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13 pages, 716 KiB

Open AccessArticle

An Analytical Protocol for the Differentiation and the Potentiometric Determination of Fluorine-Containing Fractions in Bovine Milk

by Nadia Spano, Sara Bortolu, Margherita Addis, Ilaria Langasco, Andrea Mara, Maria I. Pilo, Gavino Sanna and Pietro P. Urgeghe

Molecules 2023, 28(3), 1349; https://doi.org/10.3390/molecules28031349 - 31 Jan 2023

Cited by 3 | Viewed by 2778

Abstract

Free fluoride ions are effective in combating caries in children, and their supplementation in milk has been widely used worldwide for this purpose. Furthermore, it is known that ionic fluoride added to milk is distributed among its components, but little is known about [...] Read more.

Free fluoride ions are effective in combating caries in children, and their supplementation in milk has been widely used worldwide for this purpose. Furthermore, it is known that ionic fluoride added to milk is distributed among its components, but little is known about their quantitative relationships. This is likely due to the absence of an analytical protocol aimed at differentiating and quantifying the most important forms of fluorine present in milk. For the first time, a comprehensive protocol made up of six potentiometric methods devoted to quantifying the most important fractions of fluorine in milk (i.e., the free inorganic fluoride, the inorganic bonded fluorine, the caseins-bonded fluorine, the whey-bonded fluorine, the lipid-bonded fluorine, and the total fluorine) has been developed and tested on real samples. Four of the six methods of the procedure are original, and all have been validated in terms of limit of detection and quantification, precision, and trueness. The data obtained show that 9% of all fluorine was in ionic form, while 66.3% of total fluorine was bound to proteins and lipids, therefore unavailable for human absorption. Beyond applications in dental research, this protocol could be extended also to other foods, or used in environmental monitoring. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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15 pages, 2816 KiB

Open AccessArticle

Determination of Vitamin B3 Vitamer (Nicotinamide) and Vitamin B6 Vitamers in Human Hair Using LC-MS/MS

by Sundus M. Sallabi, Aishah Alhmoudi, Manal Alshekaili and Iltaf Shah

Molecules 2021, 26(15), 4487; https://doi.org/10.3390/molecules26154487 - 25 Jul 2021

Cited by 9 | Viewed by 5086

Abstract

Water-soluble B vitamins participate in numerous crucial metabolic reactions and are critical for maintaining our health. Vitamin B deficiencies cause many different types of diseases, such as dementia, anaemia, cardiovascular disease, neural tube defects, Crohn’s disease, celiac disease, and HIV. Vitamin B3 deficiency [...] Read more.

Water-soluble B vitamins participate in numerous crucial metabolic reactions and are critical for maintaining our health. Vitamin B deficiencies cause many different types of diseases, such as dementia, anaemia, cardiovascular disease, neural tube defects, Crohn’s disease, celiac disease, and HIV. Vitamin B3 deficiency is linked to pellagra and cancer, while niacin (or nicotinic acid) lowers low-density lipoprotein (LDL) and triglycerides in the blood and increases high-density lipoprotein (HDL). A highly sensitive and robust liquid chromatography–tandem mass spectroscopy (LC/MS-MS) method was developed to detect and quantify a vitamin B3 vitamer (nicotinamide) and vitamin B6 vitamers (pyridoxial 5′-phosphate (PLP), pyridoxal hydrochloride (PL), pyridoxamine dihydrochloride (PM), pridoxamine-5′-phosphate (PMP), and pyridoxine hydrochloride (PN)) in human hair samples of the UAE population. Forty students’ volunteers took part in the study and donated their hair samples. The analytes were extracted and then separated using a reversed-phase Poroshell EC-C18 column, eluted using two mobile phases, and quantified using LC/MS-MS system. The method was validated in human hair using parameters such as linearity, intra- and inter-day accuracy, and precision and recovery. The method was then used to detect vitamin B3 and B6 vitamers in the human hair samples. Of all the vitamin B3 and B6 vitamers tested, only nicotinamide was detected and quantified in human hair. Of the 40 samples analysed, 12 were in the range 100–200 pg/mg, 15 in the range 200–500 pg/mg, 9 in the range of 500–4000 pg/mg. The LC/MS-MS method is effective, sensitive, and robust for the detection of vitamin B3 and its vitamer nicotinamide in human hair samples. This developed hair test can be used in clinical examination to complement blood and urine tests for the long-term deficiency, detection, and quantification of nicotinamide. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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12 pages, 2817 KiB

