Chemical profiling and quantification of multiple components in Jin-Gu-Lian capsule using a multivariate data processing approach based on UHPLC-Orbitrap Exploris 240 MS and UHPLC-MS/MS

The Jin-Gu-Lian capsule, a Chinese Miao herbal compound, is widely used to treat rheumatoid arthritis. In this study, a rapid, selective, and sensitive UHPLC-Orbitrap Exploris 240 MS method was developed to analyze the chemical composition of Jin-Gu-Lian capsules. A total of 88 compounds were identified, including 23 flavonoids, 23 organic acids, 14 phenylpropanoids, 12 phenols, eight alkaloids, four terpenes, three quinones, and one ketone. Among these, 21 compounds were clearly detected based on a comparison with reference standards and selected as quality control markers. Thereafter, these compounds were simultaneously determined in the Jin-Gu-Lian capsules. The established method was successfully validated and applied for the simultaneous determination of 21 biologically active compounds in Jin-Gu-Lian capsules of 27 sample batches. Quantitative data of the analytes were analyzed using multivariate statistical analysis to determine the quality of the Jin-Gu-Lian capsules. Four compounds (JGLC6 [salidroside], JGLC8 [chlorogenic acid], JGLC12 [liriodendrin], JGLC19 [quercetin]) were identified as chemical markers for quality control of Jin-Gu-Lian capsules. Altogether, the established method was validated as a novel and efficient tool, that can be used for rapid analysis of Jin-Gu-Lian capsules. Accordingly, this study serves as a reference for scientific research on traditional Chinese and ethnic medicine.     

Qualitative and quantitative analysis of cinobufacini injection using rapid separation liquid chromatography coupled with quadrupole‐time‐of‐flight mass spectrometry and HPLC‐photodiode array detection,a feasible strategy for the quality control of Chinese medicine injections

Cinobufacini injection, prepared from the skin of Bufo bufo gargarizans Cantor, has presented its significant effects on the treatment of hepatitis and various cancers in the clinic. However, as an unclear complex chemical system, the optimization of its quality control markers has been a long‐term challenge. In present study, a feasible strategy integrated markers screening, determination, and statistical analysis was efficiently proposed, especially for the undefined Chinese medicine injections. First, rapid separation LC‐quadrupole‐TOF‐MS method was applied in the identification of 19 major compounds in the cinobufacini injection for the first time. Further, nine high‐level contents active compounds were selected as quality control markers for the quantification analysis. An acceptable and validated determination method was established in 17 batches of cinobufacini injection by HPLC‐photodiode array detection method, including linear regression relationship (r², 0.9996–1), precisions (RSD, 0.02–1.35%), repeatability (RSD, 0.05–1.97%), stability (RSD, 0.1–3.85%), and recovery (95.88–104.89%). Each analyte was detected at its maximum ultraviolet spectra wavelength. Finally, based on the quantification results, principal component analysis was performed on the quality assessment of cinobufacini injections. This three‐step strategy provides a newly feasible solution for the quality control of Chinese medicine injections.     

Determination of Halide Traces in Environmental Samples by Mass Spectrometric Isotope Dilution Analysis

Abstract

Since 1985, the Goulden large-sample extractor (GLSE) has been used to isolate a broad array of trace-organic contaminants from large volumes of water. In this study, field-applied quality control measures, including matrix and surrogate spikes and blanks, were used to monitor method performance from GLSE extraction through GC-MS analysis. The method was applied to the determination of multiple classes of pesticides isolated from 4- to 112-L filtered surface-water samples. Average recoveries of six surrogate compounds ranged from 84 ± 18% for [²H₁₀]diazinon to 15 ± 13% for 4,4′-[²H₈]DDT, the low recoveries for which were largely a result of unmonitored breakdown of this surrogate by the GC injection system. Field-matrix-spike samples were prepared by fortifying 10-L, 35-L, and 110-L filtered surface-water samples with 68 pesticides to amended concentrations of 11-to 50-ng/L each. Recoveries ranged from not detected to greater than 100%. Variability in pesticide recoveries from triplicate 10-L water samples collected at one site averaged 5.7% relative standard deviation and did not exceed 19%.     

