Earth-friendly-assessed chromatographic platforms for rapid analysis of sulfacetamide sodium and prednisolone acetate in presence of sulfanilamide impurity: Application on ophthalmic formulation and aqueous humor

Sulfacetamide sodium as an antibacterial agent is co-formulated with prednisolone acetate in Phenamide-P® ophthalmic suspension to cure the ophthalmic infections correlated with inflammatory disorders. For the first time, two innovative, sensitive, specific, and robust chromatographic platforms were developed and their conditions were optimized for rapid analysis of the aforementioned drugs either in the absence or presence of sulfanilamide; a major impurity and degradation product of sulfacetamide sodium. The first platform is HPLC/UV in which separation of the three analytes was performed on a kinetex C18 column (4.6 × 100 mm, 2.6-μm) with isocratic elution using methanol: deionized water pH 3 (adjusted with orthophosphoric acid) in a ratio of (40:60 by volume) as a green mobile phase. The flow rate of the mobile phase was 1 mL/min and the UV detector was set at 262 nm. The second platform is HPTLC/densitometry in which the three analytes were separated on silica gel 60-F₂₅₄ TLC plates using a developing system of chloroform: acetone: ammonia (6:2:0.1 by volume) and their developed bands were detected at 262 nm. The detection and quantitation limits were found in ranges of (0.01–0.03 μg/mL) and (0.03–0.09 μg/mL) for the HPLC platform while were found in ranges of (0.007–0.01 μg/band) and (0.02–0.04 μg/band) for the HPTLC platform, respectively. The suggested platforms were applied for the determination of sulfacetamide sodium and prednisolone acetate in spiked rabbit aqueous humor without interference by the matrix of aqueous humor. Greenness assessment was performed using eco-scale and GAPI tools revealing the excellent greenness of suggested platforms. According to ICH recommendations, the suggested platforms were validated producing satisfactory results within the accepted limits. The suggested platforms can be successfully applied for routine analysis of the aforementioned drugs not only in QC laboratories but also in the bioavailability centers due to the short analysis time of these platforms.     

High Performance Thin Layer Chromatographic Method for Simultaneous Estimation of Ibuprofen and Pseudoephedrine Hydrochloride

High performance thin layer chromatographic method is developed for simultaneous estimation of ibuprofen and pseudoephedrine hydrochloride in tablets. Silica gel 60F₂₅₄ plates were used as stationary phase and t.butanol: ethyl acetate: glacial acetic acid: water (7:4:2:2 v/v) as mobile phase. Wavelength selected for analysis was 254 nm. Percent estimation of ibuprofen and pseudoephedrine hydrochloride was found to be 99.56% and 98.77%, respectively. Percent recovery for both the drugs was found in the range of 98.27% to 100.91%, respectively.     

Inverse expression of Pk and Luke blood group antigens on human RBCs

BACKGROUND: Luke (LKE) is a high-frequency RBC antigen, related to the P blood group system. A LKE-negative phenotype is found in 1 to 2 percent of donors and may be associated with increased P(k). Because P(k) and similar glycolipids are receptors for shiga toxin on cell membranes, a LKE-negative phenotype could have implications for infections by Shigella dysenteriae and enterohemorrhagic Escherichia coli. STUDY DESIGN AND METHODS: Volunteer donors (n = 257) were serologically typed for LKE with a LKE MoAb, MC813-70. LKE-strong-positive, LKE-weak-positive and LKE-negative RBCs were analyzed for P(k), P, LKE, and shiga toxin binding by immunofluorescence flow cytometry, high-performance thin-layer chromatography, scanning densitometry, and high-performance thin-layer chromatography immunostaining. RESULTS: Among Iowa donors, 78.6 percent were LKE-strong-positive, 20.2 percent were LKE-weak-positive, and 1.2 percent were LKE-negative. There was an inverse expression of P(k) and LKE on RBCs. P(k) expression was increased on LKE-negative RBCs and was associated with increased shiga toxin binding. A LKE-active glycolipid was identified in the ganglioside fraction of LKE-strong-positive RBCs. CONCLUSION: A LKE-negative phenotype is associated with increased expression of P(k) on RBCs. Differences in P(k) and LKE expression may play a role in host susceptibility to infection with S. dysenteriae and E. coli.     

