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Article(id=1193877806431044062, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, articleNumber=1001-2494(2025)02-0180-04, orderNo=null, doi=10.11669/cpj.2025.02.010, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1706198400000, receivedDateStr=2024-01-26, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1762572250230, onlineDateStr=2025-11-08, pubDate=1737475200000, pubDateStr=2025-01-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1762572250230, onlineIssueDateStr=2025-11-08, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1762572250230, creator=13701087609, updateTime=1762572250230, updator=13701087609, issue=Issue{id=1193877800143777917, tenantId=1146029695717560320, journalId=1190317699101192196, year='2025', volume='60', issue='2', pageStart='109', pageEnd='206', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=0, articleOrder=1, issueType=-1, specialIssue=null, createTime=1762572248731, creator=13701087609, updateTime=1762584852274, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1193930663289123481, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1193930663289123482, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=180, endPage=183, ext={EN=ArticleExt(id=1193877806632370659, articleId=1193877806431044062, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Comparison of UHPLC-MS/MS and Chemiluminescence Microparticle Immune Assay in Determination of Vancomycin Concentration in Human Serum, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To analyze the consistency and correlation between UHPLC-MS/MS and chemiluminescence microparticle immune assay (CMIA) in measuring vancomycin concentration in serum.METHODS The concentration of vancomycin was measured by UHPLC-MS/MS and CMIA methods respectively in 95 serum samples of patients receiving vancomycin treatment. A correlation analysis and an agreement analysis were conducted for the two methods used linear regression and Bland-Altman method. RESULTS Based on the results of UHPLC-MS/MS(x) and CMIA(y), the linear regression equation was determined as follows: y=1.088x+0.174. Spearmen correlation analysis indicated a significant correlation between the results by the two methods (P<0.05), and the Spearmen correlation coefficient was 0.969 1. There was statistical difference between vancomycin serum concentrations measured by these two methods. The result was higher by CMIA compared with UHPLC-MS/MS. CONCLUSION The results of the two methods can be calculated to one another by equation, which provides reference for the formulation of clinical individualized medication plan.

, correspAuthors=Dan YAN, 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=Zhengyuan SHI, Jie XIE, Ziying LI, You JIANG, Xiqiao XU, Chunjing YANG, Yanwei FU, Dan YAN), CN=ArticleExt(id=1193877951516214141, articleId=1193877806431044062, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=超高效液相色谱-串联质谱法与化学发光微粒子免疫法测定万古霉素血药浓度的比较分析, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 比较国产QLIT-6610MD超高效液相色谱-串联质谱(UHPLC-MS/MS)法和化学发光微粒子免疫分析法(CMIA)测定万古霉素血药浓度的相关性,建立测定结果换算关系。 方法 收集接受万古霉素治疗患者的血清样本95例,分别用UHPLC-MS/MS和CMIA法进行测定,对测定结果进行回归分析,用 Spearman 法进行相关性比较,Bland-Altman法评价一致性。 结果 以UHPLC-MS/MS测定结果(x)和 CMIA 测定结果(y)所作的线性回归方程如下:y=1.088x+0.174,两组结果间存在较好的相关性(Spearmen相关系数为0.969 1)。CMIA方法的测定结果高于UHPLC-MS/MS法所测结果,差异有统计学意义(P<0.05)。 结论 2种方法测定的万古霉素血清浓度可通过方程式进行结果互算,为临床个体化用药方案的制订提供参考。

, correspAuthors=鄢丹, authorNote=null, correspAuthorsNote=
*鄢丹,男,博士,主任药师 研究方向:中西医结合临床药学 Tel:(010)63139318
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时正媛,女,博士,副主任药师 研究方向:治疗药物监测与个体化给药

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时正媛,女,博士,副主任药师 研究方向:治疗药物监测与个体化给药

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时正媛,女,博士,副主任药师 研究方向:治疗药物监测与个体化给药

