Revealing the anti-inflammatory ingredients in wine-processed Radix et Rhizoma Rhei using immobilized cysteinyl leukotriene receptor type 1 as the stationary phase.

Despite the tremendous progress of wine-processed Radix et Rhizoma Rhei (Jiudahuang, JDH) in removing toxic heat from the blood in the upper portion of the body for hundreds of years, the deep understanding of its functional material basis of the anti-inflammatory ingredients remains unclear due to the lack of high specific and efficient methods. Herein, taking Cysteinyl leukotriene receptor type 1(CysLT1R) as the target protein, we established a chromatographic method based on the immobilized CysLT1R using haloalkane dehalogenases (Halo) at the C-terminus of the receptor in one step. After careful characterization by X-ray photoelectronic spectroscopy, immune-fluorometric analysis, and chromatographic investigations, the immobilized receptor was used to screen the anti-inflammatory ingredients in JDH. Aloe-emodin, rhein, emodin, chrysophanol, and physcion were identified as the main anthraquinone exerting anti-inflammatory effects in the drug. The association constants for the five compounds to bind with the receptor were calculated as (0.30 ± 0.06)× 105, (0.35 ± 0.03)× 105, (0.46 ± 0.05)× 105, (1.05 ± 0.14)× 105, and (1.66 ± 0.17)× 105 M-1 by injection amount-dependent method. Meanwhile, hydrogen bonds were identified as the main driving force for the five compounds to bind with CysLT1R by molecular docking. Based on these results, we believe that the immobilized receptor chromatography preserves historic significance in revealing the functional material basis of the complex matrices.

Anthraquinone Removal from Cassia obtusifolia Seed Water Extract Using Baking, Stir-Frying, and Adsorption Treatments: Effects on the Chemical Composition, Physicochemical Properties of Polysaccharides, and Antioxidant Activities of the Water Extract.

Safety issues of the controversial anthraquinones from Cassia obtusifolia seed water extracts (CWEs) limit its application. This work aimed to remove the anthraquinones of CWEs by baking treatment (BT), stir-frying treatment (ST), and adsorption treatment (AT). Effects of these treatments on the chemical composition, physicochemical properties of polysaccharides, and antioxidant activities of CWEs were analyzed and compared. Results indicated that AT exhibited the best removal effect on the total anthraquinone among the three treatments. After AT, the contents of rhein, emodin, aloe-emodin, and aurantio-obtusin of the CWE were below the limit of detection. In addition, AT increased the contents of neutral sugars in CWEs in comparison to BT and ST. None of the treatments had an obvious influence on the structural characteristics of polysaccharides. However, AT decreased the antioxidant activity of CWEs due to their lower anthraquinone content. In summary, AT was considered as an efficient and simple method to remove anthraquinones, while retaining the features of polysaccharides.

Aloe-emodin Quantification using HPTLC and RP-UHPLC in Extracts and Commercial Herbal Formulations: Evaluation of Antimicrobial and Antioxidant Effects.

BACKGROUND: High-performance thin-layer chromatography (HPTLC) was developed and validated for the determination of aloe-emodin in accordance with ICH guidelines. In addition, a novel RP-UHPLC method was developed, and both methods were used to analyse the herbal extract and herbal formulation. METHODS: Separation was carried out on a silica gel 60 F254 HPTLC plate using the mobile phase Toluene: Methanol (9:1). The linearity was good across the 800-4000 ng/spot range. Validation results are within acceptable limits. The percent RSD for accuracy was 0.58-1.77, and precision was 1.10-1.97 and 1.45-1.94 for intraday and interday, respectively. The percentage of aloe-emodin found in the herbal extract and aloe vera capsule was 99.83 ± 1.19 and 99.53 ± 1.29, respectively, using this method. RESULTS: Quantification of aloe-emodin in herbal extract and herbal formulation were done using a novel UHPLC method with chromatographic conditions of orthophosphoric acid Methanol (0.1 percent OPA): Water (65:35, v/v) and pH 3, a flow rate of 1.2 ml/min, and elute detection at 254 nm. At 6.32 minutes, a sharp and symmetric peak was observed. The method developed was validated in accordance with ICH guidelines. The percent RSD numerical value of accuracy was 0.304-0.576, and the inter-day and intraday precision were 0.32-3.08 and 0.51-2.78, respectively. Herbal extract and aloe vera capsule were analysed using the new UHPLC method. Aloe-emodin percentages were reported as 100.3 ± 0.89 and 99.53 ± 1.29, respectively. CONCLUSION: The antimicrobial and anti-oxidant activities of an aloe-vera herbal formulation were studied, and the results were positive.

