Abstract
Pharmaceutical dosage forms containing gelatin are recently of high attention in regard to its porcine content. In Muslim countries, halal products assurance is strictly regulated, not only for food but also for pharmaceutical products. This study aimed to develop gelatin identification and verification method in capsule shells by performing simple simultaneous protein- and nucleic acid based-identification methods. Identification and verification performing polyacrylamide gel electrophoresis (PAGE)—coupled with principle component analysis, and polymerase-chain-reaction (PCR)-restriction fragment length polymorphism (RFLP) were carried out. Gelatin was obtained by extracting the capsules using cold acetone; the precipitate was used directly for PAGE, and was used as well for DNA extraction. The results showed that the porcine gelatin reference showed 12 major bands of peptide profile, with predominant bands at ±200 and ±100–135 kDa on 8 % Tris-glycine gel, while bovine gelatin only showed four major bands. Further, PCR-RFLP result showed two specific bands of the porcine gelatin DNA at 228 and 131 bp, whereas the bovine gelatin DNA bands occurred at 316 and 44 bp. The results demonstrated that the capsule shell samples used in this study contained the following: sample 1 was verified contain porcine gelatin, whereas the three other samples identified contain bovine gelatin. Although the simultaneous approaches seemed need to be improved, it is crucial to maintain their low cost and simple protocol.
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Abbreviations
- BGR:
-
Bovine gelatin reference
- PCA:
-
Principle component analysis
- PCR:
-
Polymerase chain reaction
- PGR:
-
Porcine gelatin reference
- RFLP:
-
Restriction fragment length polymorphism
- SDS:
-
Sodium dodecyl sulfate
- PAGE:
-
Poly acrylamide gel electrophoresis
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Acknowledgments
All authors (A. Malik, M. L. Sutantyo, I. Hapsari, A. V. Sinurat, E. M. Purwati, M. Jufri, H. Suryadi) declare that they have no conflict of interest. Funding was supported by Research Grant PUPT 2016 from Directorate for Higher Education, Republic of Indonesia to A.Malik Authors would like to thank H. J. Freisleben for English editing of this manuscript.
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Malik, A., Sutantyo, M.L., Hapsari, I. et al. Simultaneous identification and verification of gelatin type in capsule shells by electrophoresis and polymerase chain reaction. Journal of Pharmaceutical Investigation 46, 475–485 (2016). https://doi.org/10.1007/s40005-016-0245-0
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DOI: https://doi.org/10.1007/s40005-016-0245-0