Omer Ozcan , Jacquelien J. Hillebrand , Carla Beertsen , Wendy P.J. den Elzen , Annemieke C. Heijboer
{"title":"血液收集管与蛋白酶抑制剂,以防止假高肾素浓度由于冷冻活化","authors":"Omer Ozcan , Jacquelien J. Hillebrand , Carla Beertsen , Wendy P.J. den Elzen , Annemieke C. Heijboer","doi":"10.1016/j.cca.2025.120486","DOIUrl":null,"url":null,"abstract":"<div><h3>Objectives</h3><div>Exposure of plasma samples to low temperatures (between −5 and + 4 °C) leads to conversion of prorenin to its open state, which is then converted to renin by plasma proteases. This process, known as cryoactivation, results in falsely elevated renin concentrations. This study examined the effects of storage temperature, protease inhibitors, and 2-hour incubation at 37 °C on renin cryoactivation.</div></div><div><h3>Methods</h3><div>Blood samples (n = 24) were collected in four types of tubes: K<sub>2</sub>EDTA, aprotinin, trypsin inhibitor, and protease inhibitor cocktail. Renin concentrations (IDS-i10) were measured at baseline and after storage at + 4 °C (one week), −20 °C (two weeks), and −80 °C (two weeks). Samples stored at −20 °C were later incubated at 37 °C for two hours and reanalyzed. Aldosterone concentrations were measured (LC-MS/MS) and aldosterone to renin ratios (ARR) were calculated.</div></div><div><h3>Results</h3><div>Renin concentrations remained stable at −80 °C but increased significantly at + 4 °C (18–50 %) and −20 °C (61–500 %). ARR changes were minimal at −80 °C (−8 pmol/mU) but largest at −20 °C (−263 pmol/mU). Renin concentrations returned to baseline after a 2-hour incubation at 37 °C only in trypsin inhibitor and protease inhibitor cocktail tubes.</div></div><div><h3>Conclusions</h3><div>Our study shows more cryoactivation of renin in plasma aliquots stored at −20 °C compared to storage at + 4 °C. Cryoactivation of prorenin in trypsin inhibitor and protease inhibitor cocktail tubes was reversed by a 2-hours incubation at 37 °C, which may be an alternative procedure for laboratories that are not able to store renin samples in −80 °C freezers.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"577 ","pages":"Article 120486"},"PeriodicalIF":2.9000,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Blood collection tubes with protease inhibitors to prevent falsely high renin concentrations due to cryoactivation\",\"authors\":\"Omer Ozcan , Jacquelien J. Hillebrand , Carla Beertsen , Wendy P.J. den Elzen , Annemieke C. Heijboer\",\"doi\":\"10.1016/j.cca.2025.120486\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objectives</h3><div>Exposure of plasma samples to low temperatures (between −5 and + 4 °C) leads to conversion of prorenin to its open state, which is then converted to renin by plasma proteases. This process, known as cryoactivation, results in falsely elevated renin concentrations. This study examined the effects of storage temperature, protease inhibitors, and 2-hour incubation at 37 °C on renin cryoactivation.</div></div><div><h3>Methods</h3><div>Blood samples (n = 24) were collected in four types of tubes: K<sub>2</sub>EDTA, aprotinin, trypsin inhibitor, and protease inhibitor cocktail. Renin concentrations (IDS-i10) were measured at baseline and after storage at + 4 °C (one week), −20 °C (two weeks), and −80 °C (two weeks). Samples stored at −20 °C were later incubated at 37 °C for two hours and reanalyzed. Aldosterone concentrations were measured (LC-MS/MS) and aldosterone to renin ratios (ARR) were calculated.</div></div><div><h3>Results</h3><div>Renin concentrations remained stable at −80 °C but increased significantly at + 4 °C (18–50 %) and −20 °C (61–500 %). ARR changes were minimal at −80 °C (−8 pmol/mU) but largest at −20 °C (−263 pmol/mU). Renin concentrations returned to baseline after a 2-hour incubation at 37 °C only in trypsin inhibitor and protease inhibitor cocktail tubes.</div></div><div><h3>Conclusions</h3><div>Our study shows more cryoactivation of renin in plasma aliquots stored at −20 °C compared to storage at + 4 °C. Cryoactivation of prorenin in trypsin inhibitor and protease inhibitor cocktail tubes was reversed by a 2-hours incubation at 37 °C, which may be an alternative procedure for laboratories that are not able to store renin samples in −80 °C freezers.