Downloaded from British Journal of Nutrition (2015), 114, 1403–1409 doi:10.1017/S0007114515003050 © The Authors 2015 https://www.cambridge.org/core Comparison of availability and plasma clearance rates of β-hydroxy-β-methylbutyrate delivery in the free acid and calcium salt forms . IP address: John C. Fuller Jr1, Rick L. Sharp2, Hector F. Angus2, Paul Y. Khoo2 and John A. Rathmacher1,3* 1 Metabolic Technologies Inc., Iowa State University Research Park, Ames, IA 50010, USA 170.106.35.234 2Department of Kinesiology, Iowa State University, Ames, IA 50010, USA 3Department of Animal Science, Iowa State University, Ames, IA 50010, USA (Submitted 22 February 2015 – Final revision received 14 July 2015 – Accepted 17 July 2015 – First published online 16 September 2015) , on 30 Sep 2021 at 15:04:06 Abstract β-Hydroxy-β-methylbutyrate (HMB), a leucine metabolite, has long been supplemented as a Ca salt (Ca-HMB) to increase strength and performance gains with exercise and to reduce recovery time. Recently, the free acid form of HMB (HMB-FA) has become commercially available in capsule form (gelcap). The current study was conducted to compare the bioavailability of HMB using the two commercially available capsule forms of HMB-FA and Ca-HMB. We also compared the pharmacokinetics of each form when administered mixed in water. Ten human subjects (five male and five female) , subject to the Cambridge Core terms of use, available at were studied in a randomised crossover design. There was no significant sex by treatment interaction for any of the pharmacokinetic parameters measured. HMB-FA administered in capsules was more efficient than Ca-HMB capsule at HMB delivery with a 37 % increase in plasma clearance rate (74·8(SEM 4·0) v. 54·5(SEM 3·2) ml/min, P < 0·0001) and a 76 % increase in peak plasma HMB concentration (270·2(SEM 17·8) v. 153·9(SEM 17·9) μmol/l, P < 0·006), which was reached in one-third the time (P < 0·009). When HMB-FA and Ca-HMB were administered in water, the differences still favoured HMB-FA, albeit to a lesser degree. Plasma HMB with HMB-FA administered in water was greater during the early phase of absorption (up to 45 min postadministration, P < 0·05); this resulted in increased AUC during the first 60 min after administration, when compared with Ca-HMB mixed in water (P < 0·03). In conclusion, HMB-FA in capsule form improves clearance rate and availability of HMB compared with Ca-HMB in capsule form. Key words: β-Hydroxy-β-methylbutyrate: Free acid form of hydroxy-β-methylbutyrate: Calcium salt of hydroxy-β-methylbutyrate: Gelcaps: Capsules: Clearance The leucine metabolite β-hydroxy-β-methylbutyrate (HMB) has a it can persist for up to 48 h postexercise(17). These long history of use as a nutritional supplement for enhancing anabolic events are also accompanied by depletion of muscle recovery, and for increasing strength, power, aerobic performance energy stores in the form of glycogen, resulting in decreased https://www.cambridge.org/core/terms – and lean body mass with exercise(1 5). HMB improves muscle exercise intensity, and associated with increased muscle – protein balance by decreasing muscle protein breakdown and by damage(18 20). The changes have been attributed to increased – increasing muscle protein synthesis(6 11), resulting in reduced skeletal muscle AMP-activated protein kinase activity and muscle damage and faster and improved recovery(3,12,13).These reduced phosphorylation of 4E-BP1 in the mTOR pathway, benefits with HMB to improve muscle performance with vigorous as occurs with resistance exercise(16). HMB supplementation exercise have generally been achieved with the Ca salt of HMB has been shown to increase phosphorylation of 4E-BP1 in (Ca-HMB) administered in capsule form. However, as shown in human muscle(11), prompting us to hypothesise that HMB as previous studies, and confirmed by the findings of the current HMB-FA would offer a more convenient delivery form resulting study, peak plasma levels of HMB are not reached until in an improved anabolic effect of HMB on muscle protein . approximately 120 min after ingestion of Ca-HMB, which makes it synthesis and diminishing protein breakdown. https://doi.org/10.1017/S0007114515003050 inconvenient for scheduling exercise sessions to coincide with Other metabolic effects of HMB have been demonstrated peak plasma HMB levels(14,15). In this study, we show that the time in animal models. Turkeys fed HMB during the last 15 weeks to peak HMB level decreased by two-thirds when HMB was of rearing had increased bone density and strength and delivered as the free acid in capsule form (HMB-FA), thus making an increase in some plasma amino acids including the it more convenient to time peak HMB level with exercise. branched-chain amino acids(21). Tatara et al.