Open AccessArticle

Subcritical Water Extraction of Salvia miltiorrhiza

by Brahmam Kapalavavi, Ninad Doctor, Baohong Zhang and Yu Yang

Molecules 2021, 26(6), 1634; https://doi.org/10.3390/molecules26061634 - 15 Mar 2021

Cited by 8 | Viewed by 3691

Abstract

In this work, a green extraction technique, subcritical water extraction (SBWE), was employed to extract active pharmaceutical ingredients (APIs) from an important Chinese medicinal herb, Salvia miltiorrhiza (danshen), at various temperatures. The APIs included tanshinone I, tanshinone IIA, protocatechualdehyde, caffeic acid, and ferulic [...] Read more.

In this work, a green extraction technique, subcritical water extraction (SBWE), was employed to extract active pharmaceutical ingredients (APIs) from an important Chinese medicinal herb, Salvia miltiorrhiza (danshen), at various temperatures. The APIs included tanshinone I, tanshinone IIA, protocatechualdehyde, caffeic acid, and ferulic acid. Traditional herbal decoction (THD) of Salvia miltiorrhiza was also carried out for comparison purposes. Reproduction assay of herbal extracts obtained by both SBWE and THD were then conducted on Caenorhabditis elegans so that SBWE conditions could be optimized for the purpose of developing efficacious herbal medicine from Salvia miltiorrhiza. The extraction efficiency was mostly enhanced with increasing extraction temperature. The quantity of tanshinone I in the herbal extract obtained by SBWE at 150 °C was 370-fold higher than that achieved by THD extraction. Reproduction evaluation revealed that the worm reproduction rate decreased and the reproduction inhibition rate increased with elevated SBWE temperatures. Most importantly, the reproduction inhibition rate of the SBWE herbal extracts obtained at all four temperatures investigated was higher than that of traditional herbal decoction extracts. The results of this work show that there are several benefits of subcritical water extraction of medicinal herbs over other existing herbal medicine preparation techniques. Compared to THD, the thousand-year-old and yet still popular herbal preparation method used in herbal medicine, subcritical water extraction is conducted in a closed system where no loss of volatile active pharmaceutical ingredients occurs, although analyte degradation may happen at higher temperatures. Temperature optimization in SBWE makes it possible to be more efficient in extracting APIs from medicinal herbs than the THD method. Compared to other industrial processes of producing herbal medicine, subcritical water extraction eliminates toxic organic solvents. Thus, subcritical water extraction is not only environmentally friendly but also produces safer herbal medicine for patients. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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17 pages, 2727 KiB

Open AccessArticle

Cyanogenic Glycoside Analysis in American Elderberry

by Michael K. Appenteng, Ritter Krueger, Mitch C. Johnson, Harrison Ingold, Richard Bell, Andrew L. Thomas and C. Michael Greenlief

Molecules 2021, 26(5), 1384; https://doi.org/10.3390/molecules26051384 - 4 Mar 2021

Cited by 27 | Viewed by 17068

Abstract

Cyanogenic glycosides (CNGs) are naturally occurring plant molecules (nitrogenous plant secondary metabolites) which consist of an aglycone and a sugar moiety. Hydrogen cyanide (HCN) is released from these compounds following enzymatic hydrolysis causing potential toxicity issues. The presence of CNGs in American elderberry [...] Read more.

