Application of Blood Microsampling and Its Implementation of the 3Rs in Non-clinical Studies of Drugs

doi:10.12300/j.issn.1674-5817.2021.163

Abstract

Abstract:

The welfare and ethics of laboratory animals in drug development has attracted increasing attention, and how to treat animals well has become an important topic. Blood microsampling overcomes several disadvantages of conventional large-volume blood sampling methods, such as complex operation, great harm to animals and high requirement on rodent quantity, and has great advantages in animal welfare, scientificity and cost. Therefore, its application is regarded as an important practice to implement the 3Rs. This article reviewed the latest research progress of blood microsampling, and its advantages and challenges in non-clinical studies in order to promote the wider application of this new technique in this field.

Key words: Blood microsampling, Drug, Nonclinical, 3Rs principle

CLC Number:

R-332

Jiaqi CHEN, Longbao LÜ, Feiyan ZHANG, Rui LI, Yijiang LI, Lihong LI, Xiaodi ZHANG. Application of Blood Microsampling and Its Implementation of the 3Rs in Non-clinical Studies of Drugs[J]. Laboratory Animal and Comparative Medicine, 2022, 42(4): 306-312.

Figures/Tables 1

References 53

1	MACARTHUR CLARK J. The 3Rs in research: a contemporary approach to replacement, reduction and refinement[J]. Br J Nutr, 2018, 120(s1): S1-S7. DOI:10.1017/S0007114517002227 .
2	莫菲. 比较法视野中的实验动物伦理与安全法治模式: 兼谈实验动物法与中国特色动物保护法体系建设的关系[J]. 法学评论, 2021, 39(6):148-158. DOI:10.13415/j.cnki.fxpl.2021.06.013 .
	MO F. Rule of law model of experimental animal ethics and safety in perspective of comparative law[J]. Law Rev, 2021, 39(6):148-158. DOI:10.13415/j.cnki.fxpl.2021.06.013 .
3	CHAPMAN K, CHIVERS S, GLIDDON D, et al. Overcoming the barriers to the uptake of nonclinical microsampling in regulatory safety studies[J]. Drug Discov Today, 2014, 19(5):528-532. DOI:10.1016/j.drudis.2014.01.002 .
4	JONSSON O, STEFFEN A C, SUNDQUIST V S, et al. Capillary microsampling and analysis of 4-µl blood, plasma and serum samples to determine human α-synuclein elimination rate in mice[J]. Bioanalysis, 2013, 5(4):449-462. DOI:10.4155/bio.12.337 .
5	ICH. Questions and answers to ICH s 3a: note for guidance on toxicokinetics: the assessment of systemic exposure in toxicity studies focus on microsampling [EB/OL]. (2017-11-16). .
6	BEAUDETTE P, BATEMAN K P. Discovery stage pharma-cokinetics using dried blood spots[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2004, 809(1):153-158. DOI:10.1016/j.jchromb.2004.06.018 .
7	CLARK G T, HAYNES J J, BAYLISS M A J, et al. Utilization of DBS within drug discovery: development of a serial microsampling pharmacokinetic study in mice[J]. Bioanalysis, 2010, 2(8):1477-1488. DOI:10.4155/bio.10.91 .
8	BARFIELD M, SPOONER N, LAD R, et al. Application of dried blood spots combined with HPLC-MS/MS for the quantifi-cation of acetaminophen in toxicokinetic studies[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2008, 870(1):32-37. DOI:10.1016/j.jchromb.2008.05.025 .
9	COBB Z, DE VRIES R, SPOONER N, et al. In-depth study of homogeneity in DBS using two different techniques: results from the EBF DBS-microsampling consortium[J]. Bioanalysis, 2013, 5(17):2161-2169. DOI:10.4155/bio.13.171 .
10	FREY B S, DAMON D E, BADU-TAWIAH A K. Emerging trends in paper spray mass spectrometry: Microsampling, storage, direct analysis, and applications[J]. Mass Spectrom Rev, 2020, 39(4):336-370. DOI:10.1002/mas.21601 .
11	CAPIAU S, WILK L S, AALDERS M C G, et al. A novel, nondestructive, dried blood spot-based hematocrit prediction method using noncontact diffuse reflectance spectroscopy[J]. Anal Chem, 2016, 88(12):6538-6546. DOI:10.1021/acs.analchem.6b01321 .
12	DAMON D E, YIN M Z, ALLEN D M, et al. Dried blood spheroids for dry-state room temperature stabilization of microliter blood samples[J]. Anal Chem, 2018, 90(15):9353-9358. DOI:10.1021/acs.analchem.8b01962 .
13	YOUHNOVSKI N, BERGERON A, FURTADO M, et al. Pre-cut dried blood spot (PCDBS): an alternative to dried blood spot (DBS) technique to overcome hematocrit impact[J]. Rapid Commun Mass Spectrom, 2011, 25(19):2951-2958. DOI:10.1002/rcm.5182 .
14	NAKAHARA T, OTANI N, UENO T, et al. Development of a hematocrit-insensitive device to collect accurate volumes of dried blood spots without specialized skills for measuring clozapine and its metabolites as model analytes[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2018, 1087-1088:70-79. DOI:10.1016/j.jchromb.2018.04.019 .
