Chemical Fingerprint Analysis Research Articles

Chemical Fingerprint Analysis and Quantitative Analysis of Saccharides in Morindae Officinalis Radix by HPLC-ELSD.

A method based on high performance liquid chromatography with evaporative light scattering detection (HPLC-ELSD) was developed for the quantitative analysis of three active compounds and chemical fingerprint analyses of saccharides in Morindae officinalis radix. Ten batches of Morindae officinalis radix were collected from different plantations in the Guangdong region of China and used to establish the fingerprint. The samples were separated with a COSMOIL Sugar-D column (4.6 mm × 250 mm, 5 μm) by using gradient elution with water (A) and acetonitrile (B). In addition, Trapped-Ion-Mobility (tims) Time-Of-Flight (tims TOF) was used to identify saccharides of Morindae officinalis radix. Fingerprint chromatogram presented 26 common characteristic peaks in the roots of Morinda officinalis How, and the similarities were more than 0.926. In quantitative analysis, the three compounds showed good regression (r = 0.9995–0.9998) within the test ranges, and the recoveries of the method were in the range of 96.7–101.7%. The contents of sucrose, kestose and nystose in all samples were determined as 1.21–7.92%, 1.02–3.37%, and 2.38–6.55%, respectively. The developed HPLC fingerprint method is reliable and was validated for the quality control and identification of Morindae officinalis radix and can be successfully used to assess the quality of Morindae officinalis radix.

Quantitative Chemical Analysis of Fingerprints with Ion Chromatography

A single latent fingerprint consists of ∼50 μg of matter for which the anionic content often provides a differentiating characteristic between individuals. This forensic-themed, analytical chemistry experiment exploits this difference and has students employ ion-exchange chromatography to quantitatively analyze the anionic content of both authentic and synthetic fingerprints in order to identify, condemn, or exonerate “suspects”.

Quality evaluation of commercial products of Ganoderma lucidum made from its fruiting body and spore

Abstract Ganoderma lucidum (GL), also known as Reishi or Lingzhi, is a medicinal mushroom widely used in traditional and folk medicines. The extracts made from the fruiting body and spore of naturally grown GL are the most frequently used in commercial products. More than 400 compounds have been identified in GL with the triterpenoids considered to be the major active components. Large variations in the chemical components were reported in previous studies and there is no comprehensive study of the content of multiple major triterpenoids in the GL product. In addition, there is no report in the comparison of chemical profiles in different parts of GL (i.e., fruiting body and spore). Determining the chemical composition and comparing the differences between fruiting body and spore are essential for the identity, efficacy and safety of various GL products. In this study, 13 compounds (ganoderenic Acid C, ganoderic Acid C2, ganoderic Acid G, ganoderic Acid B, ganoderenic Acid B, ganoderic Acid A, ganoderic Acid H, ganoderenic Acid D, ganoderic Acid D, ganoderic Acid F, ganoderic Acid DM, ganoderol A, and ergosterol) were selected as the chemical markers. The purpose of this study is to develop an HPLC-DAD fingerprint method for quantification of these active components in GL (spore and fruiting body) and test the feasibility of using the HPLC-DAD fingerprint for quality control or identity determination of GL products. The results showed that this method could determine the levels of the major components accurately and precisely. Among the 13 components, 11 ganoderma acids were identified to be proper chemical markers for quality control of GL products, while ganoderal A was in a very low amount and ergosterol was not a specific marker in GL. The extracts of fruiting body contained more chemical compounds than those of spore, indicating that these 11 compounds could be a better chemical marker for the fruiting body than the spore. The HPLC chemical fingerprint analysis showed higher variability in the quality of GL harvest in different years, while lesser variation in batches harvested in the same year. In conclusion, an HPLC assay detecting 11 major active components and a fingerprinting method was successfully established and validated to be feasible for quality control of most commercial GL products.

Comparison of Geographical Traceability of Wild and Cultivated Macrohyporia cocos with Different Data Fusion Approaches.

