Background:: Standardization of Triphala Juice was performed by using the WHO Guidelines. The Parameters included Preliminary Analysis, Phytochemical Identification, Heavy Metal Estimation, etc. A new simple, specific, precise and accurate UV Spectrophotometric, High-Performance Liquid Methods: Chromatography and High-Performance Thin Layer Chromatography method has been developed for the Estimation of Gallic Acid in pure form. The UV- Spectrophotometric method was developed using Schimadzu 1800 UV - Visible spec-trophotometer using methanol as a solvent. The method was shown to be linear, with a detection wavelength of 273 nm for Gallic Acid. The separation was achieved on the Schimadzu Prominence-I RP-HPLC and the column used was C18 column using mobile phase consisting of mixture of Methanol: 0.1% OPA (50:50). The detection was carried out at 280 nm with a flow rate of 0.7ml/min. The retention time for Gallic Acid was found 3.89 minutes. The calibration curve was found linear (r2 = 0.999) for RP- HPLC method. The HPTLC method was developed using Aetron Sprayline instrument, Methanol as solvent and mobile phase consisting of Toluene: Ethyl Acetate: Formic Acid: Methanol (3:3:0.9:0.2). The method was found linear and the wavelength of detection for Gallic Acid was 254nm, respectively. The percentage recoveries for both methods were found in the 98.0- 102.0% range. Results:: The methods were validated in accordance with International Conference on harmoniza-tion acceptance criteria for specificity, linearity, precision, accuracy, robustness and system suita-bility. The excipients did not interfere in the determination of Gallic acid in Triphala Juice. Conclusion:: The suggested approach was effectively implemented for the quantitative determina-tion of gallic acid in Triphala juice, which would aid in quality control
Background:: There are very few methods for simultaneously determining a combined dose of SAL and KET. Objectives:: The current study aims to explore accurate, precise, simple, and cost-effective HPLC and HPTLC techniques for the simultaneous assessment of Salbutamol (SAL) and Ketotifen (KET). Methods:: The determination of Salbutamol and Ketotifen was performed by HPLC and HPTLC methods using 280 nm and 258 nm as the determination wavelength, respectively. Methanol was used to dissolve the drug for estimation in HPLC using mobile phase methanol: 10mM di-Potassium hydrogen orthophosphate in the ratio of 55:45 v/v of pH 4 at a flow rate of 1mL/min and in chloroform: toluene: methanol (7: 2: 3 v/v/v) for the estimation in HPTLC. Moreover, a statistical comparison was made between the results obtained through HPLC and HPTLC of Salbutamol (SAL) and Ketotifen (KET) using the Student’s t-test and F-test. Results:: A linear response was observed in the range of 4-24 μg/mL and 2-12 μg/mL, respectively, for SAL and KET for HPLC. R2 was found to be 0.9998 and 0.9999, respectively. For HPTLC, the linear response was observed in the concentration range of 20-120 ng/ spot and 10 - 60 ng/ spot for SAL and KET, respectively. R2 was found to be 0.9988 and 0.9998, respectively. The limit of detection (LOD) for HPLC was estimated as 0.34 μg/ml and 0.10 μg/ml for SAL and KET, respectively, and for the HPTLC method, the LOD was estimated as 4.8 μg/ml and 1.5 μg/ml, respectively. Analysing the marketed formulation by using both methods, SAL and KET within the range of 100 ± 2% were recovered. The results obtained after the estimation of the Mastifen S tablet by applying both methods were according to nominal content. Degradation studies were performed using both methods. It was found that Salbutamol was unstable in hydrolytic, oxidative and thermal degradation, whereas stable in photolytic conditions. Ketotifen was found to be stable in thermal and photolytic conditions and unstable in hydrolytic and oxidative conditions. Conclusion:: The proposed stability indicating HPLC and HPTLC methods for SAL and KET was found to be simple, accurate, and reproducible for quantitative estimation in pharmaceutical dosage form, without interference from the excipients or degradation products from the main drug component.
