{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T13:40:28Z","timestamp":1787319628812,"version":"build-2736575974"},"reference-count":59,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2019,6,29]],"date-time":"2019-06-29T00:00:00Z","timestamp":1561766400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>A method based on gas chromatography\u2013mass spectrometry (GC\u2013MS) is described for the determination of bisoprolol and atenolol in human bone. After the addition of lobivolol as internal standard, pulverized samples were incubated in acetonitrile for 1 h under ultrasounds. After adjusting the pH of the samples to 6, they were centrifuged, and the supernatants were subjected to solid phase extraction. Elution was achieved by using 3 mL of 2% ammonium hydroxide in 80:20 dichloromethane:isopropanol solution. Eluted samples were evaporated and derivatized. Chromatography was performed on a fused silica capillary column and analytes were determined in the selected-ion-monitoring (SIM) mode. The assay was validated in the range 0.1\u20130.3 ng\/mg (depending on the drug) to 150 ng\/mg, the mean absolute recoveries were 60% for bisoprolol and 106% for atenolol, the matrix effect was 69% for bisoprolol and 70% for atenolol and process efficiency was 41% for bisoprolol and 80% for atenolol. The intra- and inter-assay accuracy values were always better than 12%. The validated method was then applied to bone samples from two real forensic cases in which toxicological analysis in blood were positive for atenolol in the first case (0.65 \u00b5g\/mL) and bisoprolol in the second case (0.06 \u00b5g\/mL). Atenolol was found in bone samples from the corresponding case at the approximate concentration of 148 ng\/mg and bisoprolol was found at 8 ng\/mg.<\/jats:p>","DOI":"10.3390\/molecules24132400","type":"journal-article","created":{"date-parts":[[2019,7,1]],"date-time":"2019-07-01T03:23:59Z","timestamp":1561951439000},"page":"2400","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Development and Validation of a Method for the Analysis of Bisoprolol and Atenolol in Human Bone"],"prefix":"10.3390","volume":"24","author":[{"given":"Lucia","family":"Fernandez-Lopez","sequence":"first","affiliation":[{"name":"Legal and Forensic Medicine, School of Medicine, University of Murcia,30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manuela","family":"Pellegrini","sequence":"additional","affiliation":[{"name":"National Centre on Drug Addiction and Doping, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Concetta","family":"Rotolo","sequence":"additional","affiliation":[{"name":"National Centre on Drug Addiction and Doping, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7760-7415","authenticated-orcid":false,"given":"Aurelio","family":"Luna","sequence":"additional","affiliation":[{"name":"Legal and Forensic Medicine, School of Medicine, University of Murcia,30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Falcon","sequence":"additional","affiliation":[{"name":"Legal and Forensic Medicine, School of Medicine, University of Murcia,30100 Murcia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9787-872X","authenticated-orcid":false,"given":"Rosanna","family":"Mancini","sequence":"additional","affiliation":[{"name":"National Centre on Drug Addiction and Doping, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,6,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1097\/PAF.0b013e31818738c9","article-title":"Detection of drugs of forensic importance in post-mortem bone","volume":"30","author":"McGrath","year":"2009","journal-title":"Am. J. Forensic Med. Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1111\/j.1556-4029.2011.01994.x","article-title":"Detection of amitriptyline and citalopram and metabolites in porcine bones following extended outdoor decomposition","volume":"57","author":"Desrosiers","year":"2012","journal-title":"J. Forensic Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1039\/b609130j","article-title":"Challenges in forensic toxicology of skeletonised human remains","volume":"131","author":"Watterson","year":"2006","journal-title":"Analyst"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.forsciint.2006.03.029","article-title":"Morphine and 6-acetylmorphine concentrations in blood, brain, spinal cord, bone marrow and bone after lethal acute or chronic diacetylmorphine administration to mice","volume":"166","author":"Guillot","year":"2007","journal-title":"Forensic Sci. Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1016\/S0379-0738(01)00529-1","article-title":"Determination of opiates in postmortem bone and bone marrow","volume":"123","author":"Raikos","year":"2009","journal-title":"Forensic Sci. Int."