Poria originated from the dried sclerotium of Macrohyporia cocos is an edible traditional Chinese medicine with high economic value. Due to the significant difference in quality between wild and cultivated M. cocos, this study aimed to trace the origin of the fungus from the perspectives of wild and cultivation. In addition, there were quite limited studies about data fusion, a potential strategy, employed and discussed in the geographical traceability of M. cocos. Therefore, we traced the origin of M. cocos from the perspectives of wild and cultivation using multiple data fusion approaches. Supervised pattern recognition techniques, like partial least squares discriminant analysis (PLS-DA) and random forest, were employed in this study using. Five types of data fusion involving low-, mid-, and high-level data fusion strategies were performed. Two feature extraction approaches including the selecting variables by a random forest-based method—Boruta algorithm and producing principal components by the dimension reduction technique of principal component analysis—were considered in data fusion. The results indicate the following: (1) The difference between wild and cultivated samples did exist in terms of the content analysis of vital chemical components and fingerprint analysis. (2) Wild samples need data fusion to realize the origin traceability, and the accuracy of the validation set was 95.24%. (3) Boruta outperformed principal component analysis (PCA) in feature extraction. (4) The mid-level Boruta PLS-DA model took full advantage of information synergy and showed the best performance. This study proved that both geographical traceability and optimal identification methods of cultivated and wild samples were different, and data fusion was a potential technique in the geographical identification.

Chemical Fingerprint Analysis and Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry-Based Metabolomics Study of the Protective Effect of Buxue Yimu Granule in Medical-Induced Incomplete Abortion Rats.

Medical abortion is a common method to terminate an early pregnancy and often causes serious complications such as abnormal uterine bleeding and endometritis. Buxue Yimu granule (BYG) is a well-known traditional Chinese medicine prescription composed of five kinds of drugs and is widely used in gynecology and obstetrics. The aim of the present study was to establish the quality standard of BYG and investigate its protective effect on incomplete abortion. The chemical fingerprint of BYG was established by high performance liquid chromatography (HPLC). The major compounds of BYG were determined by ultra-performance liquid chromatography with triple quadrupole mass spectrometry. An incomplete abortion rat model was induced by intragastric administration of mifepristone (8.3 mg·kg−1) combined with misoprostol (100.0 μg·kg−1) during early pregnancy. The serum levels of human chorionic gonadotrophin (HCG), estradiol (E2), and progesterone (PG) were determined. The serum endogenous metabolites were analyzed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Multivariate analysis, including partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA), was employed to analyze the metabolic profiles, and MetaboAnalyst was used to investigate the metabolic pathways. Furthermore, hematoxylin-eosin staining (HE) was used to evaluate the histopathological changes in uterine tissue. The expression levels of VEGFA and NF-κB were detected by immunohistochemistry. The results indicated that HPLC fingerprint analysis can be successfully used to assess the quality of BYG. The medical-induced incomplete abortion rats were clearly separated from control rats, and the biochemical changes were gradually restored to normal after administration of BYG. Moreover, 19 potential biomarkers, including N-lactoylleucine, 2-piperidinone, isobutyryl-l-carnitine, eicosapentaenoylcholine, LysoPC(14:0), LysoPC(20:5), physagulin C, LysoPC(18:3), leukotriene D5, deoxycholic acid 3-glucuronide, glycine, pregnanediol 3-O-glucuronide, LysoPC(18:2), LysoPC(17:0/0:0), N-acetyl-leukotriene E4, LysoPC(18:0), platelet-activating factor, LysoPA(24:1), and LysoPC(18:1), which were mainly related to the amino acids metabolism, lipids metabolism, and bile acid biosynthesis, were identified. Consequently, BYG exerts a potential protective role in the intervention of incomplete abortion by anti-inflammatory, promote endometrial repair, and regulate the metabolic disorders.

