All reagents, reference standards and organic solvents were purchased from Merck (Darmstadt, Germany).
Reference standard
Manufactured by Merck Group
Sourced in Germany, United States
Reference standards are high-purity materials used to calibrate and validate analytical instruments and methods. They serve as points of reference to ensure the accuracy, precision, and reliability of analytical measurements.
Automatically generated - may contain errors
Lab products found in correlation
Organic solvents, by Merck Group (1 mentions)
Milli q system, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Agilent 1260 infinity autosampler, by Agilent Technologies (1 mentions)
Agilent 1260 diode array detector vl, by Agilent Technologies (1 mentions)
Zorbax sb c18, by Agilent Technologies (1 mentions)
6 protocols using reference standard
1
Merck Reagents and Solvents Protocol
2
Microwave Digestion and ICP-OES Analysis of Liver Al and Pb
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Each liver sample was microwave-digested with 8 mL nitric acid and 1 mL of 30% hydrogen peroxide. Then, the Al and Pb contents were determined by an inductively coupled plasma–optical emission spectrometer (ICP-OES, model 5100, Agilent, Santa Clara, CA) with synchronous vertical dual view (SVDV). The intensity calibration curve was developed for each set of measurements using a blank and three or more Merck Company standards (Germany). Reference standards from Merck were used to verify the accuracy and precision of the metal measurements. A quality control sample containing known concentrations of trace elements from the National Institute of Standards and Technology (NIST) was used to validate the instrument’s results.
3
Analytical Reagents and Standards Procurement
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Acetonitrile was purchased from Merck and Co. (Merck KGaA, Germany) and formic acid was obtained from Shanghai Nanxiang People Chemical Factory (China). All other solvents were of high-performance liquid chromatography grade. Reference standards were purchased from Sigma-Aldrich (St. Louis, MO, USA) and ultrapure water was prepared by Milli-Q system (Millipore Co., USA).
4
Silymarin Components Analysis by HPLC
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An Agilent 1260 Infinity High Performance Liquid Chromatography (HPLC) system equipped with an Agilent 1260 Infinity pump (G1361A), Agilent 1260 diode array detector VL (G1315D), Agilent ZORBAX SB-C18 and Agilent 1260 Infinity autosampler (G2260A) were used to analyze the content of individual silymarin components according to the reported method by (AbouZid et al., 2016b ). The mobile phase was composed of methanol/0.1% formic acid (Phase A) and water/0.1% formic acid (Phase B); 0–5 min gradient 40–45% phase A; 5–10 min isocratic 45% phase A; 10–20 min gradient 45–50% phase A; 20–25 min isocratic 50% phase A. The flow rate was 1.5 ml/min, and ambient temperature was used. The auto sampler was adjusted to inject 10 µl and quantified at 280 nm. The method offered the following retention times (minutes) for the major silymarin compounds: silychristin A (Rt = 8.9), silydianin (Rt = 10.6), silybin A (Rt = 19.6), silybin B (Rt = 21.3), isosilybin A (Rt = 25.4), and isosilybin B (Rt = 26.6). Flavonolignans identification and quantification was obtained using reference standards (Sigma-Aldrich). Total flavonolignan content is the result of the sum of the single constituents derived from HPLC analysis.
5
Identification of Thymol by TLC and GC-MS
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Colorless powder, 9 mg. The identity of thymol was confirmed by means of co-TLC with a reference standard (Sigma—Aldrich) and also by analysis of GC—MS data. In the TLC analysis, the Rf values of 14 and the reference standard were both 0.9 after elution with petroleum ether–ethyl acetate (6:4). The GC chromatogram displayed signal at 12.67 and 12.66 min, respectively, for 14 and the reference standard. GC—MS revealed the presence of [M+H]+ at m/z 150.00 for both compounds.
6
Quantification of Andrographolide in Medicinal Plants
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The reference standard procured from Sigma Company was first diluted into methanol then further serially diluted in mobile phase containing water (35%) and acetonitrile (ACN) (65%). For Sample preparation, the dried leaves powder of six different genotypes collected at 60, 90 and 120 DAP (days after plantation) were weighed 0.02 gm (20 mg) and extracted into methanol for 60 min in sonicator in 15 ml plastic tube. The tubes were transferred to water bath at 60 °C for 1 h. Samples were filtered using sterile syringe of 0.22 µm nylon membrane. From diluted filtrates, 5 µl was taken and diluted into 1 ml of 65% ACN: 35% water (Shende et al. 2016 ). The samples were stored at 4 °C in 2 ml glass vials till use. The percentage of andrographolide was calculated according to the following formula (Song et al. 2013; Shende et al. 2016 ).
Andrographolide (%) = (C × V × D)/10,000 × W; Where: C = Concentration (mg/L) of andrographolide in test solution; D = Dilution factor; V = Final make up volume (in ml) of the test solution and W = Weight (gm) of the sample used for the preparation of the test solution .
Andrographolide (%) = (C × V × D)/10,000 × W; Where: C = Concentration (mg/L) of andrographolide in test solution; D = Dilution factor; V = Final make up volume (in ml) of the test solution and W = Weight (gm) of the sample used for the preparation of the test solution .
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