Abstract
Eprinomectin (EPM) is the active pharmaceutical ingredient (API) of the pour-on finished product. The finished product also contains propylene glycol octanoate decanoate (PGOD) as an excipient and solvent, and butylated hydroxytoluene (BHT) as an antioxidant. EPM belongs to a group of compounds known as avermectins and is commercially available as a mixture of two closely related homologues, namely, EPM B1a and EPM B1b. A stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method is developed and validated, which can provide data for EPM assay and estimation of its degradation products as well as the assay of BHT from the chromatogram of a single injection of the finished product sample. The new HPLC method uses a Halo-C18 column (100 mm × 4.6 mm i.d., 2.7 µm particle size) maintained at 40 °C with 100% Water (H2O) as Mobile Phase-A and Isopropanol(IPA)/Acetonitrile(ACN) (70/30, v/v) as Mobile Phase-B. Analytes are separated by a gradient elution with a total run time of 35 min. EPM and its degradation products except 8a-oxo-B1a are detected by UV at 245 nm while 8a-oxo-B1a and BHT are detected at 280 nm.
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References
Shoop WL, Egerton JR, Eary CH et al (1996) Eprinomectin: a novel avermectin for use as a topical endectocide for cattle. Int J Parasitol 26:1237–1242. https://doi.org/10.1016/s0020-7519(96)00123-3
Holste JE, Smith LL, Hair JA et al (1997) Eprinomectin: a novel avermectin for control of lice in all classes of cattle. Vet Parasitol 73:153–161. https://doi.org/10.1016/s0304-4017(97)00063-0
Serafini S, Soares JG, Perosa CF et al (2019) Eprinomectin antiparasitic affects survival, reproduction and behavior of Folsomia candida biomarker, and its toxicity depends on the type of soil. Environ Toxicol Phar 72:103262. https://doi.org/10.1016/j.etap.2019.103262
Yoon YJ, Kim E-S, Hwang Y-S, Choi C-Y (2004) Avermectin: biochemical and molecular basis of its biosynthesis and regulation. Appl Microbiol Biot 63:626–634. https://doi.org/10.1007/s00253-003-1491-4
Batiha GE-S, Alqahtani A, Ilesanmi OB et al (2020) Avermectin derivatives, pharmacokinetics, therapeutic and toxic dosages, mechanism of action, and their biological effects. Pharm 13:196. https://doi.org/10.3390/ph13080196
Academy MSA of VM and BISMV, Dzhafarov MK, Vasilevich FI, et al (2016) Derivatives of 16-membered macrocyclic lactoneS: antiparasitic properties and interaction with gabaa receptors. Sel’skokhozyaistvennaya Biologiya 51:875–882. https://doi.org/10.15389/agrobiology.2016.6.875eng
Tang M, Hu X, Wang Y et al (2020) Ivermectin, a potential anticancer drug derived from an antiparasitic drug. Pharmacol Res 163:105207. https://doi.org/10.1016/j.phrs.2020.105207
Elhawary NM, Sorour ShSGH, El-Abasy MA et al (2017) A trial of doramectin injection and ivermectin spot-on for treatment of rabbits artificially infested with the ear mite “Psoroptes cuniculi.” Pol J Vet Sci 20:521–525. https://doi.org/10.1515/pjvs-2017-0063
Nieman CC, Floate KD, Düring R-A et al (2018) Eprinomectin from a sustained release formulation adversely affected dung breeding insects. PLoS One 13:e0201074. https://doi.org/10.1371/journal.pone.0201074
Andresen CE, Loy DD, Brick TA et al (2018) Effects of extended-release eprinomectin on productivity measures in cow–calf systems and subsequent feedlot performance and carcass characteristics of calves. Transl Animal Sci 3:txy115. https://doi.org/10.1093/tas/txy115
Lamassiaude N, Courtot E, Corset A, et al (2020) Functional investigation of conserved glutamate receptor subunits reveals a new mode of action of macrocyclic lactones in nematodes. Biorxiv 2020.12.17.423223. https://doi.org/10.1101/2020.12.17.423223
Junco M, Iglesias LE, Sagués MF et al (2021) Effect of macrocyclic lactones on nontarget coprophilic organisms: a review. Parasitol Res 120:773–783. https://doi.org/10.1007/s00436-021-07064-4
Wolstenholme AJ, Rogers AT (2005) Glutamate-gated chloride channels and the mode of action of the avermectin/milbemycin anthelmintics. Parasitology 131:S85–S95. https://doi.org/10.1017/s0031182005008218
