Agilent ICP-MS. Fundamentals of ICP-MS Analysis and Its Applications for Low Level Elemental Determination in Cannabis

Agilent ICP-MS Fundamentals of ICP-MS Analysis and Its Applications for Low Level Elemental Determination in Cannabis High Sample Matrix Tolerance Superior & Simple Interference Removal Ultra Wide Linear Range for Trace and Percent Level Elemental Concentration Determinations

Agilent products and solutions are intended to be used for cannabis quality control and safety testing in laboratories where such use is permitted under state/country law.

Benefits of ICP-MS Multi-elemental technique High sensitivity, ppb/ppt (even ppq!) for most elements Short analysis time (~ 3-4 min) Extremely linear with wide dynamic range Minimum number of interferences High Productivity Page 3

Agilent s History of Innovation in ICP-MS 1994 to 2014 1994 4500 Series introduced - World's first benchtop system. Hyperbolic profile quad, motorized torch XYZ, cool plasma 1998 First real time ICP-MS chromatographic software PlasmaChrom. T- mode reaction interface introduced 1999 4500 Series 100, 200 & 300 introduced: 1st applications-specific ICP-MS. 2000 Agilent 7500 Series introduced - 7500a, 7500i and 7500s - the next generation in ICP-MS instrumentation. 9 orders detector range 2001 Agilent 7500c launched 1 st generation ORS for high matrix samples. 2002 New digital generators and LAN control introduced. First commercial GC- ICP-MS interface. 2003 Agilent 7500cs launched 2 nd generation ORS for high purity semicon samples. 2004 Agilent 7500ce launched 2 nd generation ORS for high matrix samples. 2005 Low flow cell gas MFC s for Xe NH 3, O 2, etc added to 7500ce/cs. 2006 Agilent acquires 100% of Agilent/Yokogawa joint venture 2007 Agilent 7500cx introduced: He only mode ICP-MS 2008 High Matrix Interface developed enables 2% TDS samples to be run by ICP-MS 2009 Agilent 7700 Series introduced replaces 7500 Series. MassHunter Software introduced - common platform with other Agilent MS. ISIS-DS Discrete sampling system, for ultra high throughput analysis 2012 January Agilent 8800 Introduced The world s first triple quad ICP-MS 2013 December Over 3000 7700 ICP-MS units sold worldwide 2014 January Introduction of the 7900 ICP-MS Agilent 8800 ICP-QQQ Agilent 7500 Series Agilent 4500 Series Agilent 7700 Series Agilent 7900 Series Page 4

Current Agilent ICP-MS Portfolio Agilent 7900 Agilent 7800 Agilent 8900 (Industries 1 st Triple Quad ICP-MS!) Page 5

Why is Metals Analysis Important for Cannabis? Assure no toxic metals are present Product safety for Medical & Recreational Big Four Metals As, Cd, Pb, & Hg Determination of full elemental suite Additional elements necessary for horticulture aspects soil and potential contaminants Not just about the plant Irrigation Water Fertilizers/Added Nutrients Soil Ingredients used for Edibles Oil Extracts/Concentrates Delivery Devices Vaporizers

ICPMS with Enhanced ORS 4 Technology 2. The smallest droplets pass through the spray chamber and into the ion source - the plasma 3. The sample is desolvated and ionized in the plasma 6. The quadrupole mass spectrometer separates ions based on their mass to charge ratio. The selected ions continue on to the detector 1. The liquid sample is mixed with argon gas by the nebulizer to form an aerosol.. 4. Ions are extracted from the plasma by Sampler /Skimmer cones and extraction lenses in the interface region 5. Ion beam is deflected and then focused in Octopole. Gases can be used to remove interferences 7. Ions are measured using a discrete dynode detector providing 10(7800)-11(7900) orders of linear dynamic range. Page 7

