Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy
Polymorphic thiopurine S-methyltransferase (TPMT) is a major determinant of thiopurine toxicity. We extracted 6-thioguanine nucleotides (6-TGNs) and 6-methylmercaptopurine nucleotides (6-MMPNs) from erythrocytes with perchloric acid and converted them to 6-thioguanine (6-TG) and a 6-methylmercaptopu...
Gespeichert in:
| Veröffentlicht in: | Clinical chemistry (Baltimore, Md.) Md.), 2005-11, Vol.51 (11), p.2074-2084 |
|---|---|
| Hauptverfasser: | , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2084 |
|---|---|
| container_issue | 11 |
| container_start_page | 2074 |
| container_title | Clinical chemistry (Baltimore, Md.) |
| container_volume | 51 |
| creator | Dervieux, Thierry Meyer, Gary Barham, Robert Matsutani, Mariko Barry, Mary Boulieu, Roselyne Neri, Bruce Seidman, Ernest |
| description | Polymorphic thiopurine S-methyltransferase (TPMT) is a major determinant of thiopurine toxicity.
We extracted 6-thioguanine nucleotides (6-TGNs) and 6-methylmercaptopurine nucleotides (6-MMPNs) from erythrocytes with perchloric acid and converted them to 6-thioguanine (6-TG) and a 6-methylmercaptopurine (6-MMP) derivative during a 60-min acid hydrolysis step. The liquid chromatography system consisted of a C(18) column with an ammonium acetate-formic acid-acetonitrile buffer. 8-Bromoadenine was the internal standard. Analytes were measured with positive ionization and multiple reaction monitoring mode. With PCR-restriction fragment length polymorphism analysis and TaqMan allelic discrimination, common TPMT alleles (*1, *2, *3A, *3B, *3C) were determined in 31 792 individuals. We used perchloric acid extraction, acid hydrolysis, and HPLC with ultraviolet detection to measure erythrocyte 6-TG and 6-MMP nucleotide concentrations in 6189 patients with inflammatory bowel disease receiving azathioprine/6-mercaptopurine therapy.
Intra- and interday imprecision were 450 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for leukopenia), but an 8.2-fold lower risk for 6-MMPNs >5700 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for hepatotoxicity).
The liquid chromatography-tandem mass spectrometry method can be applied to the routine monitoring of thiopurine therapy. The association between TPMT genotype and metabolite concentrations illustrates the utility of pharmacogenetics in the management of patients undergoing treatment with thiopurines. |
| doi_str_mv | 10.1373/clinchem.2005.050831 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_17387294</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>17387294</sourcerecordid><originalsourceid>FETCH-LOGICAL-c471t-3e7fd696e1d4b4d0df830c800147c79096fbcb84d0fbdb40831660bdd850ca63</originalsourceid><addsrcrecordid>eNpdkt1u0zAUgCMEYt3gDRCykICrdHbj2MllVZWB1AKCilvLsY8bT_npbIcqe2FeA2ftNMSVZfs7n31-kuQNwXOS8exaNbZTNbTzBcb5HOe4yMizZEbyDKdFzsjzZIYxLtOSUH6RXHp_G7eUF-xlckEYYYxwMkv-bOzdYDVa1a5vZej3Th7qMd3JTkOLttJ79PMAKsRbCG5Ey042o7ce9Qat3RhimBoDoF1t-8PgbAfo66Aa6IPV4FHUoLUxUTAF_ANtIdRjE5zsvAEnPaAbiMe_pLOyC1HfRRr8AyirvrEh2myHvstgYQJ-gAL723Z7tLyXYRJP3muWbsEpeQiPD0VLTGl8lbwwsvHw-rxeJbtP693qc7r5dvNltdykinIS0gy40axkQDStqMbaFBlWBcaxiIqXuGSmUlURb0ylKzrVnDFcaV3kWEmWXSUfTtqD6-8G8EG01itoGtlBP3hBeFbwRUkj-O4_8LYfXCyuFwtCMV7QrIwQPUHK9d47MCIm2Uo3CoLFNATicQjENATiNAQx7O3ZPVQt6Kegc9cj8P4MSK9kY2IXlPVPHF-Qkj188uOJq-2-PloHwreyaaKWiOPxmBNBSHyZ0-wvGYHQ5Q</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>214002439</pqid></control><display><type>article</type><title>Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy</title><source>MEDLINE</source><source>Oxford University Press Journals All Titles (1996-Current)</source><creator>Dervieux, Thierry ; Meyer, Gary ; Barham, Robert ; Matsutani, Mariko ; Barry, Mary ; Boulieu, Roselyne ; Neri, Bruce ; Seidman, Ernest</creator><creatorcontrib>Dervieux, Thierry ; Meyer, Gary ; Barham, Robert ; Matsutani, Mariko ; Barry, Mary ; Boulieu, Roselyne ; Neri, Bruce ; Seidman, Ernest</creatorcontrib><description>Polymorphic thiopurine S-methyltransferase (TPMT) is a major determinant of thiopurine toxicity.
