Longest distance shifting: A simple and efficient approach for the alignment of shifted chromatographic peaks

The preprocessing of chromatograms, such as the alignment of retention time shifts, is often a crucial step in the proper data analysis chain. Here, an efficient approach to align shifted chromatographic signals, longest distance shifting, is presented and highlighted. The performance of this novel strategy was demonstrated by using both simulated chromatograms that covered the different kinds of retention time shifts and the real experimental chromatograms of Pudilan Xiaoyan Tablets obtained by high‐performance liquid chromatography with photodiode array detection. The averaged correlation coefficient for experimental chromatograms were in the range of 0.9517–0.9840 and the peak factor was 0.9989. As a comparison, all the chromatograms have also been aligned using correlation optimized warping and Interval Correlation Optimized Shifting algorithms. The obtained results indicate that the longest distance shifting algorithm is simpler, faster and more effective, and will be potentially suitable for the alignment of other types of signals.     

Quality evaluation of Shenmaidihuang Pills based on the chromatographic fingerprints and simultaneous determination of seven bioactive constituents

In this study, we aimed to establish a comprehensive and practical quality evaluation system for Shenmaidihuang pills. A simple and reliable high‐performance liquid chromatography coupled with photodiode array detection method was developed both for fingerprint analysis and quantitative determination. In fingerprint analysis, relative retention time and relative peak area were used to identify the common peaks in 18 samples for investigation. Twenty one peaks were selected as the common peaks to evaluate the similarities of 18 Shenmaidihuang pills samples with different manufacture dates. Furthermore, similarity analysis was applied to evaluate the similarity of samples. Hierarchical cluster analysis and principal component analysis were also performed to evaluate the variation of Shenmaidihuang pills. In quantitative analysis, linear regressions, injection precisions, recovery, repeatability and sample stability were all tested and good results were obtained to simultaneously determine the seven identified compounds, namely, 5‐hydroxymethylfurfural, morroniside, loganin, paeonol, paeoniflorin, psoralen, isopsoralen in Shenmaidihuang pills. The contents of some analytes in different batches of samples indicated significant difference, especially for 5‐hydroxymethylfurfural. So, it was concluded that the chromatographic fingerprint method obtained by high‐performance liquid chromatography coupled with photodiode array detection associated with multiple compounds determination is a powerful and meaningful tool to comprehensively conduct the quality control of Shenmaidihuang pills.     

Acetone as a greener alternative to acetonitrile in liquid chromatographic fingerprinting

A considerable amount of chemical waste from liquid chromatography analysis is generated worldwide. Acetonitrile is the most employed solvent in liquid chromatography analyses since it exhibits favorable physicochemical properties for separation and detection, but it is an unwelcome solvent from an environmental point of view. Acetone might be a much greener alternative to replace acetonitrile in reversed‐phase liquid chromatography, since both share similar physicochemical properties, but its applicability with ultraviolet absorbance‐based detectors is limited. In this work, a reference method using acetonitrile and high‐performance liquid chromatography coupled to an ultraviolet photodiode array detector coupled to a corona charged aerosol detector system was developed to fingerprint a complex sample. The possibility of effectively substituting acetonitrile with acetone was investigated. Design of experiments was adopted to maximize the number of peaks acquired in both fingerprint developments. The methods with acetonitrile or acetone were successfully optimized and proved to be statistically similar when only the number of peaks or peak capacity was taken into consideration. However, the superiority of the latter was evidenced when parameters of separation and those related to greenness were heuristically combined. A green, comprehensive, time‐ and resource‐saving approach is presented here, which is generic and applicable to other complex matrices. Furthermore, it is in line with environmental legislation and analytical trends.     

