Hplc ASSAY

Puerarin is the major bioactive ingredient in the roots of Kudzu (Radix Puerariae thomsonii), a traditional herbal that has good effects in the treatment of cardiovascular disease, antioxidants and alcohol abuse. This study was performed to develop and validate the method of quantifying puerarin in Kudzu roots; The method was applied to assay the content of puerarin in some pharmaceutical samples on the market. Chromatographic conditions were determined including Ascentis® RP C18 column (250 × 4.6mm; 5µm), detection wavelength of 250nm; 1ml/min flow rate, mobile phase methanol:water (25:75). The method was validated for the specificity, linearity ranging from 0.5 to 50µg/ml (r = 0.9998), accuracy (recovery: 96.7 to 103.65%, RSD <2%), repeatability good return (RSD <2%). The content of puerarin in roots of kudzu ranges from 0.014% to 0.762% on the dry weight.

Objective: A simple, sensitive, precise and accurate stability-indicating HPLC method has been developed and validated for determination of Azathioprine in bulk drug and in pharmaceutical dosage form in the presence of degradation products. Methods: An isocratic, reversed phase HPLC method was developed to separate the drug from the degradation products, using a Hypersil C

Objective: Development and validation of Near infrared (NIR) spectroscopic method for determination of paracetamol and its major degradation product 4-aminophenol in paracetamol tablets and show the agreement between the NIR as a greener technique and the conventional high performance liquid chromatography (HPLC) method, official in British pharmacopeia (BP). Methods: Calibration model for paracetamol and its degradation product 4-aminophenol was built by utilizing chemometric processing which is the most critical step in the development of specific and robust NIR models. It is based mainly on a partial least square regression fit on the transmission mode using paracetamol, 4-aminophenol and excipient materials of the drug products. The results obtained by NIR spectroscopy were compared with the compendial HPLC method in the BP. Results: The chosen models had a root mean square error of the cross validation (RMSECV) values of 1.38, 1.42 and coefficient of correlation (r2) of 99.1, 99.05 for paracetamol and 4-aminophenol respectively, which indicates good fitness and accuracy of the model. Conclusion: The present study showed that NIR could be used with high accuracy for determination of for parent drug and its major degradation product in paracetamol tablets. This proposed technique realizes many of green analytical aspects in developing eco-friendly analytical methods and may replace safely the conventional chromatographic technique without compromising efficacy.

Objective : Development and validation of Near infrared (NIR) spectroscopic method for determination of paracetamol and its major degradation product 4-aminophenol in paracetamol tablets and show the agreement between the NIR as a greener technique and the conventional high performance liquid chromatography (HPLC) method, official in British pharmacopeia (BP). Methods: Calibration model for paracetamol and its degradation product 4-aminophenol was built by utilizing chemometric processing which is the most critical step in the development of specific and robust NIR models. It is based mainly on a partial least square regression fit on the transmission mode using paracetamol, 4-aminophenol and excipient materials of the drug products. The results obtained by NIR spectroscopy were compared with the compendial HPLC method in the BP. Results: The chosen models had a root mean square error of the cross validation (RMSECV) values of 1.38, 1.42 and coefficient of correlation ( r 2) of 99.1...

The existing HPLC methods for the assaying of multiple markers in standardized herbal extracts use a single wavelength for quantification, which usually leads to lower assay than actual claims. It was because screening multiple phytochemical standards irrespective to their absorption maxima which drive decrease in peak are response which ultimately impact the assay. To overcome this issue, we have applied a methodology, called Mixed Standard Multi-Signal (MSMS) simultaneous detection, where multiple reference standards get extracted each with their respective λmax and quantification will be done at the same λmax in a single sequence data setup. This method was standardized using Single Signal Multi-Standards, Single Signal Single Standard, Multi-Signal Single Standard, for all individual reference standards, mixed reference standards, plant extracts, and Ayurvedic formulations. The results of our study demonstrate that higher assay reported total actives 5.04% by conventional detect...

The existing HPLC methods for the assaying of multiple markers in standardized herbal extracts use a single wavelength for quantification, which usually leads to lower assay than actual claims. It was because screening multiple phytochemical standards irrespective to their absorption maxima which drive decrease in peak are response which ultimately impact the assay. To overcome this issue, we have applied a methodology, called Mixed Standard Multi-Signal (MSMS) simultaneous detection, where multiple reference standards get extracted each with their respective λmax and quantification will be done at the same λmax in a single sequence data setup. This method was standardized using Single Signal Multi-Standards, Single Signal Single Standard, Multi-Signal Single Standard, for all individual reference standards, mixed reference standards, plant extracts, and Ayurvedic formulations. The results of our study demonstrate that higher assay reported total actives 5.04% by conventional detect...

