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Old 08-13-2010, 12:30 PM   #2
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The text below provides specifics related to the Muscle Marinade™ formula. It is in no way meant to represent a detailed discussion of all available evidence for the highlighted ingredients. Readers are encouraged to review the reference data provided at the end of this paper for additional information. While dosages of individual ingredients vary considerably across studies and not every study using a particular ingredient has been met with positive effects, consumers should feel confident that a great deal of unbiased attention has been put into the decision to include the below-discussed ingredients (as well as the exclusion of other commonly used ingredients). All those mentioned and included within the formula have been proven to be effective in human subjects with oral consumption based on peer-reviewed scientific reports and anecdotal (in the gym) evidence. Moreover, as consistently mentioned, the dosage of each ingredient within Muscle Marinade™ matches the dosage used in these clinical studies. This is indeed a novel concept in the field of sport nutrition supplements. Because the referenced studies often provide ingredients to subjects on a daily basis over the course of days or weeks (e.g., creatine, beta alanine), it should be understood that the below-discussed effects for a given ingredient may only be observed after continued use of that ingredient. It is assumed that individuals will use Muscle Marinade™ on a daily basis along with their normal nutrition and exercise training program in order to reap the product’s full benefits.

Muscle Marinade™: Energy Production, Stimulation, and Exercise Performance
Creatine—Overview


Creatine is a naturally occurring nitrogenous organic compound produced in the human body from the amino acids arginine, glycine, and methionine. While production occurs primarily in the kidney and liver, creatine is transported in the blood and taken up by other tissues (skeletal muscle primarily). To date, aside from carbohydrate, creatine is likely the most well-researched sport supplement in history. In fact, a PubMed search performed on 11/12/09 using the term “creatine and exercise” returned 3217 articles, while the term “creatine and exercise performance” returned 588 articles. Clearly, this is a well-researched ingredient and is thought to pose no adverse effects to healthy individuals (Poortmans and Francaux, 2000).

While the effects of creatine supplementation are multiple, including antioxidant activity, maintenance of neuronal health, and improved cardiac muscle performance, the effect of most interest related to exercise performance in otherwise healthy individuals is improved performance during high intensity anaerobic exercise (Hespel and Derave, 2007). Creatine aids in adenosine triphosphate (ATP) resynthesis and can lead to high intensity performance improvements as demonstrated in literally hundreds of scientific studies. Creatine supplementation has also been associated with enhanced muscle hypertrophy, which may relate to satellite cell proliferation, as well as myogenic transcription factors and insulin-like growth factor-1 signaling. Other evidence indicates that creatine could enhance muscle glycogen accumulation and glucose transporter (GLUT4) expression. Positive findings for creatine are noted for both healthy and diseased populations. Although noted in animals and not human subjects, oral creatine supplementation has been shown to significantly increase carnosine (+88%) and anserine (+40%) content in skeletal muscle, which coincides with improved resistance to contractile fatigue (Derave et al., 2008). The physiological effect of carnosine is discussed below in the section on beta alanine.

Creatine—Form

Much discussion exists related to the optimal form of creatine to be used. While creatine monohydrate is certainly the most well-researched and most common form available, other forms such as creatine ethyl ester (CEE), di-creatine malate, tri-creatine citrate, creatine gluconate, creatine taurinate, creatine pyruvate, creatine l-pyroglutamate, and “pH balanced” creatine among others are currently marketed throughout the sport supplement industry. In addition, some companies are now using an agent known as creatinol-O-phosphate within their products. Although this agent is not technically creatine, some companies proceed to falsely market it as a super-creatine. Most reports for COP were published in the late 1970s in the journal entitled Arzneimittelforschung, and most studies focus on cardiac function with COP. A PubMed search indicates that there is only one study dealing with physical performance (Nicaise, 1975) and included 50 female in-patients ranging in age from 58-96 years. Patients were treated intramuscularly and intravenously (not orally) with 2 ampoules of 500 mg each of COP. Muscular strength was then measured by having women squeeze a bulb in each hand 5 times. Results were of statistical significance but were rather meaningless from a physiological perspective (e.g., sum of 85.86 vs. 90.40 (kg/cm2)10-1 for placebo and intravenous COP, respectively; sum of 82.00 vs. 88.60 (kg/cm2)10-1 for placebo and intramuscular COP, respectively). Perhaps companies have other data to support their use of COP in their products (although they must be quite obscure, because these are not readily available via PubMed). If companies are basing their use of COP on the particular study described above and assuming that because intramuscular or intravenous COP increased hand strength to a minor extent in elderly in-patient women that oral intake of COP will lead to increased strength in young healthy men and women, they really need to reevaluate their formulation guidelines or do some real applied research using this ingredient. Inclusion of COP within a formula designed for young healthy men and women based on the data presented above is an absolute joke. Unfortunately, this is no exception in this industry.

