PURITY GRADES OF LAB REAGENTS
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1 Dr. Katarzyna Pokajewicz May 2018 PURITY GRADES OF LAB REAGENTS HOW TO CHOOSE THE RIGHT GRADE?
2 Honeywell Research Chemicals A global portfolio of more than 6,000 references for analytical and chemistry labs Worldwide known and leading brands: Fluka Riedel de Haen Burdick&Jackson Hydranal 200 year of experience in producing chemicals: Seelze, Germany Muskegon, USA Honeywell Confidential by Honeywell International Inc. All rights reserved.
3 Honeywell Confidential by Honeywell International Inc. All rights reserved.
4 4 Laboratory reagent A single chemical or a mixture of chemicals dedicated for laboratory use Lab reagent should have: Guaranteed quality parameters: specified purity and specified impurities issued on CoA Proper package and label Defined shelf life/expiration date Honeywell Confidential by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved.
5 5 Impurities in lab reagents chemical physical biological primary secondary 2018 by Honeywell International Inc. All rights reserved.
6 6 Purity of lab reagent content of proper chemical expressed in % after deducting impurities Common names for purity grade of chemicals in Eastern bloc Latin name Abbreviation National name Abbreviation purum speciale pur. spec. zvláště čistý zv. č. purissimum puriss. chemicky čistý ch. č. pro analysis p. a. pro analýzu p. a. purum pur. čistý č. technicum techn. technicky tech. Raising purity Honeywell Confidential by Honeywell International Inc. All rights reserved. Based on standards from Comecon (COuncil for Mutual ECONomic assistance) 2018 by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved.
7 7 Honeywell Confidential by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved.
8 8 No common law or obligatory standards Producer assigns the grade of chemicals at its own preference Internationally used descriptions of grades: Crud. tech., techn. Laboratory grade puriss. pure, purum C.P. (chemically pure) GR (guaranteed reagent) Synthesis grade extra pure reagent grade Pro analysis Honeywell Confidential by Honeywell International Inc. All rights reserved. Comparing chemicals only on the grade name is impossible 2018 by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved.
9 9 International standards European Pharmacopoeia (Ph. Eur.) United States Pharmacopeia (USP) ACS specifications 11-th edition contains monographies for over 500 reagents Honeywell Confidential by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved by Honeywell International Inc. All rights reserved.
10 10 ACS reagent or ACS grade is a guarantee that reagent meets some minimal parameters written in ACS specification. ACS grade is currently the worldwide standard for laboratory use by Honeywell International Inc. All rights reserved.
11 11 Honeywell s contribution to ACS (11th edition) 2018 by Honeywell International Inc. All rights reserved.
12 12 Special grades For spectroscopy UV-VIS For liquid chromatography (HPLC, LC-MS) For gas chromatography For inorganic trace analysis (ASA, ICP) For semiconductors (VLSI, ULSI grade) For DNA/RNA/peptide synthesis Anhydrous For molecular biology (PCR) For cell culture 2018 by Honeywell International Inc. All rights reserved.
13 13 Key impurities UV-VIS absorbing Volatile solvents (HS) Non-volatile GC UV-VIS spectroscopy HPLC Solid particles UV-VIS absorbing Application HPLC Water synthesis AAS/ICP LC-MS Solid particles and nonvolatiles Alkali ions Suppressing ionization Metallic cations and inorganic anions
14 14 For spectroscopy UV/VIS Requirements for the method: UV/VIS absorbing contaminants as low as possible Detailed specifications for absorbance Advantage: More sensitivity and lower detection limit Reproducible spectrum 2018 by Honeywell International Inc. All rights reserved.
15 For liquid chromatography (HPLC) 15 Degassed mobile phase Requirements for the method: High purity Degassed mobile phase No solid particles (filter 0.2 um) Low absorbance (UV-VIS detection) Low fluorescence (fluorescence detection) Low metal content and non-volatile residue (MS detection, ICP-MS) Advantage: Longer column and instrument durability Repeatable analyses Lower detection limit Better spectra No particles 2018 by Honeywell International Inc. All rights reserved.
