Cannabis sativa Species Overview

Experiment: MPGR

Sample Name HelpReplicates
Mature Leaves4
Young Leaves4

Platform: Cannabis LC/TOF MS Negative Ion

View Data Download Data Download Raw Data

+ LC-TOF Metadata
Protocol for LC-TOF
LabPIA. Daniel Jones
AnalystMary Dawn Celiz
LC systemProminence High Performace Liquid Chromatograph
Solvent DeliveryLC-20AD
System ControllerSCL-10AVP
Column OvenCTO-20A
ColumnAscentis® Express C18 Analytical HPLC column, 5 cm x 0.21 cm, 2.7 µm
Manufacturer of columnSupelco
Catalogue number53822-U
Serial numberUSMD003586
Packing Lot numberS10035
Injection volume10 uL
Flow rate0.3 mL/min
Mobile phases
A0.5 mM ammonium acetate, pH 3.85
Gradient ProfileGradient program started with 1% B- 99% A, held for one minute, increased to 30% B in 1.5 minutes, then increased to 50% B in 5.5 minutes, further increase to 70% B in four minutes, followed by 99% B in ten minutes, held at 99 % B for four minutes, decrea
Column oven temperature50?C
Sample temperature in the autosampler10?C
Inlet method nameNIH_CAN_OPT_AscExp_004
Mass SpectrometerMicromass? LCT PremierTM time-of-flight (TOF) mass spectrometer
SoftwareMassLynxTM Version 4.1
Ionization sourceElectrospray ionization (ESI)
Data acquisitionW mode, centroid
PolarityPositive and Negative
Mass Range50-1500 m/z
Aperture 1 voltage
Function 120 V
Function 240 V
Function 360 V
Function 480 V
MS method fileMDC_CAN_pos or MDC_CAN_neg
Manipulation of data
Lock massTelmisartan was used as lock mass standard.

+ MarkerLynx Metadata
Protocol for MarkerLynx
Initial Retention Time (min)0.3
Final Retention Time (min)21
Low Mass50
High Mass1500
Mass Tolerance (Da)0.05
Apex Track Peak Parameters
Peak Width at 5% Height (seconds)automatic
Peak-to-Peak Baseline Noiseautomatic
Apply smoothingYes
Collection Parameters
Marker Intensity threshold (counts)20
Mass Window (Da)0.05
Retention Time Window (min)0.2
Noise Elimintation Level10
Deisotope DataYes

+ QuanLynx Metadata
Protocol for QuanLynx
Use absolute mass windowYes
Chromatogram Mass Window (Da)0.05
Response TypeExternal (absolute)
Response UsesArea
Concentration of Standard (µM)0.25, 0.5, 1, 2.5, 5, 10, 20
Retention Time Parameters
Locate Peak UsingRetention time
Retention Time Window (min) ±0.2
Locate Peak SelectionLargest
Calibration Properties
Polynomial TypeLinear
Calibration OriginForce
Weighing Function1/X
Concentration UnitsµM
Integration Properties
Smoothing EnabledYES
Smooth Parameters
Smoothing MethodMean
Smooting Iterations1
Smoothing Width2
Apex Track EnablesYes
Apex Track Parameters
Peak-to-Peak Baseline NoiseAutomatic
Peak width at 5% heightAutomatic
Baseline Start Threshold %0
Baseline End Threshold %0.5
Sample Handling
Protocol when analyzing the samplesBefore each batch of samples was analyzed, the mass spectrometer was calibrated using 0.1% phosphoric acid. After column equilibration, two methanol injections were performed first. Standard solutions were injected followed by a blank injection. After eve
Sample handlingSamples were received as dried extract. These were dissolved in 10 mL of 9:1 v/v methanol:chloroform, then sonicated. An aliquot of 500 uL was obtained from each extract and placed in polypropylene centrifuge tubes. These were dried in a SpeedVac Concentr
Sample storage temperatureRoom temperature

Sample 1:

Sample 2:

Metabolite Search


Chemically defined metabolites in this experiment:

    [M-H]- MDC-CAN-HPLC-E-NEG-F1-313.216-17.39
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-313.213-14.7
Cannabidiolic acid
    [3M+K-2H]- MDC-CAN-HPLC-E-NEG-F1-1111.567-14.56
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-357.205-14.57
    [2M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-737.393-14.57
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-315.232-14.76
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-631.467-14.72
Cannabigerolic acid
    [3M+K-2H]- MDC-CAN-HPLC-E-NEG-F1-1117.612-14.92
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-359.222-14.95
    [2M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-741.427-14.94
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-309.183-15.9
Cannabinolic acid
    [3M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-1083.493-16.02
    [3M+K-2H]- MDC-CAN-HPLC-E-NEG-F1-1099.471-16
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-353.175-16.04
    [2M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-729.332-16.01
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-247.131-9.53
Cannabistilbene isomer 1
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-303.121-7.6
Cannabistilbene isomer 2
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-303.12-8.02
Cannflavin A or Cannflavin C
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-435.175-14.18
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-435.178-15.44
Cannflavin B
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-367.116-12.53
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-735.238-12.5
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-447.091-3.82
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-895.188-3.84
Orientin hexoside or Flavocannabiside
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-1219.283-3.89
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-609.141-3.91
Orientin rhamnoside (3.1 min); Vitexin hexoside (4.1 min)
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-1187.292-3.93
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-593.147-3.93
Propyl-4-hydroxybenzoate standard
    fragment MDC-CAN-HPLC-E-NEG-F1-137.024-7.34
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-179.071-7.33
Quercetin dihexoside
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-625.136-3.75
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-341.109-0.49
    [M+Cl]- MDC-CAN-HPLC-E-NEG-F1-377.087-0.46
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-683.224-0.49
Telmisartan standard
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-513.229-12.12
Tetrahydrocannabinolic acid - C4
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-343.19-16.41
Tetrahydrocannabiorcolic acid isomer I
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-301.144-13.76
    [2M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-625.271-13.74
    [3M+K-2H]- MDC-CAN-HPLC-E-NEG-F1-943.382-13.72
Tetrahydrocannabiorcolic acid isomer II
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-301.14-14.24
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-285.183-14.58
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-431.097-4.04
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-863.198-4.03
Vitexin rhamnoside
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-1155.299-4.14
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-577.152-4.14
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-313.215-16.48
    [2M-H]- MDC-CAN-HPLC-E-NEG-F1-627.438-16.48
Δ9-tetrahydrocannabinolic acid
    [3M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-1095.593-17.23
    [3M+K-2H]- MDC-CAN-HPLC-E-NEG-F1-1111.568-17.24
    [4M+Na+K-3H]- MDC-CAN-HPLC-E-NEG-F1-1491.747-17.22
    [M-H]- MDC-CAN-HPLC-E-NEG-F1-357.2-17.25
    [2M+Na-2H]- MDC-CAN-HPLC-E-NEG-F1-737.398-17.27