Intervention of standardized ethanol leaf extract of Annickia polycarpa, (DC.) Setten and Maas ex I.M. Turner. (Annonaceae), in Plasmodium berghei infested mice produced anti-malaria action and normalized gross hematological indices

Ethnopharmacological relevanceMalaria is a global public health burden due to large number of annual infections and casualties caused by its hematological complications. The bark of Annickia polycarpa is an effective anti-malaria agent in African traditional medicine. However, there is no standardization parameters for A. polycarpa. The anti-malaria properties of its leaf are also not known. Aim of the studyTo standardize the ethanol leaf extract of A. polycarpa (APLE) and investigate its anti-malaria properties and the effect of its treatment on hematological indices in Plasmodium berghei infected mice in the Rane's test. Materials and methodsMalaria was induced by inoculating female ICR mice with 1.0 × 107P. berghei-infected RBCs in 0.2 mL (i.p.) of blood. Treatment was commenced 3 days later with APLE 50, 200, 400 mg/kg p.o., Quinine 30 mg/kg i.m. (Standard drug) or sterile water (Negative control) once daily per group for 4 successive days. Anti-malarial activity and gross malaria indices such as hyperparasitemia, mean change in body weight and mean survival time (MST) were determined for each group. Changes in white blood cells (WBCs), red blood cells (RBCs), platelets (PLT) counts, hemoglobin (HGB) concentration, hematocrit (HCT) and mean corpuscular volume (MCV) were also measured in the healthy mice before infection as baseline and on day 3 and 8 after inoculation using complete blood count. Standardization was achieved by UHPLC-MS chemical fingerprint analysis and quantitative phytochemical tests. ResultsAPLE, standardized to its total alkaloids, phenolics and saponin contents, produced significant (P < 0.05) dose-dependent clearance of mean hyperparasitemia of 22.78 ± 0.93% with the minimum parasitemia level of 2.01 ± 0.25% achieved at 400 mg/kg p.o. on day 8. Quinine 30 mg/kg i.m. achieved a minimum parasitemia level of 6.15 ± 0.92%. Moreover, APLE (50–400 mg/kg p.o.) evoked very significant anti-malaria activity of 89.22–95.50%. Anti-malaria activity of Quinine 30 mg/kg i.m. was 86.22%. APLE also inverse dose-dependently promotes weight gain with the effect being significant (P < 0.05) at 50 mg/kg p.o. Moreover, APLE dose-dependently increased the MST of malaria infested mice with 100% survival at 400 mg/kg p.o. Quinine 30 mg/kg i.m. also produce 100% survival rate but did not promote (P > 0.05) weight gain. Hematological studies revealed the development of leukocytopenia, erythrocytosis, microcytic anemia and thrombocytopenia in the malaria infected mice which were reverted with the treatment of APLE 50–400 mg/kg p.o. or Quinine 30 mg/kg i.m. but persisted in the negative control. The UHPLC-MS fingerprint analysis of APLE led to identification of one oxoaporphine and two aporphine alkaloids (1–3). Alkaloids 1 and 3 are being reported in this plant for the first time. ConclusionThese results indicate that APLE possessed significant anti-malaria, immunomodulatory, erythropoietic and hematinic actions against malaria infection. APLE also has the ability to revoke deleterious physiological alteration produced by malaria and hence, promote clinical cure. These properties of APLE are due to its constituents especially, aporphine and oxoaporphine alkaloids.

Simultaneous Qualitative and Quantitative Evaluation of the Coptidis Rhizoma and Euodiae Fructus Herbal Pair by Using UHPLC-ESI-QTOF-MS and UHPLC-DAD.

The herbal pair of Coptidis Rhizoma (CR) and Euodiae Fructus (EF) is a classical traditional Chinese medicine formula used for treating gastro-intestinal disorders. In this study, we established a systematic method for chemical profiling and quantification analysis of the major constituents in the CR-EF herbal pair. A method of ultra high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) for qualitative analysis was developed. Sixty-five compounds, including alkaloids, phenolics, and limonoids, were identified or tentatively assigned by comparison with reference standards or literature data. The UHPLC fingerprints of 19 batches of the CR-EF herbal pair samples were obtained and the reference fingerprint chromatograms were established. Furthermore, nine compounds among 24 common peaks of fingerprints were considered as marker components, which either had high contents or significant bioactivities, were applied to quality control of the CR-EF herbal pair by quantitative analysis. This UHPLC-DAD analysis method was validated by precision, linearity, repeatability, stability, recovery, and so on. The method was simple and sensitive, and thus reliable for quantitative and chemical fingerprint analysis for the quality evaluation and control of the CR-EF herbal pair and related traditional Chinese medicines.