3 Key Benefits/Components of the Agilent ICP-MS 1) Matrix Tolerance 2) Interference Removal 3) Dynamic Range High Matrix Introduction System (HMI) Handles tough sample matrices better than any other ICP-MS. Reduces sample prep time and error Better long-term stability Octopole Reaction System He Mode He Mode effectively removes common polyatomic interferences in the samples while maintaining sensitivity. Easy to use - He collision mode only Effective for a wide range of sample types Accurate measurements Wider Dynamic Range 10(7800) 11(7900) orders dynamic range: 0.1ppt(DL) to 1,000ppm Simplifies method development Easy sample prep. Improves productivity Page 8 July 11, 2017

3 Key Benefits of Agilent ICP-MS 1) Matrix Tolerance 2) Interference Removal 3) Dynamic Range High Matrix Introduction System (HMI) Handles tough sample matrices better than any other ICP-MS. Reduces sample prep time and error Better long-term stability Octopole Reaction System He Mode He Mode effectively removes common polyatomic interferences in the samples Easy to use - He collision mode only Effective for a wide range of sample types Accurate measurements Wider Dynamic Range 10 orders dynamic range: 0.1ppt(DL) to 1,000ppm Simplifies method development Easy sample prep. Improves productivity Page 9 July 11, 2017

Introducing the Ground-Breaking Agilent UHMI(7900) UHMI gas port UHMI (ultra High Matrix Interface) much more than just a simple T-piece UHMI uses optimized gas mixing geometry and sophisticated plasma/gasflow tuning algorithm to set reproducible conditions for predictable aerosol dilution rate Page 10 July 11, 2017

The Big Four Spiked at Different Salt Content NaCl Amount 75 As [ 25 ppb ] 114 Cd [ 50 ppb ] 208 Pb [ 50 ppb ] 201 Hg [ 1 ppb ] 0% 26.9 49.2 49.7 0.85 0.5% 24.2 49.0 50.1 0.99 1% 24.8 51.5 50.2 0.93 1.5% 25.5 50.0 50.5 0.88 2% 24.6 50.0 49.7 1.03 5% 25.4 48.7 50.7 0.89 10% 22.8 46.1 49.8 0.91 25% 26.2 45.4 49.0 0.96 Average 25.1 48.7 50.0 0.93 % RSD 5% 4% 1% 6% % Recovered 100% 97% 100% 93% 0.5g / 100ml = 0.5% 1g / 100ml = 0.5% 1.5g / 100ml = 0.5% 2g / 100ml = 0.5% 5g / 100ml = 0.5% 10g / 100ml = 0.5% 25g / 100ml = 25% 11

High TDS Will Result in Ionization Suppression Ionization Suppression in 0.3% NaCl 120 100 80 60 40 20 0 Sc-45 Cr-52 Fe-56 Zn-66 Mo-95 In-115 1% HNO3 0.3%NaCl Confidentiality Label July 11, 2017 12

Cerium Oxide Formation in the Plasma The tendency to form CeO in the plasma is expressed as a ratio of the cps at 156 to the cps at 140 (CeO/Ce ratio). 156 140 Ce 16 O 140 Ce The CeO complex will break apart under hottler plasma conditions. A hotter plasma will have a lower CeO/Ce ratio. Page 13

The Importance of Oxides (CeO/Ce) CeO/Ce ratio is a performance criterion that all vendors cite, however the actual relevance of this is often overlooked. - Lower CeO/Ce ratio is the result of a hotter plasma. - A hotter plasma is the result of more electrons (joule heating) in the plasma Important factors affecting 140 Ce 16 O / 140 Ce : -Vapor loading in plasma (more water, more available Oxygen). -Plasma temperature (Higher temperature plasma more readily breaks down CeO complex). - A higher temperature plasma is the result of more electrons in the plasma. Lower CeO/Ce Ratio More CeO/Ce is expressed as the ratio of counts at mass 156 (CeO) divided by the counts at mass 140 (Ce). electrons available for ionization!!

