The target chemical (MT or DHT) was extracted from weekly-collected water samples (250 mL) from day 0 (before egg-masses were exposed) by solid phase extraction (SPE) using C18 SPE cartridges (Sep-pak Plus, Waters Ltd. UK) as per the manufacturer’s instructions. Sample extracts (double distilled water ‘blanks’, MT/DHT ‘spiked’ double distilled water and tank water samples) were eluted with analytical grade methanol (Fisher scientific, UK) and evaporated under a stream of nitrogen. Dried extracts were re-suspended in analytical grade ethanol (Hayman, UK) and stored at 4°C until further analysis using a recombinant Yeast androgen screen (YAS) [35 (link)]. Briefly, tank water (and blanks and spiked water) extracts were serially diluted in the assay plate and assayed against a DHT or MT standard curve (DHT starting concentration 1 x 10−6 M and MT 6.6 x 10−7 M) thus allowing tank water concentrations (expressed as ng DHT equivalent/L or ng MT equivalent/L) to be determined (taking in account the concentration factor of the SPE method).
Analytical grade methanol
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
Analytical grade methanol is a high-purity solvent used in various analytical and laboratory applications. It is a colorless, volatile, and flammable liquid with a characteristic odor. Analytical grade methanol meets stringent purity specifications, ensuring consistent performance in sensitive analytical procedures.
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Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Hplc grade acetonitrile, by ITW Reagents (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Lc ms grade acetonitrile, by Thermo Fisher Scientific (1 mentions)
Citric acid, by Merck Group (1 mentions)
Catechin, by Merck Group (1 mentions)
Milli q water system, by Merck Group (1 mentions)
Chloroform, by Thermo Fisher Scientific (1 mentions)
Epicatechin, by Merck Group (1 mentions)
L tryptophan, by Merck Group (1 mentions)
L phenylalanine, by Merck Group (1 mentions)
L isoleucine, by Merck Group (1 mentions)
L tyrosine, by Merck Group (1 mentions)
L leucine, by Merck Group (1 mentions)
Caftaric acid, by Biosynth (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Ammonium acetate, by Thermo Fisher Scientific (1 mentions)
Acetone, by Thermo Fisher Scientific (1 mentions)
8 protocols using analytical grade methanol
1
Quantifying Androgen Levels in Water
2
Bovine Hemin Porphyrin Extraction
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Type I bovine hemin (chloroferriprotoporphyrin IX) and formic acid were obtained from Sigma-Aldrich (USA). Hydroxyapatite and glacial acetic acid was purchased from BDH Chemical Ltd (England). DMSO (99% purity) was purchased from Merck (Germany). Analytical grade methanol was purchased from Fisher Scientific (USA) and acetonitrile HPLC grade was purchased from Panreac (Spain). H2O was deionized using Ultra Clear UV plus TM water system (SG water, Germany) to the quality of 18.2 MOhm.
3
Polyphenol Standards Acquisition Protocol
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LC-MS grade acetonitrile and formic acid, analytical grade methanol and chloroform were all purchased from Fisher Scientific (Illkirch-Graffenstaden, France). Ultrapure water was purified from a Milli-Q water system (Merck Millipore, Molsheim, France). Pure standards of catechin, epicatechin, L-phenylalanine, citric acid, L-tryptophan, L-tyrosine, L-isoleucine, L-leucine and gallic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Procyanidins B1, B2, B3 and C1 were supplied by Extrasynthese (Genay, France), procyanidin B4 and caftaric acid were obtained from Carbosynth (Compton, Berkshire, UK).
4
Quantitative Analysis of Azole Antifungals
5
Evaluation of Cathinone Metabolism
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All SCts utilised in this study were kindly provided by TicTac Communications (London, UK) and were test-purchased from the internet. All four cathinones appeared to be highly pure when checked as described by Apirakkan et al. [6 (link)]. HPLC-grade acetonitrile, analytical-grade methanol and formic acid were obtained from Fisher Scientific (Loughborough, UK). Ultra-pure water (18.2 MΩ.cm) was obtained from an Elga Purelab Flex (High Wycombe, UK). Phosphate buffer (0.5 M, pH = 7.4), Ultrapool™ human liver microsomes (HLM), NADPH regeneration system solutions A and B, and UDP Reaction Mix Solution A and B were purchased from Corning (Wolburn, MA, USA). 3′-Phosphoadenosine 5′-phosphosulphate (PAPS) was purchased from Merck (Damstadt, Germany).
6
HPLC Solvent Preparation and Filtration
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HPLC grade formic acid was purchased from Merck (Darmstadt, Germany), whereas analytical grade methanol was purchased from Fisher scientific (Leicestershire, UK), Deionized water or HPLC grade water was obtained from Millipore Sigma (Burlington, MA, USA). A 0.45 mm disposable membrane filter was purchased from Cronus Filter (UK).
