Yt Kamal

Development and validation of HPLC method for simultaneous estimation of piperine and guggulsterones in compound Unani formulation (tablets) and a nanoreservoir system

An attempt has been made to develop and validate a simultaneous HPLC method for novel approach of drug release via oil-in-water (o/w) nanoemulsion formulation and Habb-e-Khardal Unani tablet containing piperine and guggul sterones E and Z as main ingredients. Nanoemulsion was prepared by titration method using sefsol-218 as an oily phase, cremophor-EL as a surfactant, transcutol as a co-surfactant and distilled water as an aqueous phase. The formulation was optimized on the basis of thermodynamic stability and dispersibilty test. The nanoformulation was evaluated for particle size, surface morphology, electrical conductivity and viscosity determination. The in vitro dissolution was carried out by dialysis bag method. Drugs were quantified using an HPLC method developed in-house with a C(18) column as stationary phase and acetonitrile and water as mobile phase at λ(max) of 240 nm. The optimized formulation showed higher drug release, lower droplet size and less viscosity as compared with the conventional Habb-e-Khardal Unani tablet. The present study illustrated the potential of nanoemulsion dosage form in improving biopharmaceutic performance of piperine and guggul sterone. The HPLC method was also found to be quite sufficient for the routine quality control of formulations containing piperine and guggul sterone E and Z as ingredients and also for in vitro drug release studies.

FAST ANALYSIS AND VALIDATION OF RUTIN IN ANTI-PSORIATIC AYURVEDIC FORMULATION BY HPLC

777 Oil is a topically applied Ayurvedic formulation used for the effective treatment of psoriasis. The formulation is composed of leaf extract of Wrightia tinctoria and Oleum Cocus nucifera. A selective, sensitive, and reproducible HPLC method was developed for analyzing marker compound of Wrightia tinctoria (Rutin) in 777 Oil for routine standardization purpose. The chromatography was performed on Phenomenex C18 (250 × 4.6 mm, 5.0 µm particle) column using methanol-water (60:40, v/v) as mobile phase; adjusted to pH 3.0 by orthophosphoric acid. The flow rate was 1.0 mL/min. with detection at 360 nm. The values of retention times and capacity factor were 3.88 and 0.40, respectively. The calibration plot showed a good linear relationship between response curve and concentration in the range of 1.0–1000.0 µgmL−1 with regression coefficient 0.9998. The detection (LOD) and quantification (LOQ) limits were found 27.0 and 95.0 ngmL−1 respectively. The statistical analysis proved that the method was precise, reproducible, selective, and accurate for the analysis of rutin in 777 Oil. The developed HPLC method is useful for the qualitative and quantitative estimation of rutin in 777 Oil and other products of traditional systems of medicine.

Metabolic diversity in Coleus forskohlii Briq. of Indian subcontinent

New economic, easy, specific, accurate, robust, validated high performance thin layer chromatography (HPTLC) and high performance liquid chromatography methods with good range of linearity and sensitivity were developed for quantification of forskolin in ten samples collected from different regions of Indian subcontinent, which showed a large variation among samples (0.074–0.282%, w/w). Metabolic diversity of all the samples using HPTLC fingerprint method showed a total of 16 well separated spots. There is no significant metabolic diversity among the samples collected from different locations of Indian subcontinent, which was obtained from HPTLC fingerprinting. The results of locational variation showed highest content of forskolin in Bengaluru sample by both analytical methods. The validated quantification methods and fingerprint profile together can act as a good authentication tool for coleus as well as for other medicinal plants.

SIMULTANEOUS ESTIMATION OF GALLIC ACID, ELLAGIC ACID, AND ASCORBIC ACID IN EMBLICA OFFICINALIS AND IN UNANI POLYHERBAL FORMULATIONS BY VALIDATED HPLC METHOD

A simple, rapid, and economic simultaneous HPLC method was developed and validated for the quantification of Gallic acid (GA), Ellagic acid (EA), and Ascorbic acid (AA), in Emblica officinalis Linn. (aamla) and in two poly herbal Unani formulations, containing aamla as an ingredient. Separation was achieved on a reverse phase C18 (250 × 4.6 mm, 5 µm) column with a mobile phase of 0.1% orthophosphoric acid and acetonitrile in gradient elution. The flow rate was kept as 1 mL min−1 and detection was carried out at 254 nm. The proposed method was validated as per the ICH guidelines for accuracy, precision, robustness, LOD, and LOQ. The proposed method was linear over a wide range of concentration with a good regression coefficient of more than 0.99. The LOD of method was 0.035 µg mL−1, 0.31 µg mL−1, and 0.38 µg mL−1, whereas LOQ was 0.1 µg mL−1, 1.0 µg mL−1 and 1.3 µg mL−1 for ascorbic acid, ellagic acid, and gallic acid, respectively. The results demonstrated that the proposed method is a simple, reproducible, accurate, economic, and suitable method for the quality control of aamla, its formulations, as well as a large population of plants with anti-oxidant property.