Cyanogenic glycosides (CNGs) are naturally occurring plant molecules (nitrogenous plant secondary metabolites) which consist of an aglycone and a sugar moiety. Hydrogen cyanide (HCN) is released from these compounds following enzymatic hydrolysis causing potential toxicity issues. The presence of CNGs in American elderberry (AE) fruit, Sambucus nigra (subsp. canadensis), is uncertain. A sensitive, reproducible and robust LC-MS/MS method was developed and optimized for accurate identification and quantification of the intact glycoside. A complimentary picrate paper test method was modified to determine the total cyanogenic potential (TCP). TCP analysis was performed using a camera-phone and UV-Vis spectrophotometry. A method validation was conducted and the developed methods were successfully applied to the assessment of TCP and quantification of intact CNGs in different tissues of AE samples. Results showed no quantifiable trace of CNGs in commercial AE juice. Levels of CNGs found in various fruit tissues of AE cultivars studied ranged from between 0.12 and 6.38 µg/g. In pressed juice samples, the concentration range measured was 0.29–2.36 µg/mL and in seeds the levels were 0.12–2.38 µg/g. TCP was highest in the stems and green berries. Concentration levels in all tissues were generally low and at a level that poses no threat to consumers of fresh and processed AE products. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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9 pages, 2870 KiB

Open AccessArticle

In Situ Determination of Nitrate in Water Using Fourier Transform Mid-Infrared Attenuated Total Reflectance Spectroscopy Coupled with Deconvolution Algorithm

by Fangqun Gan, Ke Wu, Fei Ma and Changwen Du

Molecules 2020, 25(24), 5838; https://doi.org/10.3390/molecules25245838 - 10 Dec 2020

Cited by 21 | Viewed by 2759

Abstract

Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy has been used to determine the nitrate content in aqueous solutions. However, the conventional water deduction algorithm indicated considerable limits in the analysis of samples with low nitrate concentration. In this study, FTIR-ATR spectra of [...] Read more.

Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy has been used to determine the nitrate content in aqueous solutions. However, the conventional water deduction algorithm indicated considerable limits in the analysis of samples with low nitrate concentration. In this study, FTIR-ATR spectra of nitrate solution samples with high and low concentrations were obtained, and the spectra were then pre-processed with deconvolution curve-fitting (without water deduction) combined with partial least squares regression (PLSR) to predict the nitrate content. The results show that the typical absorption of nitrate (1200−1500 cm⁻¹) did not clearly align with the conventional algorithm of water deduction, while this absorption was obviously observed through the deconvolution algorithm. The first principal component of the spectra, which explained more than 95% variance, was linearly related to the nitrate content; the correlation coefficient (R²) of the PLSR model for the high-concentration group was 0.9578, and the ratio of the standard deviation of the prediction set to that of the calibration set (RPD) was 4.22, indicating excellent prediction performance. For the low-concentration group model, R² and RPD were 0.9865 and 3.15, respectively, which also demonstrated significantly improved prediction capability. Therefore, FTIR-ATR spectroscopy combined with deconvolution curve-fitting can be conducted to determine the nitrate content in aqueous solutions, thus facilitating rapid determination of nitrate in water bodies with varied concentrations. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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10 pages, 1965 KiB

Open AccessArticle

Simple Derivatization–Gas Chromatography–Mass Spectrometry for Fatty Acids Profiling in Soil Dissolved Organic Matter

by Neil Yohan Musadji and Claude Geffroy-Rodier

Molecules 2020, 25(22), 5278; https://doi.org/10.3390/molecules25225278 - 12 Nov 2020

Cited by 5 | Viewed by 2623

Abstract

Dissolved organic matter is an important component of the global carbon cycle that allows the distribution of carbon and nutrients. Therefore, analysis of soil dissolved organic matter helps us to better understand climate change impacts as it is the most dynamic and reactive [...] Read more.

