A high performance liquid chromatography (HPLC) method was used for determination of HbA1c. The HPLC consisted of a Waters 625 LC system together with a Waters photo-diode-array detector model 996 and a WISP 717 auto sampler for automatic injection of the samples. Millennium chromatography software was used for calculation of concentrations (Waters Associates Inc., Milford, United States). A cation exchange column Mono S HR 5/5 from Pharmacia Biotech AB, Uppsala, Sweden was used to separate HbA1c from other components in the samples [49 (link)]. HbA1c is a standard indicator of long-term glycaemic control and an HbA1c level of ≥5.6% corresponds to a fasting glucose level of ≥100 mg/dL, which is a measure of insulin resistance [50 (link)].
Millennium chromatography software
Manufactured by Waters Corporation
Sourced in United States
Millennium chromatography software is a data management and analysis tool developed by Waters Corporation. The software is designed to facilitate the collection, processing, and reporting of data generated from chromatography instruments. It provides a user-friendly interface for managing and interpreting chromatographic data.
Automatically generated - may contain errors
Lab products found in correlation
625 lc system, by Waters Corporation (1 mentions)
Dca 2000 analyzer, by Bayer (1 mentions)
Waters 717 plus autosampler, by Waters Corporation (1 mentions)
Waters 486 tunable absorbance detector, by Waters Corporation (1 mentions)
L9879, by Merck Group (1 mentions)
Nova pak, by Waters Corporation (1 mentions)
3 protocols using millennium chromatography software
1
HPLC Determination of HbA1c for Glycemic Control
2
Assessing Metabolic Biomarkers in Clinical Trials
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The serum levels of glucose, insulin, and lipids were analyzed at the University of Oklahoma Medical Center laboratory (Oklahoma City, OK, USA) and at Quest Diagnostics (Las Vegas, NV, USA) according to the manufacturer’s protocols. Serum glycated hemoglobin was analyzed with the use of a DCA 2000+ Analyzer (Bayer, Leverkusen, Germany). Insulin resistance was evaluated by HOMA-IR and was calculated as follows: [fasting insulin (mU/L) × fasting glucose (mmol/L)]/22.5 [47 (link)]. In addition, sera were stored at −80 °C for the subsequent analyses of metabolic profiling. Plasma ellagic, acid as a measure of compliance, was measured using a previously published procedure [48 (link)]. Briefly, 500 µL of plasma was treated with acetonitrile and the resulting supernatant was evaporated, reconstituted with methanol, and injected into an HPLC system. Ellagic acid was eluted using a C18 column and quantified using the Millennium Chromatography software (Waters, Milford, MA, USA).
3
Quantifying Lycopene in Liver and Egg
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LP contents in liver and egg yolk were extracted using the method of Boileau et al. (2000) (link) and analyzed by the method of Wei et al. (2001) (link) using high performance liquid chromatography (HPLC). In brief, approximately 0.1 g of liver tissue or egg yolk was homogenized thoroughly, dissolved in 6 ml of a potassium hydroxide/ethanol (1:5) solution containing 1 g/l butylated hydroxytoluene (BHT) and saponified at 60°C for 30 min. LP was extracted twice under yellow light using equal volumes of hexane (6ml) plus 2ml of distilled water. The extracts were dried and stored at −C20°C for no longer than 2 d before LP measurement by HPLC. The HPLC system included Waters 510 pumps, a Waters 717 plus auto sampler, a Waters 486 Tunable Absorbance detector, and Waters Nova-Pak (5 µm, 3.9 cm×300 mm) C18 column. The mobile phase was methanol:acetonitrile:chlorform (47:47:6, v/v/v) and the flow rate was 1.0 ml/min. The HPLC was controlled by Waters Millennium chromatography software and the lycopene peak was monitored at 472 nm. LP concentration was calculated using a calibration curve prepared with the pure LP standard (L-9879, Sigma Co., St. Louis, MO, USA).
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