Quantitative Analysis Of Alkaloids Research Articles

Qualitative and Quantitative Phytochemical Analysis of Gongronema latifolium Leaf Extract

Phytochemical compounds are secondary metabolites produced by plants in which some have medicinal uses. The aim of this study is to qualitatively and quantitatively analyze the phytochemical composition of G. latifolium leaf extract. Fresh and healthy leaves of the medicinal plant of G. latifolium leaf were harvested, washed thoroughly in distilled water and dried at room temperature for 7 days. The dried leaf was milled to fine powder. The pulverized plant was weighed and used for the phytochemical analysis. Based on qualitative analysis, the results show the presence of tannins, alkaloids, saponins, flavonoids, terpenoids, reducing sugar, cardiac glycoside, protein and carbohydrate, while steroids was absent. The quantitative analysis of alkaloids, saponins, flavonoids and tannins were done and the results obtained were expressed as a mean± SEM in each group, analysed using ANOVA followed by Dunnett’s test P<0.05 considered as significant. The quantitative analysis revealed that alkaloids, saponins, flavonoids and tannins had mean results of 9.85±1.50, 9.66±0.77, 6.83±6.25 and 4.54±6.70 mg/100mg respectively. This result shows that G. latifolium leaf extract possesses an appreciable level of phytochemicals which contributed to its efficacy in the treatment of diseases such as helminthiasis.

Research progress on the analysis of active ingredients and elements in Fritillaria ussuriensis Bulbus

Fritillaria ussuriensis Bulbus, a genuine medicinal material of Northeast China, is the dry bulb of Fritillaria ussuriensis Maxim. It contains various active ingredients, such as alkaloids, alkaloids glycosides, adenosines, polysaccharides, trace elements and so forth. It has antitussive, eliminating phlegm, antiasthmatic, antiulcer, antiplatelet aggregation, and anti-inflammatory. The qualitative and quantitative analysis of alkaloids, polysaccharides, nucleosides, and trace elements in Fritillaria ussuriensis Bulbus were reviewed, which is helpful for its cultivation and accurate application, and would provide a new choice for the treatment of coronavirus disease 2019 (COVID-19).

Evaluation of Phytochemical Constituents and Antioxidant Activities of Ricinus communis L.

Majority of the modern medicines are derived from bioactive constituents of medicinal plants. The current investigation was designed to evaluate the phytochemical constituents and antioxidant activity of methanolic extract of leaves of Ricinus communis L. Qualitative analysis of phytochemicals viz. tannins, phlobatannins, saponins, flavonoids, steroids, alkaloids, coumarin, terpenoids and cardiac glycosides and quantitative analysis of total phenolics, alkaloids, tannins and flavonoids were performed by using standard protocol. Antioxidant activity was studied through DPPH, ABTS, phosphomolybdenum and reducing power assay. To confirm the phytoconstituents of the leaf extracts, Gas chromatography-Mass spectrometry (GC-MS) method was followed. In the present study, qualitative and quantitative analysis revealed the presence of major class of phytochemicals. Quantitative analysis of alkaloids, flavonoids, tannins and phenols were 32.28 ± 0.28 mg/g, 256.29 ± 0.21 mg QE/g, 175.59 ± 0.78 mg TAE/g, 273.35 ± 2.33 mg GAE/g respectively. The extract showed a good scavenging activity with DPPH (84.22 ± 0.81 %) and the GC-MS chromatogram revealed the presence of 23 peaks with different compounds and Ricinine was found to be the major compound. The presence of these bioactive molecules in R. communis may provide the scientific evidences for the medicinal value and other biological properties.

Qualitative and quantitative analysis of alkaloids in Eurycoma longifolia by HPLC-Q-TOF-MS combined with UPLC-QQQ-MS/MS

A sensitive and efficient method was established and validated for qualitative and quantitative analysis of total alkaloids from the extract of Eurycoma longifolia by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS) combined with ultra-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS). The HPLC-Q-TOF-MS conditions are as follows: Welch Ultimate XB-C_(18) column(4.6 mm×250 mm, 5 μm) with acetonitrile(containing 0.1% formic acid)-0.1% formic acid in water as mobile phase for gradient elution. The UPLC-QQQ-MS/MS conditions are as below: Agilent Eclipse Plus C_(18) column(2.1 mm×50 mm, 1.8 μm) with acetonitrile(containing 0.1% formic acid) and 0.1% formic acid in water as mobile phase for gradient elution. MS data were collected by electrospray ionization in positive ion mode. According to the comparison with reference standards and the accurate masses of molecules, a total of 17 alkaloids in E. longifolia extract were identified by HPLC-Q-TOF-MS. The UPLC-QQQ-MS/MS quantitative analysis result of 3 alkaloids showed that the linear ranges of them were good(r≥0.999 7) and the overall recoveries ranged from 108.8%-110.2%, with RSDs of 2.9%-5.3%. The method is accurate, reliable, and efficient, which can comprehensively reflect the constituents and content of alkaloids in E. longifolia. The result can serve as a reference for further elucidating its therapeutic material basis and quality control.