Pesticide residues in groundwaters and soils of agricultural areas in the Águeda River Basin from Spain and Portugal

This research has evaluated the agricultural impact of the use of pesticides in small agricultural areas in the Águeda river basin, which straddles the Spanish-Portuguese border. Sixteen pesticides frequently used in the area, including herbicides, fungicides and insecticides and some of their degradation products, were monitored in 52 groundwater samples and 42 soil samples taken around them, using a developed multi-residual analytical method based on SPE-LC-MS. Sampling was carried out in two different seasons (winter and summer). The results indicated the presence of pesticides at several levels, both in groundwaters and soils. Thirteen of the pesticides studied were detected in one or more of the groundwater samples analysed, but only three pesticides were detected in the soil samples. Terbuthylazine, cyprodinil, tebuconazole and chlorpyrifos were the pesticides most frequently detected in groundwaters, whereas terbuthylazine, metalaxyl and tebuconazole were the sole compounds detected in soils. The distribution of the concentrations in groundwaters indicated that up to 80% and 70% of the samples collected in the summer on the Spanish and Portuguese sides, respectively, exceed the quality standards of 0.1 µg L^?1 for one or more individual compounds and, in turn, up to 64% and 40% exceed the quality standards of 0.5 µg L^?1 for all compounds. The presence of pesticide residues in the groundwaters and soils analysed may well be explained by the use of these compounds in agricultural practices.     

Evaluation of Chinese medicinal herbs fingerprinting by HPLC-DAD for the detection of toxic aristolochic acids

Aristolochic acids are known to contribute to various renal disorders; therefore, expanding the availability of analytical methodology to detect these compounds is important in order to assess the quality of Chinese herbal medicines in which they can be found. Twelve medicinal herbal samples were procured from various sources and extracted in duplicate prior to a "fingerprint" analysis using conventional HPLC-DAD. Multivariate analysis was performed on the entire chromatographed fingerprints. The resulting output was a partial least-square discriminant analysis model, which was able to evaluate the potential presence of aristolochic acids I and II as well as providing an individual herbal "fingerprint". The results of this study provide evidence that the presence of aristolochic acids contained within certain herbal extractions could be detected using a simple method, although some limitations apply to this method for quality control, since newly detected samples for aristolochic acid (positives) will need further confirmation with purity checks or MS hyphenation.     

Chionanthus virginicus L.: phytochemical analysis and quality control of herbal drug and herbal preparations

Root barks of Chionanthus virginicus L. are used in homeopathic medicines in the treatment of icterus and hepatitis. The objective of this study is to identify novel secoiridoids and lignans and to develop a simple and reliable HPLC method for the determination of oleuropein, phillyrin, total secoiridoids and total lignans for quality control and stability studies of C. virginicus herbal drug and preparations. Secoiridoids and lignans were purified by preparative HPLC. Compounds previously described were identified by HPLC according to their retention times and UV spectra. Structures of new compounds were determined by NMR. Two compounds namely excelside B and acetoxypinoresinol-4"-O-beta-D-glucoside are described for the first time in the drug. HPLC separation was performed on Symmetry C18 (Waters) by gradient elution using acetonitrile and 0.2% aqueous phosphoric acid. The method was validated for specificity, linearity, precision, accuracy, limits of detection and quantification for simultaneous determination of secoiridoids and lignans in herbal drug and herbal preparations as mother tinctures. The proposed HPLC method is linear in the range studied (r2 > or = 0.9989) for all the analytes. The method is precise with intra- and inter-day variations of less than 4%. The mean recoveries of the analytes range from 99.65 to 102.81%. The method is successfully applied to the quantification of nine compounds belonging to secoiridoids and lignans and for the stability studies of these compounds. The study allowed completing the phytochemical knowledge of C. virginicus. This simple developed assay could be used as tools for routine quality control of C. virginicus herbal drug and herbal medicinal products.     

