Malaysian Journal of Analytical Sciences, Vol 27 No 1 (2023): 87 - 95

 

DETERMINATION OF L-CARNITINE AND COENZYME Q10 ACTIVE SUBSTANCE LEVELS IN FOOD SUPPLEMENTS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

 

(Penentuan Kadar Zat Aktif L-Carnitine dan Ko-enzim Q10 dalam Suplemen Makanan Menggunakan Kromatografi Cecair Prestasi Tinggi)

 

Muhaimin1,*, Hilda Andini 2, and Henry Himawan3

 

1 Study Program of Chemistry Education,

Department of Chemistry, Islamic University of Indonesian, Indonesia

2 Diploma Program of Chemical Analysis,

Department of Chemistry, Islamic University of Indonesian, Indonesia

3 PT. Dipa Pharmalab Intersains,

Majalengka, West Java, Indonesia

 

*Corresponding author: muhaimin@uii.ac.id

 

 

Received: 23 July 2022; Accepted: 25 October 2022; Published:  22 February 2023

 

 

Abstract

This study aimed to determine the active substance levels in L-Carnitine and Coenzyme Q10 in dietary supplements using the High-Performance Liquid Chromatography (HPLC)-UV method. They are often found in dietary supplements because these ingredients can be used to maintain heart health. L-Carnitine is an active ingredient belonging to amino acids, while Coenzyme Q10 is one of the most significant lipid antioxidants, which prevents the formation of free radicals and modification of proteins, lipids, and DNA. The method used in this study is HPLC-UV because the method has good sensitivity, can use the column and solvent developer many times, the detector varies, the analysis time is generally short, easy to operate, and has relatively high accuracy and precision. The HPLC procedure for determining L-Carnitine used a UV detector with a wavelength of 225 nm using an L1 (4.6 mm x 15 cm; 5 m) column with a flow rate of 0.7 mL/min. Meanwhile, the determination of Coenzyme Q10 using the HPLC procedure used a UV detector with a wavelength of 280 nm using a C18 (4.6 mm x 15 cm, 5μm) column with a flow rate of 1.5 mL/minute. This study also reports differences in L-Carnitine and Coenzyme Q10 in dietary supplements (in capsule form) during the mixing and encapsulation processes. The results show the suitability test for the HPLC-UV system in good condition; seen from the %RSD value below 2%.  In testing the active substance Coenzyme Q10 in the mixing process is 101.07% and encapsulation is 96.77%. While the active substance L-Carnitine in the mixing process is 105.05% and encapsulation is 93.03%. The overall test results fall into the range of requirements, meaning that the product quality is in a good category.

 

Keywords: l-carnitine, coenzyme Q10, high-performance liquid chromatography, mixing, encapsulation

 

 

Abstrak

Penelitian ini untuk mengetahui kadar zat aktif L-Carnitine dan Ko-enzim Q10 pada suplemen makanan menggunakan kaedah kromatografi cecair prestasi tinggi (HPLC)-UV. Zat aktif ini sering ditemukan dalam suplemen makanan kerana bahan-bahan ini dapat digunakan untuk menjaga kesihatan jantung. L-Carnitine merupakan bahan aktif yang tergolong ke dalam asid amino, sedangkan Ko-enzim Q10 adalah salah satu antioksidan lipid yang paling signifikan, yang mencegah pembentukan radikal bebas dan modifikasi protein, lipid, dan DNA. Kaedah yang digunakan dalam penelitian ini adalah HPLC-UV karena memiliki sensitiviti yang baik, dapat menggunakan turus dan pelarut berkali-kali, pengesan pelbagai, memiliki waktu analisis yang singkat, mudah dioperasikan, dan memiliki ketepatan dan kejituan yang relatif tinggi. Prosedur HPLC untuk penentuan L-Carnitine menggunakan pengesan UV dengan panjang gelombang 225 nm menggunakan turus L1 (4.6 mm x 15 cm; 5 m) dengan kadar aliran 0.7 mL/minit. Sedangkan penentuan Ko-enzim Q10 menggunakan prosedur HPLC menggunakan pengesan UV dengan panjang gelombang 280 nm menggunakan turus C18 (4.6 mm x 15 cm, 5μm) dengan kadar aliran 1.5 mL/minit. Dalam penelitian ini, kami juga melaporkan perbezaan kadar L-Carnitine dan Koenzim Q10 dalam suplemen makanan (dalam bentuk kapsul) selama proses pencampuran dan pengkapsulan. Hasil pengujian menunjukkan bahwa uji kesesuaian sistem HPLC-UV dalam keadaan baik dilihat dari nilai %RSD dibawah 2%. Pada pengujian zat aktif Ko-enzim Q10 pada proses pencampuran adalah 101.07% dan pengkapsulan adalah 96.77%. Sedangkan zat aktif L-Carnitine pada proses pencampuran adalah 105.05% dan pengkapsulan adalah 93.03%. Hasil pengujian secara keseluruhan menepati piawaian, memberi maksud produk berada pada kategori baik.