The Cultivation of Chelidonium majus L. Increased the Total Alkaloid Content and Cytotoxic Activity Compared with Those of Wild-Grown Plants.

The effect of cultivation practises on both the phytochemical profile and biological activity of aqueous ethanol extracts of Chelidonium majus L. was studied. Extracts were prepared from aerial parts of the same plant population collected in the wild and grown under organic farming conditions. Both qualitative and quantitative analyses of alkaloids and flavonoid derivatives were performed by LC/MS methods, and the cytotoxicity of lyophilised extracts was studied in B16-F10, HepG2, and CaCo-2 cells. Coptisine was the dominant alkaloid of extracts prepared from wild-grown plants, whereas after cultivation, chelidonine was the most abundant alkaloid. The total alkaloid content was significantly increased by cultivation. Ten flavonol glycoconjugates were identified in C. majus extracts, and quantitative analysis did not reveal significant differences between extracts prepared from wild-grown and cultivated specimens. Treatment with C. majus extracts resulted in a dose-dependent increase in cytotoxicity in all three cell lines. The extracts prepared from cultivated specimens showed higher cytotoxicity than the extracts prepared from wild-grown plants. The strongest cytotoxic effect of cultivated C. majus was observed in B16-F10 cells (IC50 = 174.98 ± 1.12 µg/mL). Cultivation-induced differences in the phytochemical composition of C. majus extracts resulted in significant increases in the cytotoxic activities of the preparations.

Simple quantitative analytical methods for the determination of alkaloids from medicinal and edible plant foods using a homemade chromatographic monolithic column

A polymer-based chromatographic monolithic column was prepared via in-situ radical polymerization using tetrahydrofurfuryl methacrylate as the monomer. The homemade column was used for the separation and quantitative analysis of alkaloids, including piperine from Piper longum (fruit of Piper longum Linn.) and pepper (fruit of Piper nigrum L.), hydroxy-α-sanshool, and hydroxy-γ-sanshool from zanthoxylum (fruit of Zanthoxylum bungeanum Maxim), as well as caffeine from Wuyi rock tea. The chromatographic fractions were identified by mass spectrometry. Single factor test and orthogonal test were both carried out to optimize the extraction conditions. The method validation indicated that the accuracy represented by spiked recovery ranged in 98.89%–102.06%, the correlation coefficients in 0.99986–0.99999. These results show that the prepared monolithic column can be successfully used to quantitatively analyse alkaloids from the real medicinal and edible plant foods with reversed-phase mechanism, which can avoid the long analytical time using traditional packed C18 column. The present method is a simple, and inexpensive method for quantitatively analysing alkaloids from medicinal and edible plant foods, exhibiting good specificity and durability.

The qualitative and quantitative analyses of Gelsemium elegans

Gelsemium elegans is a traditional Chinese medicine that has been used to treat eczema, bruises, rheumatoid arthritis and skin ulcers for many years, and alkaloids are its major active and toxic constituents. This study aimed to comprehensively assess the quality of G. elegans samples including different plant parts and origins using ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry (UHPLC-PDA-QTOF/MS) and high-performance liquid chromatography coupled with UV detector (HPLC-UV). Firstly, the UHPLC-PDA-QTOF/MS approach was developed for the characterization of alkaloids in G. elegans and understanding the differences between multiple groups of samples. Based on the exact mass information, the fragmentation characteristics and the retention time of compounds, 38 alkaloids were identified or tentatively identified. 24 potential chemical markers for differentiating different plant parts of G. elegans were selected through PCA and OPLS/PLS-DA analysis. Secondly, a heatmap visualization was employed for clarifying the distribution of 24 selected alkaloids with high response in the UV. The roots, stems and leaves from Yunnan Province possess relatively consistent alkaloids composition, respectively. Most compounds in the root have a higher content than stems and leaves. Thirdly, a HPLC-UV approach was developed for quantitative analysis of three major alkaloids (gelsemine, koumine and gelsenicine) of G. elegans, and the results showed remarkable variation in the contents of these constituents. While, the contents of three alkaloids fluctuate relatively less in the stem. These results indicated that integrated chemical profiling and quantitative analysis of alkaloids in G. elegans from different plant parts and origins could be assessed by this method, which would establish the foundation for the application of G. elegans.