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1093\/jat\/bks045","article-title":"Comparison of relative distribution of ketamine and norketamine in decomposed skeletal tissues following single and repeated exposures","volume":"36","author":"Watterson","year":"2012","journal-title":"J. Anal. Toxicol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1093\/jat\/bkt078","article-title":"Discrimination between patterns of drug exposure by toxicological analysis of decomposed skeletal tissues, Part II: Amitriptyline and Citalopram","volume":"37","author":"Watterson","year":"2013","journal-title":"J. Anal. Toxicol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1093\/jat\/34.8.510","article-title":"Relative distribution of drugs in decomposed skeletal tissues","volume":"34","author":"Watterson","year":"2010","journal-title":"J. Anal. Toxicol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1093\/anatox\/35.7.452","article-title":"Relative distribution of ketamine and norketamine in skeletal tissues following various periods of decomposition","volume":"35","author":"Watterson","year":"2011","journal-title":"J. Anal. Toxicol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.forsciint.2010.08.021","article-title":"Microwave-assisted extraction in the study of the effect of dose-death interval on meperidine detection in skeletal tissues","volume":"207","author":"Watterson","year":"2011","journal-title":"Forensic Sci. Int."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.forsciint.2009.03.008","article-title":"Microwave-assisted extraction in toxicological screening of skeletal tissues","volume":"188","author":"Desrosiers","year":"2009","journal-title":"Forensic Sci. Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1002\/dta.2327","article-title":"Semi-quantitative analysis of tramadol, dextromethorphan, and metabolites in decomposed skeletal tissues by ultra performance liquid chromatography quadrupole time of flight mass spectrometry","volume":"10","author":"Cornthwaite","year":"2017","journal-title":"Drug Test. Anal."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1520\/JFS15069J","article-title":"Detection of benzodiazepines in different tissues, including bone, using a quantitative ELISA assay","volume":"46","author":"Gorczynski","year":"2001","journal-title":"J. Forensic Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1474","DOI":"10.1111\/j.1556-4029.2008.00866.x","article-title":"Utility of immunoassay in drug screening in skeletal tissues: Sampling considerations in detection of ketamine exposure in femoral bone and bone marrow following acute administration using ELISA","volume":"53","author":"VandenBoer","year":"2008","journal-title":"J. Forensic Sci."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1093\/jat\/32.8.631","article-title":"Effects of tissue type and the dose-death interval on the detection of acute ketamine exposure in bone marrow with solid-phase extraction and ELISA with liquid chromatography-tandem mass spectrometry confirmation","volume":"32","author":"Watterson","year":"2008","journal-title":"J. Anal. Toxicol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1111\/j.1556-4029.2009.00998.x","article-title":"Detection of acute diazepam exposure in bone and marrow: Influence of tissue type and the dose-death interval on sensitivity of detection by ELISA with liquid chromatography tandem mass spectrometry confirmation","volume":"54","author":"Watterson","year":"2009","journal-title":"J. Forensic Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.forsciint.2009.10.008","article-title":"Detection of acute fentanyl exposure in fresh and decomposed skeletal tissues. Part II: The effect of dose-death interval","volume":"94","author":"Lafreniere","year":"2010","journal-title":"Forensic Sci. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"548","DOI":"10.1093\/jat\/bku053","article-title":"The influence of body position and microclimate on ketamine and metabolite distribution in decomposed skeletal remains","volume":"38","author":"Cornthwaite","year":"2014","journal-title":"J. Anal. Toxicol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/jat\/bkw083","article-title":"Analysis of dextromethorphan and dextrorphan in skeletal tissues following decomposition in different microclimates condition","volume":"40","author":"Unger","year":"2016","journal-title":"J. Anal. Toxicol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.forsciint.2008.12.019","article-title":"Detection of acute fentanyl exposure in fresh and decomposed skeletal tissues","volume":"185","author":"Lafreniere","year":"2009","journal-title":"Forensic Sci. Int."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.forsciint.2004.12.017","article-title":"Determination of morphine in postmortem rabbit bone marrow and comparison with blood morphine concentrations","volume":"156","author":"Ceigniz","year":"2006","journal-title":"Forensic Sci. Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1520\/JFS2004067","article-title":"Postmortem tissue distribution of olanzapine and citalopram in a drug intoxication","volume":"50","author":"Horak","year":"2005","journal-title":"J. Forensic Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1097\/00007691-200002000-00017","article-title":"Postmortem drug analyses in bone and bone marrow","volume":"22","author":"McIntyre","year":"2000","journal-title":"Ther. Drug Monit."