Steroid Compounds Isolation from Carthamus tinctorius Linn as Antimalarial

Carthamus tinctorius Linn, also known as safflower, is a plant with the potential of being used in the production of antimalarial drugs. The purpose of this study was to isolate and identify the steroid compounds in the safflower and determine its antimalarial activity in vitro. The isolation process was conducted through extraction and chromatography methods. Then, the characterization of the isolated compounds was conducted through spectroscopic techniques which include Fourier Transform Infrared Spectroscopy (FT-IR), NMR 1-D (1H and 13C-NMR), and NMR 2-D (HMQC, HMBC, and H-H COZY) as well as comparing data with the existing literatures. In addition, the tests conducted were with variations of isolate concentrations (10, 1, 0.1, 0.01, and 0.001 μg/mL) against 3D7 strain of Plasmodium falciparum. Based on the FT-IR spectroscopic data, the steroid compounds isolated from safflowers might be stigmasterols. In addition, the isolates had -OH functional group in the region of 3431 cm-1, C-O in the region of 1053 cm-1, and Csp3-H in regions of 2960, 2934, and 2865 cm-1. The NMR 1-D data showed presence of 29 carbon atoms, while the protons were 48 in number. Furthermore, the IC50 value of the compound was 34.03 μg/mL with a percentage inhibition of 43.92% against the growth of P. falciparum. Therefore, it was classified as inactive agent in inhibiting the growth of malaria parasites, however, it could be used as a marker compound in C. tinctorius Linn extract.

Compositional Characteristics of Glucuronide Conjugates in Regional Glechoma hederacea var. longituba Herbal Extracts Using a Set of Polyphenolic Marker Compounds

Glechoma hederacea var. longituba (GHL) is one of many herbal plants widely used in hot herbal teas and in oriental prescriptions to treat various diseases. Although the beneficial effects of GHL may be influenced by differences in the composition of active constituents in the herbal extracts, there are few reports on the compositional characteristics of GHL herbal extracts to date. In this study, liquid chromatography–mass spectrometry technology was used for comparative analysis of constituents in hot-water extracts of GHL samples obtained from various cultivating provinces in South Korea. A set of marker panel consisting of nine polyphenolic compounds, including glucuronide conjugates in particular, was constructed and used to monitor the compositional characteristics in each GHL extract. Our findings show that some of the marker compounds, including rosmarinic acid, were persistently observed as major constituents in the analyses of the 22 GHL sample extracts, whereas, interestingly, other marker compounds such as polyphenol-glucuronic acid conjugates displayed dramatic differences in compositional ratios. This chromatographic approach using the marker compound panel can be applied to qualitatively and quantitatively evaluate compositional characteristics in the GHL extracts, and can also be useful for quality assays of the GHL herbal plant in medicinal and industrial fields.

QUANTITATIVE DETERMINATION OF ARTEANNUIN B – A SESQUITERPENE LACTONE FROM ARTEMISIA ANNUA

The HPLC (High performance liquid chromatography) method for the determination of arteannuin B in the herb Artemisia annua has been developed. The method is suitable for its validation characteristics, such as specificity parameters (convergence of the retention times of artianuin B in the standard and sample), suitability of the chromatographic system (separation efficiency), linearity (R2 = 0.99929 at y = 9.38x + 245.98), range of application (from 50 to 150%), accuracy in terms of repeatability (RSD = 3.45%) and correctness (Z = 100.36 ± 0.56%), determined experimentally, are within the recommended values. As a marker compound, it was proposed to use a standard sample of arteannuin B. For this, a standard sample of arteannuin B was obtained with a purity of at least 99%, which is confirmed IR, 1HNMR, 13CNMR spectral data. Organoleptic and physicochemical parameters were determined. It has been established that the presence of impurities in an amount of not more than 1% is allowed in a standard sample of arteannuin B. It was established that the content of arteannuin B in the raw material should be at least 0.2%. The development and validation HPLC-UV method for the determination of arteannuin B have to used to standardization the herb of Artemisia annua.

Profiling of 2-Acetyl-1-Pyrroline and Other Volatile Compounds in Raw and Cooked Rice of Traditional and Improved Varieties of India

Herein, optimized headspace solid phase microextraction with gas chromatography–tandem mass spectrometry (HS-SPME-GC-MS/MS) was used to estimate the 2-acetyl-1-pyrroline (2-AP) in raw and cooked rice samples of ten different traditional and improved varieties. Furthermore, HS-SPME-GC-MS-based volatile profiling was subjected to untargeted analyses to identify major odorants in raw and cooked rice samples, and to understand chemical proximities among volatile profiles. Results showed that 2-AP content was remarkably increased in cooked rice compared to raw. Among the varieties studied, Pusa-1652 (Improved Kala Namak) and Kala Namak-2 were superior in the 2-AP content than Basmati varieties. Additionally, Govind Bhog, Kala Jeera and Jeera-32 had 2-AP content equivalent to or superior to Basmati rice varieties. Altogether, 18 and 22 volatiles were identified in the raw and cooked rice samples studied, respectively. Of these, ethyl butyrate, ethyl 3-methylbutanoate, 2-undecanone, ethyl benzoate, ethyl benzeneacetate, 2-methylnaphthalene, and 1-methylnaphthalene were characteristically detected in the cooked rice. The high amount of 2-ethyl-1-hexanol was uniquely found in raw rice samples, which can be a marker compound for freshly milled rice. Along with 2-AP, butanoic acid and benzoic acid derivatives, phenylethyl alcohol, ethyl 3-hydroxybutyrate, and indole may be responsible for the overall perceived characteristic Basmati-like aroma in cooked rice.