Determination of anthraquinones in Rhamnus purshiana using high-performance liquid chromatography coupled to diode array detector and simple ultraviolet spectroscopic analysis.

A new method based on Ultraviolet spectrophotometry was developed and compared with that based on high-performance liquid chromatography for the determination and quantification of anthraquinones in the extracts of Rhamnus purshiana bark. A validated quantitative analysis of cascaroside A, cascaroside B, emodin, and aloe-emodin in these herbal products has been previously performed using high-performance liquid chromatography coupled with a diode array detector. In the high-performance liquid chromatography analysis, all the anthraquinones showed satisfactory regression (r2 > 0.98) within the test ranges, and the recovery was in the range of 94-117%. The limits of detection and quantification were 0.008-0.010 and 0.029-0.035 µg/mL, respectively. Hierarchical cluster analysis and principal component analysis showed differences in the anthraquinones determined from herbal samples. Subsequently, a simple and low-cost ultraviolet spectrophotometric methodology for the quantitative analysis of the same compounds in the extracts was applied, and all the contents were determined. A paired t-test confirmed that there were no significant differences between the two methods. Our results revealed that the developed method is simple and provides the ability to discriminate and control the quality of anthraquinones in herbal products.

The absence of genotoxicity of a mixture of aloin A and B and a commercial aloe gel beverage.

Aloe products are increasingly valued as ingredients in food supplements and as flavoring agents. The global Aloe vera market is varied, large, growing, and increasingly important in food, cosmetics, and medicines. Aloin, an anthraquinone glycoside, is one of the major components by weight of the anthraquinone derivatives of Aloe vera gel. Principal metabolites, aloe emodin and emodin, are a source of debate concerning toxic vs salutary effects, hence the accurate toxicological characterization of these compounds has become increasingly important. The purpose of this study was to determine the genotoxic profile of a stabilized Aloe vera juice product derived from the inner filet and marketed as a beverage currently sold in the European Union containing 8 to 10 ppm aloin and a mixture of purified aloin A and B. The present data confirm that a commercial stabilized Aloe vera gel intended for consumption as a juice beverage is not genotoxic. Furthermore, both aloin A and B were negative in the same assays and therefore are also not genotoxic. These results are consistent with the work of other groups and contrast with data obtained using products containing the Aloe vera latex hydroxyanthracene derivatives (HADs).

Simultaneous determination of flavonoids and anthraquinones in honey by using SPE-CE-LIF.

Based on advantages of capillary electrophoresis (CE), a new solid-phase extraction (SPE) coupled with CE has been developed for preconcentration, enrichment and determination of anthraquinones and flavonoids (rutin, emodin, quercetin, 1,8-dihydroxyanthraquinone) in honey. The environmental-friendly chitin activated after an easy processing is selected as the adsorbent to enrich analytes. Then, chitin was filled into the filter as the solid phase. To improve the extraction effect, some key parameters of extraction were optimized. Under the optional extraction conditions, the chitin showed excellent adsorption capacity and selectivity over rutin, emodin, quercetin, and 1,8-dihydroxyanthraquinone, with enrichment factors reaching 5 folds. The CE coupled with fluorescence detection was used for the detection. Results prove the method is simple, fast, and highly sensitive, with the limit of detection (LOD) is 3.00-200.0 ng/mL; the recovery is 90.0-107.0%, and relative standard deviation of (RSD) is 1.8-8.3%.