Chemical profiling and quantification of ShenKang injection,a systematic quality control strategy using ultra high performance liquid chromatography with Q Exactive hybrid quadrupole orbitrap high‐resolution accurate mass spectrometry

ShenKang injection is traditional Chinese medicine used to treat chronic renal failure in China. It is a compound preparation that consists of four herbs: Rhubarb, Salvia miltiorrhiza, Safflower and Radix Astragali . We developed an ultra high pressure liquid chromatography coupled with Q Exactive hybrid quadrupole‐orbitrap high resolution accurate mass spectrometry tandem mass spectrometry method to analyze its chemical compositions, and a total of 90 compounds were identified from ShenKang injection. Among them, 19 major compounds were unambiguously detected by comparing with reference standards. Meanwhile, 13 representative compounds selected as quality control markers were simultaneously quantified in ShenKang injection samples. Chromatographic separation was accomplished on a Waters ACQUITY HPLC® HSS C₁₈ column using gradient elution. The method was validated with respect to linearity, sensitivity, accuracy and precision, reproducibility and stability. And the validated method was successfully applied for simultaneous determination of 13 bioactive compounds in ShenKang injection from ten batches of samples by liquid chromatography with mass spectrometry. The results were analyzed by principal components analysis method, and three compounds had a significant relationship with the quality control of ShenKang injection. This research established a rapid and reliable method for the integrating quality control, including qualitation and quantification of ShenKang injection.     

气相色谱法测定车用汽油中含氧化合物和苯胺类化合物

利用中心切割技术和双毛细管色谱柱系统,采用两次进样的方式,建立了气相色谱测定车用汽油中含氧化合物和苯胺类化合物的分析方法。第一次进样分析,组分首先进入非极性DB-1色谱柱(30 m×0.32 mm×1.0μm),按沸点由低到高的顺序分离,通过电磁阀切换将沸点小于2-己酮的组分切割至强极性GS-OxyPLOT色谱柱(10 m×0.53 mm×10μm)或CP-Lowox色谱柱(10 m×0.53 mm×10μm),其余重烃组分通过阻尼柱进入FID检测器。在GS-OxyPLOT或CP-Lowox色谱柱上,烃类组分与含氧化合物分离并进入检测器检测,消除了大量的烃类组分对含氧化合物测定的影响。第二次进样分析,设定电磁阀切换时间为间-甲基苯胺从非极性色谱柱流出的时间,苯胺类化合物在GS-OxyPLOT或CP-Lowox色谱柱上与烃类和含氧化合物分离并进入检测器检测。以乙二醇二甲基醚为内标化合物进行内标法定量。实现了在一套系统上同时测定车用汽油中添加的甲基叔丁基醚(MTBE)、甲醇、甲缩醛、乙酸仲丁酯、乙酸乙酯、苯胺、邻/间/对-甲基苯胺和N-甲基苯胺的含量,各组分的检测范围为0.01%~10%(质量分数),回收率为86.0%~102.6%。该法可以为车用汽油的质量控制提供有效的检测手段。     

Fingerprint analysis of Ligusticum chuanxiong using hydrophilic interaction chromatography and reversed-phase liquid chromatography

Fingerprint analysis is considered one of the most powerful approaches to quality control in traditional Chinese medicines (TCMs). In this study, a binary chromatographic fingerprint analysis was developed using hydrophilic interaction chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) to gain more chemical information about polar compounds and weakly polar compounds. This method was used to construct a chromatographic fingerprint of Ligusticum chuanxiong. The two chromatographic methods demonstrated good precision, reproducibility, and stability, with relative standard deviations of <2% for retention time and 7% for peak area for both HILIC and RPLC separations. Data from the analysis of 14 samples by HILIC and RPLC were processed with similarity analysis, with correlation coefficients and congruence coefficients. This binary fingerprint analysis, using two chromatographic modes, is a powerful tool for characterizing the quality of samples, and can be used for the comprehensive quality control of TCMs.     

