超高效液相色谱-质谱法同时测定大麻植物中Δ9-四氢大麻酚和Δ9-四氢大麻酚酸A
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摘要
10mg样品粉末用5mL甲醇超声萃取,离心后,上清液在Waters ACQUTIY UPLC BEH C18色谱柱(2.1mm×50mm,1.7μm)上分离,以含0.1%(体积分数)甲酸的甲醇和水(体积比为87∶13)为流动相进行等度洗脱,流量为0.2mL·min-1。利用光电二极管阵列检测器在波长220nm处对Δ9-四氢大麻酚(Δ9-THC)和Δ9-四氢大麻酚酸A(Δ9-THCA-A)进行定性和定量分析,并采用质谱检测器QDa进行确证。Δ9-THC和Δ9-THCA-A的质量浓度分别在0.50~20.0mg·L-1和2.0~40.0 mg·L-1内与其峰面积呈线性关系,检出限(3S/N)分别为0.10,1.0mg·L-1。加标回收率在81.5%~97.9%之间,测定值的相对标准偏差(n=6)在0.77%~4.1%之间。采用该方法对20个大麻植物样品进行分析,由Δ9-THC和Δ9-THCA-A的测定值计算得总Δ9-THC含量略高于标准方法测定结果。
The sample powder(10 mg)was ultrasonically extracted with 5 mL of methanol.After centrifugation,the supernatant was separated on a Waters ACQUTIY UPLC BEH C18 column(2.1 mm×50 mm,1.7μm),with methanol containing 0.1%(φ)formic acid and water with volume faction of 87∶13 as mobile phase for isocratic elution at a flow rate of 0.2 mL · min-1. Δ9-Tetrahydrocannabinol(Δ9-THC)and Δ9-tetrahydrocannabinolic acid A(Δ9-THCA-A)were qualitatively and quantitatively analyzed by photodiode array detector at the wavelength of 220 nm and confirmed by mass spectrometer QDa.Linear relationships were found between peak areas and mass concentrations ofΔ9-THC andΔ9-THCA-A in the range of 0.50-20.0 mg·L-1,2.0-40.0 mg·L-1,respectively,with detection limits(3 S/N)of 0.10,1.0 mg·L-1,respectively.Values of recovery obtained by standard addition method were in the range of 81.5%-97.9%,and RSDs(n=6)were in the range of 0.77%-4.1%.The proposed method was applied to analysis of 20 cannabis plant samples,the totalΔ9-THC contents calculated with the determination results ofΔ9-THC andΔ9-THCA-A were slightly high than the determination results obtained by the standard method.
The sample powder(10 mg)was ultrasonically extracted with 5 mL of methanol.After centrifugation,the supernatant was separated on a Waters ACQUTIY UPLC BEH C18 column(2.1 mm×50 mm,1.7μm),with methanol containing 0.1%(φ)formic acid and water with volume faction of 87∶13 as mobile phase for isocratic elution at a flow rate of 0.2 mL · min-1. Δ9-Tetrahydrocannabinol(Δ9-THC)and Δ9-tetrahydrocannabinolic acid A(Δ9-THCA-A)were qualitatively and quantitatively analyzed by photodiode array detector at the wavelength of 220 nm and confirmed by mass spectrometer QDa.Linear relationships were found between peak areas and mass concentrations ofΔ9-THC andΔ9-THCA-A in the range of 0.50-20.0 mg·L-1,2.0-40.0 mg·L-1,respectively,with detection limits(3 S/N)of 0.10,1.0 mg·L-1,respectively.Values of recovery obtained by standard addition method were in the range of 81.5%-97.9%,and RSDs(n=6)were in the range of 0.77%-4.1%.The proposed method was applied to analysis of 20 cannabis plant samples,the totalΔ9-THC contents calculated with the determination results ofΔ9-THC andΔ9-THCA-A were slightly high than the determination results obtained by the standard method.
引文
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[11]GAMBARO V,ACQUA L D,FARF,et al.Determination of primary active constituents in Cannabis preparations by high-resolution gas chromatography/flame ionization detection and high-performance liquid chromatography/UV detection[J].Analytica Chimica Acta,2002,468:245-254.