Antrocaryon micraster (A. Chev. And Guillaumin) stem bark extract demonstrated anti-malaria action and normalized hematological indices in Plasmodium berghei infested mice in the Rane's test

Ethnopharmacological relevanceMalaria is caused by infection with some species of Plasmodium parasite which leads to adverse alterations in physical and hematological features of infected persons and ultimately results in death. Antrocaryon micraster is used to treat malaria in Ghanaian traditional medicine. However, there is no scientific validation of its anti-malaria properties. The plant does not also have any chemical fingerprint or standardization parameters. Aim of the studyThis study sought to evaluate the anti-malaria activity of standardized A. micraster stem bark extract (AMSBE) and its effect on mean survival time (MST) and body weight reduction of Plasmodiumberghei infested mice. And to study the effect of treatment of AMSBE on hematological indices of the P. berghei infested mice in order to partly elucidate its anti-malarial mechanism of action. Materials and methodsMalaria was induced in female ICR mice by infecting them with 0.2 mL of blood (i.p.) containing 1.0 × 107P. berghei-infested RBCs from a donor mouse and leaving them without treatment for 3 days. AMSBE or Lonart (standard control) was then orally administered at 50, 200 and 400 mg/kg or 10 mg/kg once daily for 4 consecutive days. The untreated control received sterile water. Malaria parasitemia reduction, anti-malarial activity, mean change in body weight and MST of the parasitized mice were evaluated. Furthermore, changes in white blood cells (WBCs), red blood cells (RBCs), platelets count, hemoglobin (HGB), hematocrit (HCT) and mean corpuscular volume (MCV) were also determined in the healthy animals before infection as baseline and on days 3, 5 and 8 after infection by employing complete blood count. Standardization of AMSBE was achieved by quantification of its constituents and chemical fingerprint analysis using UHPLC-MS. ResultsAdministration of AMSBE, standardized to 41.51% saponins and 234.960 ± 0.026 mg/g of GAE phenolics, produced significant (P < 0.05) reduction of parasitemia development, maximum anti-malaria activity of 46.01% (comparable to 32.53% produced by Lonart) and significantly (P < 0.05) increased body weight and MST of P. berghei infected mice compared to the untreated control. Moreover, there were significant (P > 0.05) elevation in WBCs, RBCs, HGB, HCT and platelets in the parasitized-AMSBE (especially at 400 mg/kg p.o.) treated mice compared to their baseline values. Whereas, the non-treated parasitized control recorded significant reduction (P < 0.05) in all the above-mentioned parameters compared to its baseline values. The UHPLC-MS fingerprint of AMSBE revealed four compounds with their retention times, percentage composition in their chromatograms and m/z of the molecular ions and fragments in the spectra. ConclusionsThese results show that A. micraster stem bark possessed significant anti-malaria effect and also has the ability to abolish body weight loss, leucopenia, anemia and thrombocytopenia in P. berghei infected mice leading to prolonged life span. The UHPLC-MS fingerprint developed for AMSBE can be used for rapid authentication and standardization of A. micraster specimens and herbal preparations produced from its hydroethanolic stem bark extract to ensure consistent biological activity. The results justify A. micraster's use as anti-malaria agent.

Chemical Fingerprint Analysis for Discovering Markers and Identifying Saussurea involucrata by HPLC Coupled with OPLS-DA.