Dissolved organic matter is an important component of the global carbon cycle that allows the distribution of carbon and nutrients. Therefore, analysis of soil dissolved organic matter helps us to better understand climate change impacts as it is the most dynamic and reactive fraction in terrestrial ecosystems. Its characterization at the molecular level is still challenging due to complex mixtures of hundreds of compounds at low concentration levels in percolating water. This work presents simple methods, such as thermochemolysis– or derivatization–gas chromatography, as an alternative for the analysis of fatty acids in dissolved organic matter without any purification step. The variables of the protocols were examined to optimize the processing conditions for the C₉–C₁₈ range. As a proof of concept, fatty acid distributions of soil percolating water samples from a long-term field experiment were successfully assessed. The variability of dissolved organic acid distributions was pronounced through depth profile and soil treatment but no major change in composition was observed. However, although the optimization was done from C₉ to C₁₈, detection within the C₆-C₃₂ fatty acids range was performed for all samples. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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17 pages, 5445 KiB

Open AccessArticle

Three Extraction Methods in Combination with GC×GC-TOFMS for the Detailed Investigation of Volatiles in Chinese Herbaceous Aroma-Type Baijiu

by Lulu Wang, Mengxin Gao, Zhipeng Liu, Shuang Chen and Yan Xu

Molecules 2020, 25(19), 4429; https://doi.org/10.3390/molecules25194429 - 27 Sep 2020

Cited by 30 | Viewed by 3471

Abstract

In this study, the detailed volatile compositions of Chinese herbaceous aroma-type Baijiu (HAB) were characterized by comprehensive two-dimensional gas chromatography-time of flight mass spectrometry (GC×GC-TOFMS). A total of 606 compounds were tentatively identified by similarity, mass spectral data, and retention indices, among which [...] Read more.

In this study, the detailed volatile compositions of Chinese herbaceous aroma-type Baijiu (HAB) were characterized by comprehensive two-dimensional gas chromatography-time of flight mass spectrometry (GC×GC-TOFMS). A total of 606 compounds were tentatively identified by similarity, mass spectral data, and retention indices, among which 247 compounds were positively verified by authentic standards. Esters were present in higher numbers (179), followed by aldehydes and ketones (111), and alcohols (81). In addition, there were also many terpenes (82), sulfides (37), furans (29), nitrogenous compounds (29), lactones (17), and so on. Meanwhile, the extraction effects of volatile components from different sample pretreatment methods (headspace solid-phase microextraction (HS-SPME), solid phase extraction (SPE), and stir bar sorptive extraction (SBSE)) for HAB were also revealed. The results indicated that HS-SPME has a better extraction effect on easily volatile compounds, such as alcohols and sulfides, especially for terpenes. SPE was particularly beneficial for the analysis of nitrogen-containing compounds; SBSE showed medium extraction ability for most types of compounds and was more suitable for the target analysis of trace content substances. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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14 pages, 2022 KiB

Open AccessArticle

Significantly Elevated Levels of Plasma Nicotinamide, Pyridoxal, and Pyridoxamine Phosphate Levels in Obese Emirati Population: A Cross-Sectional Study

by Ghada Rashad Ibrahim, Iltaf Shah, Salah Gariballa, Javed Yasin, James Barker and Syed Salman Ashraf

Molecules 2020, 25(17), 3932; https://doi.org/10.3390/molecules25173932 - 28 Aug 2020

Cited by 12 | Viewed by 2977

Abstract

Water-soluble vitamins like B3 (nicotinamide), B6 (pyridoxine), and B9 (folic acid) are of utmost importance in human health and disease, as they are involved in numerous critical metabolic reactions. Not surprisingly, deficiencies of these vitamins have been linked to various disease states. Unfortunately, [...] Read more.

Water-soluble vitamins like B3 (nicotinamide), B6 (pyridoxine), and B9 (folic acid) are of utmost importance in human health and disease, as they are involved in numerous critical metabolic reactions. Not surprisingly, deficiencies of these vitamins have been linked to various disease states. Unfortunately, not much is known about the physiological levels of B6 vitamers and vitamin B3 in an ethnically isolated group (such as an Emirati population), as well as their relationship with obesity. The aim of the present study was to quantify various B6 vitamers, as well as B3, in the plasma of obese and healthy Emirati populations and to examine their correlation with obesity. A sensitive and robust HPLC-MS/MS-based method was developed for the simultaneous quantitation of five physiologically relevant forms of vitamin B6, namely pyridoxal, pyridoxine, pyridoxamine, pyridoxamine phosphate, and pyridoxal phosphate, as well as nicotinamide, in human plasma. This method was used to quantify the concentrations of these vitamers in the plasma of 57 healthy and 57 obese Emirati volunteers. Our analysis showed that the plasma concentrations of nicotinamide, pyridoxal, and pyridoxamine phosphate in the obese Emirati population were significantly higher than those in healthy volunteers (p < 0.0001, p = 0.0006, and p = 0.002, respectively). No significant differences were observed for the plasma concentrations of pyridoxine and pyridoxal phosphate. Furthermore, the concentrations of some of these vitamers in healthy Emirati volunteers were significantly different than those published in the literature for Western populations, such as American and European volunteers. This initial study underscores the need to quantify micronutrients in distinct ethnic groups, as well as people suffering from chronic metabolic disorders. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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15 pages, 3112 KiB