15	LENK G, SANDKVIST S, POHANKA A, et al. A disposable sampling device to collect volume-measured DBS directly from a fingerprick onto DBS paper[J]. Bioanalysis, 2015, 7(16):2085-2094. DOI:10.4155/bio.15.134 .
16	LI W K, DOHERTY J, FAVARA S, et al. Evaluation of plasma microsampling for dried plasma spots (DPS) in quantitative LC-MS/MS bioanalysis using ritonavir as a model compound[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2015, 991:46-52. DOI:10.1016/j.jchromb.2015.03.026 .
17	LI W K, DUGYALA R, DEVINE P J, et al. Application of tail vein serial microsampling for plasma or dried plasma spots in toxicokinetic assessment in rats using acetaminophen as the model compound[J]. Biomed Chromatogr, 2020, 34(10): e4917. DOI:10.1002/bmc.4917 .
18	LI Y Y, HENION J, ABBOTT R, et al. The use of a membrane filtration device to form dried plasma spots for the quantitative determination of guanfacine in whole blood[J]. Rapid Commun Mass Spectrom, 2012, 26(10):1208-1212. DOI:10.1002/rcm.6212 .
19	STURM R, HENION J, ABBOTT R, et al. Novel membrane devices and their potential utility in blood sample collection prior to analysis of dried plasma spots[J]. Bioanalysis, 2015, 7(16):1987-2002. DOI:10.4155/bio.15.98 .
20	RYONA I, HENION J. A book-type dried plasma spot card for automated flow-through elution coupled with online SPE-LC-MS/MS bioanalysis of opioids and stimulants in blood[J]. Anal Chem, 2016, 88(22):11229-11237. DOI:10.1021/acs.analchem.6b03691 .
21	HAUSER J, LENK G, ULLAH S, et al. An autonomous microfluidic device for generating volume-defined dried plasma spots[J]. Anal Chem, 2019, 91(11):7125-7130. DOI:10.1021/acs.analchem.9b00204 .
22	DENNIFF P, SPOONER N. Volumetric absorptive micro-sampling: a dried sample collection technique for quanti-tative bioanalysis[J]. Anal Chem, 2014, 86(16):8489-8495. DOI:10.1021/ac5022562 .
23	SPOONER N, DENNIFF P, MICHIELSEN L, et al. A device for dried blood microsampling in quantitative bioanalysis: overcoming the issues associated blood hematocrit[J]. Bioanalysis, 2015, 7(6):653-659. DOI:10.4155/bio.14.310 .
24	THIRY J, EVRARD B, NYS G, et al. Sampling only ten microliters of whole blood for the quantification of poorly soluble drugs: Itraconazole as case study[J]. J Chromatogr A, 2017, 1479:161-168. DOI:10.1016/j.chroma.2016.12.009 .
25	NYS G, GALLEZ A, KOK M G M, et al. Whole blood microsampling for the quantitation of estetrol without derivatization by liquid chromatography-tandem mass spectrometry[J]. J Pharm Biomed Anal, 2017, 140:258-265. DOI:10.1016/j.jpba.2017.02.060 .
26	KITA K, NORITAKE K, MANO Y. Application of a volumetric absorptive microsampling device to a pharmacokinetic study of tacrolimus in rats: comparison with wet blood and plasma[J]. Eur J Drug Metab Pharmacokinet, 2019, 44(1):91-102. DOI:10.1007/s13318-018-0493-7 .
27	DENNIFF P, PARRY S, DOPSON W, et al. Quantitative bioanalysis of paracetamol in rats using volumetric absorptive microsampling (VAMS)[J]. J Pharm Biomed Anal, 2015, 108:61-69. DOI:10.1016/j.jpba.2015.01.052 .
28	THIRY J, KOK M G M, COLLARD L, et al. Bioavailability enhancement of itraconazole-based solid dispersions produced by hot melt extrusion in the framework of the Three Rs rule[J]. Eur J Pharm Sci, 2017, 99:1-8. DOI:10.1016/j.ejps.2016.12.001 .
29	SPREADBOROUGH M J, DAY J, JACKSON-ADDIE K, et al. Bioanalytical implementation of plasma capillary micro-sampling: small hurdles, large gains[J]. Bioanalysis, 2013, 5(12):1485-1489. DOI:10.4155/bio.13.120 .
30	VERHAEGHE T, DILLEN L, STIELTJES H, et al. The application of capillary microsampling in GLP toxicology studies[J]. Bioanalysis, 2017, 9(7):531-540. DOI:10.4155/bio-2016-0297 .
31	VERHAEGHE T, DILLEN L, STIELTJES H, et al. Comparison of toxicokinetic parameters of a drug and two metabolites following traditional and capillary microsampling in rat[J]. Bioanalysis, 2019, 11(13):1233-1242. DOI:10.4155/bio-2019-0085 .
32	JONSSON O, PALMA VILLAR R, NILSSON L B, et al. Capillary microsampling of 25 µl blood for the determination of toxicokinetic parameters in regulatory studies in animals[J]. Bioanalysis, 2012, 4(6):661-674. DOI:10.4155/bio.12.25 .
33	DILLEN L, LOOMANS T, VAN DE PERRE G, et al. Blood microsampling using capillaries for drug-exposure determi-nation in early preclinical studies: a beneficial strategy to reduce blood sample volumes[J]. Bioanalysis, 2014, 6(3):293-306. DOI:10.4155/bio.13.286 .