Yates DM, Wolstenholme AJ (2004) An ivermectin-sensitive glutamate-gated chloride channel subunit from Dirofilaria immitis. Int J Parasitol 34:1075–1081. https://doi.org/10.1016/j.ijpara.2004.04.010
Chen I, Kubo Y (2018) Ivermectin and its target molecules: shared and unique modulation mechanisms of ion channels and receptors by ivermectin. J Physiology 596:1833–1845. https://doi.org/10.1113/jp275236
Hamel D, Bosco A, Rinaldi L et al (2017) Eprinomectin pour-on (EPRINEX® Pour-on, Merial): efficacy against gastrointestinal and pulmonary nematodes and pharmacokinetics in sheep. Bmc Vet Res 13:148. https://doi.org/10.1186/s12917-017-1075-7
Hamel D, Visser M, Mayr S et al (2018) Eprinomectin pour-on: prevention of gastrointestinal and pulmonary nematode infections in sheep. Vet Parasitol 264:42–46. https://doi.org/10.1016/j.vetpar.2018.11.002
USP43–NF38-1660 (2020) United States Pharmacopeial Convention Inc., Rockville, MD, USA
Awasthi A, Razzak M, Al-Kassas R et al (2012) Separation and identification of degradation products in eprinomectin formulation using LC, LTQ FT-MS, H/D exchange, and NMR. J Pharmaceut Biomed 63:62–73. https://doi.org/10.1016/j.jpba.2011.12.030
Wang Y, Sun J, Zhang T et al (2011) Enhanced oral bioavailability of tacrolimus in rats by self-microemulsifying drug delivery systems. Drug Dev Ind Pharm 37:1225–1230. https://doi.org/10.3109/03639045.2011.565774
Salunkhe SS, Bhatia NM, Bhatia MS (2014) Implications of formulation design on lipid-based nanostructured carrier system for drug delivery to brain. Drug Deliv 23:1–11. https://doi.org/10.3109/10717544.2014.943337
Johnson W (1999) Final Report on the Safety Assessment of Propylene Glycol (PG) Dicaprylate, PG Dicaprylate/Dicaprate, PG Dicocoate, PG Dipelargonate, PG Isostearate, PG Laurate, PG Myristate, PG Oleate, PG Oleate SE, PG Dioleate, PG Dicaprate, PG Diisostearate, and PG Dilaurate. Int J Toxicol 18:35–52. https://doi.org/10.1177/109158189901800207
Mahjour M, Mauser BE, Rashidbaigi ZA, Fawzi MB (1993) Effects of propylene glycol diesters of caprylic and capric acids (Miglyol® 840) and ethanol binary systems on in vitro skin permeation of drugs. Int J Pharmaceut 95:161–169. https://doi.org/10.1016/0378-5173(93)90403-3
Gritti F, Leonardis I, Shock D et al (2010) Performance of columns packed with the new shell particles, Kinetex-C18. J Chromatogr A 1217:1589–1603. https://doi.org/10.1016/j.chroma.2009.12.079
Kirkland JJ, Schuster SA, Johnson WL, Boyes BE (2013) Fused-core particle technology in high-performance liquid chromatography: an overview. J Pharm Anal 3:303–312. https://doi.org/10.1016/j.jpha.2013.02.005
Cunliffe JM, Maloney TD (2007) Fused-core particle technology as an alternative to sub-2-μm particles to achieve high separation efficiency with low backpressure. J Sep Sci 30:3104–3109. https://doi.org/10.1002/jssc.200700260
ICH-Guidelines Q2 (R1) (2005) Validation of analytical procedures: text and methodology. https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf. Accessed 21 Oct 2019
Acknowledgements
The authors would like to thank members of Global Pharmaceutical Technical Support (GPTS) team from Boehringer-Ingelheim Animal Health (BIAH) and members of GPTS team in Pharmaron for their support during this study. They thank to Dr. Sarju Adhikari of GPTS for providing valuable suggestions and comments during manuscript preparation.
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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. This work was supported by Boehringer-Ingelheim Animal Health.
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Padivitage, N., Wang, L., Wimalasinghe, R.M. et al. Development and Validation of a Stability-Indicating RP-HPLC Method for Determination of Eprinomectin, its Degradation Products, and Butylated Hydroxytoluene in a Pour-On Finished Product. Chromatographia 84, 949–965 (2021). https://doi.org/10.1007/s10337-021-04082-3
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DOI: https://doi.org/10.1007/s10337-021-04082-3