087609, updateTime=1770795163631, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1228367205105075039, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1228367205105075040, tenantId=1146029695717560320, journalId=1227999776411738115, issueId=1228268587459216114, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=602, endPage=608, ext={EN=ArticleExt(id=1228268592106500788, articleId=1228268591108256432, tenantId=1146029695717560320, journalId=1227999776411738115, language=EN, title=Simultaneous Determination of Six Components in Toddalia Asiatica Using Quantitative Analysis of Multi-Components by Single-Marker (QAMS) Method, columnId=null, journalTitle=Pharmacy Today, columnName=null, runingTitle=null, highlight=null, articleAbstract=
OBJECTIVE

To develop a quantitative analysis of multi-components by single-marker (QAMS) method for simultaneous determination of six major components (nitidine chloride, chelerythrine, hesperidin, zanthoxylumol, dictamnine and pimpinellin) in Toddalia asiatica.

METHODS

A reversed-phase high-performance liquid chromatography (HPLC) method was adopted using a Waters Symmetry C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of 0.1% formic acid aqueous solution and acetonitrile (containing 0.1% formic acid) with gradient elution. The flow rate was set at 0.8 mL·min-1, the detection wavelength was 280 nm, and the column temperature was maintained at 30 ℃. Taking hesperidin as the reference standard, relative correction factors (RCFs) of the other five components were determined, and the contents of the six major components were calculated. The accuracy and feasibility of the QAMS method were verified by investigating the robustness of RCFs and comparing the results with those obtained by the external standard method.

RESULTS

Within the linear range, the RCFs of nitidine chloride, chelerythrine, zanthoxylumol, dictamnine and pimpinellin were 0.170, 0.353, 3.443, 1.546 and 0.664, respectively. The RCFs exhibited good robustness under different experimental conditions (RSD < 3%). Additionally, there was no significant difference between the results obtained by the QAMS method and the external standard method for the analysis of eight batches of Toddalia asiatica.

CONCLUSION

The developed QAMS method is accurate and feasible, which can be used for determining the contents of major components in Toddalia asiatica.

, correspAuthors=Lei ZHANG, 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=Yingyin ZHANG, Jingling XIE, Yahui XU, Bingxuan ZHOU, Chuwen PENG, Jinying SU, Ziming CHEN, Lei ZHANG), CN=ArticleExt(id=1228268592748229316, articleId=1228268591108256432, tenantId=1146029695717560320, journalId=1227999776411738115, language=CN, title=一测多评法同时测定飞龙掌血中6种成分含量, columnId=1228268589086606089, journalTitle=今日药学, columnName=论著, runingTitle=null, highlight=null, articleAbstract=
目的

建立同时测定飞龙掌血中6种主要成分(氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱、茴芹香豆素)含量的一测多评法(QAMS)。

方法

采用反相高效液相色谱法,色谱柱:Waters Symmetry C18高效液相色谱柱(250 mm ×4.6 mm,5 μm);流动相:0.1%甲酸水溶液-乙睛(含0.1%甲酸),梯度洗脱;流速:0.8 mL·min-1;检测波长:280 nm;柱温:30 ℃。以橙皮苷为内参物,测定了其他5个成分的相对校正因子(RCF),并计算6种主要成分的含量。通过考察RCF的耐用性和与外标法的含量测定结果进行比较,以验证QAMS法的准确性和可行性。

结果

在线性范围内,氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱和茴芹香豆素的相对校正因子分别为0.170、0.353、3.443、1.546和0.664。在不同实验条件下相对校正因子耐用性良好,8批飞龙掌血的一测多评法计算结果和外标法计算结果无统计学意义。

结论

建立的一测多评法准确可行,可用于飞龙掌血主要成分的含量测定。

, correspAuthors=张蕾, authorNote=null, correspAuthorsNote=
* 张蕾,教授,博士,研究方向:中药药效物质基础及其作用机制,E-mail:
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# 张颖茵,硕士研究生,研究方向:药物分析学

谢京玲,中药学本科生

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注:A.供试品(110 min);B.供试品(75 min);C.混合对照品溶液。1.氯化两面针碱(nitidine chloride);2.白屈菜红碱(chelerythrine);3.橙皮苷(hesperidin);4.毛两面针素(zanthoxylumol);5.白鲜碱(dictamnine);6.茴芹香豆素(pimpinellin)。

, figureFileSmall=4/+caVbDot5/Z64TI7Eokg==, figureFileBig=q2xTs0IBS+NXgMxf50Cgcw==, tableContent=null), ArticleFig(id=1228268599022908380, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min乙腈(0.1%甲酸)/%0.1%甲酸水/%
0.00694
35.02080
90.07525
100.0982
110.0982
), ArticleFig(id=1228268599110988768, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表1, caption=

