Barbaloin content of aloe (Aloe barbadensis) leaf exudates as affected by different drying techniques

Aloe (Aloe barbadensis Mill.) is commercially cultivated for its transparent leaf gel and leaf exudates. The leaf exudates collected from epidermal layer contain anthraquinone glycosides (aloins) mainly barbaloin (aloin A) and isobarbaloin (aloin B). Aloin A is used as a raw material for the production of diacylrhein, a potent drug prescribed for rheumatoid arthritis. Conventional drying of leaf exudates in open sun causes changes in physicochemical properties and altered aloin A and B composition. Various drying techniques, viz. oven drying, freeze drying, shade drying and open sun drying were employed to evaluate the qualitative and physico-chemical changes in final product of aloe leaf exudates. Freeze drying resulted in high quality dried exudates having maximum aloin A content of 54.16%. The other three drying techniques resulted in lower aloin A content in the final dried product of leaf exudates. Fresh aloe exudates contained lower amount of aloin B (4.65% w/v). Sun drying increased aloin B content to 17.73% (to the extent of 2.27 fold) in the final product compared to freeze drying. Shade drying and sun drying lowered the total aloin content by 13.2% and 8% respectively compared to freeze dried exudates. Freeze drying is the most efficient technique to obtain high quality dried aloe exudates having good textural and physicochemical property. Alternately, shade drying with proper ventilation can be employed to get acceptable final product with marginally lower (8%) total aloin content compared to freeze drying

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Not AvailableA comprehensive experiment was conducted to study the accumulation pattern and determination of three important bioactive compounds namely withaferin-A (WA), 12-deoxywithastramonolide (WO) and withanolide-A (WD) and its determination by the liquid chromatography/electrospray ionization tandem mass spectrometry (LC–ESI-MS-MS) method in root, stem, fruits and leaves of Withania somnifera. A rapid and sensitive LC–ESI-MS-MS method was developed and validated for the determination of these three important bioactive compounds, having same molecular weight. The multiplereactionmonitoringmethodwasestablishedbytwotransitionsforeachanalyteandintense transitionusedfor quantification. Separation ofthethree analyteswas achievedwithin aruntimeof 5 min on an RP-18 column using a mobile phase consisting of acetonitrile and 0.1% acetic acid in water in an isocratic condition. The developed method was validated as per the ICH guidelines. The developed method was found to be suitable for identification and quantification of WA, WO andWD indifferent plantparts suchas roots, stems, fruitsand leaves ofW.somnifera. Theaccumulation of WAwas highest in leaves samples (8.84±0.37 mg/g) and it was 2.23, 5.85 and 27.26 times higherthanitsconcentrationinfruits,stemsandroots,respectively.WOandWDcontentswerehighest (0.44±0.016 and 0.72±0.016 mg/g, respectively) in root.Not Availabl

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Not AvailableA new and rapid method for simultaneous identification and estimation of bioactive triterpenoid glycosides [asiaticoside (AS) and madecassoside (MS)] and their aglycones [asiatic acid (AA) and madecassic acid (MA)] in Centella asiatica was developed by using high-performance liquid chromatography (HPLC) coupled with triple-quadrupole mass spectrometry (MS/MS). Estimation was based on multiple reaction monitoring (MRM) using the precursor → product ion combination for determination of four analytes using Alltima C18 column (50 × 4.6 mm, 3 µm). An electrospray ionization (ESI) tandem interface in positive mode was employed prior to mass-spectrometric detection. The method was subjected to a thorough validation procedure in terms of linearity, limit of detection (LOD) and quantification (LOQ), accuracy, and precision. Six-point calibration curves were linear in the range of 50–500 ng mL−1 for ng mL−1 for AS and MS, and 25–250 ng mL−1 for AA and MA, with excellent linearity (R2 > 0.98). With the optimized conditions, the four analytes were detected accurately within 10 min. LOD and LOQ ranged from 2.5 to 5 and 10 to 15 ng mL−1, respectively. Method accuracy in terms of average recoveries of all four analytes ranged between 98.61 and 102.85 % at three spiking levels with intra- and interday precision relative standard deviation (RSD, %) of 1.01–4.62 and 1.13–4.16, respectively. The new method was successfully applied to estimate the concentration of these four bioactive compounds in extracts of C. asiatica prepared by nonpolar-to-polar solventsICA

Next Generation Sequencing and Transcriptome Analysis Predicts Biosynthetic Pathway of Sennosides from Senna (<i>Cassia angustifolia</i> Vahl.), a Non-Model Plant with Potent Laxative Properties

