The stability and bioavailability of the equivalent amounts of creatine from creatine monohydrate and Creatine Xplode Powder.

Date of test: November 2018

 

P. Kaczka, R. Jastrząb, A. Łyskowski

 

Olimp Laboratories Sp. z o.o., Research-and-Development Centre

Pustynia 84F, 39-200 Dębica,

 

 

INTRODUCTION

 

  1.  The stability and bioavailability of creatine in the gastrointestinal tract

 

Creatine monohydrate remains the most-popular form of creatine available on the market which acts anabolically within muscle tissue  Many studies have verified the pharmacokinetics and stability of this compound in different environments (MacNeil et al., 2005)(McCall & Persky, 2007)(Ganguly, Jayappa, & Dash, 2003). According to the current state of knowledge, the stomach environment (low pH, presence of digestive enzymes), as well as the digestive environment of the duodenum, are able to degrade up to several percent of consumed creatine (Purchas, Busboom, & Wilkinson, 2006), reducing its final bioavailability, as well as the expected physiological effect. One of the metabolites of creatine produced during degradation is its cyclic form - creatinine, which is not biologically active.

 

Reducing the actual bioavailability of creatine from its various chemical forms in the human gastrointestinal tract, resulting from its conversion to, inter alia, creatine, can result in a not-unfounded belief that the amount of product intake should be increased in order to ensure an actual, effective, supply of that product. Depending on the form used, varying instability of creatine in gastrointestinal tract conditions can at the same time lead to the absence of noticeable effects of creatine supplementation for some of its chemical forms, with the recommended daily supply. This has become a direct reason for the search for more-stable, and thus more-effective, forms of creatine. Currently available on the market are, i.a., malate, citrate, ethyl ester, magnesium creatine chelate, alpha-ketoglutarate, pyruvate, and buffered creatine, characterised by physico-chemical properties different from monohydrates, as well as pharmacokinetics itself (Gufford et al., 2013), (Jäger, Harris, Purpura, & Francaux, 2007). The expected final effect of the existing differences between the different chemical forms of creatine is to improve the bioavailability and efficacy of creatine supplementation for its more chemically complex forms, in the case of the monohydrate itself. However, the number of literature titles on this subject is still low and insufficient, and the very issue of the advantages of one form of creatine over another is still a subject for discussion, in both sporting and scientific circles. This was the starting point for this study, which describes the bioavailability and stability of the popular creatine monohydrate, and a product which includes 6 other chemical forms of creatine - Creatine Xplode Powder.

 

 

  1.  SHIME® Artificial Gastrointestinal Tract - an advanced research tool

 

 

The analysis of digestion and absorption processes of food components and dietary supplements under in-vitro conditions is possible from the technical point of view under static and dynamic conditions. In the first case, only the processes taking place on a specific, selected, section of the gastrointestinal tract can be analysed without the possibility of taking into account the dynamically changing conditions of the gastrointestinal tract’s lumen environment, which undoubtedly proves the disadvantages of such an analytical solution.

 

SHIME®  (Simulator of the Human Intestinal Microbial Ecosystem) is a technologically advanced device allowing the dynamic simulation of processes occurring in the gastrointestinal tract. It facilitates the experimental observation of physico-chemical, enzymatic, and microbiological changes in the gastrointestinal tract under conditions of in-vitro simulation in a strictly controlled environment. The unique design allows you to obtain data facilitating the improvement of the tested products as early as at their design stage, before they are put into production, by taking into account the processes to which they will be subjected in conditions representative of the physiology of the intestine.

 

The SHIME® system is built on the basis of a structure of interconnected reactors corresponding to individual sections of the gastrointestinal tract. The dynamic character of the system allows you to reliably track the status of selected chemical compounds, e.g. metabolites, in the gastrointestinal tract. The technical scheme of the system has been developed on the basis of scientific research. In its most-advanced form, SHIME® is a system of five reactors simulating individual stages in the digestion and absorption process. The first reactor, the stomach (ST), and the second, the small intestine (SI), are responsible for simulating the intake and initial enzymatic digestion of food. Similarly to the following reactors, simulating the colon, they are connected by a computer-controlled system of peristaltic pumps, which carry the food content with precision, according to a pre-programmed scheme. The conditions in individual reactors are subject to strict control, and have been developed on the basis of data collected in scientific publications.

 

The results from the SHIME® experiment are reproducible in vivo. This is confirmed by scientific research which has been conducted with the use of the system for over 25 years, and by numerous scientific publications in renowned journals (Molly et. al. 1994; Possemiers, et. al. 2004; Van de Wiele T., 2015).

 

 

THE PURPOSE OF THE TEST

 

An analysis of the stability and bioavailability of the equivalent amounts of creatine from its monohydrate and complex creatine product - Creatine Xplode Powder (Olimp Laboratories Sp. z o.o.).

