Search

A validated UHPLC-PDA with an ESI-MS/MS method has been developed for simultaneous estimation of six bioactive alkaloids (magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine and berberine) in the different extracts of the roots of Berberis aristata DC (Family:Berberdiaceae). It is an important medicinal herb native to Northern Himalaya and commonly known as ‘daruharidra’, ‘daruhaldi’, ‘Indian barberry’ or ‘tree turmeric’. An insight into the research literature uncovered reports on isoquinoline alkaloids like magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine, and berberine as major bioactives in B. aristata roots, possessing different pharmacological and therapeutic effects. In the present study, these aforementioned alkaloids were separated on Phenomenex Luna®, 5 µm-C8 analytical column. The HPLC-MS analysis was performed at a flow rate of 0.90 mL min−1. Each alkaloid that is resolved was characterized by precursor ions and fragment ions with electrospray ionization (ESI) source in both positive and negative ionization using scan mode. The limit of detections (LODs) were 0.087, 0.727, 0.035, 0.124, 0.782 and 0.794 μg mL−1 for magnoflorine, berbamine, columbamine, jatrorrhizine, palmatine and berberine, respectively. The proposed UHPLC-PDA method was fully validated according to international (ICH) guidelines and was found to be selective, sensitive and highly accurate for the concomitant estimation of the aforementioned symbolic bio-markers of B. aristata roots.

This study was planned to develop a simple high-performance thin-layer chromatography method for qualitative and quantitative estimation of 3-acetyl-11-keto-β-boswellic acid (AKBBA), β-boswellic acid (BBA), 3-oxo-tirucallic acid (TCA) and serratol (SRT) with HPTLC-ESI-MS/MS for characterization in Boswellia serrata Roxb. oleo gum resin extract. The method was developed with hexane-ethyl acetate-toluene-chloroform-formic acid as mobile phase. RF values observed for AKBBA, BBA, TCA and SRT were 0.42, 0.39, 0.53 and 0.72, respectively. The method was validated according to International Council for Harmonisation guidelines. The concentration range for linearity was 100–500 ng/band for AKBBA and 200–700 ng/band for the other three markers with r2 > 0.99. The method resulted in good recoveries as 101.56, 100.68, 98.64 and 103.26%. The limit of detection was noticed as 25 , 37, 54 and 38 ng/band, with a limit of quantification as 76, 114, 116 and 115 ng/band, for AKBBA, BBA, TCA and SRT, respectively. The four markers were identified and confirmed in B. serrata extract using TLC-MS by indirect profiling by LC-ESI-MS/MS and were identified as terpenoids, TCA and cembranoids: AKBBA (mass/charge (m/z) = 513.00), BBA (m/z = 455.40), 3-oxo-tirucallic acid (m/z = 455.70) and SRT (m/z = 291.25), respectively.

The objective of the present work was to develop a simple, specific, and fast high-performance thin-layer chromatographic (HPTLC) method to identify and quantify cordifolioside A, 20-β-hydroxyecdysone and columbin with HPTLC‒electrospray ionization‒tandem mass spectrometry (ESI‒MS/MS) for characterization in Tinospora cordifolia stem extracts. Chromatographic development was performed using a HPTLC aluminum plate, pre-coated with silica gel 60 F254 with hexane‒chloroform‒methanol‒formic acid as the mobile phase. Densitometric quantification for 20-β-hydroxyecdysone and cordifolioside A was performed at 254 nm and for columbin at 600 nm after derivatization with anisaldehyde‒sulfuric acid. The optimized mobile phase resulted in chromatographic separation of peaks for cordifolioside A, 20-β-hydroxyecdysone, and columbin at RF of 0.12, 0.47, and 0.86, respectively. The linear concentration range was found to be 750‒2250 ng/band for 20-β-hydroxyecdysone and cordifolioside A and 675‒1875 ng/band for columbin with (r2 > 0.99). The methodology showed good recoveries as 98.96‒101.43% for cordifolioside A, 98.15‒101.56% for 20-β-hydroxyecdysone, and 98.06‒98.80% for columbin. The limit of detection was found for columbin, 20-β-hydroxyecdysone, and cordifolioside A as 53.86 ng/band, 40.90 ng/band, and 107.05 ng/band, while the limit of quantification was found to be 163.21 ng/band, 123.94 ng/band, and 324.38 ng/band, respectively. The relative standard deviation for precision and robustness study for all the markers was found to be within 2%. Three markers were identified and confirmed in T. cordifolia stem extracts by ESI‒MS/MS. Compounds were assigned as norditerpene furan glycosides, ecdysteroids, and diterpenoid furanolactone: cordifolioside A (m/z = 527 [M + Na]+; UV λmax 221 nm), 20-β-hydroxyecdysone (m/z = 481.30 [M + H]+; UV λmax 247 nm), and columbin (m/z = 359 [M + H]+; UV λmax 210 nm). The optimized method was found accurate, reproducible, robust, and specific and can be applied for the quantification of cordifolioside A, 20-β-hydroxyecdysone, and columbin for quality control of extracts of T. cordifolia. Supplementary Information The online version contains supplementary material available at 10.1007/s00764-021-00115-7.