Separation and determination of closely related triterpenic acids by high performance thin-layer chromatography after iodine derivatization

A method based on high performance thin-layer chromatography combined with densitometry for the simultaneous determination of oleanolic and ursolic acids is described. Because of the similarity of chemical structure the prechromatographic derivatization was necessary to separate these triterpenic acids. The samples were treated by 1% iodine solution in chloroform directly on the chromatographic plate and developed with the mobile phase consisting of A (petroleum ether), B (ethyl acetate) and C (acetone) (8.2:1.8:0.1, v/v/v). After drying, the plates were sprayed with 10% (v/v) ethanol solution of sulfuric acid(VI) and heated to 120 degrees C for 3 min. Quantification was performed in absorbance/transmittance mode at a wavelength of 530 nm by using a computer-controlled densitometer Desaga CD 60. The presented method was validated for linearity, precision and accuracy. Correlation coefficient (r(2)>0.99), R.S.D. values (1.4-3.5%), detection limits as well as recovery values (98.4-103.1%) were found to be satisfactory. The method has been successfully applied in the analysis of both triterpenic acids in plant extract.     

Determination of Artemisinin in Bulk and Pharmaceutical Dosage Forms using HPTLC

A new, simple, rapid, accurate and precise HPTLC method was developed. The detector response was linear for concentrations between 100-600 ng/spot (r =0.9931). The limits of detection and quantitation were 25 ng/spot and 75 ng/spot, respectively. The recovery study was carried out by standard addition method and was found to be 99.60±0.27. Statistical analysis proved that the method was precise, accurate and reproducible, and hence was suitable for the routine analysis of artemisinin.     

HPTLC Method for the Simultaneous Estimation of Emtricitabine and Tenofovir in Tablet Dosage Form

A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the estimation of emtricitabine and tenofovir simultaneously in combined dosage form. The stationary phase used was precoated silica gel 60F 254. The mobile phase used was a mixture of chloroform: methanol (9:1 v/v). The detection of spots was carried out at 265 nm. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 200 to 1000 ng with regression coefficient of 0.9995. The proposed method can be successfully used to determine the drug content of marketed tablet formulation.     

Identification,quantification of bioactive constituents,evaluation of antioxidant and in vivo acute toxicity property from the methanol extract of Vernonia cinerea leaf extract

Context: Vernonia cinerea (L.) Less [Compositae (Asteraceae)] is used traditionally for several medical purposes such as inflammation, pain, fever, and cancer.

Objectives: The present study identified the bioactive constituents in the methanol extract of Vernonia cinerea leaf and evaluated its antioxidant activity and acute toxicity.

Methods: The identification of phytochemicals was accomplished by GC-MS and the major antioxidant phenolic compounds in the extract were quantified by HPTLC analysis. To quantify the essential elements, atomic absorption spectrophotometeric analysis was carried out. Total phenol and flavonoid content was measured by Folin-Ciocalteau reagent and 2% aluminium chloride, respectively.

Results: GC-MS analysis identified the presence of 27 phytoconstituents. The predominant phenolic compound in the extract as quantified by HPTLC was gallic acid (1.92?mg/g) followed by rutin (0.705?mg/g), quercetin (0.173?mg/g), caffeic acid (0.082?mg/g) and ferulic acid (0.033?mg/g). The following elements were quantified: Fe (0.050?ppm), Mn (0.022?ppm), Co (0.0180?ppm), Pb (0.029?ppm), Hg (3.885?ppm) and Se (4.5240?ppm). The antioxidant activity of the extract increased with increasing concentration and the correlation (r²) for all in vitro assays were satisfactory.

