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5 protocols using methanol and acetonitrile

1

Extraction and Analysis of Natural Dyes

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Indigo and alizarin (97%) were purchased from ACROS Organics; indirubin (≥98%), 9-aminoacridine (9-AA, ≥99.5%), erythrosin B (≥95%), 2-iodobenzoic acid (≥99%) and Universal MALDI matrix [1:1 mixture of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxy-cinnamic acid (α-CCA)] were obtained from Sigma-Aldrich, carminic acid (≥96%) from Fluka Analytical, lucidin from Cfm Oskar Tropitzsch GmbH (Marktredwitz), tetrahydrofuran (HPLC grade) from Roth, formic acid (99–100%), trifluoroacetic acid, ethanol, methanol and acetonitrile (all solvents HPLC gradient grade) from VWR, and TLC (thin-layer chromatography) plates (ALUGRAM Sil G) were purchased from Macherey-Nagel. Purpurin has been obtained from Aldrich’s collection of rare chemicals and rubiadin has been synthesized according to procedures of Takano et al.33 (link). Double distilled water was produced in a distillation apparatus bought from Gebr. Rettberg GmbH. An extended tabby weave with blue warps (wool fibers previously vat dyed using natural indigo) and red wefts (wool fibers previously mordant dyed using alum and cochineal) was handcrafted by a textile archaeologist. Recent animal- and plant-derived fibers dyed with synthetic indigo or E120, a food dye extract from scale insects (Dactylopius coccus Costa), were used for first tests (dyeing processes were performed as previously reported28 (link),34 (link)).
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2

Ceftriaxone Resistance in E. coli

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E. coli strains included the clinical strain Ec3536 collected from a urine sample of the community patient and provided from the MFP Laboratory collection10 (link). Its conjugative plasmid containing the blaCTX-M-15 gene was transferred by conjugation experiment in a laboratory recipient cell of E. coli K12. Consequently, the transconjugant E. coli TcK12 was resistant to CFX10 (link). Mueller-Hinton bacteria culture medium adjusted in calcium and magnesium ions (MH-CA) and microbiology consumable were purchased from Bio-Rad, France.
Ceftriaxone heptahemihydrate di-sodium salt was pourchased from Discovery Fine Chemical (UK), pharmaceutical grade, batch number: 74786.
Methanol and Acetonitrile (HPLC grade) was purchased from VWR (France).
Demineralized water was prepared at the laboratory by ion exchange (Pure Lab Option ELGA) followed by distillation (Water Still Distinction D4000).
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3

Analytical Characterization of Synthetic Cannabinoids

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MDMB-CHMICA was purchased from Chiron (Trondheim, Norway) and JWH-200-d 5 was purchased from
LGC Standards (Teddington, U.K.). Phosphate buffer (pH 6, 0.1M) was prepared in-house from disodium hydrogen orthophosphate anhydrous and sodium dihydrogen orthophosphate dihydrate from Fisher Scientific (Loughborough, U.K.) and deionised water produced from a Purite (Thame, U.K.) deionised water system.
Tertiary methyl butyl ether (tBME) and ammonium acetate were purchased from Sigma Aldrich (Gillingham, U.K.). Methanol and acetonitrile, both HPLC grade, and formic acid were obtained from VWR (Lutterworth, Leicestershire, U.K.). Blood products were purchased from the Scottish National Blood Transfusion Service (SNBTS) based at Gartnavel Hospital (Glasgow, U.K.).
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4

HPLC Reagent Preparation and Purification

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Acetonitrile and methanol (HPLC grade) and triethylamine were obtained from J.T. Baker® (Mexico City, MX, Mexico). Water was distilled and purified using Millipore Milli-Q Plus system (Bedford, MA, USA). Analytical grade phosphoric acid (H3PO4, 85%) was purchased from Merck® (Darmstadt, Germany). All other chemicals were of analytical grade with the highest purity available.
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5

Quantitative Analysis of Neurounina-1

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Ropivacaine was utilized as IS and was supplied by the United States Pharmacopeia (Rockville, MD, United States). Neurounina-1 was synthesized in house according to the previously reported study (Molinaro et al., 2013 (link)). Acetonitrile and methanol (HPLC grade) were purchased from J.T Baker (Phillipsburg, NJ, United States); ammonium acetate and hexane (analysis grade) from J.T Baker (Ecatepec, Mexico); ethyl ether (analysis grade) from Mallinckrodt (Phillipsburg, NJ, United States). The water was obtained from the purification system Synergy UV® (Millipore, Molsheim, France).
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