34	WANG B, WANG L N, BATOG A, et al. Investigation on the effect of capillary microsampling on hematologic and toxicokinetic evaluation in regulatory safety studies in mice[J]. AAPS J, 2020, 22(2):55. DOI:10.1208/s12248-020-00438-z .
35	RAJE A A, MAHAJAN V, PATHADE V V, et al. Capillary microsampling in mice: effective way to move from sparse sampling to serial sampling in pharmacokinetics profiling[J]. Xenobiotica, 2020, 50(6):663-669. DOI:10.1080/00498254. 2019.1683259 .
36	ZHU L, WANG Y, JOYCE A, et al. Fit-for-purpose validation of a ligand binding assay for toxicokinetic study using mouse serial sampling[J]. Pharm Res, 2019, 36(12):169. DOI:10.1007/s11095-019-2699-z .
37	BOWEN C L, LICEA-PEREZ H, KARLINSEY M Z, et al. A novel approach to capillary plasma microsampling for quantitative bioanalysis[J]. Bioanalysis, 2013, 5(9):1131-1135. DOI:10.4155/bio.13.58 .
38	POWLES-GLOVER N, KIRK S, WILKINSON C, et al. Assessment of toxicological effects of blood microsampling in the vehicle dosed adult rat[J]. Regul Toxicol Pharmacol, 2014, 68(3):325-331. DOI:10.1016/j.yrtph.2014.01.001 .
39	BURNETT J E. Dried blood spot sampling: practical consider-ations and recommendation for use with preclinical studies[J]. Bioanalysis, 2011, 3(10):1099-1107. DOI:10.4155/bio.11.68 .
40	DIEHL K H, HULL R, MORTON D, et al. A good practice guide to the administration of substances and removal of blood, including routes and volumes[J]. J Appl Toxicol, 2001, 21(1):15-23. DOI:10.1002/jat.727 .
41	SPARROW S S, ROBINSON S, BOLAM S, et al. Opportunities to minimise animal use in pharmaceutical regulatory general toxicology: a cross-company review[J]. Regul Toxicol Pharmacol, 2011, 61(2):222-229. DOI:10.1016/j.yrtph.2011.08.001 .
42	HOPPER L D. Automated microsampling technologies and enhancements in the 3Rs[J]. ILAR J, 2016, 57(2):166-177. DOI:10.1093/ilar/ilw020 .
43	POWLES-GLOVER N, KIRK S, JARDINE L, et al. Assessment of haematological and clinical pathology effects of blood microsampling in suckling and weaned juvenile rats[J]. Regul Toxicol Pharmacol, 2014, 69(3):425-433. DOI:10.1016/j.yrtph. 2014.05.006 .
44	NIU X Y, BEEKHUIJZEN M, SCHOONEN W, et al. Effects of capillary microsampling on toxicological endpoints in juvenile rats[J]. Toxicol Sci, 2016, 154(1):69-77. DOI:10.1093/toxsci/kfw146 .
45	MITCHARD T, KIRK S, GRANT C, et al. Investigation into the effect of microsampling on mouse fetuses and pregnant mice in the embryofetal development study design[J]. Reprod Toxicol, 2017, 67:140-145. DOI:10.1016/j.reprotox.2016.12.006 .
46	PENG S X, ROCKAFELLOW B A, SKEDZIELEWSKI T M, et al. Improved pharmacokinetic and bioavailability support of drug discovery using serial blood sampling in mice[J]. J Pharm Sci, 2009, 98(5):1877-1884. DOI:10.1002/jps.21533 .
47	JOYCE A P, WANG M M, LAWRENCE-HENDERSON R, et al. One mouse, one pharmacokinetic profile: quantitative whole blood serial sampling for biotherapeutics[J]. Pharm Res, 2014, 31(7):1823-1833. DOI:10.1007/s11095-013-1286-y .
48	KORFMACHER W, FITZGERALD M, LUO Y Y, et al. Capillary microsampling of whole blood for mouse PK studies: an easy route to serial blood sampling[J]. Bioanalysis, 2015, 7(4):449-461. DOI:10.4155/bio.14.275 .
49	PATEL N J, WICKREMSINHE E, HUI Y H, et al. Evaluation and optimization of blood micro-sampling methods: serial sampling in a cross-over design from an individual mouse[J]. J Pharm Pharm Sci, 2016, 19(4):496-510. DOI:10.18433/J3NK60 .
50	WAN K X, REIMER M T, METCHKAROVA M P, et al. Toxicokinetic evaluation of atrasentan in mice utilizing serial microsampling: validation and sample analysis in GLP study[J]. Bioanalysis, 2012, 4(11):1351-1361. DOI:10.4155/bio.12.91 .
51	SPOONER N, ANDERSON K D, SIPLE J, et al. Microsampling: considerations for its use in pharmaceutical drug discovery and development[J]. Bioanalysis, 2019, 11(10):1015-1038. DOI:10.4155/bio-2019-0041 .
52	CARON A, LELONG C, BARTELS T, et al. Clinical and anatomic pathology effects of serial blood sampling in rat toxicology studies, using conventional or microsampling methods[J]. Regul Toxicol Pharmacol, 2015, 72(3):429-439. DOI:10.1016/j.yrtph.2015.05.022 .
53	HATTORI N, TAKUMI A, SAITO K, et al. Effects of serial cervical or tail blood sampling on toxicity and toxicokinetic evaluation in rats[J]. J Toxicol Sci, 2020, 45(10):599-609. DOI:10.2131/jts.45.599 .