 

Kata kunci: L-carnitine, ko-enzim Q10, kromatografi cecair prestasi tinggi, pencampuran, pengkapsulan

 

References

1.         Manoharan, G. (2021). Quantitative determination of l-carnitine tablet formulation by a validated stability-indicating reversed-phase HPLC method. South Asian Research Journal Pharmaceutical Sciences, 3:46-51.

2.         Da̧browska, M. and Starek, M. (2014). Analytical approaches to determination of carnitine in biological materials, foods and dietary supplements. Food Chemistry, 142: 220-232.

3.         Park, J. M., Koh, J. H. and Kim, J. M. (2021). Determination of l-carnitine in infant powdered milk samples after derivatization. Food Science Animal Resource, 41:731-738.

4.         Durazzo, A., Lucarini, M., Nazhand, A., Souto, S. B., Silva, A. M., Severino, P., Souto, E. B. and Santini, A. (2020). The nutraceutical value of carnitine and its use in dietary supplements. Molecules 25: 1-19.

5.         Ahn, J. H., Kwak, B. M., Park, J. M., Kim, N. K. and Kim, J. M. (2014). Rapid determination of l-carnitine in infant and toddler formulas by liquid chromatography-tandem mass spectrometry. Korean Journal Food Science Animimal Resource, 34:749-756.

6.         Khoshkam, R. and Afshar, M. (2014). Validation of a stability-indicating RP-HPLC method for determination of l-carnitine in tablets. International Scholary Resesearch Notice, 2014: 1-7.

7.         Kakou, A., Megoulas, N. C. and Koupparis, M. A. (2005). Determination of l-carnitine in food supplement formulations using ion-pair chromatography with indirect conductimetric detection. Journal Chromatography A, 1069: 209-215.

8.         Seline, K. G. and Johein, H. (2007) The determination of l-carnitine in several food samples. Food Chemistry, 105:793-804.

9.         Gustavsen, H. S. M (2000) Bestimmung von L-carnitin und seiner Ester in Lebensmitteln. Aus dem Physiologischen Institut der Tierärztlichen Hochschule Hannover.

10.      Demarquoy, J., Georges, B., Rigault, C., Royer, M. C., Clairet, A., Soty, M., Lekounoungou, S. and Le Borgne F. (2004). Radioisotopic determination of l-carnitine content in foods commonly eaten in western countries. Food Chemistry, 86: 137-142.

11.      Bene, J., Szabo, A., Komlósi, K. and Melegh, B (2019). Mass spectrometric analysis of l-carnitine and its esters: potential biomarkers of disturbances in carnitine homeostasis. Current Molecular Medicine, 20: 336-354.

12.      Prokorátová, V., Kvasnička, F., Ševčík, R. and Voldřich, M. (2005). Capillary electrophoresis determination of carnitine in food supplements. Journal Chromatography A, 1081: 60-64.

13.      He, G.X. and  Dahl, T. (2000). Improved high-performance liquid chromatographic method for analysis of l-carnitine in pharmaceutical formulations. Journal Pharmaceutical Biomedicine Analysis, 23: 315-321.

14.      Dâbrowska, M., Sieczka, E. and Starek, M. (2012). TLC assay of l-carnitine in dietary supplements. Journal Planar Chromatography - Mod TLC, 25:450-455.

15.      Saxena, J. D., Yadav, P. and Kantharia, N. D. (2011). Coenzyme Q10: The essential nutrient. Journal Pharmaceutical Bioallied Sciences, 3: 465-466.

16.      Neergheen, V., Chalasani, A., Wainwright, L., Yubero, D., Montero, R., Artuch, R. and Hargreaves, I. (2017). Coenzyme Q10 in the treatment of mitochondrial disease. Journal Inborn Errors Metabolism Screening, 5: 1-8.