We extracted 6-thioguanine nucleotides (6-TGNs) and 6-methylmercaptopurine nucleotides (6-MMPNs) from erythrocytes with perchloric acid and converted them to 6-thioguanine (6-TG) and a 6-methylmercaptopurine (6-MMP) derivative during a 60-min acid hydrolysis step. The liquid chromatography system consisted of a C(18) column with an ammonium acetate-formic acid-acetonitrile buffer. 8-Bromoadenine was the internal standard. Analytes were measured with positive ionization and multiple reaction monitoring mode. With PCR-restriction fragment length polymorphism analysis and TaqMan allelic discrimination, common TPMT alleles (*1, *2, *3A, *3B, *3C) were determined in 31 792 individuals. We used perchloric acid extraction, acid hydrolysis, and HPLC with ultraviolet detection to measure erythrocyte 6-TG and 6-MMP nucleotide concentrations in 6189 patients with inflammatory bowel disease receiving azathioprine/6-mercaptopurine therapy.
Intra- and interday imprecision were <10% at low and high analyte concentrations. The conversion of 6-TG and 6-MMP nucleoside mono-, di-, and triphosphates was complete after hydrolysis. Allelic frequency for TPMT variant alleles ranged from 0.0063% (*3B) to 3.61% (*3A). Compared with wild types, TPMT heterozygotes had an 8.3-fold higher risk for 6-TGNs >450 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for leukopenia), but an 8.2-fold lower risk for 6-MMPNs >5700 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for hepatotoxicity).
The liquid chromatography-tandem mass spectrometry method can be applied to the routine monitoring of thiopurine therapy. The association between TPMT genotype and metabolite concentrations illustrates the utility of pharmacogenetics in the management of patients undergoing treatment with thiopurines.</description><identifier>ISSN: 0009-9147</identifier><identifier>EISSN: 1530-8561</identifier><identifier>DOI: 10.1373/clinchem.2005.050831</identifier><identifier>PMID: 16166171</identifier><identifier>CODEN: CLCHAU</identifier><language>eng</language><publisher>Washington, DC: Am Assoc Clin Chem</publisher><subject>Adult ; Ammonium ; Analytical, structural and metabolic biochemistry ; Azathioprine - blood ; Azathioprine - therapeutic use ; Biological and medical sciences ; Blood & organ donations ; Chromatography ; Chromatography, Liquid ; Cohort Studies ; Drug Therapy, Combination ; Erythrocytes ; Erythrocytes - chemistry ; Female ; Fundamental and applied biological sciences. Psychology ; Genetic Variation ; Genotype & phenotype ; Guanine Nucleotides - blood ; Hepatotoxicity ; Humans ; Hydrolysis ; Inflammatory bowel disease ; Inflammatory Bowel Diseases - blood ; Inflammatory Bowel Diseases - drug therapy ; Investigative techniques, diagnostic techniques (general aspects) ; Ionization ; Leukopenia ; Liquid chromatography ; Male ; Mass Spectrometry ; Medical sciences ; Mercaptopurine - analogs & derivatives ; Mercaptopurine - blood ; Mercaptopurine - therapeutic use ; Metabolites ; Methyltransferases - genetics ; Mutation ; Polymorphism ; Purine Nucleotides - blood ; Scientific imaging ; Thionucleotides - blood</subject><ispartof>Clinical chemistry (Baltimore, Md.), 2005-11, Vol.51 (11), p.2074-2084</ispartof><rights>2005 INIST-CNRS</rights><rights>Copyright American Association for Clinical Chemistry Nov 2005</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c471t-3e7fd696e1d4b4d0df830c800147c79096fbcb84d0fbdb40831660bdd850ca63</citedby><cites>FETCH-LOGICAL-c471t-3e7fd696e1d4b4d0df830c800147c79096fbcb84d0fbdb40831660bdd850ca63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17219694$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16166171$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dervieux, Thierry</creatorcontrib><creatorcontrib>Meyer, Gary</creatorcontrib><creatorcontrib>Barham, Robert</creatorcontrib><creatorcontrib>Matsutani, Mariko</creatorcontrib><creatorcontrib>Barry, Mary</creatorcontrib><creatorcontrib>Boulieu, Roselyne</creatorcontrib><creatorcontrib>Neri, Bruce</creatorcontrib><creatorcontrib>Seidman, Ernest</creatorcontrib><title>Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy</title><title>Clinical chemistry (Baltimore, Md.)</title><addtitle>Clin Chem</addtitle><description>Polymorphic thiopurine S-methyltransferase (TPMT) is a major determinant of thiopurine toxicity.
