Analysis was conducted using a PerkinElmer series 200 HPLC system comprising of 785 A UV–vis detector, series 200 quaternary pump and series 200 autosampler (PerkinElmer Inc., UK), Hamilton PRP-1 reversed phase column, 150 mm × 4.1 mm, 10 μm (part number: 79425) and data acquisition software (Peaksimple, version 4.09, SRI Inc., USA). Analysis of riboflavin was achieved with a run time of 5 min using the method developed by us. Isocratic conditions were used at 25 °C with the mobile phase comprising of 20% ethanol and 80% ion-pair buffer, flow rate 0.8 mL min−1, 10 μL injection volume, UV detector at 267 nm corresponding to riboflavin absorption λmax, a retention time of 3.05 min. Quantitation was achieved by reference to a calibration curve produced from riboflavin standards in PBS, pH 7.4 ± 0.2 at concentrations ranging from 0.01 to 25 μg mL−1 (r2 = 0.9996).
785 a uv vis detector
Manufactured by PerkinElmer
Sourced in United Kingdom
The 785 A UV–vis detector is a laboratory instrument used for spectroscopic analysis. It is designed to measure the absorption of ultraviolet and visible light by samples, providing information about their chemical composition and concentration.
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4 protocols using 785 a uv vis detector
1
Quantification of Riboflavin by HPLC
2
Purification and Fractionation of AsPH
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AsPH (20 mg/mL) was fractionated by centrifugal ultrafiltration (Vivaspin 20, Sartorius) using molecular weight (MW) cut-off of 5 kDa and 3 kDa, respectively, and 3 fractions were obtained: AsPH-F1 (MW > 5 kDa), AsPH-F2 (3 kDa < MW < 5 kDa), and AsPH-F3 (MW < 3 kDa). After bioactivity test (see below), the bioactive AsPH-F3 (20 mg/mL) was further separated on a Sephadex G-25 gel filtration chromatography column (1.2 cm × 150 cm), which was eluted with deionized water at a flow rate of 1 mL/min and monitored at 280 nm with a 785A UV/VIS detector (Perkin Elmer Co., Norwalk, CT, USA). The purified fraction was collected and freeze-dried as AsiPeps.
3
HPLC and Dissolution Analysis of Tablets
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Kontron analytical HPLC system with auto-sampler equipped with pump 422 was employed in this study. The HPLC detector was PerkinElmer 785A UV/Vis Detector with Powerchrome 280 software integrator. XBridge C18 with specifications 4.6 × 100 mm in length by the breadth and 3.5 μm pore size stationary phase column was used. The dissolution test was conducted using tablets dissolution test apparatus USP 30 standards. Tablet disintegration test machine by Tab-machines (Mumbai, India) was used for the disintegration test.
4
HPLC Analysis of Timolol Maleate
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HPLC analysis was conducted using a PerkinElmer series 200 HPLC system comprising of 785 A UV-vis detector, series 200 quaternary pump and series 200 autosampler (PerkinElmer Inc., UK), Ascentis C18 column, 150 mm × 4.6 mm, 5 μm (part number: 581324-U) and data acquisition software (Peaksimple, version 4.09, SRI Inc., USA). Analysis of timolol maleate was achieved with a run time of 5 min using the method adapted from (El-Kamel, 2002) . The mobile phase consisted of a mixture of methanol and triethylamine hydrochloride (45:55) under isocratic conditions, a flow rate was used at 1 mL min -1 at 30ºC and detected with a UV detector (295 nm). The retention time of timolol maleate was 2.85 min and the detection limit was 0.1 µM. Quantification of timolol maleate concentration in the samples was achieved by linear interpolation in a calibration curve of timolol maleate standards at concentrations ranging from 0.28 to 2.8 µg mL -1 .
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