微量采血法 Blood micro- sampling method	基质类型 Matrix type	基质形式 Matrix form	优点 Advantages	缺点 Disadvantages
DBS	全血	干燥	无需离心，储存和运输方面要求低	全血数据与血浆数据转化复杂，HCT效应，需要特殊的提取技术
DPS	血浆	干燥	储存和运输方面要求低	需要特殊的提取技术，可能需要离心
VAMS	全血、血浆、血清	干燥	储存和运输方面要求低，样品体积准确	全血数据与血浆数据转化复杂，血浆、血清制备程序复杂
CMS	全血、血浆、血清	液体	样品体积准确	储存和运输要求高（－20 ℃），血浆、血清制备操作不便

微量采血法 Blood micro- sampling method	基质类型 Matrix type	基质形式 Matrix form	优点 Advantages	缺点 Disadvantages
DBS	全血	干燥	无需离心，储存和运输方面要求低	全血数据与血浆数据转化复杂，HCT效应，需要特殊的提取技术
DPS	血浆	干燥	储存和运输方面要求低	需要特殊的提取技术，可能需要离心
VAMS	全血、血浆、血清	干燥	储存和运输方面要求低，样品体积准确	全血数据与血浆数据转化复杂，血浆、血清制备程序复杂
CMS	全血、血浆、血清	液体	样品体积准确	储存和运输要求高（－20 ℃），血浆、血清制备操作不便

