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Development and validation of an LC-MS/MS method for quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), THC, CBN and CBD in hair.
J Mass Spectrom. 2013 Feb; 48(2):227-33.JM

Abstract

For analysis of hair samples derived from a pilot study ('in vivo' contamination of hair by sidestream marijuana smoke), an LC-MS/MS method was developed and validated for the simultaneous quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), Δ9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD). Hair samples were extracted in methanol for 4 h under occasional shaking at room temperature, after adding THC-D(3), CBN-D(3), CBD-D(3) and THCA-A-D(3) as an in-house synthesized internal standard. The analytes were separated by gradient elution on a Luna C18 column using 0.1% HCOOH and ACN + 0.1% HCOOH. Data acquisition was performed on a QTrap 4000 in electrospray ionization-multi reaction monitoring mode. Validation was carried out according to the guidelines of the German Society of Toxicological and Forensic Chemistry (GTFCh). Limit of detection and lower limit of quantification were 2.5 pg/mg for THCA-A and 20 pg/mg for THC, CBN and CBD. A linear calibration model was applicable for all analytes over a range of 2.5 pg/mg or 20 pg/mg to 1000 pg/mg, using a weighting factor 1/x. Selectivity was shown for 12 blank hair samples from different sources. Accuracy and precision data were within the required limits for all analytes (bias between -0.2% and 6.4%, RSD between 3.7% and 11.5%). The dried hair extracts were stable over a time period of one to five days in the dark at room temperature. Processed sample stability (maximum decrease of analyte peak area below 25%) was considerably enhanced by adding 0.25% lecithin (w/v) in ACN + 0.1% HCOOH for reconstitution. Extraction efficiency for CBD was generally very low using methanol extraction. Hence, for effective extraction of CBD alkaline hydrolysis is recommended.

Authors+Show Affiliations

Institute of Forensic Medicine, Forensic Toxicology, Albertstraβe 9, 79104, Freiburg, Germany.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23378095

Citation

Roth, Nadine, et al. "Development and Validation of an LC-MS/MS Method for Quantification of Δ9-tetrahydrocannabinolic Acid a (THCA-A), THC, CBN and CBD in Hair." Journal of Mass Spectrometry : JMS, vol. 48, no. 2, 2013, pp. 227-33.
Roth N, Moosmann B, Auwärter V. Development and validation of an LC-MS/MS method for quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), THC, CBN and CBD in hair. J Mass Spectrom. 2013;48(2):227-33.
Roth, N., Moosmann, B., & Auwärter, V. (2013). Development and validation of an LC-MS/MS method for quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), THC, CBN and CBD in hair. Journal of Mass Spectrometry : JMS, 48(2), 227-33. https://doi.org/10.1002/jms.3152
Roth N, Moosmann B, Auwärter V. Development and Validation of an LC-MS/MS Method for Quantification of Δ9-tetrahydrocannabinolic Acid a (THCA-A), THC, CBN and CBD in Hair. J Mass Spectrom. 2013;48(2):227-33. PubMed PMID: 23378095.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of an LC-MS/MS method for quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), THC, CBN and CBD in hair. AU - Roth,Nadine, AU - Moosmann,Bjoern, AU - Auwärter,Volker, PY - 2012/10/05/received PY - 2012/12/05/revised PY - 2012/12/06/accepted PY - 2013/2/5/entrez PY - 2013/2/5/pubmed PY - 2013/7/3/medline SP - 227 EP - 33 JF - Journal of mass spectrometry : JMS JO - J Mass Spectrom VL - 48 IS - 2 N2 - For analysis of hair samples derived from a pilot study ('in vivo' contamination of hair by sidestream marijuana smoke), an LC-MS/MS method was developed and validated for the simultaneous quantification of Δ9-tetrahydrocannabinolic acid A (THCA-A), Δ9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD). Hair samples were extracted in methanol for 4 h under occasional shaking at room temperature, after adding THC-D(3), CBN-D(3), CBD-D(3) and THCA-A-D(3) as an in-house synthesized internal standard. The analytes were separated by gradient elution on a Luna C18 column using 0.1% HCOOH and ACN + 0.1% HCOOH. Data acquisition was performed on a QTrap 4000 in electrospray ionization-multi reaction monitoring mode. Validation was carried out according to the guidelines of the German Society of Toxicological and Forensic Chemistry (GTFCh). Limit of detection and lower limit of quantification were 2.5 pg/mg for THCA-A and 20 pg/mg for THC, CBN and CBD. A linear calibration model was applicable for all analytes over a range of 2.5 pg/mg or 20 pg/mg to 1000 pg/mg, using a weighting factor 1/x. Selectivity was shown for 12 blank hair samples from different sources. Accuracy and precision data were within the required limits for all analytes (bias between -0.2% and 6.4%, RSD between 3.7% and 11.5%). The dried hair extracts were stable over a time period of one to five days in the dark at room temperature. Processed sample stability (maximum decrease of analyte peak area below 25%) was considerably enhanced by adding 0.25% lecithin (w/v) in ACN + 0.1% HCOOH for reconstitution. Extraction efficiency for CBD was generally very low using methanol extraction. Hence, for effective extraction of CBD alkaline hydrolysis is recommended. SN - 1096-9888 UR - https://www.unboundmedicine.com/medline/citation/23378095/Development_and_validation_of_an_LC_MS/MS_method_for_quantification_of_Δ9_tetrahydrocannabinolic_acid_A__THCA_A__THC_CBN_and_CBD_in_hair_ L2 - https://doi.org/10.1002/jms.3152 DB - PRIME DP - Unbound Medicine ER -