Conclusions: V. cinerea extract has significant (p?<?0.05) antiradical activity. Hence, V. cinerea may have potential medicinal value and can be used in the formulation of pharmacological products for degenerative diseases.     

HPTLC method to study skin permeation of acylovir

A new simple, rapid and selective high performance thin layer chromatography (HPTLC) method is developed for the quantitation of acyclovir during in vitro skin permeation studies. Separation of guinea pig skin proteins and acyclovir was achieved by employing a mobile phase consisting of chloroform–methanol–ammonia (15:9:4, v/v/v) on precoated silica gel 60F254 aluminum plates. Densitometnic analysis was carried out at 255 nm. The limit of detection and quantification were 30 and 50 ng, respectively. The calibration curve was linear in the range of 10–20 μg/ml (r=0.9965). The relative standard deviation for a sample of concentration 100 μg/ml were 1.15 and 2.85 for system and method precision, respectively. Intraday and interday variation studies gave an average 0.763 and 0.463% relative standard deviation for the three levels tested. Average recoveries of 101.8 and 100.1% were recorded for two marketed preparations studied. The method was employed to optirmize topical liposomal gel formulation of acyclovir on basis of maximum skin permeation.     

灵芝和紫芝对照提取物在灵芝样品指纹图谱分析的应用

目的使用赤芝对照提取物(CZERS)、紫芝对照提取物(ZZERS)对所收集18批次灵芝药材进行高效薄层色谱(HPTLC)以及高效液相色谱(HPLC)指纹图谱分析。方法使用HPTLC指纹图谱方法,高效娃胶预制板,以氯仿-乙腈-甲醇-甲酸=13:2:0.5:0.53次展开和环己烷-乙酸乙酯-甲醇-甲酸=15:5:0.5:0.52次展开的方法分别分析灵芝三萜酸类和甾醇类成分。使用HPLC指纹图谱方法,Kromasil 100-5 C18柱(4.6mm×250 mm,5μm);流动相:A-乙腈,B-0.02%嶙酸;梯度洗脱程序:0~40min,29%→33%A,40-70 min,33%→65%A,70-105 min,65%→100%A;105-120 min,100%A;检测波长244 nm(DAD检测器);流速1.OmL·min^-1;进样量10μL;柱温25℃。结果使用对照提取物(ERS)和指纹图谱分析方法,可以区分赤芝,无柄灵芝和紫芝。不同种类和生长方式的灵芝成分存在差异。结论灵芝品种繁多,分野生和人工培养,而人工培养的培养基又有不同,导致不同灵芝个体间成分存在差异。使用特定品种的ERS和指纹图谱分析方法更加适合灵芝多成分整体质量控制的需求。     

福建灵芝高效薄层色谱鉴别及其在溯源中的应用

目的 本研究旨在建立福建灵芝高效薄层色谱方法,并对比福建产区与其他产区薄层的异同点,为灵芝药材在薄层溯源中的应用提供实验依据。方法 采用Merk HPTLC Silica gel 60预制板,氯仿-乙腈-甲醇-甲酸(13∶1.5∶0.2∶0.2)为展开剂,3%硫酸乙酸乙酯溶液显色,在紫外366 nm下检视,采用对照品比对鉴别薄层色谱条带。使用斑点条带数字化表征,对不同产区样品进行聚类分析。结果 高效薄层色谱共分离灵芝药材中17个条带,用对照品比对鉴定了其中11个成分,包括灵芝烯酸C(Rf₅=0.31)、灵芝酸C2(Rf₆=0.33)、灵芝酸I(Rf₇=0.35)、灵芝酸G(Rf₈=0.41)、灵芝酸A(Rf₉=0.44)、灵芝酸B(Rf₁₀=0.46)、灵芝内酯B(Rf₁₁=0.53)、3β,7β,15β-三羟基-11,15-二羰基-羊毛甾烷-8-烯-24→20内酯(Rf₁₂=0.56)、灵芝烯酸D(Rf