Mass Spectroscopy of the Antileukemic Drug MGBG and Related Bis(Amidinohydrazones) [‘Bis(Guanylhydrazones)’]

The first study on the mass spectroscopy of various bis(amidinohydrazones) is reported. The compounds studied included the investigational antileukemic drugs methylglyoxal bis(amidino-hydrazone) [‘methylglyoxal bis(guanylhydrazone)’, MGBG] and glyoxal bis(amidinohydrazone), as well as seven mono- and dialkylglyoxal analogs thereof. The results indicate that the free bases of these high-melting compounds are volatilized well enough to allow a facile detection of the molecular ions and to make mass spectroscopy of the underivatized compounds a suitable method for the verification of the identity of the substances. This result is of importance considering the development of novel analogs and derivatives. A compilation of electron-impact mass spectra is reported and possible fragmentation routes are outlined. The fragmentation of the various congeners appears to occur essentially similarly, the main paths involving breakage of the carbon-carbon single bond in the glyoxal moiety or breakage of either one of the nitrogen-nitrogen single bonds.     

Voltammetric detection of the α-dicarbonyl compound: Methylglyoxal as a flavoring agent in wine and beer

A simple, rapid and highly selective method for the determination of the most abundant α-dicarbonyl compound in wine and beer has been developed for the first time by employing square wave voltammetry. A novel electrochemical sensor, based on the electrodeposition of platinum nanoparticles onto single wall carbon nanotubes that were cast on a glassy carbon electrode (GCE) substrate, is presented in this paper. This modified electrode exhibited excellent catalytic activity in the electroreduction of methylglyoxal, showing much higher peak currents than those measured on an unmodified GCE. The effects of different experimental and instrumental parameters, such as solution pH and square wave frequency, were examined. The reduction peak current showed a linear range of from 0.1 × 10⁻⁶ to 100 × 10⁻⁶ M with a 0.9979 correlation coefficient; and a low detection limit of 2.8 × 10⁻⁹ M was also obtained. The proposed methodology was successfully applied to the quantitative analysis of methylglyoxal in wine and beer samples. The developed sensor possesses advantageous properties such as a high active surface area, stability, and rapid electron transfer rate, which cumulatively demonstrate high performance toward the electrocatalytic reduction and detection of methylglyoxal.     

Fourier Transform Infrared Spectroscopy of Aliphatic bis(Amidinohydrazones) and Their Deuterated Analogues

The first study on the infrared spectroscopy of the bis(amidinohydrazones) of various glyoxals is reported. The compounds studied include the antileukemic agents glyoxal bis(amidinohydrazone) and methylglyoxal bis(amidinohydrazone) (Mitoguazone) as well as seven mono-and dialkylglyoxal analogues thereof. Free bases as well as doubly protonated species (divalent salts) were investigated. Selectively deuterated analogues were also studied and were synthesized by exchanging nitrogen-bound hydrogen atoms for deuterium atoms. The effects of substituents, protonation and deuteration on the FT-IR spectra of the compounds are discussed.     

Substituent Effects in the Carbon NMR of Aliphatic bis(Amidinohydrazones) ['Bis(Guanylhydrazones)']. Construction of a set of Quantitative Empirical Rules. Unambiguous Assignment of Several Previously Unassigned Carbon Resonances

The effects of substituents on the ¹³C chemical shifts of the various carbons of aliphatic 1,2-bis(amidinohydrazones) have been systematically studied using previously published experimental data as the basis. Mathematical formulae have been constructed that describe the effects of various structural features of the molecules on the chemical shifts of the carbons and that also make possible an accurate prediction of the spectra of compounds belonging to this class. It is also shown that the effects of side chains on the chemical shifts of the two carbons of the glyoxal moiety are strictly additive. A mathematical model has been constructed that makes possible a very accurate prediction of the chemical shift of each one of the glyoxal carbons of symmetrical as well as unsymmetrical bis(amidinohydrazones). In the case of ethylmethylglyoxal bis(amidinohydrazone) free base dissolved in dimethyl sulfoxide, the theory predicts that the glyoxal carbons resonate at 157.45 ppm (the one connected to the ethyl group) and 151.21 ppm, while the experimental values are 157.30 and 151.29 ppm. This has, for the first time, made possible the unambiguous individual assignment of the resonances of the glyoxal carbons of unsymmetrical dialkylglyoxal bis(amidinohydrazones). The results also indicate that in all such compounds so far studied, that one of the glyoxal carbons that bears the longer alkyl side chain resonates more downfield than does the other one. This result is in total agreement with conclusions derived from relaxation time measurements.     

A novel capillary electrophoretic method for determining methylglyoxal and glyoxal in urine and water samples

Two non-electroactive biomarkers methylglyoxal (MGo) and glyoxal (Go) in urine and environmental water samples were determined for the first time by capillary electrophoresis with amperometric detection (CE-AD) after derivatizing with an electroactive compound 2-thiobarbituric acid. Experimental conditions of derivatization and CE-AD detection were optimized. Highly linear response was obtained for these two biomarkers over three orders of magnitude with good correlation (r² > 0.999). The limits of detection (LODs) and limits of quantitation (LOQs) of MGo and Go were 0.2 μg L⁻¹ and 1.0 μg L⁻¹, 0.5 μg L⁻¹ and 2.0 μg L⁻¹, respectively. The average recovery and relative standard deviation (RSD) were within the range of 90.9–101.3% and 0.7–2.2%, respectively. The proposed CE-AD method provides a reliable and sensitive quantitative evaluation of MGo and Go in real sample matrices by employing relatively simple and inexpensive instrument.     