Berberis aristata DC., well known as ‘Indian barberry’ or ‘tree turmeric’, (Berberidaceae) is a medicinal herb native to the Northern Himalaya. A dig in the research literature evinced different pharmacological and therapeutic effects of B. aristata and its isoquinoline alkaloids. Records on investigations for the chemical composition of the plant corroborated isoquinoline alkaloids, berberine, berbamine, palmatine, magnoflorine and jatrorrhizine as the important bioactive constituents. Recent upsurge in the demand of botanicals is a result of their scientific validation for efficacy and quality. Keeping this in view, a validated simultaneous high-performance thin-layer chromatography (HPTLC) method has been developed using silica gel 60 F254 as the stationary phase and ethyl acetate‒formic acid‒glacial acetic acid‒water (100:11:11:26, V/V) as the mobile phase. The plates were scanned densitometrically at 254 nm and 366 nm. The limits of detection and the percentage mean recoveries were 323.24, 227.00, 55.68, 88.67 and 86.56 ng/spot, and 99.35%, 99.98%, 98.55%, 99.40% and 98.92%, for berberine, berbamine, palmatine, magnoflorine and jatrorrhizine, respectively. The linearity range for berberine and berbamine was 1000‒6000 ng/spot, for palmatine 200‒700 ng/spot, for magnoflorine 500‒2000 ng/spot and for jatrorrhizine 300‒1800 ng/spot. The proposed HPTLC method offers a sensitive, specific and precise gage for the concurrent estimation of these five representative biomarkers of B. aristata and also aids in its qualitative analysis, since a fingerprint is generated, wherein the five active ingredients are resolved with other compounds present in the root.

The Ocimum genus is one of Indiaʼs prominent botanical classes of traditional medicinal culture comprising medicinally and agronomically important plants. Morphological resemblances, overlapping geographical distribution, and history of traditional nomenclature have necessitated a comprehensive qualitative report for effective quality control and removing the species ambiguity pertaining to this genus. This paper provides detailed morpho-micrometric characteristics used to differentiate between six indigenous Ocimum species of India. Among them, O. gratissimum was distinguished as the only shrub with a fleshy petiole. In green and purple forms, O. tenuiflorum leaves had serrate margins and showed no particular anatomical differences except for the anthocyanins containing epidermal cells of the latter. O. basilicum had glabrous leaves except for the veins, which were puberulous. O. filamentosum had tenuous anther filaments and was the least aromatic while O. africanum had a citrusy odour, which along with the number of xylary rows, size of mesophyll cells, and epidermal cell wall architecture, distinguished it from O. americanum. An HPTLC method was developed using experimental design and validated for quantification of multi-class compounds from terpenoic, phenolic acids, and flavonoids in Ocimum leaves. It was found linear (r 2 > 0.99) with recoveries between 95 – 100% for all compounds. The eluted bands of marker compounds were subjected to HPTLC-MS analysis as a confirmative tool. This is the first anatomical and analytical report of O. filamentosum Forssk. The obtained results could be effectively used for species identification using vegetative characters alone with the anatomical-HPTLC data backing up the former as a rapid and economical tool.