Open AccessArticle

The Potential Transformation Mechanisms of the Marker Components of Schizonepetae Spica and Its Charred Product

by Xindan Liu, Ying Zhang, Menghua Wu, Zhiguo Ma and Hui Cao

Molecules 2020, 25(16), 3749; https://doi.org/10.3390/molecules25163749 - 17 Aug 2020

Cited by 6 | Viewed by 2713

Abstract

Schizonepetae Spica (SS) is commonly used for treating colds, fevers, bloody stool and metrorrhagia in China. To treat colds and fevers, traditional Chinese medicine doctors often use raw SS, while to treat bloody stool and metrorrhagia, they usually use Schizonepetae Spica Carbonisata (SSC; [...] Read more.

Schizonepetae Spica (SS) is commonly used for treating colds, fevers, bloody stool and metrorrhagia in China. To treat colds and fevers, traditional Chinese medicine doctors often use raw SS, while to treat bloody stool and metrorrhagia, they usually use Schizonepetae Spica Carbonisata (SSC; raw SS processed by stir-frying until carbonization). However, there have been limited investigations designed to uncover the mechanism of stir-fry processing. In the present study, a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) was developed for the comprehensive analysis of the chemical profiles of SS and SSC samples. Principal component analysis of the GC-MS data demonstrated that there were 16 significant differences in volatile compounds between the SS and SSC samples. The simultaneous quantification of six nonvolatile compounds was also established based on HPLC, and remarkable differences were found between the two products. These changes were probably responsible for the various pharmacological effects of SS and SSC as well as the observed hepatotoxicity. Finally, the mechanisms could be rationalized by deducing possible reactions involved in the transformation of these marker components. This work reports a new strategy to reveal the chemical transformation of SS during stir-fry processing. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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10 pages, 1231 KiB

Open AccessArticle

Investigation of Organic Acids in Saffron Stigmas (Crocus sativus L.) Extract by Derivatization Method and Determination by GC/MS

by Laurynas Jarukas, Olga Mykhailenko, Juste Baranauskaite, Mindaugas Marksa and Liudas Ivanauskas

Molecules 2020, 25(15), 3427; https://doi.org/10.3390/molecules25153427 - 28 Jul 2020

Cited by 19 | Viewed by 3841

Abstract

The beneficial health properties of organic acids make them target compounds in multiple studies. This is the reason why developing a simple and sensitive determination and investigation method of organic acids is a priority. In this study, an effective method has been established [...] Read more.

The beneficial health properties of organic acids make them target compounds in multiple studies. This is the reason why developing a simple and sensitive determination and investigation method of organic acids is a priority. In this study, an effective method has been established for the determination of organic (lactic, glycolic, and malic) acids in saffron stigmas. N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA) was used as a derivatization reagent in gas chromatography combined with mass spectrometric detection (GC/MS). The saffron stigmas extract was evaporated to dryness with a stream of nitrogen gas. The derivatization procedure: 0.1 g of dried extract was diluted into 0.1 mL of tetrahydrofuran, then 0.1 mL MTBSTFA was orderly and successively added into a vial. Two different techniques were used to obtain the highest amount of organic acid derivatives from saffron stigmas. To the best of our knowledge, this is the first report of the quantitative and qualitative GC/MS detection of organic acids in saffron stigmas using MTBSTFA reagent, also comparing different derivatization conditions, such as time, temperature and the effect of reagent amount on derivatization process. The identification of these derivatives was performed via GC-electron impact ionization mass spectrometry in positive-ion detection mode. Under optimal conditions, excellent linearity for all organic acids was obtained with determination coefficients of R² > 0.9955. The detection limits (LODs) and quantitation limits (LOQs) ranged from 0.317 to 0.410 µg/mL and 0.085 to 1.53 µg/mL, respectively. The results showed that the highest yield of organic acids was conducted by using 0.1 mL of MTBSTFA and derivatization method with a conventional heating process at 130 °C for 90 min. This method has been successfully applied to the quantitative analysis of organic acids in saffron stigmas. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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13 pages, 691 KiB