Background: Kutki, the dried rhizome of Picrorhiza kurroa Royle ex Benth belonging to the family of Scrophulariaceae, has been utilized globally for liver ailments. Objective: Comprehensive use of kutki needs to evaluate its role as a quality control tool for dis-crimination of kutki samples, and therefore an effective HPLC fingerprinting method was estab-lished. Methods: Reverse-phase high-performance liquid chromatography with photodiode array (RP-HPLC-PDA) detection method coupled with multivariate analysis was developed, which was mod-est, consistent and, accurate for classification of 11 kutki samples including authentic Picrorhiza kurroa rhizomes from the market of Ahmedabad and Gandhinagar in Gujarat, India. Results: The method was validated for various parameters like precision, reproducibility, and stabil-ity. The lowest value of the % relative standard deviations (RSD) was 1.31%. Chromatographic fin-gerprint profiles of 11 kutki samples, including authenticated samples, were obtained by this meth-od, which showed a total of 28 peaks, and 9 peaks were important among them. Chemometric tech-niques like PCA and HCA were applied to identify the kutki samples.The Samples of kutki could be exquisitely differentiated into two clusters. Conclusion: HPLC-PDA method coupled with multivariate analysis divulged that chromatographic fingerprint analysis was reliable and effective for quality assessment and discrimination of kutki samples.
Background: Abiraterone acetate is a derivative of steroidal progesterone, used as a first-line therapy for metastatic castration of prostate cancer. Objective: The present study encompasses the design of an experiment approach for developing a simple, reliable, and rapid RP-HPLC method for the estimation of abiraterone acetate. Method: The chromatographic separation was efficiently conducted on a Hypersil Gold C18 (50 x 4.6 mm, 5 µm) HPLC column, using the mobile phase composition of acetonitrile: dibasic potassium phosphate (0.01 mM) in the ratio of 80:20 (%v/v) at pH 6.5 with an isocratic elution mode. Furthermore, the different force degradation study including hydrolysis, oxidation, thermal, and photolytic was performed for abiraterone acetate. Result: The dynamic linearity was established in the concentration range of 0.5-10 µg/mL with r2 of 0.9998. Furthermore, the limit of detection and the limit of quantitation were 0.0978 µg/mL and 0.3260 µg/mL. The degradation of abiraterone acetate was shown in both acidic (54.16 ± 0.247 after 24 hrs) and basic conditions (35.06 ± 0.458 after 24 hrs). Furthermore, the developed method was successfully employed to quantify abiraterone acetate in bulk powder and the solid dispersion did not show any change in the retention time. Conclusion: The developed method was validated according to the ICH Q2 (R1) specification, which was found to be sensitive, accurate, precise, robust, linear, and selective compared to the reported chromatographic method.
Background: The 6-mercaptopurine is an active ingredient used to treat certain types of leukemia. This drug is an immunosuppressive and antineoplastic agent that belongs to the thiopurine class. In Brazil, 6-MP is currently available only in the form of 50 mg tablets, sold as Purinethol® and manufactured by Glaxo Smith Kline. The lack of the liquid formulation’s production impedes treatment, assuming that one of its advantages is through its applicability in pediatric patients, who show the highest incidence among others. Objective: The purpose of this work was to evaluate the development and application of a reversed phase high performance liquid chromatography (HPLC) method using an Agilent 1220 Infinity® G4294B chromatograph with photodiode array detector. Methods: HPLC assays were performed on an Eclipse plus® C18 column (4.6 x 150 mm, 3.5 μm particle size) using a gradient mode mixture of acetonitrile and aqueous acetic acid solution as a mobile phase, with a flow of 1 mL.min-1 and detection at 324 nm. The method was validated by determining its selectivity, linearity, precision, accuracy, and robustness. Results: Retention time for 6-mercaptopurine was 5.12 minutes. The detector’s response was linear at concentrations from 1.6 to 2.4 μg/mL. The results of the method’s accuracy evaluation showed mean recovery of the amount of substance added to the samples of between 99.88 and 100.5%. For precision, repeatability and intermediate precision were evaluated. The repeatability showed a standard deviation of 0.0737. The intermediate precision was assessed on three different. For three days of the studies, the values of the standard deviations were less than 3%, showing repeatability and intermediate precision adequate for the analytical method in question. The limit of detection was determined as 3.6 ng/mL. The limit of quantification was determined as 12 ng/mL. The chromatographic method was robust. Conclusion: The proposed method can be applied to control the quality of 6-MP oral suspension to ensure that the required content is delivered to pediatric oncology patients.