梯度洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min乙腈(0.1%甲酸)/%0.1%甲酸水/%
0.00694
35.02080
90.07525
100.0982
110.0982
), ArticleFig(id=1228268599199069157, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
成分回归方程r线性范围/μgLOQ/ngLOD/ng
氯化两面针碱y= 8.83E+6x-10 9111.0000.018 8~0.451 21.220.41
白屈菜红碱y= 4.20E+6x-1 614.61.0000.021 4~0.513 62.750.92
橙皮苷y= 1.47E+6x+803.721.0000.18~4.320.680.28
毛两面针素y= 4.29E+5x-1 291.71.0000.165~3.962.620.85
白鲜碱y= 9.54E+5x+2 052.71.0000.106~2.5448.682.49
茴芹香豆素y = 2.22E+6x+1 771.41.0000.069 3~1.663 04.061.54
), ArticleFig(id=1228268599308121068, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表2, caption=

6个成分的回归方程、线性范围、定量限及检测限

, figureFileSmall=null, figureFileBig=null, tableContent=
成分回归方程r线性范围/μgLOQ/ngLOD/ng
氯化两面针碱y= 8.83E+6x-10 9111.0000.018 8~0.451 21.220.41
白屈菜红碱y= 4.20E+6x-1 614.61.0000.021 4~0.513 62.750.92
橙皮苷y= 1.47E+6x+803.721.0000.18~4.320.680.28
毛两面针素y= 4.29E+5x-1 291.71.0000.165~3.962.620.85
白鲜碱y= 9.54E+5x+2 052.71.0000.106~2.5448.682.49
茴芹香豆素y = 2.22E+6x+1 771.41.0000.069 3~1.663 04.061.54
), ArticleFig(id=1228268599442338806, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
成分称样量/g原有量/mg加入量/mg测得量/mg回收率/%平均回收率/%RSD/%
氯化两面针碱0.251 40.1290.0640.195102.3100.81.9
0.251 00.1290.0640.195103.5
0.250 90.1290.0640.194101.7
0.252 30.1300.1280.25598.3
0.250 10.1290.1280.25599.1
0.250 30.1290.1280.257100.2
0.251 90.1290.1920.326102.5
0.253 00.1300.1920.324101.3
0.201 50.1040.1920.29298.4
白屈菜红碱0.251 40.4880.2290.724103.2101.22.0
0.251 00.4870.2290.726104.2
0.250 90.4870.2290.721102.1
0.252 30.4890.4580.949100.3
0.250 10.4850.4580.950101.3
0.250 30.4860.4580.94199.3
0.251 90.4890.6871.193102.5
0.253 00.4910.6871.16698.2
0.201 50.3910.6871.07499.4
橙皮苷0.251 45.4032.6347.99398.398.91.4
0.251 05.3942.6347.95597.2
0.250 95.3922.6348.01199.5
0.252 35.4225.26810.770101.4
0.250 15.3755.26810.660100.3
0.250 35.3795.26810.50097.3
0.251 95.4137.90213.20098.5
0.253 05.4377.90213.19098.1
0.201 54.3307.90212.17099.2
毛两面针素0.251 40.761 70.3521.09795.397.81.8
0.251 00.760 50.3521.09996.2
0.250 90.760 20.3521.10397.3
0.252 30.764 50.7031.45698.3
0.250 10.757 80.7031.45999.7
0.250 30.758 40.7031.470101.2
0.251 90.763 31.0551.78296.6
0.253 00.766 61.0551.79497.4
0.251 50.762 01.0551.79998.2
白鲜碱0.251 41.9380.9172.869101.6100.02.2
0.251 01.9350.9172.881103.2
0.250 91.9340.9172.83598.2
0.252 31.9451.8333.75298.6
0.250 11.9281.8333.71397.3
0.250 31.9301.8333.73598.5
0.251 91.9422.7504.67499.4
0.253 01.9512.7504.707100.2
0.201 51.5542.7504.389103.1
茴芹香豆素0.251 41.4610.7032.15498.798.92.3
0.251 01.4580.7032.14597.7
0.250 91.4580.7032.13696.4
0.252 31.4661.4062.920103.4
0.250 11.4531.4062.85399.6
0.250 31.4541.4062.871100.8
0.251 91.4642.1093.52397.6
0.253 01.4702.1093.50496.5
0.201 51.1712.1093.26899.5
), ArticleFig(id=1228268600818070523, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表3, caption=

加样回收试验结果(n=9)