17.      Temova Rakuša, Ž., Kristl, A. and Roškar, R. (2020). Quantification of reduced and oxidized coenzyme Q10 in supplements and medicines by HPLC-UV. Analytical Methods, 12: 2580-2589.

18.      Choi, C. H., Kim, S. H., Shanmugam, S., Baskaran, R., Park, J. S., Yong, C. S., Choi, H. G., Yoo, B. K. and Han, K. (2010). Relative bioavailability of coenzyme Q10 in emulsion and liposome formulations. Biomolecule Theraphy, 18:99-105.

19.      Raizner, A. E. (2019). Coenzyme Q10. Methodist Debakey Cardiovasc Journal, 15: 185-191.

20.      Aaseth, J., Alexander, J. and Alehagen, U. (2021). Coenzyme Q10 supplementation – in ageing and disease. Mechanism and Ageing Development, 197: 111521.

21.      Wang, Y. and Hekimi, S. (2016). Understanding ubiquinone. Trends Cell Biology, 26:367-378.

22.      Gleize, B, Steib M, André M. and Reboul, E. (2012). Simple and fast HPLC method for simultaneous determination of retinol, tocopherols, coenzyme Q10 and carotenoids in complex samples. Food Chemistry, 134:2560-2564.

23.      Rácz, A., Vass, A., Héberger, K. and Fodor, M. (2015). Quantitative determination of coenzyme Q10 from dietary supplements by FT-NIR spectroscopy and statistical analysis. Analytical Bioanalytical Chemistry, 407: 2887-2898.

24.      Visconti, G. L., Mazzoleni, L., Rusconi, C., Grazioli, V., Roda, G., Manini, G. and Gambaro, V. (2015). Determination by UPLC/MS-MS of Coenzyme Q10 (CoQ10) in Plasma of Healthy Volunteers before and After oral intake of food supplements containing CoQ10. Journal Analytical Bioanalytical Technology, S13:011.

25.      Monakhova, Y. B., Ruge, I., Kuballa, T., Lerch, C. and Lachenmeier, D. W. (2013). Rapid determination of coenzyme Q10 in food supplements using 1H NMR spectroscopy. International Journal Vitamamin Nutrional Research, 83: 67-72.

26.      Vass, A., Deák, E. and Dernovics, M. (2015). Quantification of the reduced form of coenzyme Q10, ubiquinol, in dietary supplements with HPLC-ESI-MS/MS. Food Analytical Methods, 8: 452-458.

27.      Schou-Pedersen, A. M. V., Schemeth, D. and Lykkesfeldt, J. (2019). Determination of reduced and oxidized coenzyme Q10 in canine plasma and heart tissue by HPLC-ECD: Comparison with LC-MS/MS quantification. Antioxidants 8(8): 1-14.

28.      Tang, P. H. (2006). Determination of coenzyme Q10 in over-the-counter dietary supplements by high-performance liquid chromatography with coulometric detection. Journal AOAC International, 89: 35-39.

29.      Ün, İ., Vatansever, B., Şimşek, A. and Gören, A. C. (2016). Comparison of qNMR and HPLC-UV techniques for measurement of coenzyme Q10 in dietary supplement capsules. Journal Chemical Metrology, 10:1-10

30.      Ermer, J. and Ploss, H. J. (2005) Validation in pharmaceutical analysis: Part II: Central importance of precision to establish acceptance criteria and for verifying and improving the quality of analytical data. Journal Pharmaceutical Biomedicine Analysis, 37: 859-870.

31.      Chan, C. C., Lam, H., Lee, Y. C. and  Zhang, X. (2004). Analytical method validation and instrument performance verification. John Wiley & Sons, Inc., Hoboken, NJ, USA

32.      United States Pharmacopeia (2018). The United States Pharmacopeia, USP 41/The National Formulary, NF 36. MD: U.S. Pharmacopeial Convention, Inc., Rockville

33.      Ratnawati, N. A., Prasetya, A. T. and Rahayu, E. F. (2019). Validasi metode pengujian logam berat timbal (Pb) dengan destruksi basah menggunakan FAAS dalam sedimen sungai banjir Kanal Barat Semarang. Indonesian Journal Chemical Sciences, 8: 60-68.