The quality control of Saussurea involucrata has been greatly improved by macroscopic and microscopic identification and chemical profiling described in Chinese Pharmacopoeia since 2005. However, these methods have their own limitations, e.g., their dependence on personal experience and expertise, and it is a huge challenge to identify closely related species that share similar or identical morphological characteristics and chemical profiles. A novel and generally accepted identification strategy is urgently needed as a complement to regulations for protecting the public health interests. In this work, a comprehensive chromatographic fingerprint method was developed and tested on 43 samples from four haplotypes of S. involucrata according to DNA barcoding. Three common patterns consisting of 20, 14, and 7 common peaks were generated by frequency filters of median, upper quartile, and 100%, respectively. Based on two formerly screened patterns, S. involucrata can be effectively identified from its five easily confused snow lotus species, including the most closely related plant (S. orgaadayi) in the orthogonal partial least-squares discriminant analysis (OPLS-DA) models. The model is supported by good R and Q coefficients. In addition, different haplotypes of S. involucrata can be discriminated in the OPLS-DA model using the 20 common peaks. Among them, peaks 9, 11, 16 (zaluzanin C), and 18 (dehydrocostus lactone) have been identified as fingerprint markers of S. involucrata via S-plots and VIP values (>1). Additionally, peaks 19 and 20 were identified as linolenic acid and linoleic acid with anti-inflammatory activity, and they were isolated from the herb for the first time. Collectively, the chromatographic fingerprint of S. involucrata can be an effective and integrated method for the identification of authentic herbs from adulterant species or related plants, and discrimination of its different haplotypes provides an objective and reliable tool for quality control.

In Vitro Growth- and Encystation-Inhibitory Efficacies of Matcha Green Tea and Epigallocatechin Gallate Against Acanthameoba Castellanii.

We examined the inhibitory effect of matcha green tea (Camellia sinensis) and epigallocatechin gallate (EGCg; the most abundant catechin in tea) on the vegetative growth and encystation of Acanthamoeba castellanii T4 genotype. The sulforhodamine B (SRB) stain-based colorimetric assay and hemocytometer counting were used to determine the reduction in A. castellanii trophozoite proliferation and encystation, in response to treatment with C. sinensis or EGCg. Fourier transform infrared (FTIR) microscopy was used to analyze chemical changes in the trophozoites and cysts due to C. sinensis treatment. Hot brewed and cold brewed matcha inhibited the growth of trophozoites by >40% at a 100 % concentration. EGCg at concentrations of 50 to 500 µM significantly inhibited the trophozoite growth compared to control. Hot brewed matcha (100% concentration) also showed an 87% reduction in the rate of encystation compared to untreated control. Although 500 µM of EGCg increased the rate of encystation by 36.3%, 1000 µM reduced it by 27.7%. Both percentages were not significant compared to control. C. sinensis induced more cytotoxicity to Madin Darby canine kidney cells compared to EGCg. FTIR chemical fingerprinting analysis showed that treatment with brewed matcha significantly increased the levels of glycogen and carbohydrate in trophozoites and cysts.

Chemical Fingerprint Analysis and Simultaneous Determination of Nucleosides and Amino Acids in Kang Fu Xin Liquid by High Performance Liquid Chromatography with Diode Array Detector

Background and Objective: Kang Fu Xin liquid (KFX) is an official preparation made from the ethanol extract product from P. Americana. The present quality control method cannot control the quality of the preparation well. The aim of the present study is to establish a convenient HPLC method for multicomponents determination combined with fingerprint analysis for quality control of KFX. Methods: An HPLC-DAD method with gradient elution and detective wavelength switching program was developed to establish HPLC fingerprints of KFX, and 38 batches of KFX were compared and evaluated by similarity analysis (SA), hierarchical clustering analysis (HCA), and principal component analysis (PCA). Meanwhile, six nucleosides and three amino acids, including uracil, hypoxanthine, uric acid, adenosine, xanthine, inosine, tyrosine, phenylalanine and tryptophan in KFX were determined based on the HPLC fingerprints. Results: An HPLC method assisted with gradient elution and wavelength switching program was established and validated for multicomponents determination combined with fingerprint analysis of KFX. The results demonstrated that the similarity values of the KFX samples were more than 0.845. PCA indicated that peaks 4 (hypoxanthine), 7 (xanthine), 9 (tyrosine), 11, 13 and 17 might be the characteristic contributed components. The nine constituents in KFX, uracil, hypoxanthine, uric acid, adenosine, xanthine, inosine, tyrosine, phenylalanine and tryptophan, showed good regression (R2 &gt; 0.9997) within test ranges and the recoveries of the method for all analytes were in the range from 96.74 to 104.24%. The limits of detections and quantifications for nine constituents in DAD were less than 0.22 and 0.43 μg•mL-1, respectively. Conclusion: The qualitative analysis of chemical fingerprints and the quantitative analysis of multiple indicators provide a powerful and rational way to control the KFX quality for pharmaceutical companies.