, figureFileSmall=null, figureFileBig=null, tableContent=
成分称样量/g原有量/mg加入量/mg测得量/mg回收率/%平均回收率/%RSD/%
氯化两面针碱0.251 40.1290.0640.195102.3100.81.9
0.251 00.1290.0640.195103.5
0.250 90.1290.0640.194101.7
0.252 30.1300.1280.25598.3
0.250 10.1290.1280.25599.1
0.250 30.1290.1280.257100.2
0.251 90.1290.1920.326102.5
0.253 00.1300.1920.324101.3
0.201 50.1040.1920.29298.4
白屈菜红碱0.251 40.4880.2290.724103.2101.22.0
0.251 00.4870.2290.726104.2
0.250 90.4870.2290.721102.1
0.252 30.4890.4580.949100.3
0.250 10.4850.4580.950101.3
0.250 30.4860.4580.94199.3
0.251 90.4890.6871.193102.5
0.253 00.4910.6871.16698.2
0.201 50.3910.6871.07499.4
橙皮苷0.251 45.4032.6347.99398.398.91.4
0.251 05.3942.6347.95597.2
0.250 95.3922.6348.01199.5
0.252 35.4225.26810.770101.4
0.250 15.3755.26810.660100.3
0.250 35.3795.26810.50097.3
0.251 95.4137.90213.20098.5
0.253 05.4377.90213.19098.1
0.201 54.3307.90212.17099.2
毛两面针素0.251 40.761 70.3521.09795.397.81.8
0.251 00.760 50.3521.09996.2
0.250 90.760 20.3521.10397.3
0.252 30.764 50.7031.45698.3
0.250 10.757 80.7031.45999.7
0.250 30.758 40.7031.470101.2
0.251 90.763 31.0551.78296.6
0.253 00.766 61.0551.79497.4
0.251 50.762 01.0551.79998.2
白鲜碱0.251 41.9380.9172.869101.6100.02.2
0.251 01.9350.9172.881103.2
0.250 91.9340.9172.83598.2
0.252 31.9451.8333.75298.6
0.250 11.9281.8333.71397.3
0.250 31.9301.8333.73598.5
0.251 91.9422.7504.67499.4
0.253 01.9512.7504.707100.2
0.201 51.5542.7504.389103.1
茴芹香豆素0.251 41.4610.7032.15498.798.92.3
0.251 01.4580.7032.14597.7
0.250 91.4580.7032.13696.4
0.252 31.4661.4062.920103.4
0.250 11.4531.4062.85399.6
0.250 31.4541.4062.871100.8
0.251 91.4642.1093.52397.6
0.253 01.4702.1093.50496.5
0.201 51.1712.1093.26899.5
), ArticleFig(id=1228268600918733821, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
进样体积/μL相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
10.1750.3623.4541.5460.663
20.1710.3553.4561.5480.664
40.1700.3503.4491.5410.661
80.1690.3523.4331.5390.663
160.1680.3523.4441.5490.665
200.1670.3503.4421.5500.665
240.1670.3513.4211.5480.664
平均值0.1700.3533.4431.5460.664
RSD/%1.571.210.350.280.22
), ArticleFig(id=1228268601065533441, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表4, caption=

5个待测成分相对于橙皮苷的相对校正因子

, figureFileSmall=null, figureFileBig=null, tableContent=
进样体积/μL相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
10.1750.3623.4541.5460.663
20.1710.3553.4561.5480.664
40.1700.3503.4491.5410.661
80.1690.3523.4331.5390.663
160.1680.3523.4441.5490.665
200.1670.3503.4421.5500.665
240.1670.3513.4211.5480.664
平均值0.1700.3533.4431.5460.664
RSD/%1.571.210.350.280.22
), ArticleFig(id=1228268601162002436, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
柱温/℃相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
270.1690.3553.4301.5450.664
280.1690.3553.4371.5420.665
290.1680.3493.4711.5550.670
300.1690.3523.4331.5390.663
310.1670.3543.3941.5450.662
320.1680.3583.4261.5300.663
330.1690.3593.4271.5370.662
平均值0.1680.3553.4311.5420.664
RSD/%0.470.960.660.510.42
), ArticleFig(id=1228268601245888521, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表5, caption=

不同柱温对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
柱温/℃相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
270.1690.3553.4301.5450.664
280.1690.3553.4371.5420.665
290.1680.3493.4711.5550.670
300.1690.3523.4331.5390.663
310.1670.3543.3941.5450.662
320.1680.3583.4261.5300.663
330.1690.3593.4271.5370.662
平均值0.1680.3553.4311.5420.664
RSD/%0.470.960.660.510.42
), ArticleFig(id=1228268601308803087, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
流速/mL·min-1相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
0.70.1660.3513.4251.5350.659
0.80.1690.3523.4321.5390.663
0.90.1700.3533.4491.5420.665
1.00.1680.3503.4441.5490.674
平均值0.1680.3523.4361.5390.662
RSD/%1.020.370.320.380.94
), ArticleFig(id=1228268601405272086, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表6, caption=

不同流速对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
流速/mL·min-1相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
0.70.1660.3513.4251.5350.659
0.80.1690.3523.4321.5390.663
0.90.1700.3533.4491.5420.665
1.00.1680.3503.4441.5490.674
平均值0.1680.3523.4361.5390.662
RSD/%1.020.370.320.380.94
), ArticleFig(id=1228268601518518297, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
波长/nm相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
2790.1640.3393.5861.6010.668
2800.1690.3523.4331.5390.663
2810.1720.3683.2771.4950.650
平均值0.1680.3533.4321.5450.661
RSD/%2.404.124.513.451.43
), ArticleFig(id=1228268601669513249, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表7, caption=