Chaiqin chengqi decoction ameliorates acute pancreatitis in mice via inhibition of neuron activation-mediated acinar cell SP/NK1R signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Chaiqin chengqi decoction (CQCQD) and its derivatives have been widely used in China for the early management of patients with acute pancreatitis (AP). Numerous studies demonstrate the anti-inflammatory and anti-oxidative effects of CQCQD and derivatives, but whether these effects can be attributed to suppressing neurogenic inflammation, has never been studied. AIM OF THE STUDY: To investigate the effects of CQCQD on substance P (SP)-neurokinin 1 receptor (NK1R) based neurogenic inflammation in an experimental AP model. MATERIAL AND METHODS: For AP patients on admission, pain score was accessed by visual analog scale (VAS); the levels of serum SP and expressions of pancreatic SP and NK1R were also determined. For in vivo study, mice received 7 intraperitoneal injections of cerulein (50 µg/kg) at hourly intervals to induce AP, whilst controls received normal saline injections. In the treatment groups, CQCQD (10 g/kg, 200 µl) was intragastrically given at the third, fifth, and seventh of the cerulein injection or the NK1R antagonist CP96345 (5 mg/kg) was intraperitoneally injected 30 min before the first cerulein administration. The von Frey test was performed to evaluate pain behavior. Animals were sacrificed at 12 h from the first cerulein/saline injection for severity assessment. Pharmacology network analysis was used to identify active ingredients of CQCQD for AP and pain. In vitro, freshly isolated pancreatic acinar cells were pre-treated with CQCQD (5 mg/ml), CP96345 (1 µM), or selected active compounds of CQCQD (12.5, 25, and 50 µM) for 30 min, followed by SP incubation for another 30 min. RESULTS: The VAS score as well as the levels of serum SP and expressions of pancreatic SP-NK1R were up-regulated in moderately severe and severe patients compared with those with mild disease. CQCQD, but not CP96345, consistently and significantly ameliorated pain, pancreatic necrosis, and systemic inflammation in cerulein-induced AP as well as inhibited NK1R internalization of pancreatic acinar cells. These effects of CQCQD were associated with reduction of pancreatic SP-NK1R and neuron activity in pancreas, dorsal root ganglia, and spinal cord. Baicalin, emodin, and magnolol, the top 3 active components of CQCQD identified via pharmacology network analysis, suppressed NK1R internalization and NF-κB signal pathway activation in isolated pancreatic acinar cells. CONCLUSIONS: CQCQD ameliorated cerulein-induced AP and its associated pain via inhibiting neuron activation-mediated pancreatic acinar cell SP-NK1R signaling pathways and its active compounds baicalin, emodin, and magnolol contributed to this effect.

Quantitative determination of aloin, antioxidant activity, and toxicity of Aloe vera leaf gel products from Greece.

BACKGROUND: Aloe vera is a popular medicinal plant used widely by the cosmetic, pharmaceutical, and food industries. The A. vera leaf gel, which is used mostly for its positive effects on human health, contains over 75 different bioactive compounds, including aloin. Aloin is a toxic compound, and its content in A. vera leaf gel products depends on the different cultivation conditions and especially on leaf processing. RESULTS: In this study, A. vera leaf gel products, varied in terms of leaf processing, were analyzed using liquid chromatography for their aloin content, their antioxidant activity by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS·+ ) and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH· ) antioxidant activity assays and their toxicity against Aliivibrio fisheri and SH-SY5Y cells. In the samples processed with industrial methods and in those filtered in the lab, the content of aloin was found below the limit (0.1 mg L-1 ) of the EU legislation however, the unprocessed and unfiltered samples were found to contain more than 10 mg L-1 . Antioxidant activity was estimated to vary from 1.64 to 9.21 µmol Trolox mL-1 for DPPH· and from 0.73 to 5.14 µmol Trolox mL-1 for ABTS·+ . Toxicity values on A. fisheri, expressed as the concentration at 50% loss of initial luminescence, ranged from 0.03 to 0.09 mg mL-1 . The cytotoxic study indicated that aloin A at low concentrations (1 and 10 µg mL-1 ) protects SH-SY5Y cells from toxicity induced by hydrogen peroxide. CONCLUSIONS: Consequently, the filtration process of A. vera leaf gels, either laboratory or industrial, resulted in aloin A content below the EU legislation detection limits. © 2020 Society of Chemical Industry.

Solid phase adsorption of emodin on hydrotalcites and inorganic oxides: A preliminary study.

