Simultaneous determination of glucose, 1,5-anhydrod-glucitol and related sugar alcohols in serum by high-performance liquid chromatography with benzoic acid derivatization

Abstract

A new, simple and sensitive pre-column high-performance chromatographic method for the determination of diabetes marker d-glucose, 1,5-anhydro-d-glucitol and related compounds is reported. Sugars (d-glucose, d-galactose, d-mannose, sucrose and arabinose) were derivatized with benzoic acid (BA) at 80 degrees C for 60 min. l-Fucose, fructose, d-lactose, l-rhamnose, arabinose and ascorbic acid were not reacted. Sugar alcohols (xylitol, erythritol, mannitol, sorbitol myo-inositol) were also derivatized with BA at 80 degrees C for 60 min. The fluorescence derivatives were separated on a TSK amide 80 column (4.6 mm i.d. x 250 mm, 5 microm) with acetonitrile-50 mm acetate buffer (pH 5.6; 4:96, v/v) as the mobile phase. The detection wavelength of beizoic acid derivatives was lambda(ex) 275 nm and lambda(em) 315 nm. The detection limits of sugars were 10-80 microg/mL. The calibration graphs were linear up to 10 mg/mL. The relative standard deviations of 500 microg/mL sugars were 7.0-7.3%. The proposed method was compared with the enzymatic photometric glucose analysis method (Glucose B-Test II Wako). The correlation coefficient was 0.83 (n = 20) and y = 0.82x + 5.91, where y and x are concentrations in microg/mL obtained by the proposed pre-column HPLC and enzyme-photometric method, respectively. The detection limits of sugar alcohols were 100-1000 ng/mL. The calibration graphs were linear to 50 microg/mL and relative standard deviations of 10 microg/mL were 7.2-8.2%. The 1,5-AG data by the proposed method was also compared with the enzymatic photometric 1,5-AG analysis method (Rana AG 1,5-AG determination kit, Nihon Kayaku) and good correlation (r = 0.91, n = 20) was also obtained. The proposed method was applied to the simultaneous determination of d-glucose, 1,5-AG and related sugar alcohols in serum from healthy males.

Determination of glycemic monitoring marker 1,5-anhydroglucitol in plasma by liquid chromatography-electrospray tandem mass spectrometry

“…To reduce the contamination to the ion source of mass spectrometer, the supernatant was further diluted five times and a small volume (5 L) was injected. Unlike the existing methods including time-consuming derivatization protocols [28,29] or multiple clean-up procedures [26], the present method involves an one-step and cost-effective process.…”

“…Likewise, the results can easily be affected by endogenous interference such as glucose, conjugated bilirubin and hemoglobin [25], myo-inositol [26], maltose [27]. Therefore, other methods for plasma/serum or urine 1,5-AG measurement using chromatography techniques such as HPLC, GC/MS and LC/MS were reported [26,[28][29][30]. These methods may involve either time-consuming derivatization or laborious sample preparation procedures.…”

Simultaneous determination of two Amadori compounds in Korean red ginseng (Panax ginseng) extracts and rat plasma by high-performance anion-exchange chromatography with pulsed amperometric detection

“…Additionally, HPLC method with a refractive index detector has been tried for analyzing Amadori compounds, but it also has problems such as poor sensitivity and insufficient separation [18]. The method of pre-or post-column derivatization and UV or fluorometric detection by HPLC has been widely used for sugar analysis [19][20][21][22] and it has been applied to analyze Amadori compounds. For example, Ryu et al tried to analyze for Arg-Fru in aged garlic extract by HPLC and post-column reaction with triphenyltetrazolium chloride [23].…”

Development and validation of a liquid chromatography–tandem mass spectrometric method for the quantification of 5-thio-d-glucose in rat and human plasma

“…In general, methods for detecting monomeric sugars include differential refractive index detection [12], fluorescence detection [13], and electrochemistry detection (pulsed amperometric detection [14,15]). However, the analytical performances of these methods often yield unsatisfactory results because of the lack of selectivity or sensitivity.…”