不同波长对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
波长/nm相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
2790.1640.3393.5861.6010.668
2800.1690.3523.4331.5390.663
2810.1720.3683.2771.4950.650
平均值0.1680.3533.4321.5450.661
RSD/%2.404.124.513.451.43
), ArticleFig(id=1228268601782759462, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
仪器色谱柱相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
Waters Symmetry C180.1690.3533.4431.5460.664
岛津LC-20ATLuna 5u C18 100A0.1750.3603.3051.5110.658
Diamonsil C180.1710.3713.4511.5530.660
Waters Symmetry C180.1670.3743.5011.5630.654
安捷伦1260Luna 5u C18 100A0.1770.3513.3441.5370.673
Diamonsil C180.1640.3623.3571.5820.668
平均值0.1710.3623.4001.5490.663
RSD/%2.872.562.231.551.04
), ArticleFig(id=1228268601870839853, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表8, caption=

不同仪器和色谱柱对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
仪器色谱柱相对校正因子(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
Waters Symmetry C180.1690.3533.4431.5460.664
岛津LC-20ATLuna 5u C18 100A0.1750.3603.3051.5110.658
Diamonsil C180.1710.3713.4511.5530.660
Waters Symmetry C180.1670.3743.5011.5630.654
安捷伦1260Luna 5u C18 100A0.1770.3513.3441.5370.673
Diamonsil C180.1640.3623.3571.5820.668
平均值0.1710.3623.4001.5490.663
RSD/%2.872.562.231.551.04
), ArticleFig(id=1228268601988280371, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
流速/mL·min-1柱温/℃相对保留时间(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
0.7300.9590.9811.1261.3931.442
0.8300.9620.9861.1281.3961.448
1.0300.9600.9871.1301.3981.450
平均值0.9600.9851.1281.3961.447
RSD/%0.1600.3300.180.1800.290
0.8270.9650.9911.1261.3961.449
0.8300.9640.9831.1211.3801.429
0.8330.9560.9791.1331.3951.445
平均值0.9620.9841.1271.3901.441
RSD/%0.4200.5100.4400.5300.60
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不同流速和温度下飞龙掌血中5个待测成分相对于橙皮苷的相对保留时间

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流速/mL·min-1柱温/℃相对保留时间(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
0.7300.9590.9811.1261.3931.442
0.8300.9620.9861.1281.3961.448
1.0300.9600.9871.1301.3981.450
平均值0.9600.9851.1281.3961.447
RSD/%0.1600.3300.180.1800.290
0.8270.9650.9911.1261.3961.449
0.8300.9640.9831.1211.3801.429
0.8330.9560.9791.1331.3951.445
平均值0.9620.9841.1271.3901.441
RSD/%0.4200.5100.4400.5300.60
), ArticleFig(id=1228268602164441147, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
仪器色谱柱相对保留时间(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
岛津LC20ATWaters Symmetry C180.9620.9861.1281.3961.448
Luna 5u C18 100A0.9550.9721.1361.3731.435
Diamonsil C180.9710.9971.1411.3531.423
安捷伦1260Waters Symmetry C180.9670.9811.1161.3631.434
Luna 5u C18 100A0.9480.9831.1441.3371.423
Diamonsil C180.9750.9941.1571.3821.368
平均值0.9630.9861.1371.3671.422
RSD/%1.050.921.231.551.97
), ArticleFig(id=1228268602265104453, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表10, caption=

不同仪器和色谱柱下飞龙掌血中5个待测成分相对于橙皮苷的相对保留时间

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仪器色谱柱相对保留时间(以橙皮苷为内参物)
氯化两面针碱白屈菜红碱毛两面针素白鲜碱茴芹香豆素
岛津LC20ATWaters Symmetry C180.9620.9861.1281.3961.448
Luna 5u C18 100A0.9550.9721.1361.3731.435
Diamonsil C180.9710.9971.1411.3531.423
安捷伦1260Waters Symmetry C180.9670.9811.1161.3631.434
Luna 5u C18 100A0.9480.9831.1441.3371.423
Diamonsil C180.9750.9941.1571.3821.368
平均值0.9630.9861.1371.3671.422
RSD/%1.050.921.231.551.97
), ArticleFig(id=1228268602357379147, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
化合物S1S2S3S4
ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%
氯化两面针碱0.5140.5262.330.1640.1671.830.5730.5872.440.5620.5752.31
白屈菜红碱1.9401.9500.521.5301.5400.650.6400.6501.560.3030.3081.65
橙皮苷21.49021.450-0.1917.56017.520-0.2324.67024.630-0.1629.19029.140-0.17
毛两面针素3.0303.010-0.663.1103.090-0.647.5507.530-0.268.3008.290-0.12
白鲜碱7.7107.700-0.134.9204.9210.023.4803.450-0.865.7805.720-1.04
茴芹香豆素5.8105.800-0.175.5105.490-0.366.7806.760-0.294.3404.300-0.92
化合物S5S6S7S8
ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%
氯化两面针碱0.6770.6881.620.7830.7992.040.1320.1352.270.5730.563-1.75
白屈菜红碱0.4340.4391.150.4930.4970.810.4620.4671.080.6530.6530.00
橙皮苷28.62028.570-0.170.1390.1411.4435.44035.380-0.174.2904.2920.05
毛两面针素15.14015.1500.071.3301.310-1.500.7000.680-2.8615.04015.0500.07
白鲜碱6.4506.400-0.785.5945.591-0.052.7702.7800.367.1207.100-0.28
茴芹香豆素3.2203.200-0.621.9401.920-1.031.9801.970-0.513.1403.1600.64
), ArticleFig(id=1228268602445459535, tenantId=1146029695717560320, journalId=1227999776411738115, articleId=1228268591108256432, language=CN, label=表11, caption=