7
Standardized QDTS Herbal Granules: Extraction and Analysis
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QiDiTangShen granules, whose patent is currently under review (Patent Accepting No. 201410436459.6) from the State Food and Drug Administration of China, contains seven Chinese herbs, including Astragali Radix (Astragalus membranaceus Fisch., Inner Mongolia Autonomous Region), Rehmanniae radix praeparata (Rehmannia glutinosa Libosch., Henan Province), Euryale Semen (Euryale ferox Salisb., Hunan Province), Sophorae Tonkinensis Radix et Rhizoma(Sophora tonkinensis Gagnep., Zhejiang Province), Hirudo (Whitemania pigra Whitman, Guangdong Province), Rhei Radix et Rhizoma (Rheum palmatum L. Qinghai Province) and Baihuasheshecao (Hedyotis diffusa Willd., Fujian Province). The extract of QDTS was provided by Zhuozhoudongle Pharmaceutical Co., Ltd. (Hebei, China). The doses of QDTS and valsartan (Beijing Novartis Pharmaceutical Co., Ltd., Beijing, China) for mice were 3.37 g/kg/day and 10.29 mg/kg/day, respectively, which were acquired at a ratio with the regular human adult dosage (Gao et al., 2018 (link)).
Acetonitrile, analytical grade methanol, and formic acid (HPLC grade) were purchased from Thermo Scientific (Waltham, MA, United States). Ultra-pure water (18.2 MΩ⋅cm) was from a Milli-Q system (Millipore, Bedford, MA, United States). Polytetrafluoroethylene (PTFE) membranes of 0.45 micron used for processing samples were from ANPEL (Shanghai, China).
Acetonitrile, analytical grade methanol, and formic acid (HPLC grade) were purchased from Thermo Scientific (Waltham, MA, United States). Ultra-pure water (18.2 MΩ⋅cm) was from a Milli-Q system (Millipore, Bedford, MA, United States). Polytetrafluoroethylene (PTFE) membranes of 0.45 micron used for processing samples were from ANPEL (Shanghai, China).
8
Potato Growth and Metabolite Extraction
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Plant materials, bacterial strains, and chemicals.
S. tuberosum (DM1) and S. chacoense (Hassl.) (M6) were grown in a greenhouse at 18 to 24°C with a 16-h day length. Plants were grown in ProMix Bx General Purpose mix, fertilized with Osmocote Plus 15-9-12 (Scotts-MiracleGro, U.S.A.), and irrigated to saturation every other day until used for assays. Aphids and other pests were managed with Botaniguard ES and Molt-X (BioWorks, U.S.A.), Entrust SC (Corteva, U.S.A.), Distance IGR (Valent Biosicences, U.S.A.), Judo and Azatin (OHP Inc., U.S.A.), Avid 0.15EC (Syngenta, U.S.A.), and Compass (Bayer, U.S.A.)
Tubers and 1-month old stems were used for extraction of metabolites. LC-MS-grade water, analytical-grade methanol, ACN, and hydrochloric acid were purchased from Fisher Chemicals (Thermo Fisher Scientific, U.S.A.). P. brasiliense Pb1692 was used for all experiments in this study (Duarte et al. 2004) . Nutrient broth (NB), agar, skim milk powder, tryptone, Congo red, and NaCl were purchased from Difco Laboratories (Thermo Fisher Scientific). Bacteria were grown at 30°C under continuous shaking conditions.
S. tuberosum (DM1) and S. chacoense (Hassl.) (M6) were grown in a greenhouse at 18 to 24°C with a 16-h day length. Plants were grown in ProMix Bx General Purpose mix, fertilized with Osmocote Plus 15-9-12 (Scotts-MiracleGro, U.S.A.), and irrigated to saturation every other day until used for assays. Aphids and other pests were managed with Botaniguard ES and Molt-X (BioWorks, U.S.A.), Entrust SC (Corteva, U.S.A.), Distance IGR (Valent Biosicences, U.S.A.), Judo and Azatin (OHP Inc., U.S.A.), Avid 0.15EC (Syngenta, U.S.A.), and Compass (Bayer, U.S.A.)
Tubers and 1-month old stems were used for extraction of metabolites. LC-MS-grade water, analytical-grade methanol, ACN, and hydrochloric acid were purchased from Fisher Chemicals (Thermo Fisher Scientific, U.S.A.). P. brasiliense Pb1692 was used for all experiments in this study (Duarte et al. 2004) . Nutrient broth (NB), agar, skim milk powder, tryptone, Congo red, and NaCl were purchased from Difco Laboratories (Thermo Fisher Scientific). Bacteria were grown at 30°C under continuous shaking conditions.
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