Phytochemical Analysis and Antimicrobial Activity of Iris kashmiriana and Iris ensata Extracts against Selected Microorganisms

Infectious diseases are the second leading cause for worldwide death. Treatment of infections continues to be difficult in modern time because of the severe side effects of some drugs and the growing resistance to antibacterial agents. Over the past few decades the use of antibiotics is under threat as many commonly used antibiotics have become less effective against certain illnesses due to emergence of multi drug-resistant bacteria. In the present study the effects of 3 types of solvents, chloroform, ethyl acetate and methanol were investigated to determine the presence of various phytochemical constituent, total phenolic content, total flavonoids content and in vitro antimicrobial activity from rhizomes of Iris kashmiriana (Kashmir Iris) and Iris ensata (Japanese Iris), belong to family Iridaceae. The reason for selecting in vitro method was to minimize the usage of experimental animals. The antimicrobial activity of chloroform, ethyl acetate and methanol extract of rhizomes of Iris kashmiriana and Iris ensata were evaluated on bacterial strains of Bacillus cereus, Pseudomonas auregenosa, Proteus vulgaris and Eschirichia coli and fungal strains of Candida albicans and Aspergillus niger by agar well diffusion method. The preliminary phytochemical studies and quantitative analysis of alkaloids, phenol and flavonoids were performed by well reported method. These extracts were further subjected to TLC (Thin layer chromatography analysis). The chemical contents of the Iris kashmiriana and Iris ensata were presented as total phenolic content and total flavonoids content. Phytochemical screening of the extract showed the presence of some common compounds like phenols, terpenoids, flavonoids, carbohydrate etc. The antimicrobial potential of the plant extract was evaluated against different bacterial species which shows significant inhibitory action against all the tested bacterial and fungal strain. Methanolic extract was found to be more active than chloroform and ethyl acetate extracts. It reveals that the methanol soluble components of the plant are highly active against the above mentioned microorganism.
 Keywords: Iris kashmiriana, Iris ensata, Phytochemical constituent, Total phenolic content, Total flavonoids content, In vitro antimicrobial activity

Preliminary Phytochemical Screening, Quantitative Analysis of Alkaloids, and Antioxidant Activity of Crude Plant Extracts from Ephedra intermedia Indigenous to Balochistan.

The aim of this study was to evaluate the antioxidant activity, screening the phytogenic chemical compounds, and to assess the alkaloids present in the E. intermedia to prove its uses in Pakistani folk medicines for the treatment of asthma and bronchitis. Antioxidant activity was analyzed by using 2,2-diphenyl-1-picryl-hydrazyl-hydrate assay. Standard methods were used for the identification of cardiac glycosides, phenolic compounds, flavonoids, anthraquinones, and alkaloids. High performance liquid chromatography (HPLC) was used for quantitative purpose of ephedrine alkaloids in E. intermedia. The quantitative separation was confirmed on Shimadzu 10AVP column (Shampack) of internal diameter (id) 3.0 mm and 50 mm in length. The extract of the solute in flow rate of 1 ml/min at the wavelength 210 nm and methanolic extract showed the antioxidant activity and powerful oxygen free radicals scavenging activities and the IC50 for the E. intermedia plant was near to the reference standard ascorbic acid. The HPLC method was useful for the quantitative purpose of ephedrine (E) and pseudoephedrine (PE) used for 45 samples of one species collected from central habitat in three districts (Ziarat, Shairani, and Kalat) of Balochistan. Results showed that average alkaloid substance in E. intermedia was as follows: PE (0.209%, 0.238%, and 0.22%) and E (0.0538%, 0.0666%, and 0.0514%).

Determination of terpenoid indole alkaloids in hairy roots of Rhazya stricta (Apocynaceae) by GC-MS.

Rhazya stricta Decne. (Apocynaceae) is a medicinal plant rich in terpenoid indole alkaloids (TIAs), some of which possess important pharmacological properties. The study material including transgenic hairy root cultures have been developed and their potential for alkaloid production are being investigated. In this study, a comprehensive GC-MS method for qualitative and quantitative analysis of alkaloids from Rhazya hairy roots was developed. The composition of alkaloids was determined by using GC-MS. In quantification, the ratio between alkaloid and internal standard was based on extracted ion from total ion current (TIC) analyses. The developed method was validated. An acceptable precision with RSD ≤ 8% over a linear range of 1 to 100 µg/mL was achieved. The accuracy of the method was within 94-107%. Analysis of hairy root extracts indicated the occurrence of a total of 20 TIAs. Six of them, pleiocarpamine, fluorocarpamine, vincamine, ajmalicine and two yohimbine isomers are reported here for the first time in Rhazya. Trimethylsilyl (TMS) derivatisation of the extracts resulted in the separation of two isomers for yohimbine and also for vallesiachotamine. Clearly improved chromatographic profiles of TMS-derivatives were observed for vincanine and for minor compounds vincamine and rhazine. The results show that the present GC-MS method is reliable and well applicable for studying the variation of indole alkaloids in Rhazya samples.