Open AccessArticle

Development and Validation of a Spectrometric Method for Cd and Pb Determination in Zeolites and Safety Evaluation

by Marin Senila, Oana Cadar and Ion Miu

Molecules 2020, 25(11), 2591; https://doi.org/10.3390/molecules25112591 - 2 Jun 2020

Cited by 11 | Viewed by 2807

Abstract

An analytical method based on microwave-assisted acid digestion and atomic absorption spectrometry with graphite furnace as atomization source was developed and validated for determining trace elements (Cd and Pb) in zeolites used as dietary supplements, for their characterization and safety evaluation. The method [...] Read more.

An analytical method based on microwave-assisted acid digestion and atomic absorption spectrometry with graphite furnace as atomization source was developed and validated for determining trace elements (Cd and Pb) in zeolites used as dietary supplements, for their characterization and safety evaluation. The method was checked for the main performance parameters according to the legislation requirements in the field of dietary supplements. In all cases, the obtained performance parameters were satisfactory. The selectivity study showed no significant non-spectral matrix effect. The linearity study was conducted for the calibration curves in the range of 0–10 ng mL⁻¹ for Cd and 0–30 ng mL⁻¹ for Pb. The obtained limits of detection (LoDs) and the limits of quantification (LoQs) were sufficiently low in order to allow Pb and Cd determination in dietary supplements. For the internal quality control, certified reference materials were analysed and good recoveries were obtained. The precision study was performed in terms of repeatability and reproducibility, considering the requirements imposed by the Commission Decision (2007/333/EC) and the method fulfilled these performance parameters. Expanded measurement uncertainties were estimated to 11% for Cd and 10% for Pb. Cd and Pb content were measured in real zeolite samples and, using these data, a safety evaluation was carried out. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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16 pages, 5888 KiB

Open AccessArticle

Simultaneous Measurement of Urinary Trimethylamine (TMA) and Trimethylamine N-Oxide (TMAO) by Liquid Chromatography–Mass Spectrometry

by Xun Jia, Lucas J. Osborn and Zeneng Wang

Molecules 2020, 25(8), 1862; https://doi.org/10.3390/molecules25081862 - 17 Apr 2020

Cited by 17 | Viewed by 6547

Abstract

Trimethylamine (TMA) is a gut microbial metabolite—rendered by the enzymatic cleavage of nutrients containing a TMA moiety in their chemical structure. TMA can be oxidized as trimethylamine N-oxide (TMAO) catalyzed by hepatic flavin monooxygenases. Circulating TMAO has been demonstrated to portend a [...] Read more.

Trimethylamine (TMA) is a gut microbial metabolite—rendered by the enzymatic cleavage of nutrients containing a TMA moiety in their chemical structure. TMA can be oxidized as trimethylamine N-oxide (TMAO) catalyzed by hepatic flavin monooxygenases. Circulating TMAO has been demonstrated to portend a pro-inflammatory state, contributing to chronic diseases such as cardiovascular disease and chronic kidney disease. Consequently, TMAO serves as an excellent candidate biomarker for a variety of chronic inflammatory disorders. The highly positive correlation between plasma TMAO and urine TMAO suggests that urine TMAO has the potential to serve as a less invasive biomarker for chronic disease compared to plasma TMAO. In this study, we validated a method to simultaneously measure urine TMA and TMAO concentrations by liquid chromatography–mass spectrometry (LC/MS). Urine TMA and TMAO can be extracted by hexane/butanol under alkaline pH and transferred to the aqueous phase following acidification for LC/MS quantitation. Importantly, during sample processing, none of the nutrients with a chemical structure containing a TMA moiety were spontaneously cleaved to yield TMA. Moreover, we demonstrated that the acidification of urine prevents an increase of TMA after prolonged storage as was observed in non-acidified urine. Finally, here we demonstrated that TMAO can spontaneously degrade to TMA at a very slow rate. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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11 pages, 3127 KiB