Solid phase extraction applied to plant matrices is nowadays a well validated technique allowing the concentration and purification of selected secondary metabolites for subsequent analysis. In this short communication we screened the efficiency of 16 selected solid supports including layered structures (hydrotalcites and zirconium phosphate), magnesium oxide and hydroxide, and finally the phyllosilicates talc and bentonite for the selective concentration of the anthraquinone emodin from raw solid extracts of Polygonum cuspidatum Siebold & Zucc. (sin. Reynoutria japonica Houtt.) (Polygonaceae), commonly known as "Japanese knotweed". An ethanolic solution of sample extract from this plant was vigorously mixed with fixed quantities of each solid support. Subsequent HPLC analysis, coupled to photodiode array detection, revealed that, among the solid supports assayed, the hydrotalcite zinc aluminum oleate and magnesium oxide were largely the most effective to this concern. Both were able to extract emodin from the raw extract in percentages of 81.5 % and 92.4 %, respectively. The application of the title supports for the extraction and concentration in the solid phase of anthraquinones from raw plant extracts have been reported herein for the first time.

Fungal and plant metabolites in industrially-processed fruit juices in Nigeria.

There is scarce data on the mycotoxin profile in retailed fruit juices in Nigeria. Thirty-five industrially-processed fruit juice samples randomly purchased from retailers in Ogun state, Nigeria, were analysed for the presence of > 650 toxic fungal and plant metabolites using a liquid chromatography tandem mass spectrometric method. Only 18 metabolites, including 3-nitropropionic acid, alternariol methylether and emodin, but excluding citrinin, fumonisin B2, ochratoxin A and patulin, were detected in trace levels in at least one juice sample. Amygdalin, a plant cyanogen, was quantified (2.05-359 µg/L) in 40% of the samples. Although the levels of mycotoxins and toxic plant metabolites found in the juice may be relatively low, daily consumption of juices containing such low levels may contribute to dietary exposures to these natural chemical contaminants in consumers. Fruit juice processors should be encouraged to adhere strictly to good manufacturing practices in order to keep mycotoxins away from the final products.

Amount of Eurotium sp. in Chinese Liupao tea and its relationship with tea quality.

OBJECTIVE: Eurotium sp. are the sexual states of the genus Aspergillus, and their ascospore is a spherical closed capsule with a golden colour. The growth of Eurotium sp. during tea production is a key step in achieving the unique quality of dark tea. This study aimed to evaluate the relationship between Eurotium sp. amount and Liupao tea quality. METHODS AND RESULTS: The amounts of Eurotium sp. in 26 differently aged Liupao tea samples from several factories were studied. Indicators related to the quality of Liupao tea were investigated. The amounts of Eurotium sp. were divided into 0, 105 and 106 levels, and quantitative real-time polymerase chain reaction (qPCR) was performed. Using ultrahigh-performance liquid chromatography and high-performance liquid chromatography, the amounts of emodin and physcion were determined to be closely related to the amount of Eurotium sp. Emodin was not found or occurred in minimal amounts in all raw Liupao tea samples. By contrast, physcion was found in Liupao tea at the 106 level of Eurotium sp. Liupao tea samples with varying levels of Eurotium sp. also exhibited evident differences in aroma and chromaticity. Result of the Pearson correlation test showed that the amount of Eurotium sp. plays a key role in creating the unique quality of Liupao tea. CONCLUSION: The amount of Eurotium sp. in dark tea detected via qPCR can be used as a quantitative quality indicator for evaluating dark tea. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study provides an efficient method for identifying the different qualities of dark tea and addressing quality control issues in fermenting dark tea.

Quality control of Aloe vera (Aloe barbadensis) and Aloe ferox using band-selective quantitative heteronuclear single quantum correlation spectroscopy (bs-qHSQC).