Bank of reference samples of blank urine from livestock

From three species of livestock, bovine, ovine and porcine, samples of urine from each 10–20 different animals were collected. The animals originated from a governmental experimental farm and differed in race, age, sex and were fed with different types of feed. Fifty different samples of urine were lyophilised in units of 5 mL in sealed amber glass bottles. After lyophilisation of the samples, several quality control tests were performed. The variation in net weight of the units of urine and the lyophilisates ranged from 0.01%–1.7% and 0.1%–10%, respectively. Most values did not exceed 0.1% for urines and 1% for lyophilisates. The residual water content of the lyophilisates was determined by the Karl-Fisher titration method. The average percentage of residual water ranged from 1.0%–7.0% for bovine, from 1.3%–3.0% for porcine and from 1.6%–5.5% for ovine urine lyophilisates. Further, all different samples were analysed for the presence of anabolic compounds with a multi-residue procedure developed at the CRL. In two samples of bovine urine the presence of α-boldenone was detected and confirmed. In most porcine urines the endogenous steroids α-testosterone and α-estradiol were detected. In a number of porcine and ovine urine samples α- and β-zearalenol were detected. The origin of these compounds is the f2-toxin produced by a Fusarium fungus. In ovine samples α-testosterone and α-estradiol were detected.     

Development and Application of an LC-MS/MS Untargeted Exposomics Method with a Separated Pooled Quality Control Strategy

Pooled quality controls (QCs) are usually implemented within untargeted methods to improve the quality of datasets by removing features either not detected or not reproducible. However, this approach can be limiting in exposomics studies conducted on groups of exposed and nonexposed subjects, as compounds present at low levels only in exposed subjects can be diluted and thus not detected in the pooled QC. The aim of this work is to develop and apply an untargeted workflow for human biomonitoring in urine samples, implementing a novel separated approach for preparing pooled quality controls. An LC-MS/MS workflow was developed and applied to a case study of smoking and non-smoking subjects. Three different pooled quality controls were prepared: mixing an aliquot from every sample (QC-T), only from non-smokers (QC-NS), and only from smokers (QC-S). The feature tables were filtered using QC-T (T-feature list), QC-S, and QC-NS, separately. The last two feature lists were merged (SNS-feature list). A higher number of features was obtained with the SNS-feature list than the T-feature list, resulting in identification of a higher number of biologically significant compounds. The separated pooled QC strategy implemented can improve the nontargeted human biomonitoring for groups of exposed and nonexposed subjects.     

Chemical fingerprint analysis for quality assessment and control of Bansha herbal tea using paper spray mass spectrometry

Chemical fingerprinting methodology is an approach for quality assessment and control of herbal medicines and related products based on the holistic chemical profile obtained by various analytical techniques. This study demonstrates the first application of paper spray mass spectrometry (PS-MS) as a chemical fingerprinting methodology for tracing the origins, establishing the authenticity, and assessing the overall quality of a famous herbal product, Bansha herbal tea (BHT). A negative ion PS-MS spectrum yielded the best chemical profiling information and was most appropriate for fingerprint analysis of BHT. In addition to the identification of active ingredients, various compounds present in BHT were simultaneously detected without any sample pretreatment and chromatographic separation, providing valuable information for the quality assessment and control of this herbal product. According to the principal component analysis of the PS-MS fingerprints, two sources of commercially available BHT products made by different manufacturers were easily differentiated. Qualified and expired products from the two manufacturers were also successfully distinguished, and the consistency of the quality between the manufacturers was assessed. Our experimental data demonstrated that the PS-MS chemical fingerprinting is a simple, rapid, and robust methodology for pharmaceutical analysis, with promising prospects for quality assessment and control of herbal medicines and related products with high-throughput.     

Anionic metabolic profiling of urine from antibiotic-treated rats by capillary electrophoresis–mass spectrometry