Qualitative and quantitative analyses of alkaloids in Uncaria species by UPLC-ESI-Q-TOF/MS.

An ultra performance liquid chromatography (UPLC) coupled with quadrupole time-of-flight mass spectrometry (Q-TOF/MS) method has been optimized and established for the rapid analysis of the alkaloids in 22 samples originating from five Uncaria (U.) species. The accurate mass measurement of all the protonated molecules and subsequent fragment ions offers higher quality structural information for the interpretation of fragmentation pathways of the various groups of alkaloids. A total of 19 oxindole alkaloids, 16 indole alkaloids and 1 flavone were identified by co-chromatography of the sample extract with authentic standards, comparison of the retention time, characteristic molecular ions and fragment ions, or were tentatively identified by MS/MS determination. Moreover, the method was validated for the simultaneous quantification of the 24 components within 10.5 min. The potential chemical markers were identified for classification of the U. species samples by principal component analysis (PCA) and orthogonal partial least squared discriminant analysis (OPLS-DA). The results demonstrate the similarity and differences in alkaloids among the five U. species, which is helpful for the standardization and quality control of the medical materials of the U. Ramulus Cum Unics (URCU). Furthermore, with multivariate statistical analysis, the determined markers are more definite and useful for chemotaxonomy of the U. genus.

Tropane alkaloids of Atropa belladonna L.: in vitro production and pharmacological profile

ObjectiveThe aim of the present work was to study the production of tropane alkaloids by in vitro cultures of Atropa belladonna L. and to evaluate the anticonvulsant, antinociceptive, motor incoordination, and antioxidant activities of both in vitro and original plant extracts.BackgroundA. belladonna is a very important medicinal plant with multipurpose therapeutic effects. The yield of its alkaloid content is very low, which makes it difficult for industrial application.Materials and methodsMurashige and Skoog media were used for callus and plant differentiation induction from leaf explants of A. belladonna L. Qualitative and quantitative analysis of alkaloids was carried out using high-performance liquid chromatography. The anticonvulsant activity was screened by the pentylenetetrazole seizure test. The antinociceptive activity was evaluated by adopting the writhing test, whereas motor incoordination was evaluated using the rotarod test. In addition, antioxidant activity was estimated using the 2,2'-diphenyl-1-picrylhydrazyl radical-scavenging test.ResultsCallus and differentiated plants were successfully induced in Murashige and Skoog media supplemented with growth regulators. High-performance liquid chromatography analysis revealed the production of higher concentrations of tropane alkaloids in differentiated plants than in the original plant. Anticonvulsant and antinociceptive activities, motor incoordination, and the antioxidant effect of callus extracts were much higher than those of the original plant leaf extract.ConclusionPlant tissue culture could be considered as an efficient and alternative source of continuous supply of tropane alkaloids with potent anticonvulsant, antinociceptive, motor incoordination, and antioxidant activities. It is also a powerful tool for producing A. belladonna strain with a high tropane alkaloid content.

Identification of Sinomenine from Sinomenium actum and Its Simultaneous Quantitation by GC–MS and Non-Aqueous CE

In the present study, the qualitative and quantitative analysis of alkaloids in the stem and root of Sinomenium acutum (S. acutum) is presented by gas chromatography-mass spectrometry (GC–MS) and non-aqueous capillary electrophoresis (NACE). The extract of alkaloids in S. acutum was examined by GC–MS and the major alkaloids were identified. Sinomenine (SIN) was found as the principal alkaloid in the extracts (about 84.38%). The quantitation of SIN was then accomplished by GC–MS and NACE with diode array detection. NACE was selected in order to use a running buffer fully compatible with samples in organic solvent. Optimum separation was achieved with a fused-silica capillary column and a running buffer containing 80 mM ammonium acetate, 2.0% acetic acid and 20% (v/v) acetonitrile in methanol medium. The applied voltage was 22 kV. The different selectivity displayed by these techniques allowed different separation profiles that could be useful in phytochemical characterization of the sample. The GC–MS and NACE methods were successfully validated and applied for the quantitation of SIN in S. acutum.

Quantitative analysis of alkaloids from Cinchona bark by high-performance liquid chromatography

A procedure has been developed for the extraction and quantitative analysis of the major alkaloids from Cinchona bark using high-performance liquid chromatography. The method uses a commercial octylsilyl-bonded column, recommended for basic compounds, in the reversed-phase mode. The method is reproducible (relative standard deviation approximately 4% for each alkaloid) and gives quantitative recovery of alkaloids added to bark samples (100–103%). A variety of different types of bark have been studied successfully in the validation scheme. The method can be extended to the analysis of some minor alkaloids which are of interest in the elucidation of the biosynthetic pathways of alkaloid production.