中国临床药理学杂志

LC-MS/MS检测干血点样品中泊沙康唑浓度的方法建立及其在治疗药物监测中的应用

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陈焰 ¹^,²^,³ , 丁玺 ⁴ , 吴越 ⁵ , 范亚新 ¹^,²^,³ , 张菁 ¹^,²^,³ , 郭蓓宁 ¹^,²^,³ , 李熠 ¹^,²^,³ , 李鑫 ¹^,²^,³ , 李婉贞 ¹^,²^,³ , 刘笑芬 ¹^,²^,³

中国临床药理学杂志 | 研究方法 2026,42(11): 1573-1580

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中国临床药理学杂志 |研究方法 2026 , 42 (11) : 1573 -1580

LC-MS/MS检测干血点样品中泊沙康唑浓度的方法建立及其在治疗药物监测中的应用

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陈焰¹^,²^,³, 丁玺⁴, 吴越⁵, 范亚新¹^,²^,³, 张菁¹^,²^,³, 郭蓓宁¹^,²^,³, 李熠¹^,²^,³, 李鑫¹^,²^,³, 李婉贞¹^,²^,³, 刘笑芬¹^,²^,³

作者信息

^1.复旦大学附属华山医院抗生素研究所，上海 200040

^2.国家卫健委抗生素临床药理重点实验室，上海 200040

^3.耐药菌感染病原诊断与创新药物研发院企联合实验室，上海 200040

^4.东台市人民医院，江苏东台 224200

^5.深圳市儿童医院，广东深圳 518000

通讯作者:

刘笑芬，副研究员，硕士生导师 Tel: （021）52888189 E-mail: liuxiaofen227@163.com

作者简介:

陈焰（1999），男，初级药师，主要从事抗菌药物治疗药物监测相关研究及工作

Development of an LC-MS/MS method for determining posaconazole concentration in dried blood spot samples and its application in therapeutic drug monitoring

Yan CHEN¹^,²^,³, Xi DING⁴, Yue WU⁵, Ya-xin FAN¹^,²^,³, Jing ZHANG¹^,²^,³, Bei-ning GUO¹^,²^,³, Yi LI¹^,²^,³, Xin LI¹^,²^,³, Wan-zhen LI¹^,²^,³, Xiao-fen LIU¹^,²^,³

Affiliations

^1.Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai 200040, China

^2.Key Laboratory of Antibiotic Clinical Pharmacology of the National Health Commission, Shanghai 200040, China

^3.Joint Laboratory of Hospital & Enterprise for Pathogen Diagnosis of Drug-resistant Bacterial lnfections and lnnovative Drug R&D, Shanghai 200040, China

^4.Dongtai People’s Hospital, Dongtai 224200, Jiangsu Province, China

^5.Shenzhen Children’s Hospital, Shenzhen 518000, Guangdong Province, China

出版时间: 2026-06-17 doi: 10.13699/j.cnki.1001-6821.2026.11.012

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摘要

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目的

开发并验证一种测定干血点（DBS）样品中泊沙康唑药物浓度的液相色谱串联质谱法（LC-MS/MS），为干血点法的应用和泊沙康唑治疗药物监测（TDM）的临床实践提供一种可行的参考。

方法

干血点样本通过将含泊沙康唑的全血样本滴加到Whatman 903^®卡片上干燥获得。样本处理过程为裁取3 mm直径的圆孔得到分析样本，通过添加0.2%甲酸-50%乙腈水溶液（含内标0.50 μg·mL^-1）提取，经超声、离心，取上清直接进样检测。用LC-MS/MS测定，色谱柱选择Phonomenex Kinetex XB-C₁₈柱，梯度洗脱方式分离，质谱检测模式为正离子多反应监测，待测物泊沙康唑和内标D₅-泊沙康唑监测离子对分别为m/z 701.3 →614.4和m/z 705.3→618.4。方法学验证包括选择性、特异性、线性、残留、准确度、精密度、提取回收率、基质效应、稳定性，以及针对基于干血点分析的特殊参数验证。此外，将所开发的DBS方法应用于泊沙康唑临床TDM样本的检测，并与血浆方法进行比较，对两种方法的一致性进行评价。

结果

DBS的LC-MS/MS测定方法具有良好的选择性和线性（0.10~10.00 μg·ml^-1），无残留和交叉干扰的影响，样本处理过程分析物的回收率在70.67%~75.66%范围内，批内准确度偏差、精密度分别在-12.18%~-0.86%、3.05%~8.75%范围内，批间准确度偏差、精密度分别在-4.21%~3.49%、8.57%~12.21%范围内。6个不同来源的空白全血测试的基质效应，准确度在±15%以内、精密度均<15%，稳定性结果显示泊沙康唑在干血点样本中室温（20~25 ℃）、37 ℃、4 ℃、-20 ℃、-70 ℃不同温度条件下分别至少稳定3、3、14、30、90 d。78个临床验证样本的Bland-Altman、Passing-Bablok分析结果显示，泊沙康唑在DBS和血浆中测定结果具有较好的相关性，但具有显著偏差（P<0.001），DBS测定结果较低于血浆测定结果，平均偏差为0.33 μg·mL^-1，在低Hct中更显著。

结论

本研究开发并验证了一种简单、便捷的测定DBS中泊沙康唑浓度的LC-MS/MS方法，检测所需样本量小（全血最少10 μL）、分析时间短（2.00 min）、操作便捷、适用性高，为泊沙康唑TDM的临床实践提供一种便捷可行的参考检测方法。

关键词

泊沙康唑 / 干血点法 / 治疗药物监测 / 液相色谱串联质谱法

Abstract

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Objective

To develop and validate a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determining the concentration of posaconazole in dried blood spot (DBS) samples, providing a feasible reference for the application of the dried blood spot method and the clinical practice of posaconazole therapeutic drug monitoring (TDM).