A recently developed capillary electrophoresis (CE)-negative-ionisation mass spectrometry (MS) method was used to profile anionic metabolites in a microbial-host co-metabolism study. Urine samples from rats receiving antibiotics (penicillin G and streptomycin sulfate) for 0, 4, or 8 days were analysed. A quality control sample was measured repeatedly to monitor the performance of the applied CE-MS method. After peak alignment, relative standard deviations (RSDs) for migration time of five representative compounds were below 0.4 %, whereas RSDs for peak area were 7.9–13.5 %. Using univariate and principal component analysis of obtained urinary metabolic profiles, groups of rats receiving different antibiotic treatment could be distinguished based on 17 discriminatory compounds, of which 15 were downregulated and 2 were upregulated upon treatment. Eleven compounds remained down- or upregulated after discontinuation of the antibiotics administration, whereas a recovery effect was observed for others. Based on accurate mass, nine compounds were putatively identified; these included the microbial-mammalian co-metabolites hippuric acid and indoxyl sulfate. Some discriminatory compounds were also observed by other analytical techniques, but CE-MS uniquely revealed ten metabolites modulated by antibiotic exposure, including aconitic acid and an oxocholic acid. This clearly demonstrates the added value of CE-MS for nontargeted profiling of small anionic metabolites in biological samples.     

Analysis of pesticides and metabolites in Spanish surface waters by isotope dilution gas chromatography/mass spectrometry with previous automated solid-phase extraction Estimation of the uncertainty of the analytical results

A method based on isotope dilution gas chromatography/mass spectrometry (GC/MS) with automated solid-phase extraction (SPE) is described for the analysis of 32 pesticides and metabolites in surface waters. This approach consist in the use of nine isotopically labelled representative pesticides as internal standards, which allows high accuracy (trueness and precision) and sensitivity for most analysed compounds, as it is required for isotope dilution-based methods. Uncertainties associated with pesticide determination in real samples were estimated using quality assurance/quality control (QA/QC) data. For most pesticides expanded uncertainty was below 40%, according to the commonly established requirements for analytical results. Ninety three Spanish surface waters collected in June-July and September-November 2004 were analysed. Concentration and occurrence of pesticides were evaluated. These parameters were higher in the summer than in the autumn period. In summer four pesticides were found in more than 50% of the analysed samples and four compounds were detected above the concentration level of 1 microg/l (atrazine, terbutylazine, 3,4-dichloroaniline and fenitrothion), while in autumn percentage of detection was below 50% for all pesticides and only one compound (terbutylazine) exceeded 1 microg/l.     

Comparison of preconcentration methods for nontargeted analysis of natural waters using HPLC-HRMS: Large volume injection versus solid-phase extraction

Nontargeted analysis of water samples using liquid chromatography combined with high-resolution mass spectrometers is an emerging approach for surface water monitoring and evaluation of water treatment processes. In this study, sample preconcentration via direct, large volume injection with 500 μL and 1000 μL injection volumes was compared to SPE regarding analytical performance parameters in targeted and nontargeted workflows. In targeted analysis, the methods were evaluated in terms of LOD and intrabatch precision of the selected compounds, whereas in nontargeted analysis, the number of detected unknown compounds, the method's intra-batch precision, and the retention time versus molecular mass pattern of the detected unknowns were evaluated. In addition, a novel intensity drift correction method was developed that is not based on quality control samples and makes use of the signals obtained for continuously infused reference compounds, which are conventionally utilized for online mass drift correction. It could be demonstrated that the new correction method significantly reduced the bias introduced by instrumental drift and is important for the reliable intercomparison of different nontargeted methods. Intercomparison of results showed that the 1000 μL large volume injection method revealed the best performance in terms of precision under repeatability conditions of measurement as well as lower LODs for targeted compound analysis. In nontargeted analysis, the SPE method detected a higher number of unknown compounds but exhibited also a higher uncertainty of measurement caused by matrix effects.     

Pressurized liquid extraction followed by high-performance liquid chromatography for determination of seven active compounds in Cortex Dictamni

A new method based on pressurized liquid extraction (PLE) followed by a sensitive and specific HPLC-DAD analysis is developed for determination of seven compounds in Cortex Dictamni. The operational parameters of PLE, such as extraction solvent, extraction temperature, extraction pressure, static extraction time, flush volume and extraction cycles were optimized, using the extraction efficiencies of dictamnine, obacunone and fraxinellone as targets. The optimized procedure employed MeOH as extraction solvent, 150 degrees C of extraction temperature, 1,500 psi extraction pressure, 5 min of static extraction time, 60% flush volume and the extraction recoveries of the three compounds were nearly to 100% for only one cycle. The following HPLC analysis was performed on a reversed-phase C(18) column with methanol-water as mobile phase in gradient manner, detected at 236 and 218 nm. The limits of detection (LOD) and limits of quantification (LOQ) of the seven compounds were in the range of 0.4-15.6 ng and 1.2-38.8 ng. This assay can be readily utilized as a quality control method for Cortex Dictamni and other related medicinal plants.     