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figureFileSmall=pnvbZnUYkiNJEbqLjWEtVw==, figureFileBig=ubTJjkQUJLuuuaBJ5DBzuQ==, tableContent=null), ArticleFig(id=1193928905254994709, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877806431044062, language=EN, label=Tab.1, caption=

The dermination mass spectrum parameters of vancomycin in human serum

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Analyte m/z
(Parention)		 m/z
(Daughterion)		 T(Enrichment)
/ms		 Fragmentation
/point q		 Fragmentation
energy/%
Vancomycin 725.4 144.0 150 0.13 35
IS 690.8 144.0 200 0.14 40
), ArticleFig(id=1193928905313714966, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877806431044062, language=CN, label=表1, caption=

人血清中万古霉素检测的质谱参数

, figureFileSmall=null, figureFileBig=null, tableContent=
Analyte m/z
(Parention)		 m/z
(Daughterion)		 T(Enrichment)
/ms		 Fragmentation
/point q		 Fragmentation
energy/%
Vancomycin 725.4 144.0 150 0.13 35
IS 690.8 144.0 200 0.14 40
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超高效液相色谱-串联质谱法与化学发光微粒子免疫法测定万古霉素血药浓度的比较分析
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时正媛 1 , 谢洁 2 , 李姿颖 2 , 江游 2 , 续茜桥 1 , 杨春静 1 , 付延伟 1 , 鄢丹 3, *
中国药学杂志 | 论著 2025,60(2): 180-183
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中国药学杂志 | 论著 2025, 60(2): 180-183
超高效液相色谱-串联质谱法与化学发光微粒子免疫法测定万古霉素血药浓度的比较分析
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时正媛1, 谢洁2, 李姿颖2, 江游2, 续茜桥1, 杨春静1, 付延伟1, 鄢丹3, *
作者信息
  • 1 首都医科大学附属北京世纪坛医院药学部, 北京 100038
  • 2 中国计量科学研究院前沿计量科学中心,国家市场监管技术创新中心(质谱), 北京 100029
  • 3 首都医科大学附属北京友谊医院药学部, 北京 100050

    • 时正媛,女,博士,副主任药师 研究方向:治疗药物监测与个体化给药

通讯作者:

*鄢丹,男,博士,主任药师 研究方向:中西医结合临床药学 Tel:(010)63139318
Comparison of UHPLC-MS/MS and Chemiluminescence Microparticle Immune Assay in Determination of Vancomycin Concentration in Human Serum
Zhengyuan SHI1, Jie XIE2, Ziying LI2, You JIANG2, Xiqiao XU1, Chunjing YANG1, Yanwei FU1, Dan YAN3, *
Affiliations
  • 1 Department of Pharmacy,Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China
  • 2 Technology Innovation Center of Mass Spectrometry for State Market Regulation, Center for Advanced Measurement Science, National Institute of Metrology, Beijing 100029, China
  • 3 Department of Pharmacy, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China
出版时间: 2025-01-22 doi: 10.11669/cpj.2025.02.010
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摘要
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目的 比较国产QLIT-6610MD超高效液相色谱-串联质谱(UHPLC-MS/MS)法和化学发光微粒子免疫分析法(CMIA)测定万古霉素血药浓度的相关性,建立测定结果换算关系。 方法 收集接受万古霉素治疗患者的血清样本95例,分别用UHPLC-MS/MS和CMIA法进行测定,对测定结果进行回归分析,用 Spearman 法进行相关性比较,Bland-Altman法评价一致性。 结果 以UHPLC-MS/MS测定结果(x)和 CMIA 测定结果(y)所作的线性回归方程如下:y=1.088x+0.174,两组结果间存在较好的相关性(Spearmen相关系数为0.969 1)。CMIA方法的测定结果高于UHPLC-MS/MS法所测结果,差异有统计学意义(P<0.05)。 结论 2种方法测定的万古霉素血清浓度可通过方程式进行结果互算,为临床个体化用药方案的制订提供参考。