</div></div>\",\"PeriodicalId\":10205,\"journal\":{\"name\":\"Clinica Chimica Acta\",\"volume\":\"577 \",\"pages\":\"Article 120486\"},\"PeriodicalIF\":2.9000,\"publicationDate\":\"2025-07-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Clinica Chimica Acta\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0009898125003651\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICAL LABORATORY TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Clinica Chimica Acta","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0009898125003651","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICAL LABORATORY TECHNOLOGY","Score":null,"Total":0}
Blood collection tubes with protease inhibitors to prevent falsely high renin concentrations due to cryoactivation
Objectives
Exposure of plasma samples to low temperatures (between −5 and + 4 °C) leads to conversion of prorenin to its open state, which is then converted to renin by plasma proteases. This process, known as cryoactivation, results in falsely elevated renin concentrations. This study examined the effects of storage temperature, protease inhibitors, and 2-hour incubation at 37 °C on renin cryoactivation.
Methods
Blood samples (n = 24) were collected in four types of tubes: K2EDTA, aprotinin, trypsin inhibitor, and protease inhibitor cocktail. Renin concentrations (IDS-i10) were measured at baseline and after storage at + 4 °C (one week), −20 °C (two weeks), and −80 °C (two weeks). Samples stored at −20 °C were later incubated at 37 °C for two hours and reanalyzed. Aldosterone concentrations were measured (LC-MS/MS) and aldosterone to renin ratios (ARR) were calculated.
Results
Renin concentrations remained stable at −80 °C but increased significantly at + 4 °C (18–50 %) and −20 °C (61–500 %). ARR changes were minimal at −80 °C (−8 pmol/mU) but largest at −20 °C (−263 pmol/mU). Renin concentrations returned to baseline after a 2-hour incubation at 37 °C only in trypsin inhibitor and protease inhibitor cocktail tubes.
Conclusions
Our study shows more cryoactivation of renin in plasma aliquots stored at −20 °C compared to storage at + 4 °C. Cryoactivation of prorenin in trypsin inhibitor and protease inhibitor cocktail tubes was reversed by a 2-hours incubation at 37 °C, which may be an alternative procedure for laboratories that are not able to store renin samples in −80 °C freezers.
期刊介绍:
The Official Journal of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)
Clinica Chimica Acta is a high-quality journal which publishes original Research Communications in the field of clinical chemistry and laboratory medicine, defined as the diagnostic application of chemistry, biochemistry, immunochemistry, biochemical aspects of hematology, toxicology, and molecular biology to the study of human disease in body fluids and cells.
The objective of the journal is to publish novel information leading to a better understanding of biological mechanisms of human diseases, their prevention, diagnosis, and patient management. Reports of an applied clinical character are also welcome. Papers concerned with normal metabolic processes or with constituents of normal cells or body fluids, such as reports of experimental or clinical studies in animals, are only considered when they are clearly and directly relevant to human disease. Evaluation of commercial products have a low priority for publication, unless they are novel or represent a technological breakthrough. Studies dealing with effects of drugs and natural products and studies dealing with the redox status in various diseases are not within the journal''s scope. Development and evaluation of novel analytical methodologies where applicable to diagnostic clinical chemistry and laboratory medicine, including point-of-care testing, and topics on laboratory management and informatics will also be considered. Studies focused on emerging diagnostic technologies and (big) data analysis procedures including digitalization, mobile Health, and artificial Intelligence applied to Laboratory Medicine are also of interest.