While the more modern creatine versions are often heavily advertised so as to appear superior, there exists very little evidence that any of these creatine forms are better than creatine monohydrate, despite their substantial costs. For example, a study presented at the National Strength and Conditioning Association meeting in 2007 indicated that CEE was actually less stable than creatine monohydrate and experienced an accelerated breakdown to the byproduct known as creatinine (Child and Tallon, 2007). Other work agrees with this finding (Spillane et al., 2009). Authors comparing CEE with creatine monohydrate have concluded “when compared to creatine monohydrate, CEE was not as effective at increasing serum and muscle creatine levels or in improving body composition, muscle mass, strength, and power.” Investigators from another recent study concluded that “the half-life of CEE in blood is on the order of one minute, suggesting that CEE may hydrolyze too quickly to reach muscle cells in its ester form (Katseres et al., 2009). Collectively, these findings indicate that CEE is not a desired form of creatine to be used as a nutritional supplement.

Similar negative findings have been noted for the supposed “pH balanced” creatine known as Kre-Alkalyn® (Tallon and Child, 2007). Marketers claim that this product, unlike creatine monohydrate, is stabilized and will not undergo conversion into creatinine. To the contrary, investigators noted that the rate of creatinine formation for creatine monohydrate was <1% of the initial dose, indicating that creatine monohydrate is actually very stable under acidic conditions. Additionally, the Kre-Alkalyn® resulted in 35% greater conversion to creatinine than creatine monohydrate.

As for other creatine forms, very little research has been conducted to determine differences in either absorption or effectiveness as compared to creatine monohydrate. Therefore, at the present time, there is little to no evidence to support the use of forms other than creatine monohydrate. One recent study determined the plasma creatine appearance in men and women assigned to ingest a single dose of isomolar amounts of creatine (4.4 grams) as creatine monohydrate, tricreatine citrate, or creatine pyruvate (Jäger et al., 2007). The investigators noted that while peak concentration and area under the curve of plasma creatine was highest for creatine pyruvate, there was no difference between the estimated velocity constants of absorption or elimination between the three creatine forms. These investigators concluded that “differences in bioavailability are thought to be unlikely since absorption of creatine monohydrate is already close to 100%. The small differences in kinetics are unlikely to have any effect on muscle creatine elevation during periods of creatine loading.” This is especially true considering that oral creatine monohydrate is rapidly and efficiently absorbed, a fact established over 10 years ago (Vanakoski et al., 1998).

Despite these solid findings related to the already excellent absorption of creatine monohydrate, new products continue to be developed in an attempt to further improve creatine absorption. One such product (BIOCREAT) was recently studied and reported to yield similar adaptations in both muscle strength and lean mass as compared to a creatine+carbohydrate supplement, with no significant differences noted between the two creatine conditions (Lewing et al., 2009). Another form recently studied is polyethylene glycosylated creatine (creatine bound to polyethylene glycosylate [PEG]), hypothesized to result in increased creatine absorption and uptake into muscle cells (Herda et al., 2009). Subjects were assigned to a placebo condition, 1.80 or 3.60 grams of PEG (providing 1.25 and 2.50 grams of creatine, respectively), or 5 grams of creatine monohydrate per day for 30 days. Although the dosage of actual creatine was less in the PEG conditions, the results indicated that the creatine monohydrate condition yielded similar or better results in terms of lean body mass and performance improvement as compared to the PEG. These data reinforce the fact that creatine monohydrate, despite being considered “old school”, yields favorable results comparable or better than those observed with “new school” creatine forms.