万古霉素  /  血清  /  治疗药物监测  /  超高效液相色谱-串联质谱法  /  化学发光微粒子免疫检测法

OBJECTIVE To analyze the consistency and correlation between UHPLC-MS/MS and chemiluminescence microparticle immune assay (CMIA) in measuring vancomycin concentration in serum.METHODS The concentration of vancomycin was measured by UHPLC-MS/MS and CMIA methods respectively in 95 serum samples of patients receiving vancomycin treatment. A correlation analysis and an agreement analysis were conducted for the two methods used linear regression and Bland-Altman method. RESULTS Based on the results of UHPLC-MS/MS(x) and CMIA(y), the linear regression equation was determined as follows: y=1.088x+0.174. Spearmen correlation analysis indicated a significant correlation between the results by the two methods (P<0.05), and the Spearmen correlation coefficient was 0.969 1. There was statistical difference between vancomycin serum concentrations measured by these two methods. The result was higher by CMIA compared with UHPLC-MS/MS. CONCLUSION The results of the two methods can be calculated to one another by equation, which provides reference for the formulation of clinical individualized medication plan.

vancomycin  /  serum  /  therapeutic drug monitoring  /  UHPLC-MS/MS  /  CMIA
时正媛, 谢洁, 李姿颖, 江游, 续茜桥, 杨春静, 付延伟, 鄢丹. 超高效液相色谱-串联质谱法与化学发光微粒子免疫法测定万古霉素血药浓度的比较分析. 中国药学杂志, 2025 , 60 (2) : 180 -183 . DOI: 10.11669/cpj.2025.02.010
Zhengyuan SHI, Jie XIE, Ziying LI, You JIANG, Xiqiao XU, Chunjing YANG, Yanwei FU, Dan YAN. Comparison of UHPLC-MS/MS and Chemiluminescence Microparticle Immune Assay in Determination of Vancomycin Concentration in Human Serum[J]. Chinese Pharmaceutical Journal, 2025 , 60 (2) : 180 -183 . DOI: 10.11669/cpj.2025.02.010
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万古霉素(vancomycin)属于糖肽类抗生素,主要用于治疗耐甲氧西林金黄色葡萄球菌(MRSA)、耐甲氧西林表皮葡萄球菌(MRSE)和肠球菌等引起的严重感染[1-6]。由于其血药浓度治疗窗窄,血药浓度过高会导致严重的肾毒性和耳毒性,血药浓度过低则起不到较好的杀菌作用,甚至产生细菌耐药。
《中国万古霉素治疗药物监测指南(2020 更新版)》及国际血药浓度监测(therapeutic drug monitoring,TDM)指南均建议临床使用万古霉素时应进行TDM [7-13]。