Methods

DBS samples were obtained by dropping whole blood samples containing posaconazole onto Whatman 903^® cards and drying them. The sample processing procedure involved cutting out a round hole with a diameter of 3 mm to obtain the analytical sample. The sample was extracted by adding 0.2% formic acid-50% acetonitrile (containing an internal standard of 0.50 μg·ml^-1), followed by sonication and centrifugation. The supernatant was directly injected for detection. LC-MS/MS was used for determination. The Phonomenex Kinetex XB-C₁₈ column was selected as the chromatographic column, and gradient elution was used for separation. The mass spectrometry detection mode was positive ion multiple reaction monitoring. The monitored ion pairs for the analyte posaconazole and the internal standard D₅-posaconazole were 701.3→614.4 and 705.3→618.4, respectively. Full method validation was conducted in accordance with ICH Guideline M10 (Bioanalytical Method Validation and Study Sample Analysis), covering selectivity, specificity, linearity, carryover, accuracy, precision, extraction recovery, matrix effect, analyte stability, together with DBS-specific validation parameters including hematocrit (Hct) impact, spotting volume effect and spot homogeneity. In addition, the developed DBS method was applied to the detection of clinical TDM samples of posaconazole and compared with the plasma method, and the consistency of the two methods was evaluated.

Results

The LC-MS/MS method for the determination of posaconazole in DBS had good selectivity and linearity (0.10-10.00 μg·ml^-1), without the influence of cross-talk and carry-over. The recovery rate of the analyte during the sample processing was in the range of 70.67%-75.66%. The intra-batch accuracy and precision were in the ranges of -12.18%--0.86% and 3.05%-8.75%, respectively, and the inter-batch accuracy and precision were in the ranges of -4.21%-3.49% and 8.57%-12.21%, respectively. For the matrix effect tested with six blank whole blood samples from different sources, the accuracy was within ±15% and the precision was all <15%. The stability results showed that posaconazole was stable for at least 3, 3, 14, 30, and 90 days at room temperature (20-25 ℃), 37 ℃, 4 ℃ -20 ℃, and -70 ℃ in DBS samples, respectively. The results of Bland-Altman and Passing-Bablok analyses of 78 clinically verified samples showed that there was a good correlation between the determination results of posaconazole in dried blood spots (DBS) and plasma. However, there was a significant deviation (P<0.001). The determination results in DBS were lower than those in plasma (average difference=0.33 μg·ml^-1), and this was more significant in cases with low hematocrit (Hct).

Conclusion

A straightforward and practical LC-MS/MS method was successfully developed and fully validated for posaconazole quantitation in DBS samples. This method requires a minimal blood volume (minimum 10 μL whole blood), features a short analytical runtime (2.00 min per injection) and facilitates streamlined laboratory operation with robust practicability, which can serve as a convenient alternative analytical strategy for routine clinical TDM of posaconazole.

Key words

posaconazole / dried blood spot method / therapeutic drug monitoring / liquid chromatography-tandem mass spectrometry

引用本文

陈焰, 丁玺, 吴越, 范亚新, 张菁, 郭蓓宁, 李熠, 李鑫, 李婉贞, 刘笑芬. LC-MS/MS检测干血点样品中泊沙康唑浓度的方法建立及其在治疗药物监测中的应用. 中国临床药理学杂志, 2026 , 42 (11) : 1573 -1580 . DOI: 10.13699/j.cnki.1001-6821.2026.11.012

Yan CHEN, Xi DING, Yue WU, Ya-xin FAN, Jing ZHANG, Bei-ning GUO, Yi LI, Xin LI, Wan-zhen LI, Xiao-fen LIU. Development of an LC-MS/MS method for determining posaconazole concentration in dried blood spot samples and its application in therapeutic drug monitoring[J]. Chinese Journal of Clinical Pharmacology, 2026 , 42 (11) : 1573 -1580 . DOI: 10.13699/j.cnki.1001-6821.2026.11.012

基金

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上海申康医院发展中心临床三年行动计划(SHDC2022CRW004)
复旦大学附属华山医院临床研究基金资助(2023-YN009)

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2026年第42卷第11期

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文章信息

doi: 10.13699/j.cnki.1001-6821.2026.11.012

接收时间：2026-02-11
首发时间：2026-08-06
出版时间：2026-06-17

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收稿日期：2026-02-11

基金

上海申康医院发展中心临床三年行动计划(SHDC2022CRW004)

复旦大学附属华山医院临床研究基金资助(2023-YN009)

作者信息

^1.复旦大学附属华山医院抗生素研究所，上海 200040

^2.国家卫健委抗生素临床药理重点实验室，上海 200040

^3.耐药菌感染病原诊断与创新药物研发院企联合实验室，上海 200040

^4.东台市人民医院，江苏东台 224200

^5.深圳市儿童医院，广东深圳 518000

通讯作者:

刘笑芬，副研究员，硕士生导师 Tel: （021）52888189 E-mail: liuxiaofen227@163.com

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https://castjournals.cast.org.cn/joweb/zglcylxzz/CN/10.13699/j.cnki.1001-6821.2026.11.012

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2种不同金属材料的力学参数

科 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 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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