Another new product, CON-CRĒT (Creatine HCL) marketed by ProMera health, is also being heavily advertised as a superior form of creatine in terms of solubility and absorption in the bloodstream. However, regardless of whether or not this is true (see below for more info), the company makes no claim related to the variable of real importance—skeletal muscle creatine uptake. While the product website indicates two university studies were conducted demonstrating this enhanced absorption, no reference data are provided, and no such studies are readily available via PubMed. Therefore, it is unknown whether or not the findings being claimed by ProMera health can stand up to the scrutiny of peer review. While it is possible that we may someday see published scientific evidence in support of CON-CRĒT (to date we simply have testimonials—which are alone, next to worthless in the scientific world), the question remains as to how much greater benefit a consumer could experience using this product (or any other novel creatine product for that matter) in order to justify the significant increase in cost as compared to creatine monohydrate. Aside from this important consideration, the fact that ProMera health boldly and deceptively states on their website, “CON-CRĒT is 59-times more potent than creatine monohydrate,” and “CON-CRĒT offers steroid-like results in strength, endurance and muscle recovery” is concerning to say the least. If the first claim were true, one serving (1500 mg for a 200 pound man) of CON-CRĒT would be equal to 88,500mg of creatine monohydrate. It is also stated on the website that one serving has potency equal to 5-10 grams of creatine monohydrate. There is clearly a discrepancy here within ProMera’s own claims. Such ridiculous and contradicting statements lead me to believe that this is yet another product fueled by pure marketing and hype, not hard scientific evidence.

As alluded to above, it should be understood that even if small differences in absorption time or concentration were noted between a novel form of creatine and creatine monohydrate, the question a consumer should have is “Who cares?” What real difference does this make considering creatine monohydrate already has absorption of close to 100% (Jäger et al., 2007)? Is it really worth paying more in order to use one of these hyped up novel creatine forms only to maybe experience a 5-10% increased plasma appearance rate? The rate of appearance of creatine is irrelevant anyway considering it is intramuscular and not plasma creatine that is important. Consumers also need to keep in mind that it is not the creatine taken immediately prior to each workout that is assisting in that particular workout; rather, it is the creatine that has been taken repeatedly over time that is now built up within the muscle that can provide for a benefit. Taking the daily dosage of creatine prior to (or immediately following) exercise makes good sense simply based on the fact that creatine transport into muscle may be enhanced due to the increased blood flow (Candow and Chilibeck, 2008) and possibly the increased activity of creatine transport proteins associated with acute exercise.

Aside from acute exercise, intake of creatine along with carbohydrate (usually simple sugars at high dosage; the basis of many creatine+carbohydrate products) has been shown to enhance creatine absorption in skeletal muscle (Green et al., 1996) and may enhance the effectiveness of creatine supplementation. Therefore, if adding extra carbohydrate to the diet does not interfere with daily caloric requirements, combining creatine and carbohydrate supplementation may be something to consider. That being said, PURUS LABS™ has chosen not to include carbohydrate within Muscle Marinade™, as more emphasis is placed on actual active ingredients rather than on inexpensive fillers. After all, carbohydrates are an inexpensive and readily-available addition if one chooses to include them.

Creatine—Dosage

Oral supplementation with creatine has been reported to substantially elevate the creatine content of human skeletal muscle. The most common dosage schedule in research studies has included a “loading” phase of 20 grams per day taken in 4 dosages of 5 grams each for a period of 5-7 days. Following this, creatine saturation in skeletal muscle can be maintained at a daily dosage as low as 2-5 grams for most individuals (Preen et al., 2003), although the International Society of Sport Nutrition (ISSN) has recommended a daily intake as high as 0.1 gram/kg body mass/day (Kerksick et al., 2008). As with all dietary supplements, individual needs may vary. As mentioned above, it has been suggested that creatine ingestion proximate to resistance exercise may be more beneficial for increasing muscle mass and strength than ingestion at times distant to the exercise session, possibly due to increased blood flow and therefore increased transport of creatine to skeletal muscle (Candow and Chilibeck, 2008). Hence, inclusion of creatine within a pre workout supplement appears logical, and this is why creatine is contained within Muscle Marinade™.