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.forsciint.2018.07.004","article-title":"Determination of drugs of abuse and pharmaceuticals in skeletal tissue by UHPLC\u2013MS\/MS","volume":"290","author":"Orfanidis","year":"2018","journal-title":"Forensic Sci. Int."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.jflm.2013.11.012","article-title":"The clavicle bone as an alternative matrix in forensic toxicological analysis","volume":"22","author":"Vardakou","year":"2014","journal-title":"J. Forensic Leg. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1007\/s004140050086","article-title":"Detection of triazolam in skeletal remains buried for 4 years","volume":"110","author":"Kudo","year":"1997","journal-title":"Int. J. Leg. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1111\/1556-4029.13890","article-title":"Alprazolam and Zolpidem in Skeletal Tissue of Decomposed Body Confirms Exposure","volume":"64","author":"Orfanidis","year":"2019","journal-title":"J. Forensic Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"636","DOI":"10.1016\/j.jpba.2018.11.015","article-title":"Development and validation of a gas chromatography\u2013mass spectrometry method for opiates and cocaine in human bone","volume":"164","author":"Falcon","year":"2019","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.forsciint.2019.04.001","article-title":"Development and validation of a method for analysing of duloxetine, venlafaxine and amitriptyline in human bone","volume":"299","author":"Pellegrini","year":"2019","journal-title":"Forensic Sci. Int."},{"key":"ref_30","unstructured":"World Health Organization (2019, May 02). The Global Burden of Disease: 2004 Update. Geneva (2008). Available online: https:\/\/www.who.int\/healthinfo\/global_burden_disease\/2004_report_update\/en\/."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2224","DOI":"10.1016\/S0140-6736(12)61766-8","article-title":"A comparative risk assessment of burden of disease and injury attributable to 67 risk factors and risk factor clusters in 21 regions, 1990\u20132010: A systematic analysis for the Global Burden of Disease Study 2010","volume":"380","author":"Lim","year":"2012","journal-title":"Lancet"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.forsciint.2005.01.001","article-title":"Quantitation of atenolol, metoprolol, and propranolol in postmortem human fluid and tissue specimens via LC\/APCI-MS","volume":"156","author":"Johnson","year":"2006","journal-title":"Forensic Sci. Int."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1016\/j.jacc.2007.04.060","article-title":"Cardiovascular protection using beta-blockers: A critical review of the evidence","volume":"50","author":"Bangalore","year":"2007","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1016\/j.phrs.2018.10.001","article-title":"The European Society of Cardiology (ESC)\/European Society of Hypertension (ESH) 2018 guidelines for hypertension diagnosis and treatment: New concepts and recommendations","volume":"139","author":"Grassi","year":"2019","journal-title":"Pharmacol. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2560","DOI":"10.1001\/jama.289.19.2560","article-title":"The seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure: The JNC 7 report","volume":"289","author":"Chobanian","year":"2003","journal-title":"JAMA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1730","DOI":"10.1136\/bmj.318.7200.1730","article-title":"Beta Blockade after myocardial infarction: Systematic review and meta regression analysis","volume":"318","author":"Freemantle","year":"1999","journal-title":"BMJ"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"550","DOI":"10.7326\/0003-4819-134-7-200104030-00008","article-title":"Beta-blockers in congestive heart failure. A Bayesian meta-analysis","volume":"134","author":"Brophy","year":"2001","journal-title":"Ann. Intern. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1056\/NEJM199612053352301","article-title":"Atenolol reduced mortality and cardiovascular events after noncardiac surgery","volume":"335","author":"Mangano","year":"1996","journal-title":"N. Engl. J. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1097\/01.PSYPHR.0000436763.15959.dc","article-title":"Beta-Blockers: Off-Label Use in Psychiatric Disorders","volume":"48","author":"Newman","year":"2013","journal-title":"Psychopharm Rev."},{"key":"ref_40","unstructured":"World Anti-Doping Agency (2019, May 02). Prohibited List. The World Anti-Doping Code International Standard. Available online: https:\/\/www.wada-ama.org\/."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Wiysonge, C.S., Bradley, H.A., Volmink, J., Mayosi, B.M., and Opie, L.H. (2017). Beta-blockers for hypertension. Cochrane Database Syst. Rev., 1.","DOI":"10.1002\/14651858.CD002003.pub5"},{"key":"ref_42","unstructured":"Berenson, A. (The New York Times, 2005). Big drug makers see sales decline with their image, The New York Times."