The existing HPLC methods for the assaying of multiple markers in standardized herbal extracts use a single wavelength for quantification, which usually leads to lower assay than actual claims. It was because screening multiple phytochemical standards irrespective to their absorption maxima which drive decrease in peak are response which ultimately impact the assay. To overcome this issue, we have applied a methodology, called Mixed Standard Multi-Signal (MSMS) simultaneous detection, where multiple reference standards get extracted each with their respective λ max and quantification will be done at the same λ max in a single sequence data setup. This method was standardized using Single Signal Multi-Standards, Single Signal Single Standard, Multi-Signal Single Standard, for all individual reference standards, mixed reference standards, plant extracts, and Ayurvedic formulations. The results of our study demonstrate that higher assay reported total actives 5.04% by conventional detection and for current proposed method 13.81% with shorter span of analysis time and lesser solvent consumption. It helps to decide the actual standardization label claim on product which drives better commercial costings, and dosage regimen claims.

An UV-spectrophotometric method for simultaneous determination of amlodipine and captopril has been developed and validated. Simultaneous equation method is applied for the quantification of both drugs, which is based upon the measurement of absorbance at two wavelengths, l max = 236 and 213 nm for amlodipine and captopril, respectively, in 0.1 N HCl solutions. The calibration curves for amlodipine and captopril are linear in concentration ranges of 0.195-100 and 0.39-100 mg/mL, respectively. Accuracy of the method is 98 and 99% for amlodipine and captopril, respectively. The inter-day precision for amlodipine and captopril in terms of RSD is 10 and 8%, while the intra-day RSD is 6 and 9%, respectively. Sensitivity of the method in term of the limit of detection for amlodipine and captopril is 0.195 and 0.39 mg, respectively. The method gives 98% reproducible results.

Two new simple, selective economical and reproducible UV spectrophotometeric method simultaneous estimation of two component drug mixture of Amlodipine besylate (AB)) and hydrochlorothiazide (HCT) in pharmaceutical preparation, blood sample and urine has been developed. The first method depends on first derivative method at 245 and 238 nm for AB and HCT also in plasma and urine respectively. The linearity over the concentration ranges 2.5-17.5µg/ml for both the drugs AB and HCT. The second method is based on two wavelength method, which uses the difference of absorbance value at 271 nm and 395 nm for estimation of Hydrochlorothiazide (HCT) and absorbance at 363 nm for amlodipine besylate (AB). In formulation, plasma and urine Methanol: water (50:50v/v) was used as solvent, in which Amlodipine besylate and Hydrochlorothiazide shows linearity in the range of 2.5-17.5 µg/ml for both the drugs respectively. Standard deviation was &lt;1.5 in the assay of tablets. This method validated an...

A simple, sensitive and accurate spectrophotometric method has been developed for the determination of Acebrophylline in bulk drug and capsule. The λ max of the Acebrophyllline was found to be 274 nm. The method showed high sensitivity with linearity in the range of 2 to 20μg/ml, coefficient regression was found to be 0.9994. The regression of the curve was found to be Y = 0.0187X-0.0062. Percent relative standard deviation value is below 2.0 for intraday and interday precision indicated that method is highly precised. The percentage recovery value (average 100.1458 percentages), indicated the accuracy and specificity of the method. The proposed method will be suitable for the analysis of Acebrophylline in bulk and pharmaceutical formulation.

The bioassay, first order derivative UV spectrophotometry and chromatographic methods for assaying fluconazole capsules were compared. They have shown great advantages over the earlier published methods. Using the first order derivative, the UV spectrophotometry method does not suffer interference of excipients. Validation parameters such as linearity, precision, accuracy, limit of detection and limit of quantitation were determined. All methods were linear and reliable within acceptable limits for antibiotic pharmaceutical preparations being accurate, precise and reproducible. The application of each method as a routine analysis should be investigated considering cost, simplicity, equipment, solvents, speed, and application to large or small workloads.

The aim of the present research was broadly focused on the estimation of carbamazepine in bulk and pharmaceutical dosage form by using two UV-Spectrophotometric methods namely, Zero order UV spectrophotometry (Method-1) and Area under the curve UV spectrophotometry (Method-2). The Zero order UV Spectrophotometric method was based on the measurement drug absorbance at wave length of 284 nm, which was its wavelength of maximum absorbance .The Area under the curve method, was based on the calculation of area occupied by the UV absorbance curve between 278-290 nm. The solvent employed for both methods was 50% v/v ethanol. In the estimation of Carbamazepine, both the methods showed linearity in the range of 2-12µg/ml. The correlation coefficient was ≥ 0.999. The precision for both the methods was ≤2% RSD. The accuracy was performed by using percentage recovery studies of standard drug spiked at 50,100 and 150% of the test concentration and the values obtained were within the limits. The developed methods were applied for the assay of the drug in its respective dosage forms. The assay of pharmaceuticals dosage form was found to be within limits. All the results were satisfactory, the developed methods can be routinely used for the analysis of the drugs in both bulk and dosage forms.