Simultaneous extraction of polycyclic aromatic hydrocarbons and polychlorinated biphenyls in agricultural soils by pressurized liquid extraction and determination by gas chromatography coupled to tandem mass spectrometry

A simple and rapid method based on pressurized liquid extraction has been validated for the simultaneous extraction of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) from agricultural soil samples. Effective extraction was carried out in less than 17 min for all the studied compounds, and good recoveries were obtained for PAHs and PCBs, ranging from 70% to 112%, when blank samples were spiked at 2.5 μg kg⁻¹, except for naphthalene with recoveries close to 40%. The separation and determination were performed by gas chromatography coupled to tandem mass spectrometry using a triple quadrupole mass analyzer. The target compounds were detected by electron impact with selected reaction monitoring, and mass spectrometric conditions were optimized in order to increase selectivity and sensitivity. The developed method was validated, and matrix-matched calibration was used for quantification purposes. Repeatability and interday precision ranged from 0.9% to 16.8% and from 1.6% to 22.3%, respectively. Limits of quantification ranged from 0.07 to 2.50 μg kg⁻¹. The proposed method was applied to the analysis of agricultural soil samples collected from Almeria (Spain), and PAHs and PCBs were detected in some samples at concentrations ranging from 0.1 to 210 μg kg⁻¹.     

Comparative Analysis of Chemical Constituents in Different Parts of Lotus by UPLC and QToF-MS

Six parts of lotus (seeds, leaves, plumule, stamens, receptacles and rhizome nodes) are herbal medicines that are listed in the Chinese Pharmacopoeia. Their indications and functions have been confirmed by a long history of clinical practice. To fully understand the material basis of clinical applications, UPLC-QToF-MS combined with the UNIFI platform and multivariate statistical analysis was used in this study. As a result, a total of 171 compounds were detected and characterized from the six parts, and 23 robust biomarkers were discovered. The method can be used as a standard protocol for the direct identification and prediction of the six parts of lotus. Meanwhile, these discoveries are valuable for improving the quality control method of herbal medicines. Most importantly, this was the first time that alkaloids were detected in the stamen, and terpenoids were detected in the cored seed. The stamen is a noteworthy part because it contains the greatest diversity of flavonoids and terpenoids, but research on the stamen is rather limited.     

Optimization and validation of a methodology based on solvent extraction and liquid chromatography for the simultaneous determination of several polyphenolic families in fruit juices

A solvent extraction procedure of freeze-dried aliquots followed by the analysis of phenolic compounds by reversed-phase high-performance liquid chromatography (RP-HPLC) with photodiode array detection (DAD) has been developed for the analysis of polyphenolic compounds in fruit juices. This methodology is focussed on the characterization of fruit juices, mainly for quality control purposes. The effects of experimental variables, such as solvent composition and volume and time and temperature on extraction, have been studied. A unique gradient program for the separation of several phenolic classes (hydroquinones, hydroxybenzoic acids, flavan-3-oles, hydroxycinnamic acids, coumarins, flavanones, flavones, dihydrochalcones and flavonols) has been optimized, using standards of 55 commercially available phenolic compounds present in fruits, as well as representative real extracts from fruit juices. All phenolic compounds showed a high repeatability within-day (n=5) and between days (n=3) in peak area (RSD<8%) and excellent stability of their retention times. High precision was also observed in calibration slopes (RSD<8%). Detection limits ranged between 0.005 and 0.03 microg/mL for the different detected polyphenols. Complete recoveries (98-100%) were obtained for the majority of the phenolic structures of all representative phenolic families present in fruits. The method was successfully employed to measure diverse phenolic families in juices from 18 different fruits and consequently could be used for evaluate the quality of fruit juices.