},{"key":"ref_43","unstructured":"Agencia Espa\u00f1ola de Medicamentos y Productos Sanitarios (2019, May 02). Observatorio de uso d Medicamentos. Utilizaci\u00f3n de Medicamentos Antihipertensivos en Espa\u00f1a Durante el Periodo 2010\u20132017. Available online: https:\/\/www.aemps.gob.es\/en\/medicamentosUsoHumano\/observatorio\/informes-publicados\/informes-antihipertensivos-espana-2010-2017.htm."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1080\/15563650.2018.1533727","article-title":"2017 Annual Report of the American Association of Poison Control Centers\u2019 National Poison Data System (NPDS): 35th Annual Report","volume":"56","author":"Gummin","year":"2018","journal-title":"Clin. Toxicol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1093\/jat\/19.6.519","article-title":"Distribution of metoprolol enantiomers in a fatal overdose","volume":"19","author":"Mozayani","year":"1995","journal-title":"J. Anal. Toxicol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/S0379-0738(03)00056-2","article-title":"An autopsy case of combined drug intoxication involving verapamil, metoprolol and digoxin","volume":"133","author":"Kinoshita","year":"2003","journal-title":"Forensic Sci. Int."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1097\/00000433-200003000-00009","article-title":"An unusual death by propranolol ingestion","volume":"21","author":"Fucci","year":"2000","journal-title":"Am. J. Forensic Med. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/S0196-0644(85)81081-7","article-title":"Beta blocker overdose with propranolol and with atenolol","volume":"14","author":"Weinstein","year":"1985","journal-title":"Ann. Emerg. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1111\/j.1365-2796.2001.00892.x","article-title":"Fatal adverse drug events: The paradox of drug treatment","volume":"250","author":"Buajordet","year":"2001","journal-title":"J. Intern. Med."},{"key":"ref_50","first-page":"447","article-title":"Therapeutic and toxic blood concentrations of more than 800 drugs and other xenobiotics","volume":"58","author":"Schulz","year":"2003","journal-title":"Pharmazie"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/jat\/bkx076","article-title":"Pilot-reported beta-blockers identified by forensic toxicology analysis of postmortem specimens","volume":"42","author":"Canfield","year":"2017","journal-title":"J. Anal. Toxicol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1007\/s12024-014-9588-x","article-title":"Nails are a potential alternative matrix to hair for drug analysis in general unknown screenings by liquid-chromatography quadrupole time-of-flight mass spectrometry","volume":"10","author":"Krumbiegel","year":"2014","journal-title":"Forensic Sci. Med. Pathol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1002\/dta.1507","article-title":"Drug concentrations in post-mortem femoral blood compared with therapeutic concentrations in plasma","volume":"6","author":"Launiainen","year":"2014","journal-title":"Drug Test. Anal."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.jchromb.2006.11.030","article-title":"Simultaneous determination of 6 beta-blockers, 3 calcium-channel antagonists, 4 angiotensin-II antagonists and 1 antiarrhytmic drug in post-mortem whole blood by automated solid phase extraction and liquid chromatography mass spectrometry: Method development and robustness testing by experimental design","volume":"850","author":"Kristoffersen","year":"2007","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1517\/17425255.2014.937702","article-title":"Pharmacokinetics of selective \u03b21-adrenergic blocking agents: Prescribing implications","volume":"10","author":"Jankovic","year":"2014","journal-title":"Expert Opin. Drug Metab. Toxicol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"S131","DOI":"10.2215\/CJN.04151206","article-title":"Normal bone anatomy and physiology","volume":"3","author":"Clarke","year":"2008","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_57","first-page":"5442","article-title":"Update of Standard Practices for New Method Validation in Forensic Toxicology","volume":"23","author":"Wille","year":"2017","journal-title":"Curr. Pharm. Des."},{"key":"ref_58","first-page":"5455","article-title":"Method Development in Forensic Toxicology","volume":"23","author":"Peters","year":"2017","journal-title":"Curr. Pharm. Des."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.1021\/ac020361s","article-title":"Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS\/MS","volume":"75","author":"Matuszewski","year":"2003","journal-title":"Anal. Chem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/13\/2400\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:02:21Z","timestamp":1760187741000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/24\/13\/2400"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,6,29]]},"references-count":59,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2019,7]]}},"alternative-id":["molecules24132400"],"URL":"https:\/\/doi.org\/10.3390\/molecules24132400","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,6,29]]}}}