飞龙掌血中6种成分含量测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物S1S2S3S4
ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%
氯化两面针碱0.5140.5262.330.1640.1671.830.5730.5872.440.5620.5752.31
白屈菜红碱1.9401.9500.521.5301.5400.650.6400.6501.560.3030.3081.65
橙皮苷21.49021.450-0.1917.56017.520-0.2324.67024.630-0.1629.19029.140-0.17
毛两面针素3.0303.010-0.663.1103.090-0.647.5507.530-0.268.3008.290-0.12
白鲜碱7.7107.700-0.134.9204.9210.023.4803.450-0.865.7805.720-1.04
茴芹香豆素5.8105.800-0.175.5105.490-0.366.7806.760-0.294.3404.300-0.92
化合物S5S6S7S8
ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%ESM/mg·g-1QAMS/mg·g-1RE/%
氯化两面针碱0.6770.6881.620.7830.7992.040.1320.1352.270.5730.563-1.75
白屈菜红碱0.4340.4391.150.4930.4970.810.4620.4671.080.6530.6530.00
橙皮苷28.62028.570-0.170.1390.1411.4435.44035.380-0.174.2904.2920.05
毛两面针素15.14015.1500.071.3301.310-1.500.7000.680-2.8615.04015.0500.07
白鲜碱6.4506.400-0.785.5945.591-0.052.7702.7800.367.1207.100-0.28
茴芹香豆素3.2203.200-0.621.9401.920-1.031.9801.970-0.513.1403.1600.64
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一测多评法同时测定飞龙掌血中6种成分含量
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张颖茵 1, 2 , 谢京玲 1 , 徐雅慧 1 , 周冰旋 1 , 彭楚雯 1 , 苏金莹 1 , 陈梓铭 1 , 张蕾 1, *
今日药学 | 论著 2025,35(8): 602-608
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今日药学 | 论著 2025, 35(8): 602-608
一测多评法同时测定飞龙掌血中6种成分含量
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张颖茵1, 2, 谢京玲1, 徐雅慧1, 周冰旋1, 彭楚雯1, 苏金莹1, 陈梓铭1, 张蕾1, *
作者信息
  • 1.广州中医药大学,广东 广州 510006
  • 2.广州华商学院,广东 广州 511300

通讯作者:

* 张蕾,教授,博士,研究方向:中药药效物质基础及其作用机制,E-mail:
Simultaneous Determination of Six Components in Toddalia Asiatica Using Quantitative Analysis of Multi-Components by Single-Marker (QAMS) Method
Yingyin ZHANG1, 2, Jingling XIE1, Yahui XU1, Bingxuan ZHOU1, Chuwen PENG1, Jinying SU1, Ziming CHEN1, Lei ZHANG1, *
Affiliations
  • 1.Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China
  • 2.Guangzhou Huashang College, Guangzhou, Guangdong 511300, China
出版时间: 2025-08-25 doi: 10.12048/j.issn.1674-229X.2025.08.008
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目的

建立同时测定飞龙掌血中6种主要成分(氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱、茴芹香豆素)含量的一测多评法(QAMS)。

方法

采用反相高效液相色谱法,色谱柱:Waters Symmetry C18高效液相色谱柱(250 mm ×4.6 mm,5 μm);流动相:0.1%甲酸水溶液-乙睛(含0.1%甲酸),梯度洗脱;流速:0.8 mL·min-1;检测波长:280 nm;柱温:30 ℃。以橙皮苷为内参物,测定了其他5个成分的相对校正因子(RCF),并计算6种主要成分的含量。通过考察RCF的耐用性和与外标法的含量测定结果进行比较,以验证QAMS法的准确性和可行性。

结果

在线性范围内,氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱和茴芹香豆素的相对校正因子分别为0.170、0.353、3.443、1.546和0.664。在不同实验条件下相对校正因子耐用性良好,8批飞龙掌血的一测多评法计算结果和外标法计算结果无统计学意义。

结论

建立的一测多评法准确可行,可用于飞龙掌血主要成分的含量测定。

飞龙掌血  /  含量测定  /  一测多评  /  高效液相色谱  /  相对校正因子
OBJECTIVE

To develop a quantitative analysis of multi-components by single-marker (QAMS) method for simultaneous determination of six major components (nitidine chloride, chelerythrine, hesperidin, zanthoxylumol, dictamnine and pimpinellin) in Toddalia asiatica.