TDM的发展过程中会用到不同的检测技术,而不同的方法得到的结果会有一定的差异。目前临床检测万古霉素常用的方法为免疫法,该方法操作简单,但其专属性不强[14-16]。超高效液相色谱-串联质谱(UHPLC-MS/MS)法对于人员和环境的要求较高,目前主要用于科学研究,该方法的优势在于准确性、灵敏度高,越来越受到临床治疗药物检测领域的重视[17-19]。QLIT-6610MD UHPLC-MS/MS联用仪是国内自主研发仪器,在保持准确定量的基础上精简了质量分析器结构,同时具有离子阱质量分析器的高灵敏度优势。本研究采用UHPLC-MS/MS建立了万古霉素血药浓度的检测方法,并与化学发光微粒子免疫检测法(chemiluminescent microparticle immunoassay,CMIA)进行对比,为国产质谱仪器的开发和临床应用提供方法学参考。
1 材料与方法
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1.1 仪器与试剂
QLIT-6610MD UHPLC-MS/MS(皖械注准 20222220215,安徽砺沐医疗器械有限公司);化学发光微粒子免疫分析仪(ARCHITECT i1000SR,德国Abbott Gmbh & Co. KG 公司)。AX-105 1 /10万分析天平(瑞士Mettler Toledo 公司);Vortex-Genie2 涡旋混合器(美国Scientific Industries 公司);3K15低温高速离心机(美国Sigma Aldrich公司);Milli-Q纯水机(美国Millipore公司)。
万古霉素对照品(批号:425992,北京百灵威公司);万古霉素检测试剂盒(注册证号:鲁械注准20212400421,山东英盛生物技术有限公司);甲醇、乙腈均为质谱级;甲酸、乙酸铵均为色谱级;Abbott i1000自动免疫分析仪配套微粒子化学发光法商品试剂盒(德国Abbott GmbH & Co.KG公司); 注射用盐酸万古霉素(礼来苏州制药有限公司);水为超纯水。
1.2 血清样本采集
空白血清样本由体检健康人的空白血清混匀得到。UHPLC-MS/MS法和CMIA比对用样本的采集,选取在我院住院患者 95例,男67例,女28 例,年龄8~92岁,平均75岁。原发疾病包括肺部感染、脓毒症、恶性肿瘤、心功能不全、呼吸心脏骤停、呼吸衰竭、乳糜性胸腔积液、纵隔脓肿等,临床均有体温升高和白细胞升高等感染症状。所有患者均使用注射用盐酸万古霉素治疗。用药第4天,给药前30 min采集血样,进行首次谷浓度监测; 药物输注结束后立即采集血样,检测峰浓度。本研究已通过首都医科大学附属北京世纪坛医院伦理审查,编号:sjtkyll-lx-2021(21),由患者或者家属签署知情同意书。
1.3 溶液配制
精确称取万古霉素对照品1.0 mg,溶于1 mL超纯水中,得到1 mg·mL-1储备液,用超纯水将储备液依次稀释成质量浓度为1.56、3.13、6.25、12.5、25.0、50.0、100、500 mg·L-1的梯度溶液。以上溶液均于-20 ℃避光储存。
1.4 样本前处理
血清样本或校准品或质控品在室温下解冻,取100 μL至 1.5 mL离心管中,加入350 μL提取液,涡旋振荡混匀 5 min,以12 000 r ·min-1离心 5 min 后,取 100 μL 上清液于 1.5 mL 离心管中,加入 100 μL 超纯水混匀,转移至进样小瓶中,放入液质联用仪进样分析。
1.5 色谱质谱条件
色谱条件[20]: 采用 Kinetex C18色谱柱(3 mm×50 mm,2.6 μm),以体积分数0.1%甲酸水为流动相 A,体积分数0.1%甲酸甲醇为流动相 B,采用梯度洗脱:0~1 min 10%B,1.01~3 min 99%B,3.01~5 min 10%B。流速0.5 mL·min-1,柱温40 ℃,进样体积 5 μL,所得万古霉素与万古霉素内标色谱图(图1)。
质谱条件[20]:电喷雾离子源(ESI),正离子模式,子离子扫描(QTPIS);电喷雾电压5 000 V,离子源温度300 ℃,大气压接口温度300 ℃,鞘气压强2.96×105Pa,辅助气压强1.38×105 Pa,反吹气压强5.52×103 Pa。其他质谱参数列于表1。
1.6 CMIA法
采用试剂盒对样本血清中万古霉素进行定量检测,其信号值与万古霉素含量成反比。检测范围为3.0~100.0 μg·mL-1,方法定量限为≤3 μg·mL-1。
1.7 两种方法检测比对
比对样本分别采用雅培公司 ARCHITECT i1000 SR 微粒子化学发光免疫分析仪和 UHPLC-MS/MS法两种方法检测,严格按照雅培i1000 试剂说明书和 UHPLC-MS/MS法检测步骤操作,在当日质控合格的前提下,进行样本检测。2种方法共分析了95个样品。用Bland-Altman图、Passing-Bablok回归和Spearmen检验评估2种方法的一致性和相关系数。配对样本t检验或Wilcoxon检验用于比较2种方法测量的平均值。