Ellagic acid is a popular antioxidant dietary supplement. It is a natural phenolic compound common to multiple botanical sources. The ellagic acid based dietary supplements are manufactured by hydrolysis of ellagitannins from Punica granatum. However, use of ellagic acid from sources other than P. granatum is not uncommon. Currently, there is no robust analytical methodologies for quantification and confirmation of the enriched ellagic acid (›40%) source in commercial products. Therefore, we have developed and validated ultra-fast liquid chromatography - photodiode array detector and ultra-fast liquid chromatography - tandem mass spectrometry methods for quantification and source identification of ellagic acid in commercial products. The results of the study confirm punicalin A-B, punicalagin A and B as positive markers and catechin and chebulinic acid as adulteration markers.

Osteoarthritis (OA) is an aging disorder characterized by degenerated cartilage and sub-chondral bone alteration in affected knee joints. Globally, millions of people suffer from this disease. However, there is a lack of safe and promising therapeutics, making the exploration and development of leads from natural sources urgent. Accordingly, food as medicine may be the most suitable approach for the treatment of this degenerative disease. Herein, we elucidated the protective role of Spinacia oleracea extract (SOE) in an anterior cruciate ligament transection (ACLT) model of osteoarthritis as a mimic of the human condition. ACL transection was done in the tibio-femoral joints of rats. SOE was orally administered at the dosage of 125 and 250 mg kg−1 day−1 for four weeks. It was shown that the animals with SOE treatment had better joint morphology than the ACLT animals, as evident by the shiny appearance of their cartilage. Hematoxylin and safranin-o staining showed that the number of chondrocytes was significantly reduced in the OA model, which was prevented with SOE treatment. The reduction in the cartilage thickness was well observed by toluidine blue staining. The reduced stain by safranin-o and toluidine blue, indicated proteoglycan loss in the ACLT-induced osteoarthritis model. The proteoglycan content and cartilage thickness were restored in the SOE group upon treatment at an SOE dosage of 125 and 250 mg kg−1 day−1. The micro-CT parameters of subchondral bone (SCB) and cartilage degradation markers in the serum corroborated our findings of the protective effects of SOE. In summary, our study suggests that SOE has therapeutic potential, which if taken regularly as a food supplement, can have beneficial effects.

Kashmir saffron (Crocus sativus L.), also known as Indian saffron, is an important Asian medicinal plant with protective therapeutic applications in brain health. The main bioactive in Kashmir or Indian Saffron (KCS) and its extract (CSE) are apocarotenoids picrocrocin (PIC) and safranal (SAF) with carotenoids, crocetin esters (crocins), and crocetins. The ultra-fast liquid chromatography(UFLC)- photodiode array standardization confirmed the presence of biomarkers PIC, trans-4-GG-crocin (T4C), trans-3-Gg-crocin (T3C), cis-4-GG-crocin (C4C), trans-2-gg-crocin (T2C), trans-crocetin (TCT), and SAF in CSE. This study’s objectives were to develop and validate a sensitive and rapid UFLC-tandem mass spectrometry method for PIC and SAF along T4C and TCT in rat plasma with internal standards (IS). The calibration curves were linear (R2 &gt, 0.990), with the lower limit of quantification (LLOQ) as 10 ng/mL. The UFLC-MS/MS assay-based precision (RSD, &lt, 15%) and accuracy (RE, −11.03–9.96) on analytical quality control (QC) levels were well within the acceptance criteria with excellent recoveries (91.18–106.86%) in plasma samples. The method was applied to investigate the in vivo pharmacokinetic parameters after oral administration of 40 mg/kg CSE in the rats (n = 6). The active metabolite TCT and T4C, PIC, SAF were quantified for the first time with T3C, C4C, T2C by this validated bioanalytical method, which will be useful for preclinical/clinical trials of CSE as a potential neuroprotective dietary supplement.