METHODS

A reversed-phase high-performance liquid chromatography (HPLC) method was adopted using a Waters Symmetry C18 column (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of 0.1% formic acid aqueous solution and acetonitrile (containing 0.1% formic acid) with gradient elution. The flow rate was set at 0.8 mL·min-1, the detection wavelength was 280 nm, and the column temperature was maintained at 30 ℃. Taking hesperidin as the reference standard, relative correction factors (RCFs) of the other five components were determined, and the contents of the six major components were calculated. The accuracy and feasibility of the QAMS method were verified by investigating the robustness of RCFs and comparing the results with those obtained by the external standard method.

RESULTS

Within the linear range, the RCFs of nitidine chloride, chelerythrine, zanthoxylumol, dictamnine and pimpinellin were 0.170, 0.353, 3.443, 1.546 and 0.664, respectively. The RCFs exhibited good robustness under different experimental conditions (RSD < 3%). Additionally, there was no significant difference between the results obtained by the QAMS method and the external standard method for the analysis of eight batches of Toddalia asiatica.

CONCLUSION

The developed QAMS method is accurate and feasible, which can be used for determining the contents of major components in Toddalia asiatica.

Toddalia asiatica  /  quantitative determination  /  quantitative analysis of multi-components by single-marker (QAMS)  /  high-performance liquid chromatography (HPLC)  /  relative correction factor (RCF)
张颖茵, 谢京玲, 徐雅慧, 周冰旋, 彭楚雯, 苏金莹, 陈梓铭, 张蕾. 一测多评法同时测定飞龙掌血中6种成分含量. 今日药学, 2025 , 35 (8) : 602 -608 . DOI: 10.12048/j.issn.1674-229X.2025.08.008
Yingyin ZHANG, Jingling XIE, Yahui XU, Bingxuan ZHOU, Chuwen PENG, Jinying SU, Ziming CHEN, Lei ZHANG. Simultaneous Determination of Six Components in Toddalia Asiatica Using Quantitative Analysis of Multi-Components by Single-Marker (QAMS) Method[J]. Pharmacy Today, 2025 , 35 (8) : 602 -608 . DOI: 10.12048/j.issn.1674-229X.2025.08.008
飞龙掌血[Toddalia asiatica(L.)Lam.]是芸香科植物飞龙掌血的根或根皮,主要分布于西南及陕西、浙江、湖北、湖南、广东、海南、广西等地,又名飞龙斩血、见血飞、黄大金根、血棒头、簕钩等,为我国西南地区少数民族的传统中药材[1]。飞龙掌血性味辛苦,性温,有毒,主要用于风湿痹痛、胃痛、疮痈肿毒等,临床上常用于治疗风湿痹痛、跌打损伤、吐血、衄血、经闭、痛经等疾病[2]。作为治疗风湿类疾病的中药苗医药,飞龙掌血的成方制剂主要有龙钻通痹颗粒、痛可舒酊、龙掌口含液等[3]。飞龙掌血中主要富含香豆素类和生物碱类成分,目前飞龙掌血主要成分的HPLC含量测定方法多数采用外标法[4-7],但是该方法必须具备充足的对照品,对照品稀缺、不稳定或者价格昂贵等将直接影响多成分含量测定方法的应用,而一测多评法(QAMS)的出现则解决了这个问题。