In the present study, band-selective quantitative heteronuclear single quantum correlation spectroscopy (bs-qHSQC) was applied for the quality control of the two Aloe species present in the European Pharmacopeia. After development and validation of a complete spectral range (csr-) qHSQC assay, a specific pulse program with selective excitation was applied and the measuring time was reduced from 135 to 32 min, while maintaining the same resolution. This bs-qHSQC method (method I) showed slightly higher RSD values compared to the csr-qHSQC method (maximum RSD of 2.80%), but better recovery rates in comparison to the assay of the Pharmacopeia (97.3% for Aloe vera and 96.6% for Aloe ferox). Apart from quantifying the total anthranoid content, the method moreover allows the quantitation of aloin among other aloin derivatives, such as 7-hydroxyaloin, as well as the differentiation of the two investigated species. Additionally, a second bs-qHSQC method (method II) for the fast (4 min) determination of the aloin A/B ratio was developed and compared to 13C qNMR measurements. Showing the same results in much less analysis time, the latter approach contributes to a general problem in natural product chemistry, the co-occurrence of structurally similar compounds and their analysis in complex matrices, e.g. plant extracts.

Simultaneous Quantitation of Five Bioactive Ingredients in Raw and Processed Fallopia multiflora by Employing UHPLC-Q-TOF-MS.

Fallopia multiflora is used for treatment of premature graying hair and blood deficiency. In this study, a quantitative method was developed for determination of five bioactive components (emodin, 2,3,5,4'-tetrahydroxy-stilbene- 2-Ο-ß-d-glucoside, emodin-8-O-ß-d-glucopyranoside, ω-hydroxyemodin and kaempferol) in raw and processed F. multiflora by using ultra-high performance liquid chromatography (UHPLC)-quadrupole time-of-flight mass spectrometry-based method. The sample handling procedure was optimized. Chromatographic separation was carried out on a Thermo Syncronis AQ-C18 UHPLC column with mobile phase consisting of 0.01% aqueous formic acid and acetonitrile. The method was interrogated in terms of linearity, precision, stability and recovery tests. All calibration curves displayed good linearity (R2 > 0.9992). The limit of detection and limit of quantification of these components ranged from 0.01 to 0.03 µg/mL and from 0.03 to 0.07 µg/mL, respectively. The average recoveries of these components were from 98.2 to 102.9% with relative standard deviation values from 0.8 to 2.9% for F. multiflora. The developed method can be applied to quality control of raw and processed F. multiflora.

CYP3A Activation and Glutathione Depletion Aggravate Emodin-Induced Liver Injury.

1,3,8-Trihydroxy-6-methylanthraquinone (emodin), a widely existing natural product in herbal medicines, has been reported to be hepatotoxic, but the exact underlying mechanism is still not fully understood. The objective of the present study was to evaluate the role of CYP3A and glutathione (GSH) in emodin-induced liver injury. Primary human hepatocytes were exposed to emodin with and without addition of CYP3A inducer/inhibitor and GSH synthesis inhibitor. It was found that emodin-mediated cytotoxicity increased when CYP3A was activated and GSH was depleted. Hepatotoxicity induced by emodin in rats by activation/inhibition of CYP3A and depletion of GSH was further investigated. Administration of emodin in combination with l-buthionine sulfoximine (BSO) or dexamethasone (DEX) resulted in aggravated liver injury, whereas pretreatment with ketoconazole (KTZ) suppressed the side effects caused by emodin. In addition, plasma exposure of emodin and its glucuronide metabolite were measured by ultraperformance liquid chromatography triple quadrupole mass spectrometry. Emodin and its glucuronide were lower in BSO-, DEX-, and KTZ- co-treated rats compared with those administered with emodin alone. In conclusion, these mentioned results suggested that CYP3A induction and GSH depletion might be involved in hepatotoxicity induced by emodin. This study may help to understand the risk factors and the mechanism of hepatotoxicity of emodin in humans.

Rapid quantification of aloin A and B in aloe plants and aloe-containing beverages, and pharmaceutical preparations by microchip capillary electrophoresis with laser induced fluorescence detection.

A microchip capillary electrophoresis coupled with laser induced fluorescence detection method for the fast determination of aloin was developed and comprehensively applied for the quantification of aloin A and B present in seven aloe plant species, 42 aloin-containing crude drugs, ten aloe pharmaceutical preparations, and four aloe gel-containing functional foods. The excitation and emission wavelengths for detection of both aloins were set at 473 and 520 nm, respectively. Sample analysis on a 35 mm length of glass microchip channel was completed within 40 s. An interference study indicated that the other main anthraquinones present in the samples did not interrupt with the target aloins detection, demonstrating the good selectivity of this method. It is demonstrated that this method is fast, facile, and specific for determination of aloin A and B from matrix samples which can be applied to the quality control of a wide varieties of aloe species and aloe-derived products.