Background: The combination of doxycycline (DOXY) and rifampicin (RIF) is recom-mended as a treatment therapy for brucellosis by the World Health Organization. Objective: The aim of the current study was to develop and validate the stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method for the analysis of a combina-tion of doxycycline & rifampicin. Methods: The RP-HPLC method was developed and validated to estimate the doxycycline and ri-fampicin combination as per ICH guidelines. The drug combination solution was exposed to differ-ent stress conditions: acidic, basic, photo-oxidation, and oxidation. Results: The method was found linear in the range of 2 -10μg/mL for both the drugs with a reten-tion time of 3.5 min for doxycycline and 6.5 min for rifampicin at lambda maximum of 350 nm. The RP-HPLC method was precise and accurate with %RSD < 2%. The intra-day and inter-day precisions were calculated and found within the acceptable range of 5%. Both drugs demonstrated good stability in the mobile phase after 6 hours. The LOD and LOQ of doxycycline and rifampicin were 100 ng/mL & 200ng/mL and 150ng/mL & 500ng/mL, respectively. The forced degradation of the combination of drug solutions was performed. The degraded drug peaks were well-resolved from the peaks of drugs. The percentage encapsulation efficiency of doxycycline and rifampicin in the nanoparticle system was assessed by utilizing the validated RP-HPLC method and found >60% (DOXY) and >70% (RIF). Conclusion: The developed RP-HPLC method of DOXY-RIF combination was rapid, accurate, precise, and stability-indicating. The method can be appropriately applied todetecte drugs in the na-noparticulate system.
Aims: This study aimed to develop a new, fast and sustainable method by high-performance liquid chromatography (HPLC) for simultaneous determination of ascorbic acid and nicotinamide in the cosmetic emulsion. Background: Nicotinamide (NIC) and ascorbic acid (AA) are powerful antioxidants. AA presents excellent reducing power and protects the cell from oxidation. NIC is a precursor of NADPH and NADH, substances that present a strong reduction power, resulting in a huge antioxidant capacity. Objective: A new, fast and sustainable method using HPLC was developed and validated for simul-taneous quantification of AA and NIC in the cosmetic emulsion. Method: For this purpose, purified water with 0.01 % of trifluoracetic acid and ethanol (95:5, v/v), Symmetry Shield column (4.6 x 250 mm, 5 μm), 20 μL, 1.7 mL min-1 at 254 nm were used. The method was validated according to the ICH, AOAC, and ANVISA, following parameters of lineari-ty, detection and quantification limits, selectivity, accuracy, precision, and robustness. Result: The method was fast (2.7 min for AA and 3.2 min for NIC), linear between 20 and 80 μg mL-1 (r = 0.9991 for AA and r = 0.9999 for NIC), precise (RSD< 5 % for AA and NIC), accurate (RSD 0.53 % for AA and 1.02 % for NIC), and selective for the emulsion base, and also robust to small changes in flow rate, injection volume, and purified water source. Conclusions: This work shows an ecologically alternative for the quantification of AA and NIC in the study of cosmetic emulsion by HPLC, which contemplates the requirements of the current green and sustainable analytical chemistry.