QAMS是由王智民等[8]在2006年提出的适合中医药特点又能解决中药对照品瓶颈问题的多指标含量测定方法。该法是在相对应的线性范围内,对所需待测样之间进行计算得到其相关的内在函数关系和比例关系,选择1个性质稳定、保留时间合适、价格低廉和含量大等的成分作为内参物,经方法学考察,建立内参物与其他多种成分间的相对校正因子,从而实现多个待测成分的同时测定,是一种适用于待测成分的对照品难以获得、稳定性差、成本价格高等中药的多成分含量测定,现已被广泛应用于多种中药质量评价的研究中[9-12],并且在2015年版中国药典开始,QAMS法的应用拓展至饮片、提取物及制剂[13,14]。氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素为飞龙掌血中的主要有效成分,能够反映药材质量优劣,也是体内外分析中检测频率较高的指标成分[15]。故本文采用高效液相色谱法,首次以橙皮苷为内参物,建立QAMS法同时测定不同产地飞龙掌血中这6种主要化学成分的含量,并通过耐用性试验考察了QAMS法在飞龙掌血质量控制上的可行性和可推广性。
LC-20AT岛津高效液相色谱仪,日本岛津公司;Agilent1260高效液相系统,美国安捷伦科技有限公司;KQ-250E超声波清洗器,昆山市超声仪器有限公司;SHZ-D(Ⅲ)(四氟)循环水真空泵,上海力辰邦西仪器科技有限公司;QUINTIX125D-ICN型十万分之一分析天平,赛多利斯斯泰帝(上海)贸易有限公司。RADWAG AS 62.R2 PLUS精密分析电子天平、JP-060型超声波清洗器,昆山市超声仪器有限公司;TG16G型台式高速离心机,广州贝拓科学技术有限公司;HC-2000粉碎机,永康市天祺盛世工贸有限公司;LC-LX-HR165A台式高速冷冻离心机,上海力辰邦西仪器科技有限公司。Waters Symmetry C18色谱柱(4.6 mm×250 mm,5 μm),沃特世科技(上海)有限公司;Luna 5u C18 100A色谱柱(250 mm×4.6 mm,5 μm),美国飞诺美公司;Diamonsil C18色谱柱(250 mm×4.6 mm,5 μm),北京迪马科技有限公司。甲醇(分析纯),天津市科密欧化学试剂有限公司;甲醇和乙腈(色谱纯),西格玛奥德里奇贸易有限公司。
氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素对照品均购自上海诗丹德生物技术有限公司,纯度≥98%;8批次不同产地飞龙掌血药材饮片(S1-湖南;S2-四川;S3-湖北;S4-浙江;S5-云南;S6-梅州;S7-贺州;S8-河池)购于各大医药公司,经广州中医药大学彭光天教授鉴定芸香科植物飞龙掌血[Toddalia asiatica(L.)Lam.]的根茎。
精密称取氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素对照品适量,经甲醇溶解、定容及稀释后制备成含氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素质量浓度分别为18.8、21.4、180、165、106.4、69.3 μg·mL-1的混合对照品溶液。
精密称取干燥的药材粉末(40目)0.5 g,置于250 mL具塞锥形瓶中,精密加入甲醇溶液50 mL,密塞,称定重量,超声提取30 min,放冷,再次称重,用甲醇补足减失重量,摇匀,于转速13 000 r·min-1下离心5 min,取上清即得供试品溶液。
色谱柱:Waters Symmetry C18(4.6 mm×250 mm,5 μm);流动相:0.1%甲酸水溶液-乙睛(含0.1%甲酸),梯度洗脱;检测波长:280 nm;流速:0.8 mL·min-1;柱温:30 ℃;进样量:10 μL。见表1
按“2.2”项下色谱条件,分别进样混合对照品溶液和供试品溶液,见图1。样品中的6个待测成分色谱峰形及分离效果均良好,符合分析要求。
精密吸取混合对照品溶液1.0、2.0、4.0、8.0、16.0、20.0、24.0 μL,按“2.2”项色谱条件测定,记录峰面积。以对照品质量(μg)为横坐标(x),峰面积为纵坐标(y)绘制标准曲线,得到各成分的标准曲线回归方程、相关系数(r)及线性范围。取混合对照品溶液,经甲醇逐级稀释后,以信噪比分别约为10和3估算定量限(LOQ)和检测限(LOD)。结果见表2
精密吸取混合对照品溶液10 μL,连续进样6次,记录氯化两面针碱、白屈菜红碱、橙皮昔、毛两面针素、白鲜碱、苗芹香豆素的色谱峰面积,并计算RSD。结果显示氯化两面针碱、白屈菜红碱、橙皮昔、毛两面针素、白鲜碱和茴芹香豆素峰面积的RSD分别为0.79%、0.63%、0.83%、0.92%、0.89%、0.88%,结果表明仪器精密度良好。
精密称取S1样品粉末,按“2.1.2”项下方法制备供试品溶液,分别在制备好后的0、2、4、8、12、24 h按“2.2”项下色谱条件进样分析,记录峰面积,并计算RSD。结果表明氯化两面针碱、白屈菜红碱、橙皮昔、毛两面针素、白鲜碱和茴芹香豆素峰面积的RSD分别为2.4%、1.4%、0.87%、2.8%、0.86%和0.69%,表明所制备的供试品溶液在24 h内稳定性良好。
精密称取S1样品粉末(40目)6份,按“2.1.2”项下方法制备供试品溶液,按“2.2”项下色谱条件进样分析,记录峰面积,并计算含量的RSD分别为2.2%、1.1%、2.5%、1.9%、0.97%和2.7%,表明该法的重复性良好。
精密称取S1样品粉末(40目)9份,每份约0.25 g,分别按照约1∶0.5、1∶1和1∶ 1.5精密加入对照品适量,按“2.1.2”项下操作制备低、中、高浓度的供试品溶液,每个浓度3样本分析,计算回收率、平均回收率及RSD。结构见表3,氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素的平均回收率分别为100.8%、101.2%、98.9%、97.8%、100.0%和98.9%,RSD分别为1.9%、2.0%、1.4%、1.8%,2.2%和2.3%,表明方法的准确度良好。
