Tracking the Counterfeit and substandard of Ciclosporin Capsules by High Performance Liquid Chromatography.

Badr Aljohani1, Faisal Al Otaibi2, Essam Ghazaly1, David Perrett1, Atholl Johnston1. 1Queen Mary School of Medicine and Dentistry, London, UK, 2Medical Research Center, Medical Services Of Ministry Of Interior, Riyadh, Saudi Arabia

Background: Substandard medicines do not contain the correct amount of active ingredients or meet the standard (innovator\'s) preparation\'s requirements for quality, safety and efficacy. Counterfeit medicines may also be substandard but are purposefully mislabeled intentionally or illegally mislabeled and sold as original brands with wrong ingredients and/or strength. They may have the wrong manufacturer\'s name or the country of origin. Counterfeits may include both branded and generic products. Using such drugs may result in causing harmful effect to the patients. Substandard and counterfeit medicines are a worldwide problem. It affects both developed and developing countries.

Methods: This study was developed, validated, and applied using high performance liquid chromatography (HPLC) to quantify ciclosporin in capsules. Branded and generic ciclosporin products obtained from different countries in Europe, Asia, Africa and South America. Dissolution tests were performed on the capsules using Apparatus 2 (Pharmatest, Germany) under the following conditions: temp: 37.5 ± 0.5°C,: 50 rpm, 500mL deionised water, time of sampling were 5, 10, 15, 20, 30, 45, 60, 90, 120 minutes. Separation carried out using C18 column 5 µm, (4.6 x 250 mm, ACE 5) held at 50 ± 0.3°C. Flow rate 0.7 mL/min using isocratic mobile phase consisted of water and acetonitrile (30 + 70%) and 0.03% trifluroacetic acid over 25 minutes runtime.

Results: Intra-day imprecision for ciclosporin across the standard range was <1% and the inter-day imprecision < 4%. Accuracy of the assay was within ± 13% of the true value at standard curve concentrations range from 25 to 500 mg/L of ciclosporin. All brands obtained from Saudi Arabia (Sa), Turkey (TK), Pakistan (Pak), Jordan (Jor), Egypt (Egy) and two generics Colombia (Col) and Iran (Ir) showed more than 80% of ciclosporin after 90 minutes (99, 100, 94, 84, 92, 85 and 97%) respectively. One generic from Morocco (M), showed less than the minimum percentage of labelled amount 54%. One or more impurities were detected in all capsules. These impurities are currently under investigation in order to identify their nature and their effect on ciclosporin pharmacokinetics. Relative to the brand (TK), statistical analysis showed significant differences (p<0.0001) of the mean percentage content between brand and generic, the confidence interval 95% range for the brands (Egy, Jor, Pak, Sa) were (79-108), (71-98), (81-110), (85-117), respectively, and (73-100), (83-114), (45-62) for the generics (Col), (Ir) and (M) respectively.

Conclusion: Based on these results we conclude that most of ciclosporin preparations contained ciclosporin amount which meet the USP requirements. However, one preparation was substandard and contained unidentified impurities.