Open AccessArticle

In Situ Raman Investigation of TiO₂ Nanotube Array-Based Ultraviolet Photodetectors: Effects of Nanotube Length

by Yanyu Ren, Xiumin Shi, Pengcheng Xia, Shuang Li, Mingyang Lv, Yunxin Wang and Zhu Mao

Molecules 2020, 25(8), 1854; https://doi.org/10.3390/molecules25081854 - 17 Apr 2020

Cited by 5 | Viewed by 2907

Abstract

TiO₂ nanotube arrays (TNAs) with tube lengths of 4, 6, and 7 μm were prepared via two-step anodization. Thereafter, ultraviolet (UV) photodetectors (PDs) with Au/TiO₂/Au structures were prepared using these TNAs with different tube lengths. The effects of TNA length [...] Read more.

TiO₂ nanotube arrays (TNAs) with tube lengths of 4, 6, and 7 μm were prepared via two-step anodization. Thereafter, ultraviolet (UV) photodetectors (PDs) with Au/TiO₂/Au structures were prepared using these TNAs with different tube lengths. The effects of TNA length and device area on the performance of the device were investigated using in situ Raman spectroscopy. The maximum laser/dark current ratio was achieved by using a TNA with a size of 1 × 1 cm² and a length of 7 μm, under a 532 nm laser. In addition, when the device was irradiated with a higher energy laser (325 nm), the UV Raman spectrum was found to be more sensitive than the visible Raman spectrum. At 325 nm, the laser/dark current ratio was nearly 24 times higher than that under a 532 nm laser. Six phonon modes of anatase TNAs were observed, at 144, 199, 395, 514, and 635 cm⁻¹, which were assigned to the E_g(1), E_g(2), B_1g(1), A_1g/B_1g(2), and E_g(3) modes, respectively. The strong low-frequency band at 144 cm⁻¹ was caused by the O-Ti-O bending vibration and is a characteristic band of anatase. The results show that the performance of TNA-based PDs is length-dependent. Surface-enhanced Raman scattering signals of 4-mercaptobenzoic acid (4-MBA) molecules were also observed on the TNA surface. This result indicates that the length-dependent performance may be derived from an increase in the specific surface area of the TNA. In addition, the strong absorption of UV light by the TNAs caused a blueshift of the E_g(1) mode. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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Review

Jump to: Research

61 pages, 14417 KiB

Open AccessFeature PaperReview

Fluorescent Probes for Live Cell Thiol Detection

by Shenggang Wang, Yue Huang and Xiangming Guan

Molecules 2021, 26(12), 3575; https://doi.org/10.3390/molecules26123575 - 11 Jun 2021

Cited by 34 | Viewed by 5899

Abstract

Thiols play vital and irreplaceable roles in the biological system. Abnormality of thiol levels has been linked with various diseases and biological disorders. Thiols are known to distribute unevenly and change dynamically in the biological system. Methods that can determine thiols’ concentration and [...] Read more.