Two‐way and three‐way approaches to ultra high performance liquid chromatography–photodiode array dataset for the quantitative resolution of a two‐component mixture containing ciprofloxacin and ornidazole

Two‐way and three‐way calibration models were applied to ultra high performance liquid chromatography with photodiode array data with coeluted peaks in the same wavelength and time regions for the simultaneous quantitation of ciprofloxacin and ornidazole in tablets. The chromatographic data cube (tensor) was obtained by recording chromatographic spectra of the standard and sample solutions containing ciprofloxacin and ornidazole with sulfadiazine as an internal standard as a function of time and wavelength. Parallel factor analysis and trilinear partial least squares were used as three‐way calibrations for the decomposition of the tensor, whereas three‐way unfolded partial least squares was applied as a two‐way calibration to the unfolded dataset obtained from the data array of ultra high performance liquid chromatography with photodiode array detection. The validity and ability of two‐way and three‐way analysis methods were tested by analyzing validation samples: synthetic mixture, interday and intraday samples, and standard addition samples. Results obtained from two‐way and three‐way calibrations were compared to those provided by traditional ultra high performance liquid chromatography. The proposed methods, parallel factor analysis, trilinear partial least squares, unfolded partial least squares, and traditional ultra high performance liquid chromatography were successfully applied to the quantitative estimation of the solid dosage form containing ciprofloxacin and ornidazole.     

Fast determination of four active compounds in Sanqi Panax Notoginseng Injection samples by high‐performance liquid chromatography with a chemometric method

The application of chemometric methods could decrease the requirements of separation and simplify time‐consuming pretreatment and experimental optimization. In this report, the accumulated projection method was developed and utilized for the rapid simultaneous quantification of four active components (notoginsenoside R1, ginsenosides Rg1, ginsenosides Re, and ginsenosides Rb1) in Sanqi Panax Notoginseng Injection samples. The proposed method is based on the three‐dimensional fingerprint spectra obtained from high‐performance liquid chromatography coupled with photodiode array detection. Although the chromatograms consisted of overlapping peaks, retention time shifts, and unknown interference, all established models showed good linearity (R > 0.9869) within test ranges. The relative standard deviation for intra‐ and interday precision of the four compounds did not exceed 2.4 and 4.7%, respectively, and the overall recovery was 91.2–106.8%. Compared with N‐way principal component analysis, our method provides more satisfactory results, which indicate that the proposed approach is simple, fast, and reliable for the determination of the four analytes in Sanqi Panax Notoginseng Injection samples.     

Rapid Determination of Costunolide and Dehydrocostuslactone in Human Plasma Sample and Chinese Patent Medicine Xiang Sha Yang Wei Capsule Using HPLC‐DAD Coupled with Second‐order Calibration

A novel methodology that combines high performance liquid chromatography with photodiode‐array detector (HPLC‐DAD) coupled with second‐order calibration method based on alternating trilinear decomposition (ATLD) algorithm was used in determination of the effective constituents such as costunolide and dehydrocostuslactone, in plasma sample and Chinese patent medicine Xiang Sha Yang Wei (XSYW) capsule. Complicated systems such as plasma and Chinese patent medicine which have intricate components are tedious to isolate and purify. The problem that chromatographic peaks are heavily overlapped among the analytes and interferents from the background matrices can be resolved, and the satisfactory quantification results have been gained with the help of the ATLD algorithm which utilized "mathematical separation" instead of partial "physical or chemical separation". Meanwhile, HPLC‐MS/MS method was used to validate the accuracy of the proposed determination method.     

Up‐and‐down shaker‐assisted ionic liquid‐based dispersive liquid—liquid microextraction of benzophenone‐type ultraviolet filters

Sun protection is an important part of our lives. UV filters are widely used to absorb solar radiation in sunscreens. However, excess UV filters constitute persistent groups of organic micropollutants present in the environment. An environmentally friendly ionic‐liquid‐based up‐and‐down shaker‐assisted dispersive liquid?liquid microextraction device combined with ultra‐performance liquid chromatography coupled with photodiode‐array detection has been developed to preconcentrate three UV filters (benzophenone, 2‐hydroxy‐4‐methoxybenzophenone, 2,2′‐dihydroxy‐4‐methoxybenzophenone) from field water samples. In this method, the optimal conditions for the proposed extraction method were: 40 μL [C₈MIM][PF₆] as extraction solvent and 200 μL methanol as disperser solvent were used to extract the UV filters. After up‐and‐down shaking for 3 min, the aqueous solution was centrifuged at 5000 rpm speed, then using microtube to collect the settled extraction solvent and using ultra‐performance liquid chromatography for further analysis. Quantification results indicated that the linear range was 2–1000 ng/mL. The LOD of this method was in the range 0.2–1.3 ng/mL with r² ≥ 0.9993. The relative recovery in studies of different types of field water samples was in the range 92–120%, and the RSD was 2.3–7.1%. The proposed method was also applied to the analysis of field samples.     