Background Ocimum genus, known as Tulsi or Basil, is a prominent botanical class in Asian culture, especially in India. The leaves have immunomodulatory, antioxidant, stress-relieving, and adaptogenic roles in traditional and modern medicine, with prominent usage in herbal teas and nutraceuticals. Objective An high-performance liquid chromatography–photodiode array (HPLC-PDA) method was developed and validated for quantification of vicenin-2, orientin, cynaroside, betulinic acid, genistein with syringic acid, rosmarinic acid, eugenol, carnosic acid, oleanolic acid, ursolic acid, luteolin, and apigenin and was confirmed using a novel electrospray ionisation-mass spectrometry (ESI-MS/MS) method in the Ocimum genus samples. Method The methodology parameters were developed on an reverse phase (RP) C18 column with a gradient elution of 1 mL/min flow rate for 0.1% o-phosphoric acid and acetonitrile at 210 and 340 nm wavelengths. Results The validation data for 13 bioactive compounds showed good linearity (r2 > 0.99) with sensitive LOD (0.034–0.684 µg/mL) and LOQ (0.100–2.068 µg/mL) with recoveries (83.66–101.53%). The results of the quantification were found to be precise (RSD, Conclusions The validated HPLC-PDA and LC-ESI-MS/MS method was found to be selective and suitable for analyzing 13 compounds in O. tenuiflorum and 12 cultivars from the Ocimum genus as a quality control tool. This method can be used in routine analysis as an inexpensive alternative to advanced techniques. Highlights This work is the first to report for vicenin-2, orientin, cynaroside, betulinic acid, and genistein, with simultaneous analysis of eight bioactive compounds in the Ocimum genus.

Osteoarthritis (OA) is an aging disorder characterized by degenerated cartilage and sub-chondral bone alteration in affected knee joints. Globally, millions of people suffer from this disease. However, there is a lack of safe and promising therapeutics, making the exploration and development of leads from natural sources urgent. Accordingly, food as medicine may be the most suitable approach for the treatment of this degenerative disease. Herein, we elucidated the protective role of Spinacia oleracea extract (SOE) in an anterior cruciate ligament transection (ACLT) model of osteoarthritis as a mimic of the human condition. ACL transection was done in the tibio-femoral joints of rats. SOE was orally administered at the dosage of 125 and 250 mg kg-1 day-1 for four weeks. It was shown that the animals with SOE treatment had better joint morphology than the ACLT animals, as evident by the shiny appearance of their cartilage. Hematoxylin and safranin-o staining showed that the number of chondrocytes was significantly reduced in the OA model, which was prevented with SOE treatment. The reduction in the cartilage thickness was well observed by toluidine blue staining. The reduced stain by safranin-o and toluidine blue, indicated proteoglycan loss in the ACLT-induced osteoarthritis model. The proteoglycan content and cartilage thickness were restored in the SOE group upon treatment at an SOE dosage of 125 and 250 mg kg-1 day-1. The micro-CT parameters of subchondral bone (SCB) and cartilage degradation markers in the serum corroborated our findings of the protective effects of SOE. In summary, our study suggests that SOE has therapeutic potential, which if taken regularly as a food supplement, can have beneficial effects.

Ethnopharmacological relevance Dalbergia sissoo DC. (Indian rosewood or Sheesham) is a traditional medicinal plant, reported since time immemorial for its analgesic, anti-nociceptive, anti-inflammatory, and immuno-modulatory properties. D. sissoo DC (DS). is being used traditionally to cure joint inflammation and joint pain. Aim To study the potential of DS leaves and its derived novel compound CAFG to treat the clinical symptoms of osteoarthritis (OA) and its underlying mechanism. Methods The chemical profile of DS extract (DSE) with isoflavonoids and isoflvaonoid glycosides from the DS was established by UHPLC-PDA and UHPLC-MS/MS. Monosodium iodoacetate (MIA) was injected into the knee joint to develop the OA model in rats. DSE was given orally for 28 days daily at 250 and 500 mg.kg−1day−1. For in-vitro experiments, chondrocytes isolated from joint articular cartilage were negatively induced with interleukin-1β (IL-1β) and CAFG was given to the cells as a co-treatment. Results Chondrocytes undergo apoptosis following inflammation and proteoglycan synthesis affected in MIA injected knees. DSE administration prevented these effects as assessed by H&E and Toluidine blue staining. Micro-CT analysis showed that subchondral bone loss was restored. DSE decreased elevated serum levels of cartilage-bone degradation (CTX-I, CTX-II, and COMP), inflammation markers IL-1β, and matrix-degrading MMP-3 and 13. The effects of IL-1β on gene expression of chondrocytes were reversed by CAFG treatment at 1 μM. Conclusion Data showed that DSE protected joint cartilage and deterioration in subchondral bone in vivo while in in-vitro, its active ingredient CAFG prevented interleukin-1β induced effects and inhibited OA. This finding suggest that DSE and CAFG could be used as a possible therapeutic to treat osteoarthritis.