Holistic quality evaluation of Qingwen Baidu Decoction and its anti-inflammatory effects

Ethnopharmacological relevanceQingwen Baidu Decoction (QBD), a famous traditional Chinese medicine prescription with heat-clearing and detoxifying efficacies, is widely used in the treatment of inflammatory diseases. However, due to lack of holistic quality evaluation research, the further study on the detailed molecular mechanisms of action are still insufficient. Aim of the studyThis study aimed to evaluate the overall quality of QBD and to explore the anti-inflammatory effects and associated intracellular signaling pathways. Materials and methodsa comprehensive method of chemical fingerprint analysis and simultaneous multi-component quantification was firstly developed by high performance liquid chromatography with diode array detector (HPLC-DAD). Similarity analysis, principal component analysis and hierarchical cluster analysis with heatmap were also applied to screen out the markers components in QBD samples. Moreover, its anti-inflammatory effects and mechanisms were further investigated by survival analysis, hematoxylin-eosin staining (H&E), neutrophil observation, quantitative real-time PCR analysis (qRT-PCR), Western blotting and confocal microscopy. ResultsTwenty-one characteristic peaks from 11 herbs were chemically identified in the chromatographic fingerprint. Fifteen quantitative markers from 11 herbs, such as baicalin, wogonoside, geniposidic acid, oxypaeoniflora and so on, were screened out with the aid of chemometrics to further quantitatively assess the quality of QBD. The results of survival analysis, H&E and neutrophil observation in zebrafish inflammatory models consistently showed that QBD exerts potent anti-inflammatory effects in a dose-dependent manner. Additionally, QBD inhibited the activation of NF-κB and STAT3 signal pathways in LPS-induced zebrafish and RAW 264.7 macrophage cells. ConclusionCollectively, our investigations firstly described the chemical profile of QBD and its possible mechanism of anti-inflammation, which provides a preferred strategy for monitoring the overall quality of QBD and supports its clinical application in treating inflammation-related diseases.

Chemical fingerprint analysis of Gardenia jasminoides fruit by high-performance liquid chromatography

A high performance liquid chromatography-ultraviolet detector-electrospray ionization tandem mass spectrometry (HPLC/UV/ESI/MSn) method was used for the chromatographic fingerprint analysis of Gardenia jasminoides fruit, Fructus Gardeniae (zhi-zi in Chinese). Thirteen batches of samples collected from 8 provinces were analyzed to standardize the fingerprint of zhi-zi. The thirteen peaks in the fingerprints of all the 13 batches of samples with reasonable height and good resolution were assigned as "characteristic peaks". Seven "characteristic peaks" were identified by comparing their retention time and MS (mass spectrum) with those of the reference substances for the first time. The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine software was used to evaluate the similarities among the 13 batches of zhi-zi samples. The results indicated that samples from different origins had similar HPLC fingerprint and the method could be applied for the quality control of zhi-zi. In addition, the fingerprint can be used to analyze the differences of zhi-zi samples with different harvest time and different parts of Gardenia jasminoides including calyx, root, fruit (zhi-zi), leaf and stem. The results revealed that the chemical compounds in some samples picked in November varied significantly from samples in September and October. The content of geniposide in zhi-zi picked in September and October was much higher than that picked in November. Calyx, leaf, stem and root of Gardenia jasminoides had similar chemical components that were significantly different from those of zhi-zi.