Fingerprint Analysis of Euonymus alatus (Thuhb) siebold by LC–DAD and LC–ESI–MS

A liquid chromatography coupled with photodiode array detection and electrospray ionization tandem mass spectrometry method was first developed for a chemical fingerprint analysis of Euonymus alatus (Thuhb) siebold (EAS) and rapid identification of major compounds in the fingerprints. Fingerprint profiles were found to be consistent for the herbs acquired from different locations, but the relative abundance of peaks was varied. Twelve peaks were chosen as the common peaks. Quercetin and rutin were detected by comparing the retention times, MS and UV spectra with the standards. The relative retention time and relative peak area of the 12 peaks in the fingerprint were calculated by setting the quercetin as the reference peak. The experimental data were used for similarity calculation and hierarchical clustering analysis. By comparing the UV and MS spectra data with those of the authentic standards and literature, five main peaks in the fingerprints were identified. Finally, five medicinal portions of the herb (leaf, fruit, stem, pterygium and root) were also analyzed by this method. It was found that there were similar chemical components in different parts of this herb but the contents were very different. The developed fingerprint assay was specific and could be readily utilized for comprehensive evaluation of EAS, as well as to distinguish different medicinal portions.     

Determination of residues of UV filters in natural waters by solid-phase extraction coupled to liquid chromatography-photodiode array detection and gas chromatography-mass spectrometry

This study describes a procedure for the enrichment, separation and quantification of four major UV filters in natural waters. Solid-phase extraction combined with liquid chromatography and photodiode array detection (LC-UV-DAD), and gas chromatography with mass spectroscopy (GC-MS) were employed for the analyses. LC of the four compounds with surfactant-modified hydro-organic eluents gave satisfactory resolution of overlapping peaks. In GC, a significant improvement of the detection limits was attained, but only three compounds could be detected. The method was validated for, and applied to, various water samples prone to UV filter accumulation due to recreational activities. Recoveries from real samples were 86-99% with LOQs as low as 0.5 ng/l depending on the sample volume and the analytical procedure.     

超高效液相色谱法检测土壤中的多环芳烃

采用二极管阵列(PDA)检测器,建立了超高效液相色谱(UPLC)定性定量分析土壤中16种多环芳烃(PAHs)的方法。并将该方法与传统高效液相色谱(HPLC)的分析性能进行了详细的比较。研究结果表明,采用UPLC法分析16种PAHs具有分析速度快(13.5 min)、检出限低(2～20 pg)、灵敏度高等优点。     

Quality assessment of crude and processed ginger by high‐performance liquid chromatography with diode array detection and mass spectrometry combined with chemometrics

A sensitive, simple, and validated high‐performance liquid chromatography with diode array detection and mass spectrometry detection method was developed for three ginger‐based traditional Chinese herbal drugs, Zingiberis Rhizoma, Zingiberis Rhizome Preparatum, and Zingiberis Rhizome Carbonisata. Chemometrics methods, such as principal component analysis, hierarchical cluster analysis, and analysis of variance, were also employed in the data analysis. The results clearly revealed significant differences among Zingiberis Rhizoma, Zingiberis Rhizome Preparatum, and Zingiberis Rhizome Carbonisata, indicating variations in their chemical compositions during the processing, which may elucidate the relationship of the thermal treatment with the change of the constituents and interpret their different clinical uses. Furthermore, the sample consistency of Zingiberis Rhizoma, Zingiberis Rhizome Preparatum, and Zingiberis Rhizome Carbonisata can also be visualized by high‐performance liquid chromatography with diode array detection and mass spectrometry analysis followed by principal component analysis/hierarchical cluster analysis. The comprehensive strategy of liquid chromatography with mass spectrometry analysis coupled with chemometrics should be useful in quality assurance for ginger‐based herbal drugs and other herbal medicines.     