"Curcumin (CUR)" is the principal active phytoconstituent present in Curcuma longa (CL), also known as Turmeric, is a popular natural product used in food and dietary supplements industries. For economic advantage, CUR is manufactured synthetically. The synthetic curcumin (SC) could be mislabeled, mistaken, or mixed with natural origin CL or CL extract (CLE) or CL products for replenishing CUR. The study aimed to differentiate CLE and SC by targeting CIMP-1,i.e. (1E,4Z)-5-hydroxy-1-(3-hydroxy-4-methoxyphenyl) hexa-1,4-dien-3-one by HPLC-PDA (photodiode array) and HPTLC-DS (densitometry) based on unique patterns. The validated HPLC-PDA method for CIMP-1 and CUR in SC showed robustness and sensitivity up to 1% adulteration with recovery, precision, and linearity of compounds as per guidelines. All four compounds were identified and confirmed by ESI-MS/MS. In this research, the presence of Boron (B) found as a qualitative indicator of SC (> 250.0 mg/kg) and CLE (< 2.0 mg/kg) by ICP-MS. Further, this HPLC-PDA method was successfully applied for sixteen samples of CLE procured across India, out of which four samples showed the presence of synthetically origin curcumin. This research is the first report of simple, lab-based methods for profiling of CUR based on natural or synthetic origin and identification of SC.

In today's era of increased standards of lifestyle and life expectancy, there has been a constant demand for supplements by consumers. Nutraceuticals are among the supplements in demand. Although there is a big opportunity for the nutraceutical business, there are no uniform regulatory requirements in different regions. Nations are looking to the nutraceutical sector to help keep their populations healthy and safe by introducing certain rules and regulations. Generally, developed countries have regulations in place, but there are some countries, such as those in the Asia Pacific regions or in Association of Southeast Asian Nations (ASEAN) countries, that have not yet fine-tuned their regulations for nutraceutical products. The ASEAN countries involve highly commercialized markets such as Singapore, Thailand, Malaysia, and the Philippines. The overall nutraceutical market of ASEAN countries is growing at a compound annual growth rate of ∼8.4%. About 40% of the ASEAN population consumes nutraceuticals on a daily basis. ASEAN countries are forming harmonized regulations for dietary supplements. This could be a big opportunity for manufacturers to introduce their products into the ASEAN market. A special unit of the Traditional Medicine and Health Supplements Product Working Group (TMHA PWG) helps manufacturers understand the regulatory procedures of these countries. Despite countries' own special requirements, manufacturers can follow the standards and harmonized guidelines put forth by TMHA PWG. The aim of this review is to introduce the regulatory procedure and requirements for international business developers to launch any new nutraceutical products into the ASEAN market.

Cows affected with retained placenta are at a higher risk of developing puerperal metritis. Retained fetal membranes (RFM) in cattle have adverse effect on fertility and production. Understanding the pathophysiology and causes of RFM is important for managing this disease. The hormonal processes that lead to normal placental expulsion or the separation are multifactorial and begin before parturition. A variety of risk factor, including early or induced parturition, dystocia, hormonal imbalance, and immunosuppression can interrupt these normal processes and result in retention of the placenta. Current research does not support the efficacy of much commonly practiced treatment for RFM. Systematic administration of antibiotics can be beneficial for the treating metritis after RFM, but antibiotic administration has not been shown to significantly improve future reproduction in the cows with RFM. The Polyherbal oral therapy is protects cattle from puerperal disease and enhance placental release after delivery. Herbal therapy is not costly and it having the minimum side effect. Herbal remedies bear a high potential to treat postpartum uterine diseases in cows. Various herbs are reported to for retention of fetal membrane. Key Words: Cattle, placental detachment, Causes.