Chemical fingerprint and quantitative analysis of Cistanche deserticola by HPLC-DAD-ESI-MS

Chemical fingerprint and quantitative analysis of Cistanche deserticola by HPLC-DAD-ESI-MS

Quantitative and chemical fingerprint analysis for quality control of Zingiber zerumbet based on HPTLC combined with chemometric methods

A simple, reliable high-performance thin-layer chromatography (HPTLC) method was developed for chemical fingerprinting of Zingiber zerumbet and quantitative estimation of zerumbone. Thirty-six batches of Z. zerumbet were collected from five eco-regions of eastern India. Zerumbone content varied from 52.4 to 214.6 mg/g (dry weight) in methanolic extract of Z. zerumbet rhizomes. Zerumbone content was in the following order: moist deciduous forests of the lower Gangetic plains > Brahmaputra valley evergreen forest > Odisha semi-evergreen forests > Sundarbans freshwater swamp forests > moist deciduous forest of the eastern highlands. Relative Standard Deviation (RSD) of the relative peak areas (RPA) and relative retention times (RRT) of eight characteristic peaks in repeatability and stability test were <3%, and the fingerprinting method was confirmed to be suitable for Z. zerumbet rhizomes. Chemometric approaches like hierarchical cluster analysis (HCA) and principal component analysis (PCA) were employed to classify Z. zerumbet samples based upon their eco-region. Consistent results were achieved showing Z. zerumbet samples could be effectively grouped according to their eco-region. The PCA loading plots identified three probable chemical markers, which might be useful in discriminating the samples. This combinative approach could be used for quality assessment of Z. zerumbet and for the formulations containing zerumbone.

HPTLC and UPLC-MS/MS Methods for Quality Control Analysis of Itrifal Formulations of Unani System of Medicine.

Ultra-performance LC (UPLC)-tandem MS (MS/MS) and high-performance TLC (HPTLC) assay methods were developed for chemical fingerprinting and quantitative analysis of bioactive constituents of a certain "Itrifal formulation," a traditional kind of Unani medicine. In the present investigation, HPTLC and UPLC-MS/MS methods were developed and validated for the detection and quantification of major metabolites present in itrifal formulation. The metabolites present in the formulation were separated using modern chromatographic techniques, and a quantitative analysis was performed. Analytical performance of the proposed HPTLC and UPLC-MS/MS methods was validated as per the defined guidelines with respect to linearity, accuracy, precision, robustness, and specificity. The developed UPLC-MS/MS and HPTLC methods were used for quantification of gallic acid, tannic acid, catechin, and quercetin. All four constituents were quantified by UPLC-MS/MS, while two constituents were quantified by HPTLC in the commercial itrifal formulation. The calibration plot was found to be linear, accurate, precise, robust, and specific for both HPTLC and UPLC-MS/MS. The present methods were successfully applied for analysis of the given markers in itrifal formulations. The same can be used for QC and stability testing of itrifal formulations.

Analysis of the similarities and differences between Auclandia and Vladimirae rhizomes by chemical profiling and chemometric analysis

Ethnopharmacological relevanceAucklandiae Radix (AR) and Vladimiriae Radix (VR), as traditional Chinese medicine, have been included in many editions of Chinese Pharmacopoeia with similar efficacy such as promoting qi and relieving pain, which are used to treat chest, hypochondriac, abdominal fullness and pain, diarrhea and tenesmus. In most conditions, VR is used to be a substitute of AR or a local habit. However, whether VR could substitute for AR to play a same role in the formulation and clinical applications needs further study. Aim of the studyIn this study, similarities and differences between AR and VR would be assessed, and possible reasons that may influence the efficacy of the AR and VR would be explained from the perspective of chemical composition. Materials and methodsHPLC-PDA was used to obtain the data of 10 batches of AR and VR, and to establish chemical fingerprint and chemometric analysis. UPLC-ESI-Q-TOF-MS was used to identify the structure of chemical compounds which contributed to the differences between AR and VR. ResultsThe chemical fingerprint analysis results showed that 20 peaks in common for AR and 26 peaks in common for VR both presented a good similarity (>0.9), and 15 peaks in common for AR and VR also showed a good similarity (>0.9). Nevertheless, chemometric showed AR was distinct from VR and three chemical compounds, which leading to their differences, were identified by UPLC-ESI-Q-TOF-MS. The three chemical compounds were 3β-acetoxy-11β-guaia-4 (15),10 (14)-diene-12,6α-olide, 10α,14-epoxy-11β-guaia-4 (15)-ene-12,6α-olide and costunolide, respectively. ConclusionIn general, AR and VR were highly similar, but their differences were deserved to be paid attention to. This research could provide reference for quality control and set a foundation for clinical applications of AR and VR.