Beta alanine

Beta alanine, also referred to as 3-aminopropanoic acid, is a non-proteinogenic amino acid. Although initially discovered over 100 years ago, research with beta alanine pertaining to exercise performance in human subjects is relatively new, with the first scientific paper published just a few years ago. The plasma concentration of beta alanine is significantly and rapidly elevated following oral intake of beta alanine ranging from 20-40 mg/kg body mass (Harris et al., 2006). Moreover, the muscle carnosine (beta-alanyl-l-histidine) concentration, comprised of both beta alanine and histidine, is significantly increased when beta alanine is provided at a dosage of 3-6 grams per day (Harris et al., 2006). Carnosine helps to stabilize muscular pH by acting as a buffer for hydrogen ions that are released as a result of high intensity exercise. While not all studies have reported positive findings, the majority of work involving beta alanine supplementation indicates a significant performance-enhancing effect with regards to high intensity exercise.

One concern expressed in relation to beta alanine is the mild “prickling/tingling” sensation often felt soon after ingestion (e.g., as soon as 15 minutes and often lasting up to 60 minutes). This is referred to as parethesia, and is thought to be caused by beta-alanine binding to nerve receptors and causing them to fire. While this is well-tolerated by some users, others would prefer not to feel this prickling/tingling. In a study involving acute ingestion of beta alanine at dosages of 10, 20, and 40 mg/kg body mass, extreme tingling was noted with the 40 mg/kg body mass dosage, while only moderate tingling was experienced with the 20 mg/kg body mass dosage (Harris et al., 2006). Moreover, the increase in plasma beta alanine from the 10 to 20 mg/kg body mass dose was 6-8 fold, while the increase from 20-40 mg/kg body mass was only 2.2 fold. Peak plasma concentration of beta alanine occurred within 30-40 minutes following acute ingestion, and a subsequent study indicates that chronic supplementation (e.g., 15 days) does not affect this. Additionally, less beta alanine is lost in the urine following a 20 vs. 40 mg/kg body mass dosage. Therefore, based on the relatively small further increase in plasma beta alanine following ingestion of a single dosage from 20 to 40 mg/kg body mass, the fact that dosages as low as 2 grams per day have been found to be efficacious in scientific investigations (Van Thienen et al., 2009), and the fact that higher dosages of beta alanine lead to greater parethesia, Muscle Marinade™ contains a dosage of beta alanine equivalent to 25 mg/kg body mass for an 80 kg man. This dosage should minimize profound parethesia and is close to the dosage previously reported to increase muscle carnosine content by ~40% following four weeks of ingestion (Harris et al., 2006).

As discussed above in the section on creatine, although noted in animals and not human subjects, it has been reported that creatine intake alone results in enhanced muscle carnosine content (Derave et al., 2008). Considering this evidence, using an adequate dosage of creatine along with beta alanine may justify using a slightly lower dosage of beta alanine. As with creatine, it has been suggested that beta alanine uptake into skeletal muscle to form carnosine may be enhanced by carbohydrate intake due to the insulin response from such feeding. Again, users may add carbohydrate as they see fit.

Betaine

Betaine (chemically known as 2-(Trimethylammonio) ethanoic acid, hydroxide, inner salt) is an osmolyte (i.e., protects the cells against dehydration), an antioxidant agent, as well as a methyl group donor serving a chief purpose of lowering homocysteine (Olthof and Verhoef, 2005), a known risk factor for cardiovascular disease (Boushey et al., 1995). The B-vitamins folic acid (B9), B12, and B6 are often used for this same purpose of lowering homocysteine. As a methyl group donor, betaine has a potential effect on creatine biosynthesis by providing a methyl group to guanidinoacetate via methionine that can synthesize creatine in skeletal muscle (du Vigneaud et al., 1946).

In regards to exercise performance, a few studies have been conducted over the past few years using betaine (anhydrous form). The dosage of betaine in these studies has been 2.5 grams per day. Muscle endurance (Hoffman et al., 2009) as well as muscular power and force (Maresh et al., 2007) have been reported to increase following 14 days of betaine supplementation. Mechanistically, betaine may improve exercise performance by providing antioxidant activity, maintaining cellular hydration, and increasing blood flow, the latter possibly mediated by the effect betaine has on increasing NO (unpublished data). Although betaine is relatively new to the sport nutrition market, PURUS LABS™ believes that this ingredient has promise as a sport supplement. For this reason it is included within Muscle Marinade™ at the proper, researchsupported dosage.
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