Thiols play vital and irreplaceable roles in the biological system. Abnormality of thiol levels has been linked with various diseases and biological disorders. Thiols are known to distribute unevenly and change dynamically in the biological system. Methods that can determine thiols’ concentration and distribution in live cells are in high demand. In the last two decades, fluorescent probes have emerged as a powerful tool for achieving that goal for the simplicity, high sensitivity, and capability of visualizing the analytes in live cells in a non-invasive way. They also enable the determination of intracellular distribution and dynamitic movement of thiols in the intact native environments. This review focuses on some of the major strategies/mechanisms being used for detecting GSH, Cys/Hcy, and other thiols in live cells via fluorescent probes, and how they are applied at the cellular and subcellular levels. The sensing mechanisms (for GSH and Cys/Hcy) and bio-applications of the probes are illustrated followed by a summary of probes for selectively detecting cellular and subcellular thiols. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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25 pages, 7537 KiB

Open AccessReview

Trends in the Hydrogen−Deuterium Exchange at the Carbon Centers. Preparation of Internal Standards for Quantitative Analysis by LC-MS

by Paulina Grocholska and Remigiusz Bąchor

Molecules 2021, 26(10), 2989; https://doi.org/10.3390/molecules26102989 - 18 May 2021

Cited by 21 | Viewed by 6530

Abstract

The application of internal standards in quantitative and qualitative bioanalysis is a commonly used procedure. They are usually isotopically labeled analogs of the analyte, used in quantitative LC-MS analysis. Usually, ²H, ¹³C, ¹⁵N and ¹⁸O isotopes are used. The [...] Read more.

The application of internal standards in quantitative and qualitative bioanalysis is a commonly used procedure. They are usually isotopically labeled analogs of the analyte, used in quantitative LC-MS analysis. Usually, ²H, ¹³C, ¹⁵N and ¹⁸O isotopes are used. The synthesis of deuterated isotopologues is relatively inexpensive, however, due to the isotopic effect of deuterium and the lack of isotopologue co-elution, usually they are not considered as good internal standards for LC-MS quantification. On the other hand, the preparation of ¹³C, ¹⁵N and ¹⁸O containing standards of drugs and their metabolites requires a complicated multistep de novo synthesis, starting from the isotopically labeled substrates, which are usually expensive. Therefore, there is a strong need for the development of low-cost methods for isotope-labeled standard preparations for quantitative analysis by LC-MS. The presented review concentrates on the preparation of deuterium-labeled standards by hydrogen−deuterium exchange reactions at the carbon centers. Recent advances in the development of the methods of isotopologues preparation and their application in quantitative analysis by LC-MS are evaluated. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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26 pages, 2675 KiB

Open AccessReview

Molecular Spectroscopic Markers of Abnormal Protein Aggregation

by Natalia Wilkosz, Michał Czaja, Sara Seweryn, Katarzyna Skirlińska-Nosek, Marek Szymonski, Ewelina Lipiec and Kamila Sofińska

Molecules 2020, 25(11), 2498; https://doi.org/10.3390/molecules25112498 - 27 May 2020

Cited by 23 | Viewed by 4956

Abstract

Abnormal protein aggregation has been intensively studied for over 40 years and broadly discussed in the literature due to its significant role in neurodegenerative diseases etiology. Structural reorganization and conformational changes of the secondary structure upon the aggregation determine aggregation pathways and cytotoxicity [...] Read more.

Abnormal protein aggregation has been intensively studied for over 40 years and broadly discussed in the literature due to its significant role in neurodegenerative diseases etiology. Structural reorganization and conformational changes of the secondary structure upon the aggregation determine aggregation pathways and cytotoxicity of the aggregates, and therefore, numerous analytical techniques are employed for a deep investigation into the secondary structure of abnormal protein aggregates. Molecular spectroscopies, including Raman and infrared ones, are routinely applied in such studies. Recently, the nanoscale spatial resolution of tip-enhanced Raman and infrared nanospectroscopies, as well as the high sensitivity of the surface-enhanced Raman spectroscopy, have brought new insights into our knowledge of abnormal protein aggregation. In this review, we order and summarize all nano- and micro-spectroscopic marker bands related to abnormal aggregation. Each part presents the physical principles of each particular spectroscopic technique listed above and a concise description of all spectral markers detected with these techniques in the spectra of neurodegenerative proteins and their model systems. Finally, a section concerning the application of multivariate data analysis for extraction of the spectral marker bands is included. Full article

(This article belongs to the Special Issue 25th Anniversary of Molecules—Recent Advances in Analytical Chemistry)

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