Quality and Safety of Medicines
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1 Falsified lamivudine/zidovudine/nevirapine tablets: rapid identification using X-ray fluorescence technique The X-ray fluorescence (XRF) technique was used to determine bromine (Br) concentration in lamivudine/zidovudine/nevirapine (Zidolam-N ) tablets from fourteen batches drawn from different sources. Results showed that Br concentration in falsified drugs was significantly higher than that in the authentic drugs. It was also found that Br concentration in the paper liner of the bottle caps containing the falsified drugs was elevated when compared to the liner from the container holding the authentic drug. In conclusion, falsified (Zidolam-N ) can be rapidly identified using the handheld, portable XRF instrument. On 22 September 2011, the World Health Organization (WHO) announced the discovery of falsified Zidolam-N tablets in Kenya. These were labelled as manufactured and supplied by Hetero, India (batch No. E100766). Later, medicines labelled as Hetero batches A9351, A9366 and E were also confirmed to be falsified. Hetero further declared that batches E and E were never supplied to Kenya and that the quantities declared as batches A9351, A9357 and A9366 exceeded those actually manfactured. Falsified medicines labelled as batches A9366 and E were sent to the Kenya National Drug Quality Control Laboratory and to Hetero for examination. The test results were not conclusive in tracking the source of the falsification. Given this situation, WHO requested the China National Institutes for Food and Drug Control (NIFDC) to develop a rapid detection method to differentiate the authentic from the falsified medicines. The following is a description of testing methods and results. Instrument used Thermo Scientific Model Nition XL3t XRF Analyzer. Manufacturer: Thermo NITION Analyzen LLC.Model:XL3t970.Serial: Testing Method Instrument setting Plastic mode, 50 kv excitation voltage, main filter measurement time set to 60s, error of measurement display set to ±1σ standard deviation (68.3% confidence). Tablet testing Six tablets were randomly taken from each sample batch. For each test, two tablets were used and three tests were performed for one batch and tests were repeated the next day. YIN Li-hui 1 LI Jun-qing 2 CHEN Jin-quan 2 WANG Jun 2 ZHANG Xue-bo 1 YANG Mei 1 ZHU Li 1 ZHAO Yu 1 XIAO Xin-yue 1 JIN Shao-hong 1 1 National Institutes for Food and Drug Control, Beijing, China. 2 Dongying Institute for Drug Control, Dongying, Shandong Province, China. 213
2 WHO Drug Information Vol. 27, No. 3, 2013 Sample Information Sample source Batch No. Sample No. Kenya, transfered to WHO A9366 A (Original package with E B bottle) E C Kenya A9366 D (Nude tablets E E without bottle) E F Hetero, India A9366 G (Nude tablets E H without bottle) E I Hetero, India Control sample not for sale A9351 J (Original package with bottle) A9357 K WHO (originated Kenya) A9366 L (Original package E M with bottle) E N Packaging material The bottle bottoms, bottle caps and the paper cap liner were determined. Optimization of testing conditions Testing conditions for bromine in tablets (See tables 1 and 2) Condition a Instrument setting: Plastic mode, thickness correction enabled, main filter set to 60s measurement time, error of measurement set to display ±1σ standard deviation. Condition b Instrument setting: Plastic mode, thickness correction disabled, main filter set to 60s measurement time, error of measurement set to display ±2σ standard deviation (95.5% confidence). Condition c Instrument setting: Plastic mode, thickness correction disabled, main filter set to 90s measurement time, error of measurement set to display ±1σ standard deviation. Results Results of the packaging materials of samples tested are shown in Table