(22) also demon- The anabolic stimulus of exercise results in an increase in strated increased growth and bone density, and increased muscle protein synthesis in as little as 1 h postexercise(16), and growth hormone (GH) and insulin-like growth factor-1 (IGF-1), Abbreviations: Ca-HMB, calcium salt of HMB; HMB, β-hydroxy-β-methylbutyrate; HMB-FA, free acid form of HMB. * Corresponding author: Dr J. A. Rathmacher, fax +1 515 296 0908, email [email protected] Downloaded from 1404 J. C. Fuller et al. in offspring of sows fed HMB for the final 2 weeks of preg- HPLC analysis, and were given with 355 ml of water. Although https://www.cambridge.org/core nancy; increased bone density and plasma amino acid con- the pH of the free acid itself is <3, this is similar to the pH of the centrations in pigs with fundectomy-induced osteopaenia(23); stomach. For the treatments used in this study, the free acid and improved somatotrophic axis and accelerated bone meta- was given either in capsule form, a buffered gel form or as a bolism in lambs fed HMB for the first 21 d after birth(24). The dilute solution in water. In contrast, the Ca-HMB is neutral to improvement in somatotrophic axis function was further slightly alkaline when in solution. None of the participants demonstrated in rats when chronic HMB administration was reported any oesophageal or gastrointestinal irritations with any shown to stimulate the GH/IGF-1 axis(25). Recently, HMB sup- of the treatment forms of HMB. For the capsule treatments, plementation was shown to enhance the exercise-stimulated samples of the capsules were sent to an independent laboratory . IP address: increase in GH and IGF-1 response in humans, similar to what and tested for dissolution times (Eurofins). Because of the the previous studies in animals had demonstrated(26). nature of the various treatments, the study could not be fully The pharmacokinetics of orally ingested Ca-HMB in blinded. 170.106.35.234 capsules were originally described by Vukovich et al.(14) and showed a time to peak plasma level of HMB of 2 h after a 1 g dosage. Recently, Fuller et al.(15) demonstrated that Study design HMB-FA gel form increased plasma HMB concentrations , on All studies were conducted after a 10- to 12-h overnight fast. 30 Sep 2021 at 15:04:06 in a shorter period of time (30–40 min) with a greater Subjects arrived at the laboratory in the morning, and an blood clearance rate of HMB compared with the Ca-HMB intravenous catheter was inserted into a forearm vein using capsule. A recent 12-week exercise study by Wilson et al.(27) sterile procedures. A preingestion blood sample, of approxi- used these favourable advantages to test the efficacy of mately 7 ml, was drawn. The treatment was then consumed HMB-FA administered 30 min before exercise in a periodised over approximately a 2 min period, and additional blood sam- training programme and showed increased strength, lean body ples (7 ml) were taken at 2, 5, 10, 15, 25, 35, 45, 60, 90, 120, 180, , subject to the Cambridge Core terms of use, available at mass gain and muscle power in a highly trained athlete 360 (6 h), 720 (12 h) and 1440 (24 h) min after the ingestion of population(27), demonstrating the importance of dosage timing the supplement; 24 h urine collection was also obtained. Plasma and peak levels of HMB for maximisation of exercise and urine samples were stored at −80°C until later analyses of benefits. The current human study was thus designed to HMB. Serum and blood samples at times 0 and 1440 min were compare the pharmacokinetics of the current commercially analysed by a commercial laboratory (LabCorp.) for glucose, available delivery form of Ca-HMB in a capsule and compared uric acid, blood urea nitrogen, creatinine, Na, K, Cl, Ca, CO ,P, this with the newly designed delivery form of HMB-FA in a 2 protein, albumin, globulin, total bilirubin, alkaline phosphatase, gelcap. Our results confirmed the previous findings of Fuller estimated glomerular filtration rate (eGFR), creatine kinase, et al.(15) and showed that HMB-FA had superior bioavailability lactate dehydrogenase, aspartate aminotransferase, alanine in humans as assessed by blood levels, and improved aminotransferase, γ-glutamyl transpeptidase, Fe, TAG, HDL, clearance. LDL, VLDL and cholesterol; a complete blood count was also performed. Subjects completed a brief health questionnaire to report any physical symptoms (such as nausea, headache and Methods so on) they may have experienced during the experiment. https://www.cambridge.org/core/terms Human subjects After the 180 min blood sampling, each subject was provided with a standardised lunch (1673·6–2092 kJ (400–500 kcal)), We performed a randomised crossover study consisting of which was consumed at approximately 240 min postingestion; five men and five women (aged 21–32 years).