 

MATERIALS AND METHODS

 

The applied model of creatine bioavailability analysis was based on procedures which have been validated and widely recognised as reliable, reproducing the natural processes of nutrient digestion under in-vitro conditions (INFOGest®, ProDigest®). The procedures were extended to include detailed data on gastrointestinal motility and the duration of the stay of digestive content in specific sections of the tract. In this manner, it was possible to obtain an extremely reliable representation of the passage of the tested products through the human gastrointestinal tract.

 

A portion of 3 g of creatine was used in the study, referring to its pure form, obtained by the supply of a larger portion of the tested products - 5.8 g for Creatine Xplode Powder (Olimp Laboratories Sp. z o.o.) and 3.4 g of creatine monohydrate (Creatine monohydrate ≥98%, Sigma-Aldrich).

 

The study used a gastric medium prepared from commercially available nutrients, a solution of pepsin (2%) suspended in 5.0 mM of HCl (stored at -20 °C until its use in the experiment), and a solution of pancreatic juices, together with pancreatin.

 

A pH control in the reactors simulating particular sections of the gastrointestinal tract was performed automatically with the use of 0.5 M of HCl and 0.5 M of NaOH.

 

Samples for analysis (5 mL) were collected every 20 minutes from all reactors (stomach (ST), duodenum (DU), and small intestine (SI), containing gastric content at the given stage of the experiment. The collected samples were stored at -20 °C until the level of the selected chemical compounds was determined.

 

The amount of creatine and creatinine in the tested samples was determined using high-performance liquid chromatography (HPLC).

 

The test and analysis conditions were the same for creatine monohydrate and Creatine Xplode Powder.

 

The applied research model did not take into account the absorption of creatine, but only its physico-chemical dynamics in the environment of the human gastrointestinal tract.

 

 

RESULTS

When referring to “stomach”, “duodenum”, and “small intestine” it is always in connection with the SJIME® artificial gastrointestinal bioreactor, which faithfully reflects the physiological conditions of these parts of the human gastrointestinal tract.

 

The creatine contained in Creatine Xplode Powder reached higher concentrations in the stomach (within 4 h) and small intestine (within 2 h 20 min) and had greater chemical stability (a lower conversion to creatinine) than the equivalent amounts of creatine from creatine monohydrate.

 

STOMACH

 

 Creatine

 

1)      During the entire 240 minutes of Creatine Xplode Powder and creatine monohydrate stayed in the stomach, the following values of the creatine ratio (the anabolic effect on muscle tissue) to inactive creatinine could be observed for the tested products

 

 Creatine

 

In the case of Creatine Xplode Powder, this value was greater at each time point of the sample analysis and was between 58.3% and 13.7% compared to creatine monohydrate (as shown in the table below).

 

 Creatine

 

2)      During the entire 240 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the stomach, the following values of inactive creatinine from the conversion of creatine contained in them could be observed for the tested products

 

 Creatine

 

In the case of creatine monohydrate, this value was greater at each time point of the sample analysis, and was between 47% and 6% compared to Creatine Xplode Powder (as shown in the Figure below).

 

 Creatine

 

PASSAGE FROM THE STOMACH TO THE SMALL INTESTINE (DUODENUM)

 

1) At the time point (140 min; duodenum) of the Creatine Xplode Powder and the creatine monohydrate’s passage from the stomach to the intestine, 17% more creatine could be observed for Creatine Xplode Powder compared to creatine from creatine monohydrate.

 

 Creatine

 

Taking into account the above, the determined value of the creatine (the anabolic effects on muscle tissue) to the inactive creatinine ratio for the tested products was as follows

 

 Creatine

 

This value was 2.5 times higher for Creatine Xplode Powder than for creatine monohydrate.

 

 

Analysing the amount of inactive creatinine formed during the passage from the stomach to the small

intestine, the following amounts of inactive creatinine were determined

 

 Creatine

 

The amount of creatinine marked for creatine monohydrate was 2.1 higher than for Creatine Xplode Powder.

 

THE SMALL INTESTINE

 

The creatine contained in Creatine Xplode Powder reached higher concentrations in the small intestine (within 2 h 20 min) and had greater chemical stability (a lower conversion to creatinine) than the equivalent amounts of creatine from creatine monohydrate. In addition, creatine from monohydrate also showed a very-rapid decrease in creatine concentration after 100 minutes in the small-intestine environment. This effect is not observed with Creatine Xplode Powder.

 

 Creatine

 

1)      During the entire 140 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the small intestine, the following values of the creatine ratio (the anabolic effect on muscle tissue) to inactive creatinine could be observed for the tested products

 

 Creatine

 

In the case of Creatine Xplode Powder, this value was greater at each time point of the sample analysis, and was between 16.3% and 9.2% compared to creatine monohydrate, as shown in the Figure below.

 

 Creatine

 

2)      During the entire 140 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the small intestine, the following values of inactive creatinine from the conversion of creatine contained in them could be observed for the tested products

 

 Creatine

 

In the case of creatine monohydrate, this value was greater at each time point of the sample analysis and was between 14% and 3% compared to Creatine Xplode Powder (as shown in the Figure below).