3 Table 1. Results of tablet samples tested Sample Measured concentration of Br (mg/kg) No. Condition a Condition b Condition c A 13±1 15±1 16±1 B 20±1 23±1 25±1 C 34±1 43±2 42±1 D 16±1 19±1 19±1 E 14±1 18±1 18±1 F 8±1 10±1 10±1 G <3 <3 <3 H <3 <3 <3 I <3 <3 <3 J <3 <3 <3 K <3 <3 <3 L 12±1 17±1 15±1 M 71±1 89±2 90±1 N 44±1 54±2 51±1 Table 2. Results of tablet samples tested on different days Sample Br (mg/kg) No. Day 1 Day 2 A 17±1 16±1 B 24±1 24±1 C 43±1 42±1 D 19±1 20±1 E 17±1 16±1 F 9±1 9±1 G <3 <3 H <3 <3 I <3 <3 J <3 <3 K <3 <3 L 17±1 17±1 M 89±1 88±1 N 57±1 57±1 215
4 WHO Drug Information Vol. 27, No. 3, 2013 Table 3. Results of packaging material samples tested Sample No. Bottle bottoms Bottle caps Paper cap liner A <3 <3 56±2 B <3 < C <3 <3 94±2 J <3 <3 <3 K <3 <3 <3 L <3 <3 42±1 M <3 <3 56±2 N <3 <3 87±2 Discussion As seen from the data reported, Br concentrations detected in samples A, B, C, D, E, F, L, M and N are much higher than those in samples G, H, I, J and K which are below the instrument detection limit. Note: When the instrument is set in plastic mode and the total testing time on the main filter is 30s, the detection limit for Br is 3 ppm. Instrument repeatability was good. Relative average deviations of the test results on the two consecutive days are all smaller than 6%. Br was not detected in inner packages of samples D, E, F, G, H and I. Br was not detected in the bottom, cap, and side wall of the plastic bottles containing samples A, B, C, J, K, L, M and N. Br was not detected in the paper liner of the bottle cap of control samples J and K. However, the levels of Br detected in the paper liner of the bottle cap of suspected counterfeit samples A, B, C, L, M and N is relatively high. Conclusion An XRF technique was succssfuly used for rapid and non-destructive measurement of Br concentration in samples of tablets of medicines. A handheld XRF instrument employed in this study is simple to use and offers good repeatability and sensitivity for Br in medicinal drugs. It is suitable for rapid, positive authentication of lamivudine/ zidovudine/nevirapine (Zidolam-N ) tablets using Br measurement as an indicator. 216
5 References 1. Chou J, Clement G, Bursavich B et al. Rapid detection of toxic metals in non-crushed oyster shells by portable X-ray fluorescence spectrometry. Environmental pollution 2010; 158(6): Gupta S, Deep K, Jain L et al. X-ray fluorescence (XRF) set-up with a low power X-ray tube. Appl Radiat Isot 2010; 68(10): Smolek S, Streli C, Zoeger N et al. Improved micro x-ray fluorescence spectro-meter for light element analysis. Rev Sci Instrum 2010;81(5): Anderson DL. Analysis of beverages for Hg, As, Pb, and Cd with a field portable X-ray fluorescence analyzer. J AOAC Int 2010;93(2): Luo Li-qiang Zhan Xiu-chu, Li Guo-hui. X-ray fluorescence spectrometer. Beijing, Chemical Industry Press 2008;5. 6. Tsuyumoto I, Maruyama Y. X-ray fluorescence analysis of hexavalent chromium using Kβ satellite peak observed as counterpart of X-ray absorption nearedge structure pre-edge peak. Anal Chem 2011; 83(19): Aranda PR, Moyano S, Martinez LD, De Vito IE. Determination of trace chromium(vi) in drinking water using X-ray fluorescence spectrometry after solid-phase extraction. Anal Bioanal Chem 2010;398(2): Yin Li-hui,Li Jun-qing,et al.rapid screening of Cr in vacant capsule by X-ray fluorescence elemental analysis technology. Chin J Pharm Anal 2012;32(6): Yin Li-hui,Li Jun-qing,et al.rapid detection on Cr in gelatin by X-ray fluorescence elemental analysis technology. Chin J Pharm Anal 2012;32(7): Li Jun-qing, Yin Li-hui, et al.preliminary discusion on X-ray fluorescence elemental analysis technology the rapid determination of Pb and As elements in cosmetics. Chin J Pharm Anal 2012;32(7):
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