Background: Levothyroxine is a synthetic thyroid hormone that is chemically identical to Thyroxine (T4), which is secreted by the follicular cells of the thyroid gland. Levothyroxine is used to treat deficiency of thyroid hormone and prevent the recurrence of thyroid cancer. Levothyroxine is present endogenously in the human body. Method: It requires a treated matrix for the preparation of calibration curve standard and quality control samples. The method was developed using LC-MS/MS and validated in human charcoal stripped serum. Charcoal stripped matrix was used for the preparation of Calibration curve standards and Quality control samples. The method involves Solid-Phase Extraction technique. Levothyroxine D3 is used as an internal standard (ISTD). Result: Chromatographic separation was achieved using reversed phase analytical column Gemini NX-C18 110Å, 3μm (50x3.6) mm. Mobile phase consisted of acetonitrile and water in a ratio of 70:30 with 150μL of formic acid in 1000 mL of the mobile phase. Mobile phase achieved shorter run-time of 0.9 minute due to the use of Ultra-high performance liquid chromatography (UHPLC). Positive electro-spray ionization technique detected MRM ion pair transitions 777.60→731.65 for Levothyroxine and 780.70→734.6 for Levothyroxine- D3 (ISTD) were used. AB SCIEX Triple Quad™ API-4000 LC-MS/MS system and the bioanalytical method with 10ng/mL as the limit of quantification have been applied successfully to pharmacokinetics studies. Conclusion: Chromatographic separation was achieved using reversed phase analytical column Gemini NX-C18 110Å, 3μm (50x3.6) mm. Mobile phase consisted of acetonitrile and water in a ratio of 70:30 with 150μL of formic acid in 1000 mL of the mobile phase. Mobile phase achieved shorter run-time of 0.9 minutes due to the use of Ultra-high performance liquid chromatography (UHPLC). AB SCIEX Triple Quad™ API-4000 LC-MS/MS system and the bioanalytical method with 10ng/mL as the limit of quantification have been applied successfully to pharmacokinetics studies.
Background: Analysis of experimental retention data upon several variants of argentation liquid chromatographic separations of different mixtures of the same lipid class into their molecular species was made to estimate new parameters of their π-complexes for every unsaturated fatty acid residue as its silver ion cluster. Methods: Planar reversed-phase liquid chromatography, both in the presence and absence of silver ions as well as adsorption argentation liquid chromatography was applied for the separation of complex rac-1,2-diacylglycerol samples from three plant sources (cocoa butter, poppy seed, and linseed oils). Results: Every value of the argentation liquid chromatographic separation selectivity for any lipid molecular species upon both planar and column variants of reversed-phase fractionation of different complex samples from native sources into their molecular components is described by additive relative polarity levels of their fatty acid residues. These levels are always connected with equivalent lipophilicity values for every lipid molecule and its potential chemical variations during all variants of reversed-phase liquid chromatography in the presence of silver ion clusters. Conclusion: New parameters for several fatty acid residues of major native polyunsaturated lipid samples may be reflected by different coordination numbers of single silver atoms of its triangular pyramidal nanoclusters. Both hydrophobicity and total polarity levels of the coordination complexes of different lipid molecular species upon their adsorption argentation liquid chromatography may also be quantitatively estimated by their fixed methylene unit variations of these molecular species for two centigrade lipid scales.
Abstract: The quality of drugs is a major concern for drug regulatory authorities and other stake-holders across the globe. Recently, drug regulatory authorities across the globe are facing a chal-lenge in controlling the purity of cardiovascular (CVS) drugs for human use, especially drugs from the Angiotensin Receptor Blocker family, such as Valsartan. The present article aims to provide comprehensive knowledge on how pharmacopeias worldwide play a key role in ensuring the quality of active pharmaceutical ingredients (API) and finished pharmaceutical products (FPPs). In this ar-ticle, the focus is on comprehensive information regarding pharmaceutical impurities, separation strategies, relevant regulatory guidelines to control impurities, and their acceptable limits, particu-larly with respect to cardiovascular active drug substances and drug formulations for human use.
Aims: This study aims to determine the quantitative prasugrel (PG) and its all possible process-related impurities. Background: To the best of our knowledge, very few analytical methods are available in the literature for monitoring process related impurities and degradation products of PG in bulk drug substance/ active pharmaceutical ingredient (API). Objective: The objective of this study is the separation of Prasugrel and its all possible process-related impurities viz., desacetyl prasugrel-tautomeric forms, intermediates including desacetyl impurity existing in its keto-enol form and positional tautomer impurities with degradation products. Method: A simple and robust HPLC-UV method having Zorbax XDB C18 column (15 cm x 4.6 mm) 3.5μm particle size column was used. Result: Prasugrel and its process related impurities were separated as well as analyzed in pharmaceutical samples' biological matrices. Conclusion: RP-LC method was developed for quantitative determination of PG and related substantial impurities were found to be highly specific, sensitive and precise. The major oxidative degradant was identified as PG desacetyl IMPs (keto-enol and positional tautomer) and hydroxyl IMP.