Simultaneous qualitative and quantitative analysis of flavonoids and alkaloids from the leaves of Nelumbo nucifera Gaertn. using high‐performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry

A reliable method, combining qualitative analysis by high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry and quantitative assessment by high‐performance liquid chromatography with photodiode array detection, has been developed to simultaneously analyze flavonoids and alkaloids in lotus leaf extracts. In the qualitative analysis, a total of 30 compounds, including 12 flavonoids, 16 alkaloids, and two proanthocyanidins, were identified. The fragmentation behaviors of four types of flavone glycoside and three types of alkaloid are summarized. The mass spectra of four representative components, quercetin 3‐O‐glucuronide, norcoclaurine, nuciferine, and neferine, are shown to illustrate their fragmentation pathways. Five pairs of isomers were detected and three of them were distinguished by comparing the elution order with reference substances and the mass spectrometry data with reported data. In the quantitative analysis, 30 lotus leaf samples from different regions were analyzed to investigate the proportion of eight representative compounds. Quercetin 3‐O‐glucuronide was found to be the predominant constituent of lotus leaf extracts. For further discrimination among the samples, hierarchical cluster analysis, and principal component analysis, based on the areas of the eight quantitative peaks, were carried out.     

Rapid identification of triterpenoid sulfates and hydroxy fatty acids including two new constituents from Tydemania expeditionis by liquid chromatography-mass spectrometry

Tydemania expeditionis Weber-van Bosse (Udoteaceae) is a weakly calcified green alga. In the present paper, liquid chromatography coupled with photodiode array detection and electrospray mass spectrometry was developed to identify the fingerprint components. A total of four triterpenoid sulfates and three hydroxy fatty acids in the ethyl acetate fraction of the crude extract were structurally characterized on the basis of retention time, online UV spectrum, and mass fragmentation pattern. Furthermore, a detailed liquid chromatography-mass spectrometry analysis revealed two new hydroxy fatty acids, which were then prepared and characterized by extensive nuclear magnetic resonance (NMR) analyses. The proposed method provides a scientific and technical platform for the rapid identification of triterpenoid sulfates and hydroxy fatty acids in similar marine algae and terrestrial plants.     

Qualitative and Quantitative Analysis of Alkaloids in Cortex Phellodendri by HPLC-ESI-MS/MS and HPLC-DAD

A combined method of high performance liquid chromatograph-elecrtrospray-ionization mass spectrometer(HPLC-ESI-MS/MS) coupled with a photodiode array detector(HPLC-DAD) and principal component analysis(PCA) was applied to the qualitative and quantitative analyses of alkaloids in Cortex Phellodendri(CP) samples, and to the differentiation of two species of CP, Cortex Phellodendri Chinensis(CPC) and Cortex Phellodendri Amurensis(CPA). Twenty-two peaks appeared in the HPLC-MS base peak chromatogram of CP detec...     

Determination of total phthalates in edible oils by high‐performance liquid chromatography coupled with photodiode array detection

The previously reported procedure for the determination of the total phthalate in fatty food involved the extraction of phthalates using chloroform/methanol followed by the removal of the solvents before alkaline hydrolysis requiring 20 h and derivatization of phthalic acid. In this study, a phase‐transfer catalyst (tetrabutylammonium chloride) was used in the liquid–liquid heterogeneous hydrolysis of phthalates in oil matrix shortening the reaction time to within 25 min. The resulting phthalic acid in the hydrolysate was extracted by a novel molecular complex based dispersive liquid–liquid microextraction method coupled with back‐extraction before high‐performance liquid chromatography coupled with photodiode array detection. Under the optimal experimental conditions, the linearity of the method was in the range of 0.5–12 nmol/g with the correlation coefficients (r) >0.997. The detection limit (S/N = 3) was 0.11 nmol/g. Intraday and interday repeatability values expressed as relative standard deviation were 3.9 and 7.1%, respectively. The recovery rates ranged from 82.4 to 99.0%. The developed method was successfully applied for the analysis of total phthalate in seven edible oils.     