2 结果
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2. 1 UHPLC-MS/MS法测定人血清中万古霉素浓度的方法学评价
2.1.1 专属性
取6份不同来源的空白血清,加入一定浓度万古霉素标准溶液,所配制成的模拟生物样本经过前处理得到的色谱图,和实际临床样本得到的色谱图进行比对。结果表明,在选用条件下,血清中内源性物质不干扰被测物和内标的测定,专属性良好。
2.1.2 标准曲线制备
以不同浓度标准品为横坐标,万古霉素与内标去甲万古霉素色谱峰的峰面积比值为纵坐标,用加权最小二乘法进行线性回归运算,线性范围为1.56~50 μg·mL-1,r2≥0.99。
2.1.3 方法的准确度和精密度
日内、日间相对标准偏差(RSD)<15%,准确度范围在 95.97%~97.74%。
2.2 检测结果比对
比较2种方法,95例样本中有3例低于定量下限,92例纳入统计。UHPLC-MS/MS法测得的值(x)为(20.14±11.80) μg·mL-1,CMIA法测得的值(y)为(22.23±13.19) μg·mL-1,Wilcoxon配对检验显示CMIA法的结果高于UHPLC-MS/MS法。Passing-Bablok回归方程为y=1.088x+0.174(图2),Spearmen相关系数为0.969 1。经Bland-Altman分析,2种方法产生的偏倚为2.093,95%可信区间为(1.575 1~2.610 6) μg·mL-1 (图3)。
3 讨论
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利用先进的检测技术进行TDM,可以提高药物的疗效并减低不良反应的发生,推动了个体化精准用药的发展。本研究利用我国自主研发的质谱仪建立的万古霉素检测方法快速简便,并与CMIA法进行比较,二者相关性良好,拟合方程相关系数在0.9以上; CMIA法检测结果高于质谱方法,存在约2.093的偏倚。有文献报道比较各种免疫法与质谱法测定血浆中万古霉素浓度的相关性,发现检测结果分别存不同程度的偏倚[21]。这与本研究结果相似,可能由于免疫学方法测定过程中,万古霉素的代谢产物有一部分可能会影响到检测特异性。本研究共收集95例样本,其中3例质谱检测低于定量下限,是由于临床给药为去甲万古霉素,而免疫法不能区分,因此研究结果基于92例样本的比对分析,结果还需要进一步扩大样本量证实。
对于万古霉素的临床应用,前期已基于CMIA法的检测结果积累了大量的个体化用药经验,并取得了良好的预后效果[22-23]。在给药剂量制订过程中,要考虑到不同检测方法的结果不同,对于新建立的方法不应代替原方法指导个体化用药方案的制订。本研究利用2种方法间的线性相关性,将 CMIA法测定结果换算成 UHPLC-MS/MS方法估算值,估算结果可以供临床医生借鉴,从而为患者提供更精准的万古霉素的个体化治疗方案,同时为国产检测仪器的推广使用提供实验基础。
基金
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  • 国家重点研发计划-重大科学仪器设备开发专项资助(2020YFF01014606)
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doi: 10.11669/cpj.2025.02.010
  • 接收时间：2024-01-26
  • 首发时间：2025-11-08
  • 出版时间：2025-01-22
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  • 收稿日期：2024-01-26
基金
国家重点研发计划-重大科学仪器设备开发专项资助(2020YFF01014606)
作者信息
    1 首都医科大学附属北京世纪坛医院药学部, 北京 100038
    2 中国计量科学研究院前沿计量科学中心,国家市场监管技术创新中心(质谱), 北京 100029
    3 首都医科大学附属北京友谊医院药学部, 北京 100050

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*鄢丹,男,博士,主任药师 研究方向:中西医结合临床药学 Tel:(010)63139318
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
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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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