Aims: This study aimed to develop a high-performance liquid chromatography (HPLC) technique for estimating paracetamol glucuronide and paracetamol sulphate in the urine samples of preterm neonates. Background: Validated methods exist for estimating the principal metabolites of paracetamol in older children and those with liver disease. Here, we have developed and validated a simple technique for estimating the same in urine samples of preterm neonates. Objective: The study aims to develop and validate a simple, reliable, and accurate HPLC technique for estimating urinary paracetamol glucuronide and paracetamol sulphate metabolites. Methods: Preterm neonates of either sex diagnosed with patent ductus arteriosus (PDA) receiving paracetamol intravenously at the dose of 15 mg/kg every six hours were recruited. We ran the samples under standardized chromatographic conditions and using various dilutions of the calibration standards. Measures of assay selectivity, linearity, accuracy, and precision were estimated. Results: We observed that the peaks for paracetamol glucuronide and paracetamol sulphate were distinguished from those of the drug-free urine samples. The results for both metabolites revealed good reproducibility, with a percent coefficient of variation (% CV) of 4.3 and 4.9 for the slope for paracetamol glucuronide and paracetamol sulphate, respectively. Similarly, we observed good linearity, as indicated by the correlation coefficients of 0.99 for the metabolites. The validation assays revealed that the method is linear, accurate, and precise over the defined concentration ranges. Conclusion: We demonstrated that HPLC has good accuracy, reliability, and precision, and it can be used for estimating the principal metabolites from urine samples in neonates for defining the ontogeny of conjugation enzymes and in paracetamol overdose.
Objective: To address the separation of interfering potential impurities associated with the drug is always a daunting task. We present the method validation and quantitative determination of sulfadoxine (SUL), an anti-malarial drug with the most important interfering impurities present in pharmaceutical dosages and bulk samples using HPLC-UV method. Methods: The UV detection was obtained at 270 nm and SUL is separated on Sunfire C18 (25 cm x 4.6 mm x 5 μm) column at 45°C with a flow rate of 1.0 mL/min in a mobile phase (CH3COOH: CH3CN). The stress testing (acidic/basic/oxidative) was performed using HPLC for SUL and its impurities, showing the highly efficient separation peaks between degradant and drug products. Results: The developed method was found to be highly accurate and sensitive in regulation with ICH guidelines. Also, it was found to be free from interference from degradation products which allows the stability indicating capability of developed HPLC-UV method for SUL for validation in bulk drugs. Conclusion: The main advantages of the present method; (a) Separation achieved in 30 minutes, (b) MS compatible mobile phase renders this developed method can be directly adapted to LC-MS without any major modifications in the near future, and (c) separation of twelve impurities on Sunfire C18 column. The CFs (correction factors) had been calculated for all the impurities. It was found to be 1.6 (IMP IX), 1.70 (IMP XI) and in between 0.8-1.3 for all other impurities. The LOD of the developed method for all the analytes was in the range of 0.05 to 0.11 μg/mL and the LOQ values were in the range of 0.17 to 0.36 μg/mL.
Aim: To develop RP-HPLC method for the simultaneous estimation of methotrexate (MTX) and minocycline (MNC). Background: Different HPLC methods were reported for the estimation of MTX/MNC individually, but there is no report for the simultaneous estimation of both MTX and MNC in a simple method. Objective: The objective of the developed method is to utilize the method for the estimation of MTX/MNC in different pharmaceutical formulations and in biological fluids Methods: An HPLC method for the estimation of Methotrexate (MTX) and Minocycline (MNC) relevance to the evaluation of nanoparticulate formulations has been developed and validated. Chromatographic estimation was achieved using the mobile phase composition of sodium acetate buffer and acetonitrile (70:30% v/v) at pH 4.0 at a flow rate of 0.2 mL/min at 307 nm. Results: The calibration curve for MTX and MNC was found to be linear at nanogram (5 to 25 ng.mL-1) and microgram (5 to 25 μg.mL-1) levels at correlation coefficient range of 0.98 to 0.99 for both MTX/MNC. The lower limit of detection and limit of quantification were found to be 0.026 ng.mL-1 and 0.079 ng.mL-1 for MTX and MNC, respectively. The percentage relative standard deviation for validation parameters of both drugs was found to be less than 6.5%. The amount of MTX and MNC present within the nanoparticles was found to be MTX (0.84 mg/mL) and MNC (0.61 mg/mL). The in vitro release showed an immediate release pattern for MTX (64.95±2.08%) and MNC (90.90±1.78%) within 12 h. Conclusion: The developed analytical method for the simultaneous estimation of MTX and MNC was found to be simple, affordable, dynamic, low cost, rapid and easy to perform with good repeatability. This method is also time consuming, since the peaks were obtained within a moderate analysis time.