Adhatoda vasica (L.) Nees (Acanthaceae) is a well known plant drug in Ayurvedic medicine, used for the treatment of different types of diseases and disorders. The tissue cultures of this plant species were established on Murashige and Skoog culture medium by using various concentrations of growth hormones (0.5-5.0 mg/L α-napththaleneacetic acid + 0.5-3.5 mg/L 6-benzylaminopurine). The concentration of vasicine acetate was found higher (0.51  0.519%) in six week old callus than in vivo (0.47  0.556%) plant material (roots). The antioxidant activity was evaluated by using DPPH-radical scavenging, reducing power and superoxide anion scavenging models. We found that 2-acetyl benzyl amine showed maximum antioxidant activity against various antioxidant models, including DPPH-radical scavenging (40.59 ± 0.774%), reducing power (461.55 ± 0.402%) and superoxide anion scavenging (52.54 ± 0.553%) than other isolated alkaloids at 10-30 μM concentrations. Our observed results suggest that the pyrroloquinazoline alkaloids of A. vasica demonstrated strong antioxidant activity, which could be used as potent drugs against different types of oxidative stresses.

Simple and rapid assay method is primary prerequisite in quality assurance department for any pharmaceutical dosage form. When two drugs are combined in a single dosage form, it becomes difficult to estimate them simultaneously with required accuracy and precision. Here we have proposed a simple and rapid UV spectrophotometric assay method for simultaneous determination of amlodipine besylate and valsartan in both bulk drug and commercial dosage forms. Method employed was Zero absorbance method where amlodipine and valsartan were determined at 363 nm and 290 nm respectively. At 363 nm valsartan don't have absorption and at 290 nm amlodipine has zero absorption. Linearity was established at 8 µg/mL -80 µg/mL for amlodipine (y= 0.012X + 0.002 R 2 = 0.999) and 4 µg/mL -300 µg/ml for Vasartan (y= 0.002X + 0.017 R 2 = 0.999). Method was found to be accurate and precise with %RSD<2 for all the validation parameters employed. Method was found to be very sensitive with LOD 2.4 µg/mL and 3.6 µg/mL whereas LOQ was 7.5 µg/mL and 11 µg/mL for Amlodipine besylate and Valsartan respectively. As method uses methanol as a solvent to dissolve, it is cost effective for routine analysis in quality assurance department for determination of these drugs individually or simultaneously.

The present study describes development and validation of simple, rapid, sensitive and stability indicating highperformance liquid chromatographic assay method for simultaneous determination of Amlodipine besylate and Valsartan in bulk as well as in commercial formulation. Analytical separation was achieved with RP Waters Symmetry C18 Column [150×4.6 mm] using a combination of methanol and potassium dihydrogen phosphate buffer [0.01 M] pH 2.5, in ratio 60:40 v/v with flow rate of 1 mL min −1 . The retention times for Amlodipine and Valsartan were found to be 4.6 and 7.6 min respectively. Both the drugs were exposed to thermal, photolytic, hydrolytic, and oxidative stress conditions individually as well as in combination, subsequently samples were analyzed by the proposed method. Detection was done by PDA detector at 238 nm. Method was found to be very sensitive as LOD found to be 20 ng/mL and 44 ng/mL for Amlodipine besylate and Valsartan respectively. The method was found to be specific and stability indicating as no interfering peaks of degradation compounds and excipients were noticed. The developed method was validated with respect to linearity, accuracy, precision, specificity, robustness as per International Conference on Harmonization (ICH) guidelines. The proposed method hence useful for the application in quality control laboratories for quantitative analysis of both the drug individually or in combination, as it is simple and rapid method with excellent accuracy and precision.

Amlodipine, a dihydropyridine calcium channel blocker, and valsartan, an angiotensin II receptor blocker, are co-formulated in a single-dose combination for the treatment of hypertension. The combination is used by patients whose blood pressure is not adequately controlled on either component monotherapy. This work describes a simple, sensitive, and reliable spectrofluorimetric method for the simultaneous determination of the two antihypertensive drugs; amlodipine besylate (AML) and valsartan (VAL) in their combined tablets. The method involved measurement of the native fluorescence at 455 nm (λ Ex 360 nm) and 378 nm (λ Ex 245 nm) for AML and VAL, respectively. Analytical performance of the proposed spectrofluorimetric procedure was statistically validated with respect to linearity, ranges, precision, accuracy, selectivity, robustness, detection, and quantification limits. Regression analysis showed good correlation between fluorescence intensity and concentration over the concentration ranges 0.2-3.6 and 0.008-0.080 µg mL −1 for AML and VAL, respectively. The limits of detection were 0.025 and 0.0012 µg mL −1 for AML and VAL, respectively. The proposed method was successfully applied for the assay of the two drugs in their combined pharmaceutical tablets with recoveries not less than 98.85%. No interference was observed from common pharmaceutical additives. The results were favourably compared with those obtained by a reference spectrophotometric method.