We extracted 6-thioguanine nucleotides (6-TGNs) and 6-methylmercaptopurine nucleotides (6-MMPNs) from erythrocytes with perchloric acid and converted them to 6-thioguanine (6-TG) and a 6-methylmercaptopurine (6-MMP) derivative during a 60-min acid hydrolysis step. The liquid chromatography system consisted of a C(18) column with an ammonium acetate-formic acid-acetonitrile buffer. 8-Bromoadenine was the internal standard. Analytes were measured with positive ionization and multiple reaction monitoring mode. With PCR-restriction fragment length polymorphism analysis and TaqMan allelic discrimination, common TPMT alleles (*1, *2, *3A, *3B, *3C) were determined in 31 792 individuals. We used perchloric acid extraction, acid hydrolysis, and HPLC with ultraviolet detection to measure erythrocyte 6-TG and 6-MMP nucleotide concentrations in 6189 patients with inflammatory bowel disease receiving azathioprine/6-mercaptopurine therapy.
Intra- and interday imprecision were <10% at low and high analyte concentrations. The conversion of 6-TG and 6-MMP nucleoside mono-, di-, and triphosphates was complete after hydrolysis. Allelic frequency for TPMT variant alleles ranged from 0.0063% (*3B) to 3.61% (*3A). Compared with wild types, TPMT heterozygotes had an 8.3-fold higher risk for 6-TGNs >450 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for leukopenia), but an 8.2-fold lower risk for 6-MMPNs >5700 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for hepatotoxicity).
The liquid chromatography-tandem mass spectrometry method can be applied to the routine monitoring of thiopurine therapy. The association between TPMT genotype and metabolite concentrations illustrates the utility of pharmacogenetics in the management of patients undergoing treatment with thiopurines.</description><subject>Adult</subject><subject>Ammonium</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Azathioprine - blood</subject><subject>Azathioprine - therapeutic use</subject><subject>Biological and medical sciences</subject><subject>Blood & organ donations</subject><subject>Chromatography</subject><subject>Chromatography, Liquid</subject><subject>Cohort Studies</subject><subject>Drug Therapy, Combination</subject><subject>Erythrocytes</subject><subject>Erythrocytes - chemistry</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic Variation</subject><subject>Genotype & phenotype</subject><subject>Guanine Nucleotides - blood</subject><subject>Hepatotoxicity</subject><subject>Humans</subject><subject>Hydrolysis</subject><subject>Inflammatory bowel disease</subject><subject>Inflammatory Bowel Diseases - blood</subject><subject>Inflammatory Bowel Diseases - drug therapy</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Ionization</subject><subject>Leukopenia</subject><subject>Liquid chromatography</subject><subject>Male</subject><subject>Mass Spectrometry</subject><subject>Medical sciences</subject><subject>Mercaptopurine - analogs & derivatives</subject><subject>Mercaptopurine - blood</subject><subject>Mercaptopurine - therapeutic use</subject><subject>Metabolites</subject><subject>Methyltransferases - genetics</subject><subject>Mutation</subject><subject>Polymorphism</subject><subject>Purine Nucleotides - blood</subject><subject>Scientific imaging</subject><subject>Thionucleotides - blood</subject><issn>0009-9147</issn><issn>1530-8561</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkt1u0zAUgCMEYt3gDRCykICrdHbj2MllVZWB1AKCilvLsY8bT_npbIcqe2FeA2ftNMSVZfs7n31-kuQNwXOS8exaNbZTNbTzBcb5HOe4yMizZEbyDKdFzsjzZIYxLtOSUH6RXHp_G7eUF-xlckEYYYxwMkv-bOzdYDVa1a5vZej3Th7qMd3JTkOLttJ79PMAKsRbCG5Ey042o7ce9Qat3RhimBoDoF1t-8PgbAfo66Aa6IPV4FHUoLUxUTAF_ANtIdRjE5zsvAEnPaAbiMe_pLOyC1HfRRr8AyirvrEh2myHvstgYQJ-gAL723Z7tLyXYRJP3muWbsEpeQiPD0VLTGl8lbwwsvHw-rxeJbtP693qc7r5dvNltdykinIS0gy40axkQDStqMbaFBlWBcaxiIqXuGSmUlURb0ylKzrVnDFcaV3kWEmWXSUfTtqD6-8G8EG01itoGtlBP3hBeFbwRUkj-O4_8LYfXCyuFwtCMV7QrIwQPUHK9d47MCIm2Uo3CoLFNATicQjENATiNAQx7O3ZPVQt6Kegc9cj8P4MSK9kY2IXlPVPHF-Qkj188uOJq-2-PloHwreyaaKWiOPxmBNBSHyZ0-wvGYHQ5Q</recordid><startdate>20051101</startdate><enddate>20051101</enddate><creator>Dervieux, Thierry</creator><creator>Meyer, Gary</creator><creator>Barham, Robert</creator><creator>Matsutani, Mariko</creator><creator>Barry, Mary</creator><creator>Boulieu, Roselyne</creator><creator>Neri, Bruce</creator><creator>Seidman, Ernest</creator><general>Am Assoc Clin Chem</general><general>American Association for Clinical Chemistry</general><general>Oxford University Press</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>4U-</scope><scope>7QO</scope><scope>7RV</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>S0X</scope></search><sort><creationdate>20051101</creationdate><title>Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy</title><author>Dervieux, Thierry ; Meyer, Gary ; Barham, Robert ; Matsutani, Mariko ; Barry, Mary ; Boulieu, Roselyne ; Neri, Bruce ; Seidman, Ernest</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c471t-3e7fd696e1d4b4d0df830c800147c79096fbcb84d0fbdb40831660bdd850ca63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Adult</topic><topic>Ammonium</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Azathioprine - blood</topic><topic>Azathioprine - therapeutic use</topic><topic>Biological and medical sciences</topic><topic>Blood & organ donations</topic><topic>Chromatography</topic><topic>Chromatography, Liquid</topic><topic>Cohort Studies</topic><topic>Drug Therapy, Combination</topic><topic>Erythrocytes</topic><topic>Erythrocytes - chemistry</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic Variation</topic><topic>Genotype & phenotype</topic><topic>Guanine Nucleotides - blood</topic><topic>Hepatotoxicity</topic><topic>Humans</topic><topic>Hydrolysis</topic><topic>Inflammatory bowel disease</topic><topic>Inflammatory Bowel Diseases - blood</topic><topic>Inflammatory Bowel Diseases - drug therapy</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Ionization</topic><topic>Leukopenia</topic><topic>Liquid chromatography</topic><topic>Male</topic><topic>Mass Spectrometry</topic><topic>Medical sciences</topic><topic>Mercaptopurine - analogs & derivatives</topic><topic>Mercaptopurine - blood</topic><topic>Mercaptopurine - therapeutic use</topic><topic>Metabolites</topic><topic>Methyltransferases - genetics</topic><topic>Mutation</topic><topic>Polymorphism</topic><topic>Purine Nucleotides - blood</topic><topic>Scientific imaging</topic><topic>Thionucleotides - blood</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dervieux, Thierry</creatorcontrib><creatorcontrib>Meyer, Gary</creatorcontrib><creatorcontrib>Barham, Robert</creatorcontrib><creatorcontrib>Matsutani, Mariko</creatorcontrib><creatorcontrib>Barry, Mary</creatorcontrib><creatorcontrib>Boulieu, Roselyne</creatorcontrib><creatorcontrib>Neri, Bruce</creatorcontrib><creatorcontrib>Seidman, Ernest</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>University Readers</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>SIRS Editorial</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dervieux, Thierry</au><au>Meyer, Gary</au><au>Barham, Robert</au><au>Matsutani, Mariko</au><au>Barry, Mary</au><au>Boulieu, Roselyne</au><au>Neri, Bruce</au><au>Seidman, Ernest</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2005-11-01</date><risdate>2005</risdate><volume>51</volume><issue>11</issue><spage>2074</spage><epage>2084</epage><pages>2074-2084</pages><issn>0009-9147</issn><eissn>1530-8561</eissn><coden>CLCHAU</coden><abstract>Polymorphic thiopurine S-methyltransferase (TPMT) is a major determinant of thiopurine toxicity.