Quantitative and Chemical Fingerprint Analysis of Desmodium styracifolium by High-Performance Liquid Chromatography Combined with Chemometrics.

In this study, a valid and comprehensive evaluation method for assessing the quality of Desmodium styracifolium (Osb.) Merr has been established, based on analysis of high-performance liquid chromatography fingerprint combined with the similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), discriminant analysis (DA) and the quantitative analysis multi-components by single marker (QAMS) method. Eleven peaks of the common model were obtained and analyzed using SA, HCA, PCA and DA analysis. These methods indicated a similar conclusion that 31 batches of D. styracifolium samples were categorized into two clusters basically coincident with their geographical regions of origin. Four peaks were identified as schaftoside, isoorientin, isoschaftoside and isovitexin. Schaftoside was selected as the internal standard, and the relative correction factors between schaftoside and the other three flavonoids were calculated using the QAMS method. The accuracy of the QAMS method was verified by comparing with the results calculated by the external standard method. No significant difference between the two methods was found. In conclusion, the established methods were scientifically applied in the quality evaluation of D. styracifolium.

Chemical Analysis and Imaging of Fingerprints by Air-flow Assisted Desorption Electrospray Ionization Mass Spectrometry

Fingerprint analysis is of great significance in forensic sciences. Compared with existing fingerprint analytical methods, mass spectrometry-based methods can not only identify chemical components in fingerprints, but also obtain fingerprint imaging. In this study four kinds of fingerprints, including sweat, inkpad, sunscreen, and liquid foundation, were analyzed by air-flow assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI). AFADESI was employed with air flow of 45 L min−1, and 5 μL min−1 acetonitrile was used as spray solvent at spray voltage of 7000 V. Positive ion full scan mode (100−1000 Da) was chosen. The results showed that AFADESI-MSI method could not only obtain chemical information of various endogenous and exogenous substances in fingerprints, but also obtain high resolution images of fingerprints. In addition, overlapped fingerprints were distinguished according to the typical chemical information in fingerprints. As a new fingerprint analysis method, AFADESI-MSI would be widely used in forensic scientific research and practical applications.

Discrimination of the Geographical Origin of the Lateral Roots of Aconitum carmichaelii Using the Fingerprint, Multicomponent Quantification, and Chemometric Methods.

Fuzi is a well-known traditional Chinese medicine developed from the lateral roots of Aconitum carmichaelii Debx. It is rich in alkaloids that display a wide variety of bioactivities, and it has a strong cardiotoxicity and neurotoxicity. In order to discriminate the geographical origin and evaluate the quality of this medicine, a method based on high-performance liquid chromatography (HPLC) was developed for multicomponent quantification and chemical fingerprint analysis. The measured results of 32 batches of Fuzi from three different regions were evaluated by chemometric analysis, including similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA), and linear discriminant analysis (LDA). The content of six representative alkaloids of Fuzi (benzoylmesaconine, benzoylhypaconine, benzoylaconine, mesaconitine, hypaconitine, and aconitine) were varied by geographical origin, and the content ratios of the benzoylmesaconine/mesaconitine and diester-type/monoester-type diterpenoid alkaloids may be potential traits for classifying the geographical origin of the medicine. In the HPLC fingerprint similarity analysis, the Fuzi from Jiangyou, Sichuan, was distinguished from the Fuzi from Butuo, Sichuan, and the Fuzi from Yunnan. Based on the HCA and PCA analyses of the content of the six representative alkaloids, all of the batches were classified into two categories, which were closely related to the plants’ geographical origins. The Fuzi samples from Jiangyou were placed into one category, while the Fuzi samples from Butuo and Yunnan were put into another category. The LDA analysis provided an efficient and satisfactory prediction model for differentiating the Fuzi samples from the above-mentioned three geographical origins. Thus, the content of the six representative alkaloids and the fingerprint similarity values were useful markers for differentiating the geographical origin of the Fuzi samples.