Exploring the Q-marker of "sweat soaking method" processed radix Wikstroemia indica: Based on the "effect-toxicity-chemicals" study.

BACKGROUND: Radix Wikstroemia indica (RWI), named "Liao Ge Wang" in Chinese, is a kind of toxic Chinese herbal medicine (CHM) commonly used in Miao nationality of South China. "Sweat soaking method" processed RWI could effectively decrease its toxicity and preserve therapeutic effect. However, the underlying mechanism of processing is still not clear, and the Q-markers database for processed RWI has not been established. PURPOSE: Our study is to investigate and establish the quality evaluation system and potential Q-markers based on "effect-toxicity-chemicals" relationship of RWI for quality/safety assessment of "sweat soaking method" processing. METHODS: The variation of RWI in efficacy and toxicity before and after processing was investigated by pharmacological and toxicological studies. Cytotoxicity test was used to screen the cytotoxicity of components in RWI. The material basis in ethanol extract of raw and processed RWI was studied by UPLC-Q-TOF/MS. And the potential Q-markers were analyzed and predicted according to "effect-toxicity-chemical" relationship. RESULTS: RWI was processed by "sweat soaking method", which could preserve efficacy and reduce toxicity. Raw RWI and processed RWI did not show significant difference on the antinociceptive and anti-inflammatory effect, however, the injury of liver and kidney by processed RWI was much weaker than that by raw RWI. The 20 compounds were identified from the ethanol extract of raw product and processed product of RWI using UPLC-Q-TOF/MS, including daphnoretin, emodin, triumbelletin, dibutyl phthalate, Methyl Paraben, YH-10 + OH and matairesinol, arctigenin, kaempferol and physcion. Furthermore, 3 diterpenoids (YH-10, YH-12 and YH-15) were proved to possess the high toxicity and decreased by 48%, 44% and 65%, respectively, which could be regarded as the potential Q-markers for quality/safety assessment of "sweat soaking method" processed RWI. CONCLUSION: A Q-marker database of processed RWI by "sweat soaking method" was established according to the results and relationship of "effect-toxicity-chemicals", which provided a scientific evidence for processing methods, mechanism and the clinical application of RWI, also provided experimental results to explore the application of Q-marker in CHM.

[Research on period of validity of emodin standard solution and chrysophanol standard solution].

In order to study the storage and period of validity of emodin standard solution and chrysophanol standard solution in this study, the content of emodin and chrysophanol was determined by HPLC through classical constant temperature test, and the change rule of the content of the standard solution was studied, which could be applied to standardize the management of the standard substance of traditional Chinese medicine. The results showed that the content of emodin and chrysophanol standard solution matched with the first order reaction rule. Under the storage condition of 10 °C, the change rate constant of emodin and chrysophanol were Ke=4.661 7×10⁻7 and Kc=4.438 9×10⁻7, respectivedy; and the period of validity of emodin standard solution and chrysophanol standard solution were 1 806 d and 1 896 d respectively. The determination and standardization of the period of validity of the standard solution will not only help to reduce the loss of the standard substance and save the cost of drug testing, but also help to standardize the use of the standard substance, which will contrite to obtain more accurate and satisfactory experimental results, and provide a basis for the setting of the storage period and standardized management of the reference solution of Chinese medicine.

An optimized protocol for anthraquinones isolation from Rhamnus frangula L.

Different from works described in the literature, which use expansive analytical methods to separation of anthraquinones derivatives (AQs), this communication reported a simple and inexpensive methodology to get them. In this way, the expensive commercial AQs: Chrysophanol, physcione and emodine were extracted from plant material (Rhamnus frangula L.) and isolated by classical column chromatography technique under optimised binary mobile phase gradients (CHCl3 : AcOEt(a), a = 1 to 5%) in excellent yields.

Formulation of an aloe-based product according to Iranian traditional medicine and development of its analysis method.