<div><p>Senna (<i>Cassia angustifolia</i> Vahl.) is a world’s natural laxative medicinal plant. Laxative properties are due to sennosides (anthraquinone glycosides) natural products. However, little genetic information is available for this species, especially concerning the biosynthetic pathways of sennosides. We present here the transcriptome sequencing of young and mature leaf tissue of <i>Cassia angustifolia</i> using Illumina MiSeq platform that resulted in a total of 6.34 Gb of raw nucleotide sequence. The sequence assembly resulted in 42230 and 37174 transcripts with an average length of 1119 bp and 1467 bp for young and mature leaf, respectively. The transcripts were annotated using NCBI BLAST with ‘green plant database (txid 33090)’, Swiss Prot, Kyoto Encylcopedia of Genes & Genomes (KEGG), Cluster of Orthologous Gene (COG) and Gene Ontology (GO). Out of the total transcripts, 40138 (95.0%) and 36349 (97.7%) from young and mature leaf, respectively, were annotated by BLASTX against green plant database of NCBI. We used InterProscan to see protein similarity at domain level, a total of 34031 (young leaf) and 32077 (mature leaf) transcripts were annotated against the Pfam domains. All transcripts from young and mature leaf were assigned to 191 KEGG pathways. There were 166 and 159 CDS, respectively, from young and mature leaf involved in metabolism of terpenoids and polyketides. Many CDS encoding enzymes leading to biosynthesis of sennosides were identified. A total of 10,763 CDS differentially expressing in both young and mature leaf libraries of which 2,343 (21.7%) CDS were up-regulated in young compared to mature leaf. Several differentially expressed genes found functionally associated with sennoside biosynthesis. CDS encoding for many CYPs and TF families were identified having probable roles in metabolism of primary as well as secondary metabolites. We developed SSR markers for molecular breeding of senna. We have identified a set of putative genes involved in various secondary metabolite pathways, especially those related to the synthesis of sennosides which will serve as an important platform for public information about gene expression, genomics, and functional genomics in senna.</p></div

Assembly and CDS statistics of Illumina sequencing of <i>Cassia angustifolia</i> in the leaf transcriptome.

<p>Assembly and CDS statistics of Illumina sequencing of <i>Cassia angustifolia</i> in the leaf transcriptome.</p

Statistics of SSRs identified using MISA in the leaf transcriptome of <i>Cassia angustifolia</i>.

<p>Statistics of SSRs identified using MISA in the leaf transcriptome of <i>Cassia angustifolia</i>.</p

Mapping of the genes expressing differentially in young vs mature leaf on the putative sennoside biosynthetic pathway in <i>Cassia angustifolia</i>.

<p>DAHPS:3-deoxy-7-phosphoheptulonate synthase[EC:2.5.1.54], DHQS: 3-dehydroquinate synthase[EC:4.2.3.4],DHQS/SDH: 3-dehydroquinate dehydratase / shikimate dehydrogenase[EC:4.2.1.10 1.1.1.25], SMK: shikimate kinase[EC:2.7.1.71], EPSP Synthase: 3-phosphoshikimate 1-carboxyvinyltransferase/enolpyruvylshikimate phosphate synthase [EC:2.5.1.19], chorismate synthase [EC:4.2.3.5] menF: isochorismate synthase [EC:5.4.4.2], menF/menD/menH/menC or PHYLLO: isochorismate synthase / 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate synthase / 2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase / O-succinylbenzoate synthase[EC:5.4.4.2 2.2.1.9 4.2.99.20 4.2.1.113], menE: Succinylbenzoic acid-CoA ligase/acyl-activating enzyme 14 [EC:6.2.1.26], menB: naphthoate synthase[EC:4.1.3.36], DXPS: 1-deoxy-D-xylulose-5-phosphate synthase[EC:2.2.1.7], DXR: 1-deoxy-D-xylulose-5-phosphate[EC:1.1.1.267], ISPD: 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase [EC:2.7.7.60], CDPMEK:4-diphosphocytidyl-2-C-methyl-D-erythritol kinase[EC:2.7.1.148], ISPF: 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase[EC:4.6.1.12], HDS: (E)-4-hydroxy-3-methylbut-2-enyl-diphosphate synthase[EC:1.17.7.1], HDR: 4-hydroxy-3-methylbut-2-enyl diphosphate reductase[EC:1.17.1.2], ACCP Transferase: acetyl-CoA C-acetyltransferase [EC:2.3.1.9], HMGS:hydroxymethylglutaryl-CoA synthase [EC:2.3.3.10], HMGR: hydroxymethylglutaryl-CoA reductase (NADPH) [EC:1.1.1.34], MK: mevalonate kinase[EC:2.7.1.36], PMK: phosphomevalonate kinase [EC:2.7.4.2], MPD: diphosphomevalonate decarboxylase[EC:4.1.1.33], IPPS: isopentenyl-diphosphate delta-isomerase[EC:5.3.3.2], PKS: Polyketide Synthase, PKC:Polyketide Cyclase, UGT:UDP-Glucosyl Transferase; Y_ID:Young leaf CDS ID number for enzyme, M_ID:Mature leaf CDS ID number for enzyme, Y_FPKM: Young leaf CDS FPKM value, M_FPKM: Mature leaf CDS FPKM value, Log Fold Change: Log of change in folds of expression of CDS in young compared to matured leaf transcripts.</p

Variation in the total sennoside content (%) with ontogeny of the leaves.

<p>Variation in the total sennoside content (%) with ontogeny of the leaves.</p

Clusters of orthologous groups (COG) functional classification of CDS in the young and mature leaf transcriptome of <i>Cassia angustifolia</i>.

<p>Clusters of orthologous groups (COG) functional classification of CDS in the young and mature leaf transcriptome of <i>Cassia angustifolia</i>.</p

Heat map of top 100 differentially expressed genes in young and mature leaf transcriptome of <i>Cassia angustifolia</i>.

<p>Heat map of top 100 differentially expressed genes in young and mature leaf transcriptome of <i>Cassia angustifolia</i>.</p