16 UV-VIS detection 16 Impurities absorbing UV-VIS light: Raising baseline in gradient elution mode (baseline drift) UV Abs. Gradient elution contaminated solvent Less sensitivity and higher uncertainty of the method Gradient elution Isocratic elution Retention time Reagent grade HPLC grade 2018 by Honeywell International Inc. All rights reserved. Method: HPLC Chromatograms at 200 nm for acetonitrile. A Waters 2695 HPLC with a B&J OD5 (C- 18) (#9575) 25cm X 4.6mm, 5um column was used for this experiment. The flow rate was 1mL/min. Gradient water/acetonitrile.
17 UV Absorbance (abs) UV Absorbance at 225nm (abs. units) Comparison of acetonitrile: reagent grade and HPLC grade 17 0,99 0,49-0, Wavelength (nm) Lot Higher UV absorbance in range nm indicates organic impurities Low and consistent UV absorbance results for different batches of HPLC grade- enable achieving higher sensitivity and good repeatability 2018 by Honeywell International Inc. All rights reserved.
18 18 MS detection Impurities can: destroy equipment suppress/alter ionization in the source create adducts Very high purity is required and very low impurities like: UV-VIS absorbing non-volatiles alkali metal ions and other compounds impairing ionization 2018 by Honeywell International Inc. All rights reserved.
19 19 ( and many metal ions) 2018 by Honeywell International Inc. All rights reserved.
20 20 Purity grades of HPLC solvents from Honeywell 20 LC-MS Ultra CHROMASOLV LC-MS CHROMASOLV CHROMASOLV Plus CHROMASOLV gradient CHROMASOLV 2018 by Honeywell International Inc. All rights reserved.
21 21 Borosilicate glass bottles Intens. 800 LCMS Ultra UVMSneg_RD1_01_3235.d: -MS, min #( ), -Constant Bkgrnd LC-MS Ultra grade (borosilicate glass) Chrom Gradient Grade UVMSneg_RD1_04_3232.d: -MS, min #( ), -Constant Bkgrnd Sodium formate adducts Gradient grade (amber glass) m/z Chromasolv LC-MS Ultra Alkali ions leaching from the glass
22 22 Modern techniques quality really matters! Problem: UV mode both solvents gives the same result and the same sensitivity MS mode invisible to UV impurities impair ionization and as a result sensitivity is worse Intens. Intens. x10 5 Competitor R (green) LC-MS Ultra Acetonitrile (blue) LC-MS Ultra Acetonitrile (magenta) 15 H 3 C 1.5 H 3 C CH 3 H Competitor R (orange) CH 3 H 10 H 1.0 H CH2 CH2 5 HO 0.5 HO Vitamin D 3 Vitamin D Time [min] Vitamin Chromatogram, Vitamin Chromatogram, 254 D3 254 nm D3 Test_BB1_03_6848.d: UV 254 nm D3 Test_BB1_02_6847.d: UV nm Vitamin Test_BB1_01_6849.d: UV Chromatogram, Vitamin D3 Test_BB1_02_6850.d: UV Chromatogram, 254 nm Vitamin D3 Test SpS_BB1_06_6857.d: UV Chromatogram, 254 nm Vitamin D3 Test SpS_BB1_05_6856.d: UV Chromatogram, 254 nm Vitamin D3 Test SpS_BB1_04_6855.d: UV Chromatogram, 254 nm Time [min] Vitamin D3 Test_BB1_03_6848.d: BPC All MS Vitamin D3 Test_BB1_02_6847.d: BPC All MS Vitamin D3 Test_BB1_01_6849.d: BPC All MS Vitamin D3 Test_BB1_02_6850.d: BPC All MS Vitamin D3 Test SpS_BB1_06_6857.d: BPC All MS Vitamin D3 Test SpS_BB1_05_6856.d: BPC All MS Vitamin D3 Test SpS_BB1_04_6855.d: BPC All MS Vitamin D3 Test SpS_BB1_03_6854.d: BPC All MS Advances in modern instrumentation and analytical chemistry force us to create new grades of reagents with higher purity and tougher specifications
23 ICP-MS detection For speciation analysis 23 For analyses by LC-ICP-MS Solvent line TraceSELECT Extremely pure the group combines both features of HPLC and ICP grades Metal traces at ppb level or lower