[12]DUSSY F E,HAMBERG C,LUGINBHL M,et al.Isolation of Delta9-THCA-A from hemp and analytical aspects concerning the determination of Delta9-THC in cannabis products[J].Forensic Science International,2005,149:3-10.
[13]PESCHEL W,POLITI M.1 H NMR and HPLC/DAD for Cannabis sativa L.chemotype distinction,extract profiling and specification[J].Talanta,2015,140:150-165.
[14]LILIAN V T,NAYARA A S,RAYZA R T,et al.Isomeric separation of cannabinoids by UPLC combined with ionic mobility mass spectrometry[J].International Journal of Mass Spectrometry,2016,10:1-10.
[15]GA/T 1008.3-2013常见毒品的气相色谱、气相色谱-质谱检验方法第3部分:大麻中三种成分[S].
[16]ST/NAR/40 Recommended methods for the identification and analysis of Cannabis and cannabis products[S].
[2]TEOTIA A K,PAL R.Analysis of drugs and poisons of Forensic interest-A review[J].Journal of Forensic Medicine and Toxicology,2009,26(1):49-53.
[3]TASCHWER M,SCHMID M G.Determination of the relative percentage distribution of THCA andΔ9-THC in herbal cannabis seized in Austria-Impact of different storage temperatures on stability[J].Forensic Science International,2015,254:167-171.
[4]TROFIN I G,DABIJA G,VAIREANU D I,et al.The influence of long-term storage conditions on the stability of cannabinoids derived from cannabis resin[J].Revista Chimmie,2012,63(4):422-427.
[5]TROFIN I G,VLAD C C,DABIJA G,et al.Influence of storage conditions on the chemical potency of herbal Cannabis[J].Revista Chimie,2011,62(6):639-645.
[6]LINDHOLST C.Long term stability of cannabis resin and cannabis extracts[J].Australian Journal Forensic Science,2010,42(3):181-190.
[7]PELLEGRINI M,MARCHEI E,PACIFICI R,et al.A rapid and simple procedure for the determination of cannabinoids in hemp food products by gas chromatography-mass spectrometry[J].Journal of Pharmaceutical Biomedical Analysis,2005,36(5):939-946.
[8]AMBACH L,PENITSCHKA F,BROILLET A,et al.Simultaneous quantification of delta-9-THC,THCacid A,CBN and CBD in seized drugs using HPLCDAD[J].Forensic Science International,2014,243:107-111.
[9]ROTH N,MOOSMANN B,AUWRTER V.Development and validation of an LC-MS/MS method for quantification ofΔ9-tetrahydrocannabinolic acid A(THCA-A),THC,CBN and CBD in hair[J].Journal of Mass Spectrom,2013,48:227-233.
[10]BACKER B D,DEBRUS B,LEBRUN P,et al.Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material[J].Journal of Chromatography B,2009,877:4115-4124.
[11]GAMBARO V,ACQUA L D,FARF,et al.Determination of primary active constituents in Cannabis preparations by high-resolution gas chromatography/flame ionization detection and high-performance liquid chromatography/UV detection[J].Analytica Chimica Acta,2002,468:245-254.
[12]DUSSY F E,HAMBERG C,LUGINBHL M,et al.Isolation of Delta9-THCA-A from hemp and analytical aspects concerning the determination of Delta9-THC in cannabis products[J].Forensic Science International,2005,149:3-10.
[13]PESCHEL W,POLITI M.1 H NMR and HPLC/DAD for Cannabis sativa L.chemotype distinction,extract profiling and specification[J].Talanta,2015,140:150-165.
[14]LILIAN V T,NAYARA A S,RAYZA R T,et al.Isomeric separation of cannabinoids by UPLC combined with ionic mobility mass spectrometry[J].International Journal of Mass Spectrometry,2016,10:1-10.
[15]GA/T 1008.3-2013常见毒品的气相色谱、气相色谱-质谱检验方法第3部分:大麻中三种成分[S].
[16]ST/NAR/40 Recommended methods for the identification and analysis of Cannabis and cannabis products[S].