Ionization Suppression as a Function of CeO With With CeO CeO @ @ 1.0%, 1.7%, suppression only at 10-25% 20-50%

Sample Introduction HMI Sample Introduction is: Low-flow (typically 0.15mL/min) Temperature stabilized (Peltier cooled spray chamber) Now features HMI(7800 shown) UHMI(7900), allowing auto setup of plasma conditions and much higher matrix tolerance

Polyatomic Interference Formation 75 40 Ar 35 Cl 75 As Page 18

Troublesome Region of the Periodic Table: Polyatomics Interfernces from Ar, O, Cl, C, Na, Mg, Ca. Page 19

Principle of Helium Collision Mode and Kinetic Energy Discrimination (KED) Polyatomic ions Analyte ions Energy distribution of analyte and interfering polyatomic ions with the same mass Polyatomic ions Bias voltage rejects low energy (polyatomic) ions Analyte ions Energy Energy At cell entrance, analyte and polyatomic ion energies overlap. Energy spread of both groups of ions is narrow, due to ShieldTorch System Cell Entrance Energy loss from each collision with a He atom is the same for analyte and polyatomic ion, but polyatomics are bigger and so collide more often Cell Exit By cell exit, ion energies no longer overlap; polyatomics are rejected using a bias voltage step. Analyte ions have enough residual energy to get over step; polyatomics don t (energy discrimination) Page 20

Acid Matrices and IPA in NoGas Mode (HNO 3 + HCl + H 2 SO 4 + IPA) 2E5 cps CO 2 SO, SOH ClO ArC ClO ArO, CaO SO 2, S 2, ArN 2 H, SO 2 H Unspiked 5% HNO 3 + 5% HCl + 1% H 2 SO 4 + 1% IPA Matrix Ar 2, Ca 2, ArCa, S 2 O, SO 3 Unspiked Matrix ALL peaks are due to polyatomic interferences ArN ArCl CO 2 H ArC S 2, SO 2 Cl 2 ArOH, CaOH ArCO, ArCN ArS, Cl 2 Ar 2 ArCl Br, Ar 2 H Br, Ar 2 H SN ClO, NaS CaO, NaCl CaO ClN 2, CaOH, ArNa CaO, NaClH NaCl S 2, SO 2 45 50 55 60 Mass 65 70 75 80 No Gas Mode Cl 2 H ArS, Cl 2 ArS Ar 2 Page 21 July 11, 2017

Acid Matrices and IPA in He Mode (HNO 3 + HCl + H 2 SO 4 + IPA) 2E5 cps Unspiked 5% HNO 3 + 5% HCl + 1% H 2 SO 4 + 1% IPA Matrix ALL polyatomic interferences are removed in He Mode (same cell conditions) He Mode Page 22

Acid Matrices and IPA in He Mode + 10ppb spike Consistent sensitivity and perfect template match for all elements 2E5 cps 10ppb Spike in 5% HNO 3 + 5% HCl + 1% H 2 SO 4 + 1% IPA Matrix Consistent high sensitivity for all isotopes of all elements in He Mode Good signal for all spike elements in 10ppb spike. Perfect template fit for all elements ALL isotopes available for quantification / confirmation No residual interferences and no loss of analyte signal by reaction 45 50 55 60 Mass 65 70 75 80 He Mode Page 23 July 11, 2017

Elimination of ArO Interference on Fe-56 Pag

Cal 0, 1, 10 ppb Arsenic in 1%HNO3/0.5% HCl (ArCl interference on m/z 75) Page 25