The Standardized Wood Extract of Albizia myriophylla: Its Potential as an Active Ingredient in an Anti-inflammatory Herbal Gel Formulation

Albizia myriophylla has been used in Thai folk medicine for treating inflammation-related diseases. The wood of this medicinal plant is traditionally used as a single herbal drug in the form of an aqueous decoction and as a component in several Thai herbal formulations for the remedy of fever, sore throat, and aphthous ulcers. This study aimed to evaluate in vivo the anti-inflammatory potential and possible mechanism of action of the standardized wood extract of A. myriophylla as well as to investigate the anti-inflammatory activity and physicochemical properties of the developed herbal gel formulation containing standardized wood extract of A. myriophylla. Results of quantitative HPLC analysis demonstrated that the standardized wood extract of A. myriophylla contained 22.95 mg/g of 8-methoxy-7,3′,4′-trihydroxyflavone, a bioactive marker compound of A. myriophylla. The standardized wood extract of A. myriophylla (1% w/v) exhibited remarkable inhibition (54.4 – 80.3%) in the croton oil model of topical inflammation at all assessment times, comparable to standard indomethacin (55.3 – 63.6%). Real-time quantitative reverse transcription-polymerase chain reaction was performed to clarify the anti-inflammatory mechanism of standardized wood extract of A. myriophylla, and the result showed that this standardized extract decreased lipopolysaccharide-induced nitric oxide synthase mRNA levels in a dose-dependent manner. The developed herbal gel containing standardized wood extract of A. myriophylla (1% w/w) had good physicochemical characteristics and exhibited potent inhibition (51.4 – 77.8%) of inflammation in a rat ear edema model at all assessment times, comparable to indomethacin gel (33.3 – 40.5%). The notable anti-inflammatory activity of standardized wood extract of A. myriophylla and its developed herbal gel formulation indicates their potential application as natural anti-inflammatory agents.

EFFECT OF STERILIZATION USING GAMMA RADIATION ON THE PHYSICOCHEMICAL PROPERTIES OF GEL CONTAINING ALOE VERA POWDER

Objective: This study aims to prepare sterile gel containing aloe vera (AV) powder and evaluate its physicochemical properties after sterilization by gamma radiation. Methods: The gel was prepared using carbomer as stabilizer, and sterilized by gamma radiation. The physical stability was evaluated including organoleptic, pH, viscosity and sterilization. Furthermore, malic acid concentration was determined as a marker compound contained in the gel. Results: The gel was successfully prepared containing 20% AV powder and 25% carbomer. The physical properties including organoleptic, pH and viscosity were not significantly changed after sterilization, and also stable even after 28 storage days. Meanwhile, malic acid concentration before and after sterilization were 47.2 mg/ml and 43.9 mg/ml, respectively. This showed the physicochemical properties were not significantly different after sterilization. Conclusion: Gamma radiation is suitable to sterilize gel containing AV powder.

Investigation of the long-term stability of various tinctures belonging to the lamiaceae family by HPLC and spectrophotometry method

The aim of the current study was to analyze the stability of rosmarinic acid in ethanolic tinctures of lemon balm (Melissa officinalis L.), oregano (Origanum vulgare L.), peppermint (Mentha x piperita), rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.), and thyme (Thymus vulgaris L.). High-performance liquid chromatography with diode-array detection (HPLC–DAD) was employed to monitor the concentration of the marker compound over a six month period. Furthermore, the tinctures were also evaluated for caffeic acid, total phenolic content, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. We observed that the concentration of rosmarinic acid in tincture stored in closed amber glasses at ambient temperature decreased significantly during 6-month storage. Furthermore, our study squarely confirms the fact that a part of rosmarinic acid is converted to caffeic acid. The tested tinctures can be listed in the following order according to the greatest stability of the marker compound: rosemary > peppermint > oregano > lemon balm > thyme > sage. The results of the study indicated a linear relationship between DPPH values and total phenolic (R2 = 0.92) or rosmarinic acid (R2 = 0.85) contents.

Detection of puerarin from Pueraria mirifica tuber and its formulated cosmetic products

Pueraria mirifica (PM) has traditionally been used to relief postmenopausal symptoms. Recently, its extract has been developed into various cosmetic products to promote skin rejuvenation and youthfulness. This study investigated the phytochemicals of PM tuber and compared between the tuber flesh and its outer peel. Puerarin which is one of the major isoflavones and being considered as the marker compound was used to determine the presence of PM extract in local cosmetic products. Puerarin could be ionized by a mass spectrometer at both negative and positive modes. The peak ionized at the negative mode showed to have a narrower peak width (0.2 min) and higher signal-to-noise ratio (30) for pueararin (1 mg/L). The results also found PM extract contained many C- and O-glycosylated isoflavones, especially from its peel extract. This explains the peel extract showed to have four times higher antiradical activity than those of flesh extract. Puerarin from the cosmetic products was recovered via successive methanolic sonication and followed by liquid-liquid extraction using ethyl acetate. Puerarin was successfully partitioned from the highly complex chemical mixture of cosmetic products with the recovery ranged from 89.1 % to 115 %. Hence, isoflavones was found to be higher at the outer peels than its tuber flesh. A simple and reliable method has been developed to analyse the presence of PM extract in cosmetic products based on the detection of puerarin after successive extraction via methanolic sonication and ethyl acetate partition.