Background: Metformin is the first-line drug to enhance glycemic control of type 2 DIABETES Mellitus (DM2) patients. Some reported methods to determine plasma metformin by HPLC-UV are not sensitive enough. Other methods require long extraction processes. Objective: The objective of this study was to develop and validate a simple and rapid analytical method to determine plasma metformin by HPLC-UV for application in a population pharmacokinetic study. Methods: Analyte was extracted from plasma by a simple protein precipitation technique using trichloroacetic acid (15%, w/v) as the precipitating agent. Plasma samples were analyzed using a C18 column (3.0 x 150 mm, 3.5 μm) under isocratic elution with 30 mM sodium hexansulfonate (pH 5) and acetonitrile (95: 5, v,v). Results: The limit of quantification (LOQ) was 0.1 μg mL-1 and the calibration curve was linear up to 4 μg mL-1 with a correlation coefficient >0.99. The mean recovery for metformin using this extraction procedure was 84.4 - 86.6%. The intra- and inter-day coefficients of variation and percent error values of the assayed method were <20% and <15% for LOQ and QCs, respectively. Metformin was stable in plasma samples by subjecting it to three freeze-thaw cycles and storing it up to 60 days at -80°C. This method was applied to determine plasma metformin concentrations in patients with type 2 diabetes mellitus treated with this drug. Conclusion: The HPLC-UV method developed is selective, accurate and precise for the quantification of metformin in plasma samples, since sample processing is fast and simple, in addition to being applicable in pharmacokinetic studies.
Background: Sulfated Polyborate, a novel inorganic material primarily designed as a catalyst, has shown properties such as high solubility in organic solvents, low U.V. cut-off, and pKa ≈2.0, which suggests its potential as a mobile phase buffer for reverse-phase liquid chromatography. Objective: This study aims to substantiate the role of Sulfated Polyborate as mobile phase buffer for reverse-phase liquid chromatographic analysis of basic drugs with high pKa values viz. Bisoprolol fumarate, Timolol maleate, Verapamil hydrochloride, and Carvedilol. Methods: Solubilities, U.V. cut-offs, and pKa of Sulfated Polyborate were first experimentally confirmed. The behaviour of Sulfated Polyborate as mobile phase buffer at pH 3.0 was ascertained by varying the buffer concentration, flow rates, and percent organic modifier for elution of the four basic drugs on a non-end capped octyl silyl (C8) column. Similarly, the study was performed with KH2PO4 as a reference buffer. The column performance and conductometric measurements ascertained the impact of Sulfated Polyborate on the stationary phase. Results: Sulfated Polyborate and KH2PO4 buffers showed correlation coefficients of 0.99 and 1.00 for analyte retention factors for variation of buffer concentration and organic modifier composition, respectively. Peak symmetries and the number of theoretical plates were improved from > 2.0 to < 2.0 and ≈1000 to ≈3000, respectively, for variation in buffer concentrations. Similar Van Deemter plots indicated equivalency of Sulfated Polyborate and KH2PO4 buffers. The column performance and conductometric measurements depicted no adsorption on the stationary phase. Conclusion: The present study demonstrates Sulfated Polyborate as a novel buffer for analytes with higher pKa on reverse-phase liquid chromatography.