24 For GC (gas chromatography) 24 Mobile phase is gas, not liquid mixture of solvents. Different problems with contaminants. Reagents are used for sample preparation and for injection. Thus solvents' impurities are then the constituents of the sample. They are not delivered continuously as in HPLC. Impurities are undergoing chromatographic separation on GC column and their peaks are visible on chromatograms. Impurities can be multiply enriched during sample preparation If retention time of impurities is close to of our analytes problem! Direct injection GC Headspace
25 25 GC grade solvents (for pesticide residue analysis) Developed specifically for residue analysis of pesticides Suitable for the analysis of polychlorinated biphenyls (PCBs) Tested for suitability in pesticide residue analysis against 5 ng/l lindane or heptachlor Distinguishing GC grade Very low content of non-volatile matter (max %) improves column lifetime Meet requirements for both extraction steps and for GC injection Consistently high purity for wide range of GC applications
26 Intensity (ECD Counts) Intensity (ECD Counts) Comparison of methanol reagent grade and GC grade Time (min) Note the solvent front interference and many large contamination peaks in standard grade methanol are absent in the purified methanol. Common impurities in methanol: 2-butanone (MEK), 3-pentanone, N, N-dimethyl acetamide, and 2,3-butanedione. These impurities are not detected by GC-MS in purified methanol Time (min) Method: 200mL of methanol was concentrated (400x) and exchanged into hexane. The data was obtained on an Agilent 6890N GC with an ECD detector and a Restek Rtx-5 30 meter column. Sample injection = 5 microliters. The initial oven temperature was held at 200C for 15 minutes then increased by 10C/min until 250C, then held for 10 minutes.
27 Inorganic trace analysis Requirements for AAS/ICP reagents : High purity versus metals and ionic traces High quality packaging from proper organic polymer TraceSELECT line Purified by subboiling distillation Up to 80 cation and anion impurities tested High quality package: HDPE for TraceSELECT PFA (fluoropolymer) bottles fortraceselect Ultra
28 28
29 Water Concentration (ppm) Lot-to-lot variation Lower grades can be very good against one parameter in specific lot, but we do not have guarantee it will repeated in next batch. Only higher grades with tighter specification on specific parameter give us such guarantee and lot-to lot consistency Lot Reagent grade and HPLC grade acetonitrile
30 Packaging Safe storing and handling Ensures product stability Isolation from light, air Inert Not dissolving the package material and its contaminants (bottles, drums, caps, septa) High quality package is very important for high quality reagents and significant part of production cost Ensures special function Fixanal VOLPAC Moisture absorption
31 General rules - summary 31 There are different grades of lab reagents but there is one universal and the best one for all kinds of applications. Each technique has different requirements. Application test in product specification confirms that product was tested for this technique. The higher purity, tighter specification as well as unusual package the higher the cost of the lab reagent. When choosing lab reagent one needs to analyze specification for parameters important in the application. There is no need to overpay irrelevant parameters. The lower the grade the higher the lot-to-lot variability and less repeatability in results*. Technical grades can be used in fact therefore for teaching classes and for some limited applications. In research/analytical work one needs to choose at least: reagent grade, pro analysis, puriss. or ACS grade. Only ACS grade is really comparable this one is good to choose for scientific work - to enhance reproducibility of published studies.
32 Honeywell Confidential by Honeywell International Inc. All rights reserved. Thank you
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