The assay of the different tablet preparation was performed using HPLC, and the HPLC system used was a Beckman Gold system, prepared with a detector set at 254 nm, and a UV-vis spectrophotometer was used to evaluate the maximum absorption wavelength for both medicines. Standard solutions of approximately 50 mg pyridoxine hydrochloride and 25 mg meclizine hydrochloride were precisely weighed and dissolved in a 100 mL mobile phase. Sample solutions tablets were ground and mixed well. The amount is equivalent to 50 mg pyridoxine hydrochloride, and 25 mg meclizine hydrochloride was weighed and moved to a 100 mL volumetric flask. A mobile phase was added, and the solution was shaken for 15 min. The volume was made up to 100 mL with a mobile phase, and the solution was centrifugated. The column packed with a C18 silica gel, a flow rate of 1.5 mL/min, and a mobile phase (water, methanol, and acetonitrile (4 : 3 : 2)) was used. The resulting solutions were shaken for 30 min, and the volume was made up with the mobile phase. The concentration was determined in triplicate, and the % of the assay was determined.
Gold system
Manufactured by Beckman Coulter
The Gold system is a laboratory equipment product offered by Beckman Coulter. It is designed to perform specific functions within a laboratory setting. The core function of the Gold system is to provide accurate and reliable measurements, but a detailed description of its intended use cannot be provided in an unbiased and factual manner without the risk of extrapolation.
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Lab products found in correlation
4 protocols using gold system
1
Quantitative Analysis of Pyridoxine and Meclizine in Tablets
2
Enzymatic Synthesis of GlcA-Xylose Disaccharide
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The GlcA-β1,4-xylose-β-MU disaccharide (B4GAT1 product) was added to 10 mM of UDP-Xyl in 50 mM sodium acetate buffer at pH 5.5 and with 10 mM MgCl2, 10 mM MnCl2, 0.5% TX-100 and LARGEdTM attached to metal-affinity resin and incubated for 48 hr at 37°C with rotation. The sample was then run over a LC18 column (4.6 × 250 mm Supelcosil LC-18 column (Supelco)) with Buffer A (50 mM ammonium formate pH 4.0) and Buffer B (80% acetonitrile in buffer A) using a 16% B isocratic run at 1 ml/min on a Beckman Gold system. The elution of MU derivatives was monitored by fluorescence detection (325 nm for excitation, and 380 nm for emission). The trisaccharide peak was collected and lyophilized. The lyophilized sample was then brought up in 10 mM UDP-GlcA in 50 mM MOPS buffer pH 6.0, 10 mM MgCl2, 10 mM MnCl2 and 0.5% TX-100 and incubated for 48 hr at 37°C with rotation. It was again run on a C18 column with 16% B isocratic run. The product peak fraction was then collected and lyophilized. The dried sample was brought up in Milli-Q water (500 µl) and lyophilized. This procedure was repeated a total of three times. The last time the sample was brought up in Milli-Q water and the product was quantitated using a standard curve of GlcA-β-MU. This sample was used for NMR studies.
3
GlcNAc Transferase Activity Assay
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The test B4GAT1dTM for GlcNAc transferase activity Gal-β1,4-GlcNAc-β-MU (0.1 mM) was used as acceptor. The 50 µl enzyme reactions were carried out as described previously (Sasaki et al., 1997 (link)) at 37°C, with 5 mM UDP-GlcNAc in 0.1 M cocodylate buffer (pH 7.0) supplemented with 20 mM MnCl2, 5 mM ATP and 0.25 µg B4GAT1dTM enzyme. The reaction was terminated by adding 25 µl of 0.1 M EDTA and boiling for 5 min. The supernatant was analyzed using a LC18 column (4.6 × 250 mm Supelcosil LC-18 column (Supelco)) with Buffer A (50 mM ammonium formate pH 4.0) and Buffer B (80% acetonitrile in buffer A), using a 16% B isocratic run at 1 ml/min using Beckman Gold system.
4
Quantification of Synaptosomal Adenosine Release
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Adenosine release was assayed essentially as previously described [38 (link)]. Briefly, hippocampal synaptosomes were prepared using Percoll/sucrose gradients [39 (link)] and resuspended in Krebs–HEPES solution (in mM: NaCl 113, KCl 3, KH2PO4 1.2, MgSO4 1.2, CaCl2 2.5, NaHCO3 25, glucose 10, HEPES 15, pH 7.4, 37 °C). Hippocampal synaptosomes (1.2–1.4 mg protein/mL) were then incubated at 37 °C for 5 min before adding different amounts of a concentrated KCl solution (5 M) to end up with final K+ concentrations of 3 (control, no additionally added K+), 10, 15, 30 or 60 mM. After 5 min, the mixtures were centrifuged at 14,000× g for 10 min at 4 °C, and the supernatant was used for the analysis of the extracellular levels of adenosine.
The separation and quantification of adenosine and its metabolites was carried out by HPLC, as previously described [40 (link)], employing a LiChroCart-RT125-4 C-18 reverse-phase column (particle size, 5 μm) combined with a UV detector set to 254 nm (Gold System, Beckman). The mobile phase was KH2PO4 (100 mM) and methanol (85/15 v/v%) at pH 6.50 with a flow rate of 1 mL/min and an injection loop volume of 50 μL. The identification and quantification of adenosine and its metabolites was achieved by calculating the peak areas then converted to concentration values (expressed as nmol/mg protein) by calibration with standards ranging from 0.1 to 50 μM.
The separation and quantification of adenosine and its metabolites was carried out by HPLC, as previously described [40 (link)], employing a LiChroCart-RT125-4 C-18 reverse-phase column (particle size, 5 μm) combined with a UV detector set to 254 nm (Gold System, Beckman). The mobile phase was KH2PO4 (100 mM) and methanol (85/15 v/v%) at pH 6.50 with a flow rate of 1 mL/min and an injection loop volume of 50 μL. The identification and quantification of adenosine and its metabolites was achieved by calculating the peak areas then converted to concentration values (expressed as nmol/mg protein) by calibration with standards ranging from 0.1 to 50 μM.
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