Background: To prevent vitamin deficiencies or to restore their levels, multi-vitamin formulations are indicated for the elderly, adults, children, and infants. Objective: To provide a valid high-throughput method for simultaneous determination of vitamins in multi-vitamin formulations. Methods: A high-throughput UPLC-ESI-MS method was developed and validated for simultaneous determination of vitamins B1, B2, B3, B5, B6, B7, and B9. Analytes were eluted on an Acquity UPLC® BEH C18 1.7 μm, 2.1 x 50 mm column at 40 ± 5°C. Mobile phase containing acetonitrile (0.1% formic acid) and water (0.1% formic acid) in 30:70% ratio was pumped at 300 μL/min under isocratic control. Protonated ions of vitamin B1, B2, B3, B5, B6, B7, and B9 were monitored in single ion recording mode, using an electrospray ionization probe in positive mode. Results: The m/z ratios of positive ions of vitamin B1, B2, B3, B5, B6, B7, and B9 were 265.1, 377.2, 122.95, 220, 169.98, 244.9, and 442.1, respectively. The calibration curve of different linearity range (ng/ml) was prepared for each vitamin. Linearity range for vitamin B1, B2, B3, B5, B6, B7, and B9 were 60-1000, 25-1000, 75-5000, 30-1000, 30/1000, 25/1000 and 30-900 25-1000 ng/mL, respectively. Coefficient of variation for intra-day and inter-day precision for vitamin B1, B2, B3, B5, B6, B7, and B9 at the middle and higher limit of quantitation were less than 15%. Conclusion: The method was successfully developed and validated, and three different brands of multi-vitamin tablets were assayed for water-soluble vitamins.
Background: Carotenoids are natural plant pigments and precursors of vitamin A, which provide a health benefit, protect against chronic degenerative diseases, and contribute to immune functions. Utilization and identification of foods with a high content of carotenoids received greater attention nowadays. Objective: The present study was aimed at evaluating total carotenoids content (TCC) and carotene composition of unconventional leafy vegetables growing in Bangladesh. Materials and Methods: The samples were collected from different locations of Bangladesh and mixed together to ensure sample representativeness. Acetone–petroleum ether extraction followed by spectrophotometric measurement was utilized for quantification of TCC. High-performance liquid chromatography (HPLC- DAD) was used to separate and quantify carotene components. Results and Discussion: The TCC values ranged from 129.38 μg/100 g edible portion (EP) in Helencha to 12803.31 μg/100 g EP in Roktodrone. The study findings also revealed that the selected plant samples exhibited high content of β-carotene ranging from 120.99 μg/100 g EP in Parul to 11301.16 μg/100 g EP in Roktodrone. It was also observed that most of the vegetables analyzed did not have α-carotene, lycopene, β-cryptoxanthin or it was in such a small quantity that could not be detected with HPLC. Conclusion: It can be suggested that regular intake of these vegetables may provide pro-vitamin A. In order to address the vitamin A deficiency and to maintain biodiversity, the analysis of carotenoids composition of these unconventional plant foods is to be initiated and make it available to the mass population. It would also fill up the data gap in the existing food composition table of Bangladesh.
Background:Eugenol is the main constituent of clove essential oil. Past studies have found that clove oil has diverse uses in the pharmaceutical field due to its antioxidant, antibacterial and anesthetic properties.Objective:This work compares the performance of different extraction methods and factors and identifies the effect of the treatments on oil yields and eugenol content.Materials and Methods:Maceration, Hydro distillation, microwave-assisted extraction (MAE), and Soxhlet were performed. The best technique was identified according to yield and content. Further studies were conducted to examine the effects of different factors, such as solvent types (ethanol and methanol) and sample-to-solvent ratio (1:10 and 1:15). HPLC UV-Vis was utilized in the analysis of eugenol concentration.Results and Discussion:Soxhlet extraction provided the highest yield (39.98%) and eugenol content (15.83%), compared to other methods. The results observed from several Soxhlet extraction factors showed that there is no significant difference between the different factors. In the meantime, methanol 1:15 provided the greatest amount of yields (57.83%) and eugenol content (22.21%). In this regard, the higher ratio resulted in higher eugenol content.Conclusion:The results obtained are less comparable because the processing time, the working solvent, and the separation technique were carried out differently for each method. In the meantime, as there is no past study that compared the selected methods and factors, this study’s findings will contribute substantially to fill the gap in this field.