A rapid and efficient extraction method based on industrial MCM‐41‐miniaturized matrix solid‐phase dispersion extraction with response surface methodology for simultaneous quantification of six flavonoids in Pollen typhae by ultra‐high‐performance liquid chromatography

An industrial MCM‐41‐miniaturized matrix solid‐phase dispersion extraction coupled with response surface methodology was explored to determine L‐epicatechin, typhaneoside, isorhamnetin‐3‐O‐neohespeidoside, naringenin, kaempferol, and isorhamnetin in Pollen typhae by ultra‐high performance liquid chromatography connected to a photodiode array detection. Several variables were optimized in detail, including mesh number of sieve, type of adsorbent, mass ratio of sample to adsorbent, grinding time, methanol concentration, and elution volume. Central composite design was applied to optimize the best conditions for the maximum yields of the total flavonoids. The results displayed a good linear relationship (R > 0.9992) and the recoveries ranged from 92.9 to 103% (RSD < 4.53%) of the six flavonoids. The optimal method with high efficiency and low consumption was obviously better than heating reflux and ultrasonic extraction. It was proven that the developed industrial MCM‐41‐miniaturized matrix solid‐phase dispersion extraction coupled with simple ultra‐high performance liquid chromatography method could be a rapid and efficient tool for extraction and determination of flavonoids in natural products.     

Development of the Fingerprints for the Quality Evaluation of Scutellariae Radix by HPLC-DAD and LC-MS-MS

A high-performance liquid chromatography coupled with photodiode array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS²) method was firstly developed for a chemical fingerprint analysis of Scutellariae Radix and rapid identification of major compounds in the fingerprints. The experimental data for chromatography were used for hierarchical clustering analysis and similarity calculation, and those for UV and MS spectra were applied for identifying characteristic peaks. Twenty samples including S. baicalensis Georgi., S. viscidula Bge. and S. amoena C. H. Wright were classified into three groups. By comparing the UV and MS spectra data with those of the authentic standards and literature, 20 main peaks in the fingerprints were identified for the first time. The developed fingerprint assay was specific and could be readily utilized for comprehensive evaluation of Scutellariae Radix.     

Fast determination of anthocyanins and free pelargonidin in fruits,fruit juices,and fruit wines by high‐performance liquid chromatography using a core–shell column

Anthocyanins are water‐soluble pigments that are liable for colors ranging from red to blue of most fruits, vegetables, and flowers. A novel and fast method was developed for the determination of five anthocyanins and free pelargonidin by high‐performance liquid chromatography coupled to photodiode array detection. A 10% formic acid and acetonitrile mixture was employed as mobile phase in the gradient elution mode. Mobile phase composition, column temperature, flow rate, injection volume, and column conditioning time were optimized by employing a stepwise strategy. Using a C₁₈ core–shell column (100 × 4.6 mm, 2.7 μm), the separation of six analytes was accomplished in less than 9.5 min with a run‐to‐run analysis time of 19 min. The developed method was validated with respect to linearity (r > 0.9999), limit of detection, limit of quantification, intra‐/interday precision (<2%), accuracy (98.6–104.4%), and specificity. Afterwards, the method was applied to the determination of anthocyanins present in 15 different samples including fruits, fruit juices, and fruit wines.     

Semi-automated alignment and quantification of peaks using parallel factor analysis for comprehensive two-dimensional liquid chromatography–diode array detector data sets

Parallel factor analysis was used to quantify the relative concentrations of peaks within four-way comprehensive two dimensional liquid chromatography–diode array detector data sets. Since parallel factor analysis requires that the retention times of peaks between each injection are reproducible, a semi-automated alignment method was developed that utilizes the spectra of the compounds to independently align the peaks without the need for a reference injection. Peak alignment is achieved by shifting the optimized chromatographic component profiles from a three-way parallel factor analysis model applied to each injection. To ensure accurate shifting, components are matched up based on their spectral signature and the position of the peak in both chromatographic dimensions. The degree of shift, for each peak, is determined by calculating the distance between the median data point of the respective dimension (in either the second or first chromatographic dimension) and the maximum data point of the peak furthest from the median. All peaks that were matched to this peak are then aligned to this common retention data point. Target analyte recoveries for four simulated data sets were within 2% of 100% recovery in all cases. Two different experimental data sets were also evaluated. Precision of quantification of two spectrally similar and partially coeluting peaks present in urine was as good as or better than 4%. Good results were also obtained for a challenging analysis of phenytoin in waste water effluent, where the results of the semi-automated alignment method agreed with the reference LC–LC MS/MS method within the precision of the methods.