Whatman no 1 chromatography paper

Individual samples of green and blue ash freeze-dried extracts (100 g) were each mixed with 200 g of polyvinylpolypyrrolidone (PVPP) powder (Sigma). The homogeneous mixture of either sample was then placed on top of a PVPP packed 4 L Buchner funnel. Elution of individual extracts was carried out at a slow rate initially with 5 L of water followed by 5 L aliquots of increasing concentrations (20%, 50%, 70% and 100%) of aqueous ethanol to produce five fractions. Each fraction was concentrated under vacuum and chromatographed one dimensionally on Whatman No.1 chromatography paper using one or more solvent systems: BAW (n-butanol-acetic acid-water, 4:1:5, upper phase), water, or acetic acid-water (15:85). Bands detected by absorbance/fluorescence under short wave light (254 nm) and long wave light (366 nm) were identified, cut and eluted with methanol. The samples were evaporated to dryness, dissolved in a minimum volume of methanol, and placed onto a PVPP column from which they were eluted sequentially with the following solvent systems: (i) CH₂Cl₂-EtOH-MeCOEt-Me₂CO (1:1:1:1), (ii) EtOH-MeCOEt-Me₂CO-H₂O (1:1:1:1) and (iii) EtOH-H₂O (1:1). Final clean up and purification of the compounds was achieved on a Sephadex LH-20 column (1 cm × 50 cm), using methanol as the eluting solvent.

All the chemicals were of analytical grade and purchased from Merck (Darmstadt, Germany)). Doubly distilled water was used throughout. Stock solutions containing 10 and 100 mg L⁻¹ of different metals (Bi(III), Cd(II), Pb(II), Sn(II), Zn(II), In(III), Cu(II), and Tl(I)) were prepared from 1000 mg L⁻¹ standard solutions after appropriate dilution with water. A stock 2.0 mol L⁻¹ acetate buffer (pH 4.5) was prepared from sodium acetate and hydrochloric acid.
The papers used were: Macherey-Nagel chromatography paper MN 261, Whatman No. 1 chromatography paper, and Whatman No. 42 filter paper. The marker pens that were investigated were commercial and purchased from a local stationery stop: Staedtler permanent Lumocolor waterproof 0.4 mm (s) black, the Edding 300 permanent marker water-resistant 1.5–3 mm, the Grand Paint Marker Olejowy paint marker GR-25 1.8 mm, the Edding 780 0.8 mm (black), and the BIC Marking Pro ultra-resistant permanent marker 1.1 mm. Graphite inks (Loctite EDAG PF 407A or Loctite EDAG 423SS) were a gift from Henkel Belgium.

Whatman No. 1 chromatography paper, bovine serum albumin (BSA), rabbit IgG, anti-rabbit IgG (alkaline phosphatase conjugated), anti-rabbit IgG (fluorescein isothiocyanate conjugated), 3,3′,5,5′-tetramethylbenzidine (TMB) (99%), BCIP®/NBT tablets, Tween® 20, 10 × phosphate buffered saline (PBS), and potassium periodate were purchased from Sigma-Aldrich (Oakville, ON, Canada), and used as received without further purification. Recombinant rat TNF-α, rat anti-TNF-α antibody, and horseradish peroxidase (HRP)-conjugated streptavidin were purchased from Abcam (Toronto, ON). Biotinylated anti-Rat TNF-α was purchased from BioLegend (San Diego, CA). The white LED and polyolefin (PO) were purchased from Digi-Key Cooperation (Thief River Falls, MN). The Arduino UNO microcontroller and the 16 × 2 LCD were purchased from RobotShop Inc. (Mirabel, QC). The Pyralux® (LF7062) copper-coated polymide film was obtained as sample from DuPont (Research Triangle Park, NC, USA). Ferric chloride was purchased for etching the copper from MG Chemicals (Burlington, ON, Canada). The RGB color sensor (TCS34725) was purchased from Adafruit (New York, NY, USA). The scotch plastic thermal laminating pouches were purchased from 3 M (Milton, ON, Canada).

Acetone was used as the initial mobile phase in the development of ascending paper chromatography (APC) using strips of Whatman No. 1 chromatography paper (12 cm length and 1 cm width) (Whatman International Ltd., Maidstone, Kent, UK). The radiochemical yield (RCY) of ^99mTc-ORL-NC was evaluated using acetone as a mobile phase to determine the free ^99mTcO₄⁻ percent. One spot of the reaction was applied 2 cm away from the base of the strip, which then developed in a jar containing a few drops of acetone. After developing, the strip was dried and sliced into 1 cm pieces and then counted using a NaI (Tl) γ-ray scintillation counter. The ^99mTc-ORL-NC complex was calculated using the formula below.
Radiopharmaceutical purity (RCP):