High Concentration Elements and Trace Elements: Linear Dynamic Range Page 27

Largest Analytical Range of any ICP-MS Calibration ranges Hg (10 200ppt) NoGas Mode As (10 200 ppt) He Mode Se (10 200 ppt) He Mode Na (0.05 1000 ppm) He Mode These 4 plots were obtained under the same analytical conditions on the 7800 only the gas mode (NoGas for Hg) changed Hg As As Overall calibration range 10ppt (Hg, As, Se) to 1000 ppm (Na) in a single method - without attenuating ion transmission to increase working range 10 ppt Mercury 10 ppt Arsenic Na Typically, ICP-MS cannot measure above 200ppm Na without changing quad resolution or ion lens settings Hg Hg LOD on 7800 is about 2ppt 7800 can QUANTITATE at 10ppt! Agilent ICP-MS can do both of the above in the same run! Se Se 10 ppt Selenium Na 1000 ppm Sodium Good fit at 0.2ppm Page 28

7800 Nutritional Supplement Samples #1 -Major constituents can be determined at any dilution. -Trace toxics measureable at even the highest dilution. *A Single Helium mode is used across the entire mass range. Nutritional Supplements Very Similar in Composition to Edible Cannabis Products Page 29

7800 Nutritional Supplement Samples #2 -Concentrations across all dilutions for cations the same. -Agilent actually measures Fe at 56. -All elements using main isotope or on mass isotope. -Excellent sensitivity across all trace/toxic element dilutions (As, Cd, Pb). Page 30

Unique Performance of the 7800 & 7900 Better matrix tolerance than any other ICP-MS Higher plasma temperature (lower CeO/Ce ratio) under standard conditions than any other system 7800 with HMI: ~2-3% TDS & 7900 with UHMI: ~25% TDS Best performance with Helium cell gas eliminates need for reaction gases in all common applications 7800/7900 ORS 4 removes polyatomic interferences in complex matrices while maintaining excellent sensitivity. Wider dynamic range than any other quadrupole ICP-MS 7800: 10 orders of Magnitude (~1000s ppm) (3 ms Integration) 7900: 11 orders of Magnitude (~Percent) (0.1 ms Integration spnp) Page 31

Analysis of Cannabis - Sample Preparation 1. Sample is crushed and homogenized 2. Aliquot of sample is weighed (~100-500 mg) 3. Addition of HNO 3 & HCl (to stabilize elements such as Hg) 4. Samples are digested using microwave digestion procedure 5. After digestion and cooling, digested material is brought to volume with Type 1 H 2 O for ICP-MS analysis. 6. Using the Agilent ICP-MS with HMI technology, samples are directly analyzed with no additional dilution required (minimizes potential for contamination from pipette tips and additional reagents)

Analysis of Cannabis Digestates by Agilent ICP-MS Calibration Curves (1%HNO 3 /0.5%HCl)

ICP-MS Results for Cannabis Digestates (ppb) Including Continuing Calibration QC and Spike Recoveries QC True Values 10.0 ug/l for As, Cd, & Pb, 1.0 ug/l for Hg Spike True Values 10.0 mg/l for As, Cd, & Pb, 0.1 mg/l for Hg

Agilent Mass Hunter Software Automatically Collects Quick Scan Data with Each Individual Sample Analysis. Provides elemental information for additional elements, even though they have not been included in the initial calibration standards.

I. Matrix Tolerance I. High Matrix Interface (HMI(7800) & UHMI(7900)) I. Aerosol dilution increases plasma temperature II. More efficient matrix breakdown and ionization efficiency III. Reduction of Polyatomics Begins in the Plasma on the Agilent II. Interference Removal (ORS 4 ) I. Low flow of non reactive gas (He), Small Volume Octopole provides high energy collisions resulting in polyatomic interference removal for multiple matrices during all analyses. II. ICP-MS Advantages Review and Summary Linear Dynamic Range (10 (7800) 11 (7900) Orders LDR) I. ICP-MS easily provides sub ppb down to ppt detection levels II. Large linear dynamic range allows for the quantitation of high concentration analytes, no dilution, single analysis, simple.

Potential for Future Applications New Industry, prepare for the future! HPLC ICP-MS Speciation Analysis As, Se, Cr.. GC ICP-MS Metals Determination in Organic Comp ppb/ppt Determination of P, S in pesticides

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