No abstract

Background Amigrain®, the market formulation of the ternary mixture of analgin, caffeine, and ergotamine is used for the symptomatic treatment of migraine. Objectives The aim of the work is to develop and validate an HPLC- DAD method and a novel spectrophotometric method for simultaneous determination of analgin, caffeine and ergotamine in their pharmaceutical formulation. Method The HPLC separation of the ternary mixture was carried out using an Inertsil -C8 column and a gradient elution of mobile phase composed of “acetonitrile and ammonium format buffer (pH 4.2)”, the UV detection for analgin and caffeine was carried out at λ  = 280, 254 nm and fluorometric detection for ergotamine was carried out at λ exc =310 nm, λ emm =360 nm .The two spectrophotometric methods were double divisor ratio spectra derivative (DDRD) and ratio dual wavelength (RDW) methods. The first method was used for determination of ergotamine at 355 nm and caffeine at 268 nm by the third and first derivative. The second one was based on using amplitude difference for the determination of caffeine and analgin. Results HPLC and spectrophotometric methods were applied over the concentration ranges of 50–400, 25–200, and 0.5–10 μg/mL and 10 –35, 2 –30, and 10 –70 μg/mL for analgin, caffeine and ergotamine for the two methods. Conclusion The proposed methods were successfully applied for the determination of the cited drugs in their pharmaceutical formulation and the obtained results were statistically compared with those of the reported methods without any significant difference. Highlights The developed HPLC-DAD method has a high sensitivity for ergotamine. The spectrophotometric methods offer novelty, green solvent usage, and economic cost.

Two accurate, sensitive, precise and selective HPLC and stability-indicating TLC methods were developed for the simultaneous determination of camylofin-2HCl and paracetamol. Forced acid, alkali and oxidative degradation of camylofin-2HCl were tried where complete degradation was achieved using 5 N HCl. HPLC method was developed to determine the mixture of the two drugs using Zorbax NH 2 column and a mobile phase of 0.5% triethylamine and pH 3.0 adjusted with 0.1% phosphoric acid and methanol (70:30 v/v) over concentration ranges of 3-90 and 10-95 µg/mL for camylofin-2HCl and paracetamol, respectively.TLC method was used for the separation of camylofin from its acid degradate and paracetamol using chloroform-methanol-acetone-conc. ammonia (8:2:2:0.1, by volume) as developing system and band scanning at 254 nm over concentration ranges of 5-40 µg/band for camylofin-2HCl and 0.1-0.5 µg/band for paracetamol. The validation of two methods was carried out according to ICH guideline. Accuracy ranged between 98.47 and 100.67% for the two methods with acceptable precision RSD% ranging between 0.66 and 1.47%.