 

 Creatine

 

CONCLUSIONS

 

    1.  THE STOMACH

 

The creatine contained in Creatine Xplode Powder reached higher concentrations in the stomach (within 4 h) and had greater chemical stability (a lower conversion to creatinine) than the equivalent amounts of creatine from creatine monohydrate.

 

a)     During the entire 240 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the stomach, at each time point a clear advantage could be observed, in the range from 58.3% to 13.7%, of the value of the creatine ratio (the anabolic effects on muscle tissue) to the inactive creatine for Creatine Xplode Powder compared to creatine monohydrate

 

b)     During the entire 240 minutes of the Creatine Xplode Powder and Creatine Monohydrate stay in the stomach, at each time point a clear advantage could be observed in the level of inactive creatine, of which there was less in the range from 47% to 6% for Creatine Xplode Powder compared to creatine monohydrate

 

  1.  PASSAGE FROM THE STOMACH TO THE SMALL INTESTINE (DUODENUM)

 

The creatine contained in Creatine Xplode Powder reached higher concentrations in the duodenum and had a greater chemical stability (a lower conversion to creatinine) than the equivalent amounts of creatine from creatine monohydrate. In particular for Creatine Xplode Powder, compared to creatine monohydrate, the following could be observed:

a)       17% higher creatine concentration,

b)      2.5 times greater ratio of creatine to inactive creatinine,

c)       over two times less biologically inactive creatinine

  1. THE SMALL INTESTINE

 

The creatine contained in Creatine Xplode Powder reached higher concentrations in the small intestine (within 2 h 20 min), at each time point, and had a greater chemical stability (a lower conversion to creatinine) than the equivalent amounts of creatine from creatine monohydrate. In addition, creatine from monohydrate also showed a very-rapid decrease in creatine concentration after 100 minutes in the small intestine environment. This effect was not observed with Creatine Xplode Powder

a)       During the entire 140 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the small intestine at each time point a clear advantage could be observed, in the range from 16.3% to 9.2%, of the value of creatine ratio (the anabolic effects on muscle tissue) to inactive creatine for Creatine Xplode Powder compared to creatine monohydrate

b)      During the entire 140 minutes of the Creatine Xplode Powder and creatine monohydrate stay in the stomach, at each time point a clear advantage could be observed in the level of inactive creatine, of which there is less in the range from 14% to 3% for Creatine Xplode Powder compared to creatine monohydrate

 

BIBLIOGRAPHY:

Ganguly, S., Jayappa, S., & Dash, A. K. (2003). Evaluation of the stability of creatine in solution prepared from effervescent creatine formulations. AAPS PharmSciTech, 4(2), E25. https://doi.org/10.1208/pt040225

Gufford, B. T., Ezell, E. L., Robinson, D. H., Miller, D. W., Miller, N. J., Gu, X., & Vennerstrom, J. L. (2013). PH-dependent stability of creatine ethyl ester: Relevance to oral absorption. Journal of Dietary Supplements. https://doi.org/10.3109/19390211.2013.822453

Jäger, R., Harris, R. C., Purpura, M., & Francaux, M. (2007). Comparison of new forms of creatine in raising plasma creatine levels. https://doi.org/10.1186/1550-2783-4

MACNEIL, L., HILL, L., MACDONALD, D., KEEFE, L., CORMIER, J., BURKE, D., & SMITHPALMER, T. (2005). Analysis of creatine, creatinine, creatine-d3 and creatinine-d3 in urine, plasma, and red blood cells by HPLC and GC–MS to follow the fate of ingested creatine-d3. Journal of Chromatography B, 827(2), 210–215. https://doi.org/10.1016/j.jchromb.2005.09.011

McCall, W., & Persky, A. M. (2007). Pharmacokinetics of creatine. Sub-Cellular Biochemistry, 46, 261 – 273. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/18652081

Purchas, R. W., Busboom, J. R., & Wilkinson, B. H. P. (2006). Changes in the forms of iron and in concentrations of taurine, carnosine, coenzyme Q10, and creatine in beef longissimus muscle with cooking and simulated stomach and duodenal digestion. Meat Science, 74(3), 443–449. https://doi.org/10.1016/j.meatsci.2006.03.015

Molly, K., Vande Woestyne, M., De Smet, I. & Verstraete, W. (1994) Validation of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) Reactor Using Microorganism-associated Activities, Microbial Ecology in Health and Disease, 7:4, 191- 200, DOI: 10.3109/08910609409141354

Possemiers, S., Verthé, K., Uyttendaele, S., Verstraete, W., (2004). PCR-DGGE-based quantification of stability of the microbial community in a Simulator of the Human Intestinal Microbial  Ecosystem, FEMS Microbiology Ecology. 49 (3), 495-507, https://doi.org/10.1016/j.femsec.2004.05.002

Van de Wiele T., Van den Abbeele P., Ossieur W., Possemiers S., Marzorati M. (2015) The Simulator of the Human Intestinal Microbial Ecosystem (SHIME®). In: Verhoeckx K. et al. (eds) The Impact of Food Bioactives on Health. Springer, Cham. , https://doi.org/10.1007/978-3-319-16104-4_27