We extracted 6-thioguanine nucleotides (6-TGNs) and 6-methylmercaptopurine nucleotides (6-MMPNs) from erythrocytes with perchloric acid and converted them to 6-thioguanine (6-TG) and a 6-methylmercaptopurine (6-MMP) derivative during a 60-min acid hydrolysis step. The liquid chromatography system consisted of a C(18) column with an ammonium acetate-formic acid-acetonitrile buffer. 8-Bromoadenine was the internal standard. Analytes were measured with positive ionization and multiple reaction monitoring mode. With PCR-restriction fragment length polymorphism analysis and TaqMan allelic discrimination, common TPMT alleles (*1, *2, *3A, *3B, *3C) were determined in 31 792 individuals. We used perchloric acid extraction, acid hydrolysis, and HPLC with ultraviolet detection to measure erythrocyte 6-TG and 6-MMP nucleotide concentrations in 6189 patients with inflammatory bowel disease receiving azathioprine/6-mercaptopurine therapy.
Intra- and interday imprecision were <10% at low and high analyte concentrations. The conversion of 6-TG and 6-MMP nucleoside mono-, di-, and triphosphates was complete after hydrolysis. Allelic frequency for TPMT variant alleles ranged from 0.0063% (*3B) to 3.61% (*3A). Compared with wild types, TPMT heterozygotes had an 8.3-fold higher risk for 6-TGNs >450 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for leukopenia), but an 8.2-fold lower risk for 6-MMPNs >5700 pmol/8 x 10(8) erythrocytes (concentration associated with increased risk for hepatotoxicity).
The liquid chromatography-tandem mass spectrometry method can be applied to the routine monitoring of thiopurine therapy. The association between TPMT genotype and metabolite concentrations illustrates the utility of pharmacogenetics in the management of patients undergoing treatment with thiopurines.</abstract><cop>Washington, DC</cop><pub>Am Assoc Clin Chem</pub><pmid>16166171</pmid><doi>10.1373/clinchem.2005.050831</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0009-9147 |
| ispartof | Clinical chemistry (Baltimore, Md.), 2005-11, Vol.51 (11), p.2074-2084 |
| issn | 0009-9147 1530-8561 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_17387294 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current) |
| subjects | Adult Ammonium Analytical, structural and metabolic biochemistry Azathioprine - blood Azathioprine - therapeutic use Biological and medical sciences Blood & organ donations Chromatography Chromatography, Liquid Cohort Studies Drug Therapy, Combination Erythrocytes Erythrocytes - chemistry Female Fundamental and applied biological sciences. Psychology Genetic Variation Genotype & phenotype Guanine Nucleotides - blood Hepatotoxicity Humans Hydrolysis Inflammatory bowel disease Inflammatory Bowel Diseases - blood Inflammatory Bowel Diseases - drug therapy Investigative techniques, diagnostic techniques (general aspects) Ionization Leukopenia Liquid chromatography Male Mass Spectrometry Medical sciences Mercaptopurine - analogs & derivatives Mercaptopurine - blood Mercaptopurine - therapeutic use Metabolites Methyltransferases - genetics Mutation Polymorphism Purine Nucleotides - blood Scientific imaging Thionucleotides - blood |
| title | Liquid Chromatography-Tandem Mass Spectrometry Analysis of Erythrocyte Thiopurine Nucleotides and Effect of Thiopurine Methyltransferase Gene Variants on These Metabolites in Patients Receiving Azathioprine/6-Mercaptopurine Therapy |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-15T15%3A33%3A12IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Liquid%20Chromatography-Tandem%20Mass%20Spectrometry%20Analysis%20of%20Erythrocyte%20Thiopurine%20Nucleotides%20and%20Effect%20of%20Thiopurine%20Methyltransferase%20Gene%20Variants%20on%20These%20Metabolites%20in%20Patients%20Receiving%20Azathioprine/6-Mercaptopurine%20Therapy&rft.jtitle=Clinical%20chemistry%20(Baltimore,%20Md.)&rft.au=Dervieux,%20Thierry&rft.date=2005-11-01&rft.volume=51&rft.issue=11&rft.spage=2074&rft.epage=2084&rft.pages=2074-2084&rft.issn=0009-9147&rft.eissn=1530-8561&rft.coden=CLCHAU&rft_id=info:doi/10.1373/clinchem.2005.050831&rft_dat=%3Cproquest_cross%3E17387294%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=214002439&rft_id=info:pmid/16166171&rfr_iscdi=true |