2,4-二硝基苯肼衍生-高效液相色谱测定大气细粒子中二羰基类化合物

以2,4-二硝基苯肼作为衍生化试剂,采用高效液相色谱法定量检测了大气细粒子(PM2.5)中二羰基类化合物。以乙二醛和甲基乙二醛为目标化合物,对二羰基类化合物采样条件(采集时间,流速等)、样品处理及分析条件进行了优化。研究表明,利用3mL衍生化溶液(含0.25mmol/LHCl和30μL饱和DNPH-乙腈溶液)和3mL乙腈可有效提取滤膜上的二羰基类化合物;本方法在滤膜采样12h内的采样及洗脱效果较好。利用此方法对上海市宝山区上海大学和崇明东平国家森林公园大气细粒子(PM2.5)中二羰基化合物进行了检测。     

涂布2,4-二硝基苯肼的环形溶蚀器/滤膜系统和高效液相色谱法检测大气中二羰基化合物

建立了环形溶蚀器/滤膜系统(Annular denuder/filter pack system)和2,4-二硝基苯肼(DNPH)-高效液相色谱法(HPLC)采集和检测大气中气相和颗粒相二羰基化合物的方法。DNPH作为吸附剂分别涂布在环形溶蚀器的内壁和3层滤膜上,当大气样品经过环形溶蚀器时,含有气相二羰基化合物的气体吸附到环形溶蚀管内壁上与DN-PH发生反应,而颗粒相部分穿过环形溶蚀管,采集到滤膜上。样品经乙腈洗脱、浓缩后,采用HPLC进行分析。根据不同的采样流速、采样时间和DNPH的涂布量采集到的二羰基化合物的浓度,确定的最佳采样条件为:采样流速4 L/min,采样时间4~5 h,DNPH浓度0.47 g/L。使用Tedlar bag验证环形溶蚀器乙二醛和甲基乙二醛的采集效率(分别为82%和85%)。利用此方法对实际大气中的二羰基化合物进行了检测。     

Analysis of methylglyoxal in water and biological matrices by capillary zone electrophoresis with diode array detection

We describe a new method for the determination of methylglyoxal in water and biological matrices, using o-phenylenediamine as derivatizing agent and solid-phase extraction followed by capillary zone electrophoresis with diode array detection. 25 mM sodium phosphate running buffers at pH 2.2, 30 kV, and 25 degrees C allowed the best instrumental conditions for the optimum separation of methylglyoxal in a suitable analytical time (< 10 min), using an uncoated fused-silica capillary of 75 microm inner diameter and an effective length of 45.1 cm with an extended light path and the wavelength set to 200 nm. Under optimized instrumental conditions, good reproducibility of the migration time (< 1.1%), precision (< 5%), an excellent linear dynamic range from 0.1 to 3.6 mg/L (r(2) = 0.9997), and low limits of detection (7.2 microg/L) were obtained for methylglyoxal measurements, using the internal standard methodology. Assays on laboratory-spiked tap and ground water samples allowed a remarkable accuracy, presenting yields of 95.0 +/- 4.3 and 94.0 +/- 1.1%, respectively, and good performance to determine methylglyoxal in beer and yeast cells suspensions matrices was also obtained at trace level. The present methodology is a cost-effective alternative for routine quality control analysis, showing to be reliable, sensitive, and with a low sample volume requirement to monitor methylglyoxal in water and biological matrices.     

Use of capillary electrophoresis to evaluate protective effects of methylglyoxal scavengers on the activity of creatine kinase

Methylglyoxal (MGO) is a highly reactive alpha-oxoaldehyde formed endogenously in numerous enzymatic and nonenzymatic reactions. The reactions between MGO and various amino residues in proteins not only result in inactivation of enzymes, but also lead to the formation of different detrimental advanced glycation endproducts (AGEs). Recently, it was reported that creatine kinase (CK, EC 2.7.3.2) activity could be reduced or even lost under incubation with MGO in vitro. In this study, an efficient CE analytical method was developed for the evaluation of CK activity. Based on this CE method, the inhibitory effect of MGO on CK activity was confirmed. Several MGO scavengers such as aminoguanidine (AG) and some thiols showed obvious protective effects on CK activity against MGO. Furthermore, tiopronin (TP), a hepatoprotective drug, was found for the first time to counteract MGO-induced inhibition of CK activity in CK reaction. Meanwhile, TP also retained adenosine diphosphate (ADP) generation level in plasma treated with MGO, which implies that this drug may have potential protective effect on other enzymes which are associated with adenine nucleotide metabolism. Besides, the established CE approach can be utilized as a model for screening effective MGO scavengers by monitoring CK-catalyzed conversion between adenosine triphosphate and ADP.     

The structural modification of DNA nucleosides by nonenzymatic glycation: an <Emphasis Type="Italic">in vitro</Emphasis> study based on the reactions of glyoxal and methylglyoxal with 2′-deoxyguanosine

Methylglyoxal and glyoxal are generated from the oxidation of carbohydrates and lipids, and like d-glucose have been shown to nonenzymatically react with proteins to form advanced glycation end products (AGEs). AGEs can occur both in vitro and in vivo, and these compounds have been shown to exacerbate many of the long-term complications of diabetes. Earlier studies in our laboratory reported d-glucose, d-galactose, and d/l-glyceraldehyde formed AGEs with nucleosides. The objective of this study was to focus on purines and pyrimidines and to analyze these DNA nucleoside derived AGE adducts with glyoxal or methylglyoxal using a combination of analytical techniques. Studies using UV and fluorescence spectroscopy along with mass spectrometry provided for a thorough analysis of the nucleoside AGEs and demonstrated that methylglyoxal and glyoxal reacted with 2′-deoxyguanosine via the classic Amadori pathway, and did not react appreciably with 2′-deoxyadenosine, 2′-deoxythymidine, and 2′-deoxycytidine. Additional findings revealed that methylglyoxal was more reactive than glyoxal. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.