Sarcostemma viminale R. Br. (family-asclepiadaceae) is known as the “soma” plant in India. It is rich in fatty acids and hydrocarbons and used traditionally with different medicinal use. In the present study first time systemic phytochemical profile of fatty acid was established for S. Viminale. The major fatty acids isolated and identified were1-Hexadecene (C16H32), Hexadecanoic acid (C16H32O2), Octadecanoic acid (C18H36O2), 9-Octadecenoic acid (C18H34O2), 1- Docosene (C22H44). Also eight major compounds were isolated and identified as Eicosane, Docosane, 1-Octadecene, Nonacosane, α- amyrin, β- amyrin, β-Sitosterol and Gallic acid. The structure elucidation was carried out using combination of Gas Chromatographic- Mass Spectroscophy (GC-MS), Fourier transform infrared spectroscopy (FT-IR), and Carbon Nuclear magnetic resonance (13C NMR) data. This investigation is also supported with standardized phytochemical studies and moderate anti-bacterial activity of the plant.

The major volatile constituents of leaves and flowers of Cymbopogon martini from the volatile oil obtained by steam distillation were identified by GC/MS. Five constituents were identified from the volatile oil of leaves and flowers, which constituted about 82.49 and 75.63% of the total amount, respectively. A monoterpene, piperitone (6.00%), was identified in the flowers of C. martini; in addition, flowers were found to contain more olefinic terpenes, namely geraniol (69.63%), compared with leaves (53.41%). Leaves contain bicyclic monoterpene, nerol (24.76%) and alpha-pinene (4.32%). Anthelmintic activity of these oils was evaluated on adult Indian earthworms Pheretima posthuma and results showed that the volatile oil of C. martini flower required less time to cause paralysis and death of the earthworms.

Thespesia populnea Sol. ex Correa (Malvaceae), an indigenous tree species in India, is of interest to researchers because traditionally its heartwood is used in the treatment of ulcer and colic pain.To validate its folk use in the treatment of ulcerative colitis (UC).Mice were administered intrarectal DNBS and then treated with different plant extracts (100 and 200 mg/kg), 30 min before and 24 and 48 h after DNBS infusion. Colonic mucosal injury was assessed by macroscopic and histological examination. Furthermore, malondialdehyde (MDA), myeloperoxidase (MPO), protease, and hemoglobin (Hb) contents were measured in tissue and blood samples.Administration of various extracts ameliorated macroscopic and microscopic scores which were altered due to DNBS treatment in mice. Hb concentration in blood was restored significantly by the aqueous extract to 17.20 ± 0.5, which was reduced to 13.80 ± 0.5 after treatment with DNBS. MDA level was increased to 10.82 nm/mg and 10.25 nm/ml in tissue and blood, respectively, due to DNBS treatment which was reduced to 2.69 nm/mg and 3.59 nm/ml in tissue and blood, respectively, by aqueous extract treatment. Similarly, MPO level was increased to 412 U/mg and 404 U/ml in tissue and blood, respectively, which was significantly reduced to 205 U/mg and 219 U/ml in tissue and blood, respectively, by aqueous extract treatment. Aqueous extract significantly reduced protease activity which was markedly increased in DNBS-treated animals.Aqueous extract of heartwood of T. populnea is effective in the treatment of UC.

Dhaka Univ. J. Pharm. Sci. 6(1): 55-57, 2007 (June) The full text is of this article is available at the Dhaka Univ. J. Pharm. Sci. website

Methanol extracts from the leaves, stems and roots of Mentha piperita and Lantana camara were investigated for their anthelmintic activity against Pheritima posthuma. Each extract was studied in the bioassay at 20 mg/ml, which involved determination of time of paralysis and time of death of the worms. Both the extracts of both plants exhibited considerable anthelmintic activities, and the order of sensitivity of the extracts to the worms was stems > roots > leaves for the M. piperita and stems > leaves >roots for the L. camara. The methanol extracts of stems of both M. piperita and L. camara were found to be the most active. Albendazole (20 mg/ml) and distilled water were included in the assay as standard reference drug and control, respectively. Key words: Mentha piperita, Lantana camara, Pheritima posthuma, In vitro anthelmintic activity. Dhaka Univ. J. Pharm. Sci. Vol.5(1-2) 2006 The full text is of this article is available at the Dhaka Univ. J. Pharm. Sci. website