BACKGROUND: Currently, people are more interested to traditional medicine. The traditional formulations should be converted to modern drug delivery systems to be more acceptable for the patients. In the present investigation, a poly herbal medicine "Ayarij-e-Faiqra" (AF) based on Iranian traditional medicine (ITM) has been formulated and its quality control parameters have been developed. METHODS: The main ingredients of AF including barks of Cinnamomum zeylanicum Blume and Cinnamomum cassia J. Presl, the rhizomes of Nardostachys jatamansi DC., the fruits of Piper cubeba L.f., the flowers of Rosa damascena Herrm., the oleo gum resin of Pistacia terebinthus L. and Aloe spp. dried juice were powdered and used for preparing seven tablet formulations of the herbal mixture. Flowability of the different formulated powders was examined and the best formulations were selected (F6&F7). The tablets were prepared from the selected formulations compared according to the physical characteristics and finally, F7 was selected and coated. Physicochemical characters of core and coated AF tablets were determined and the HPLC method for quantitation of aloin as a marker of tablets was selected and verified according to selectivity, linearity, precision, recovery, LOD and LOQ. RESULTS: The results showed that core and coated AF tablets were in agreement with USP requirements for herbal drugs. They had acceptable appearance, disintegration time, friability, hardness, dissolution behavior, weight variation and content uniformity. The amount of aloin in tablets was found 123.1 mg/tab. The HPLC method for aloin determination in AF tablets was verified according to selectivity, linearity (5-500 µg/ml, r2:0.9999), precision (RSD: 1.62%), recovery (108.0%), LOD & LOQ (0.0053 & 0.0161 µg/ml). CONCLUSIONS: The formulated tablets could be a good substitute for powder and capsules of AF in ITM clinics with a feasible and precise method for its quality control. Ayarij-e-Faiqra formulation.

Antiplasmodial potential and quantification of aloin and aloe-emodin in Aloe vera collected from different climatic regions of India.

BACKGROUND: In this study, Aloe vera samples were collected from different climatic regions of India. Quantitative HPTLC (high performance thin layer chromatography) analysis of important anthraquinones aloin and aloe-emodin and antiplasmodial activity of crude aqueous extracts was done to estimate the effects of these constituents on antiplasmodial potential of the plant. METHODS: HPTLC system equipped with a sample applicator Linomat V with CAMAG sample syringe, twin rough plate development chamber (20 x 10 cm), TLC Scanner 3 and integration software WINCATS 1.4.8 was used for analysis of aloin and aloe-emodin amount. The antiplasmodial activity of plant extracts was assessed against a chloroquine (CQ) sensitive strain of P. falciparum (MRC-2). Minimum Inhibitory Concentration (MIC) of aqueous extracts of selected samples was determined according to the World Health Organization (WHO) recommended method that was based on assessing the inhibition of schizont maturation in a 96-well microtitre plate. EC (effective concentration) values of different samples were observed to predict antiplasmodial potential of the plant in terms of their climatic zones. RESULTS: A maximum quantity of aloin and aloe-emodin i.e. 0.45 and 0.27 mg/g respectively was observed from the 12 samples of Aloe vera. The inhibited parasite growth with EC50 values ranging from 0.289 to 1056 µg/ml. The antiplasmodial EC50 value of positive control Chloroquine was observed 0.034 µg/ml and EC50 values showed by aloin and aloe-emodin was 67 µg/ml and 22 µg/ml respectively. A positive correlation was reported between aloin and aloe-emodin. Antiplasmodial activity was increased with increase in the concentration of aloin and aloe-emodin. The quantity of aloin and aloe-emodin was decreased with rise in temperature hence it was negatively correlated with temperature. CONCLUSIONS: The extracts of Aloe vera collected from colder climatic regions showed good antiplasmodial activity and also showed the presence of higher amount of aloin and aloe-emodin in comparison to collected from warmer climatic sites. Study showed significant correlation between quantities of both the anthraquinones used as marker compounds and EC50 values of the different Aloe vera extracts. Although, both the anthraquinones showed less antiplasmodial potential in comparison to crude extracts of different Aloe vera samples. Diverse climatic factors affect the quantity of tested compounds and antiplasmodial potential of the plant in different Aloe vera samples.