[1]	Committee of Experts on Medical Animal Experiments, Chinese Research Hospital Association. Guidelines for the Selection of Animal Models and Preclinical Drug Trials for Spontaneous Intracerebral Hemorrhage (2024 Edition) [J]. Laboratory Animal and Comparative Medicine, 2024, 44(1): 3-30.
[2]	Shuwu XIE, Ruling SHEN, Jinxing LIN, Chun FAN. Progress in Establishment and Application of Laboratory Animal Models Related to Development of Male Infertility Drugs [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 504-511.
[3]	Liping FENG, Qi ZHU, Jinxing LIN. Current Status and Reflection on the Study of Welfare for Laboratory Fish [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 524-530.
[4]	Jiahui YU, Qian GONG, Lenan ZHUANG. Animal Models of Pulmonary Arterial Hypertension and Their Application in Drug Research [J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 381-397.
[5]	Miaomiao GONG, Ligui ZHOU, Jumei ZHAO, Changhong SHI. Application of Gastric Cancer Organoids in Precision Medicine Research [J]. Laboratory Animal and Comparative Medicine, 2022, 42(3): 248-254.
[6]	ZHOU Fei, WANG Hao-an. Review of Spontaneous and Drug-related Islet Fibrosis in Rats [J]. Laboratory Animal and Comparative Medicine, 2019, 39(3): 249-252.
[7]	GE Xiao-mei, ZHANG Yi-xin, XIE Fu-bo, LIU Ji-bin, YANG Lei, QU Ying-ying, GU Ying, Li Xue-ting, YANG Wei-min, LIU Xi-peng, ZHOU He, QIANG Fu-lin. Establishment and Characterization of Patient Derived Gastric Cancer Cell Lines [J]. Laboratory Animal and Comparative Medicine, 2017, 37(4): 257-265.
[8]	WEN Fu-li, ZHANG Jia, ZHENG He-ping, MA Lei, ZHANG Shi-lan, WANG Shou-kun. Effects of Different Drugs and Disinfectants on Sporulation of Coccidian Oocysts in Rabbit [J]. Laboratory Animal and Comparative Medicine, 2016, 36(4): 301-306.
[9]	GU Yun-hao, CAO Chen-jie, HU Bi-yuan, WANG Jun, HAN Dong-dong, XU Ai-hua. Establishment of S180 Tumor Multidrug Resistance Mouse Model by Increasing PFC and Observation on Stability [J]. Laboratory Animal and Comparative Medicine, 2015, 35(5): 367-373.
[10]	PENG Xiu-hua, CHEN Li-xiang, REN Xiao-nan, SHI Bi-sheng, XU Chun-hua, ZHOU Wen-jiang, ZHOU Xiao-hui. Establishment and Preliminary Evaluation of Hepatitis B Virus Transfection Mouse Model by Using Hydrodynamic Injection [J]. Laboratory Animal and Comparative Medicine, 2015, 35(1): 1-5.
[11]	XU Chun-hua, YANG Lei, TANG Xu-zhen, HU Gang, GENG Qin, OU YANG Ke-dong, XIE Fu-bo, WANG Ke, QIN Xiao-ran, LIU Ji-bin, YANG Wei-min, TAO Wei-kang, ZHANG Yi-xin, ZHOU He. Initially Establishment and Characterization of Patient Derived Gastric Cancer Xenograft Models [J]. Laboratory Animal and Comparative Medicine, 2014, 34(4): 259-265.
[12]	LIN Dan, ZHENG Jin-hua, ZHUANG Hua, SHI Ji-jing, LIN Yue-lin, WANG Zhu-gang, KUANG Ying. Generation of Trp53 Gene Knockout Mouse and Its Application in Carcinogenicity Assessment of Pharmaceuticals [J]. Laboratory Animal and Comparative Medicine, 2013, 33(6): 418-424.
[13]	OU-YANG Ke-dong, LIU Ji-bin, WANG Ke, HU Gang, GU Ying, XIE Fu-bo, ZHAO Qiang, ZHANG Ya-hua, YANG Wei-min, WENG Dan-yi, ZHANG Yi-xin, QIN Xiao-ran. Preliminary Research in Establishment, Characterization, and Application of Human Primary Esophageal Cancer Xenograft Models [J]. Laboratory Animal and Comparative Medicine, 2013, 33(2): 90-98.
[14]	ZHANG Wei1,ZHANG Zhong-guang2,XU Chong-hui3,OUYANG Shao-lun3,ZHAO Yong1,Hu Hai-xun1,LI Fu-rong1,LIN Yi-li1,ZHONG Nv-qi1,ZHUANG Shao-hui1,ZOU Yi-hai2. Acute Toxicity of Melamine and Detoxification of San Jin Tang(三金汤)in Mice [J]. , 2010, 30(3): 188-192.
[15]	ZHOU Wei-Mrn1，SHENG Hong-Qiang2, CHEN Wen-Bin3, CAI Yue-Qin1，CHEN Min-Li1. Rodent Models of Colorectal Tumor and Evaluation System of Drugs [J]. , 2010, 30(1): 60-67.