取“2.3.2”项下数据,以橙皮苷为内参物,按公式fsi = fs/fi =(As×mi)/(Ai×ms)分别计算氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱、茴芹香豆素的相对校正因子(式中As为内参物S的峰面积,ms为内参物的质量浓度,Ai为待测成分的峰面积,mi为待测成分的质量/浓度)。结果见表4
取“2.1.1”项下混合对照溶液,分别在柱温27 ℃、28 ℃、29 ℃、30 ℃、31 ℃、32 ℃、33 ℃下测定相对校正因子,考察柱温对相对校正因子的影响,结果见表5,氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱和茴芹香豆素的相对校正因子的RSD分别为0.47%,0.96%,0.66%,0.51%和0.42%,表明在设定柱温(30±3)℃范围内对温度的耐用性良好。
取“2.1.1”项下混合对照溶液,分别流动相流速为0.7、0.8、0.9、1.0 mL·min-1时测定相对校正因子,考察流速对相对校正因子的影响,结果见表6,氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱、茴芹香豆素的相对校正因子的RSD分别为1.02%、0.37%、0.32%、0.38%、0.94%,显示均无明显影响。
取“2.1.1”项混合对照溶液,分别在波长为278、279、280、281和282 nm下测定相对校正因子,考察检测波长微小变化对相对校正因子的影响,结果显示,在(280±2)nm波长范围内,氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱和茴芹香豆素相对校正因子的RSD多数>5%,而在(280±1)nm波长范围内,上述5个化合物的相对校正因子的RSD分别为2.40%、4.12%、4.51%、3.45%和1.43%,结果见表7
取“2.1.1”项混合对照溶液,分别考察在岛津LC-20AT高效液相色谱仪、Agilent 1260液相色谱仪和3种色谱柱Waters Symmetry C18色谱柱(4.6 mm×250 mm,5 μm),Luna C18 100A色谱柱(250 mm×4.6 mm,5 μm),Diamonsil C18色谱柱(250 mm×4.6 mm,5 μm),上的相对校正因子,考察不同仪器和色谱柱对相对校正因子的影响,结果见表8,相对校正因子在不同的色谱柱和仪器下均无显著差异,具有较好的适用性。
QAMS法待测成分色谱峰用相对保留值法及各色谱峰的紫外吸收特征来准确地定位。本研究考察了相对保留时间(RtR = tR(待测物)/tR(内参物))在不同柱温、流速、色谱柱和色谱仪器下的重现性,结果见表9~10。表明氯化两面针碱、白屈菜红碱、毛两面针素、白鲜碱和茴芹香豆素相对保留时间的RSD均小于2.0%,说明各成分的相对保留时间相对稳定,可选用相对保留值作为目标成分色谱峰的定位指标。
取8批不同产地的飞龙掌血药材粉末,按“2.1.2”项下制备供试品溶液,按“2.2”色谱条件下进样分析,分别采用外标法(ESM)和QAMS法计算飞龙掌血中氯化两面针碱、白屈菜红碱、橙皮苷、毛两面针素、白鲜碱和茴芹香豆素的含量,并计算相对误差(RE),结果见表11,两种方法所得飞龙掌血中6种成分含量之间无统计学意义,表明QAMS法能够准确测定飞龙掌血中6种成分的含量。
本研究参考课题组前期研究内容[16,17],在此基础上建立QAMS法同时测定飞龙掌血中6个成分,方法学考察表明该方法准确可行。相对校正因子和相对保留时间的耐用性实验结果表明其在不同仪器、不同色谱柱、不同流速、不同柱温条件下均具有良好重现性和适用性,因此选用相对保留值作为目标成分色谱峰的定位指标和含量计算参数。不同产地8批药材的测定结果与标准曲线法所测值无统计学意义,说明建立不同成分之间的相对校正因子具有较高准确度。本研究通过QAMS技术可实现仅以橙皮苷为对照品,便可以同时测定飞龙掌血中的多个指标成分,为飞龙掌血的质量控制提供了更便捷的质量评价方法。
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doi: 10.12048/j.issn.1674-229X.2025.08.008
  • 接收时间:2024-09-20
  • 首发时间:2026-02-11
  • 出版时间:2025-08-25
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  • 收稿日期:2024-09-20
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广东省基础与应用基础省企联合基金-面上项目(2023A1515220188)
作者信息
    1.广州中医药大学,广东 广州 510006
    2.广州华商学院,广东 广州 511300

通讯作者:

* 张蕾,教授,博士,研究方向:中药药效物质基础及其作用机制,E-mail:
参考文献
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https://castjournals.cast.org.cn/joweb/jryx/CN/10.12048/j.issn.1674-229X.2025.08.008
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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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