Quality control and in vitro antioxidant potential of Coriandrum sativum Linn.

Background: Coriandrum sativum Linn., commonly known as coriander, is a well-known spice and drug in India. It has various health-related benefits and used in various Unani formulations. In this present study, quality assessment of coriander fruits was carried out by studying anatomical characters, physicochemical tests, and chemoprofiling using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectroscopy (GC-MS) along with in vitro antioxidant potential. Materials and Methods: Standardization was carried out as per the pharmacopeial guidelines. Estimation of heavy metals, pesticides, and aflatoxins was carried out to ascertain the presence of any contaminant in the sample. Chemoprofiling was achieved by thin layer chromatography (TLC) by optimizing the mobile phase for different extracts. The most of the pharmacological activities of coriander are based on volatile oil constituents. Hence, GC-MS profiling was also carried out using hexane-soluble fraction of hydro-alcoholic extract. The total phenolic contents and in vitro antioxidant efficacy were determined using previously established methods. Results: The quality control and anatomical studies were very valuable for the identification whereas good antioxidant potential was observed when compared to ascorbic acid. The drug was found free of contaminant when analyzed for pesticides and aflatoxins whereas heavy metals were found under reported limits. Conclusion: The work embodied in this present research can be utilized for the identification and the quality control of the coriander fruit.

A high performance thin layer chromatographic method for the estimation of colchicine in different formulations

Background: Colchicine is a main alkaloid present in bitter and sweet variety of colchicum (Colchicum luteum Baker), which have been reported to possess anti-rheumatic, anti-gout, and anticancer potential. Colchicum is an important ingredient of several Unani and Herbal formulations. Quantification of colchicine will play a great role in quality control of these formulations. Hence, a high-performance thin layer chromatographic (TLC) method has been developed for the analysis of colchicine in Unani formulations of various dosage forms such as hubb (tablet) and capsules. Materials and Methods: The samples were applied on aluminum TLC plates precoated with silica gel 60-F254 and developed using mobile phase toluene-dichloromethane-methanol in equal proportions. Quantification was done by densitometric scanning at 350 nm, which showed a linear response in the range of 50–500 ng/spot. The developed method was validated as per the International Conference on Harmonization guidelines for linearity, precision, accuracy, specificity, robustness, limit of detection, and limit of quantification. Results and Conclusion: The developed method was applied for quantitative estimation of colchicine in different Unani and Herbal formulations. The method was found simple, selective, accurate with a wide range of linearity, hence suitable for the quality control of different formulations and varieties of colchicum with respect to colchicine content.

Matrix solid-phase dispersion extraction and quantification of alpinetin in amomum seed using validated HPLC and HPTLC methods

Alpinetin is a flavonoidal constituent of seeds of Amomum subulatum Roxb., recently reported to possess vasorelaxant and antiHIV activities. Simple, accurate and precise HPLC and HPTLC methods were developed for the analysis of alpinetin in A. subulatum seed extracts and extraction technique was optimized to get maximum yield using conventional, ultrasonic and matrix solid phase dispersion extraction. HPLC was performed on a C18 column with methanol and water (70:30, v/v) as mobile phase at a flow rate of 1.0 ml/min whereas HPTLC on silica aluminum sheet (60F254) using toluene, dichloromethane and ethyl acetate as solvent system. A sharp peak was obtained for alpinetin at a retention time (Rt) of 5.7 min by HPLC and retardation factor (Rf) of 0.48 by HPTLC. Both methods were validated as per the ICH guidelines and the content of alpinetin was estimated in different extracts. Matrix solid phase dispersion technique was found most suitable for extracting alpinetin as compared to other techniques. Validation data are indicative of good precision and accuracy and proved the reliability of the methods.

Standardization of Unani polyherbal formulation, Qurse-e-Hummaz: A comprehensive approach

Background: An increase in the awareness about the advantages of the traditional system of medicines has led to the commercialization of the formulations used for the treatments. Manufacture of these medicines to meet this increasing demand has resulted in a decline in their quality, primarily due to a lack of adequate regulations pertaining to this sector of medicine. Hence, it is necessary to come up with a systematic approach to develop well-designed methodologies for the standardization of polyherbal formulations which are used in traditional systems of medicine. Materials and Methods: Qurs-e-Hummaz formulations were prepared by a qualified “Hakim” (Unani medical practitioner) of Faculty of Unani Medicine, Hamdard University, as per the formula and instruction given in National Formulary of Unani Medicine. Results: Various quality control parameters such as organoleptic evaluations (color, odor, taste, and consistency), physicochemical evaluations (loss on drying, disintegration time, moisture content, total ash, acid insoluble ash, water soluble ash, pH of 1 and 10% solution, extractive values, water soluble matter, alcohol-soluble matter, and total phenolic content) and thin layer chromatography fingerprint profiling have been carried out in triplicate. The evaluation of contaminants such as heavy metals, aflatoxins, pesticide residues, and microbial contamination has also been carried out in the formulation. Conclusion: Help in maintaining the quality and batch to batch consistency of many important polyherbal formulations.