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This study examined the relationship between collagen content as determined by hydroxyproline assay with other measures of connective tissue, shear force and tenderness in lamb muscle. Samples were taken from both m. longissimus lumborum (LL, loin) and the m. semimembranosus (SM, topside) of 99 lambs. Sensory tenderness and compression of the LL were not correlated to any measure of collagen or connective tissue, with one exception where compression was correlated (r. =. -. 0.28; P

Collagens are highly abundant mammalian proteins that contain a high content of hydroxylated amino acids such as hydroxyproline. We have exploited the high hydroxyproline content of collagen and developed a method for hydroxyproline quantification as a measure of collagen content in muscle samples. The novel method utilizes a highly selective and sensitive method of multiple reaction monitoring (MRM) by mass spectrometry. The analytical method is simple, rapid (5min), convenient (no derivatization), precise (17% RSD), accurate (90-108%), sensitive (4.88nmol/L) and linear (R(2)0.999) over three orders of magnitude (5-5000nmol/L).

The high molecular weight glycosaminoglycan hyaluronan (HA) is an essential component of the extracellular matrix (ECM), however, the link between HA regulation and development of the adipocyte ECM, which is essential for differentiation, remains undefined. Hyaluronan synthase gene expression, HA synthetic rate and molecular weight during differentiation of 3T3-L1 pre-adipocytes were compared to undifferentiated 3T3-L1 pre-adipocytes and non-adipogenic NIH/3T3 fibroblasts. In the 3T3-L1 pre-adipocytes, the predominant genes associated with HA metabolism were found to be HA synthase-2 (Has-2) and hyaluronidase-2 (Hyal-2) demonstrating a co-regulation of expression which was stimulated by adipogenic induction consequently resulting in increased synthesis of high molecular weight HA (>10 MDa) and its simultaneous degradation. Accumulation of HA correlated positively with cell number, although synthetic rate was inversely related suggesting a regulatory feedback mechanism. Within 24 h post-induction, pre-adipocytes responded with a higher HA synthetic rate and later, accumulated cytoplasmic lipid. In contrast, undifferentiated pre-adipocytes had a reduced HA synthetic rate during clonal expansion and did not accumulate lipid. HA was continuously and rapidly metabolised throughout 3T3-L1 adipogenesis, where terminal differentiation coincided with the increased generation of low molecular weight, angiogenic HA fragments, a likely prerequisite for concurrent neovascularisation of adipose tissue. This study has highlighted a relationship between HA metabolism and adipocyte differentiation, suggesting that the balance between the formation and regulation of the adipocyte extracellular matrix is finely coordinated in a growth phase-specific dependent manner.

Multiple reaction monitoring (MRM) mass spectrometry may be regarded as the gold standard methodology for quantitative mass spectrometry and has been adopted for the analysis of small molecules especially within the pharmaceutical industry. It can also be applied to the analysis of peptides and proteins and to the measurement of the basic building blocks of proteins, amino acids. Here, we describe the application of MRM mass spectrometry to the measurement of hydroxyproline after acid hydrolysis of various animal tissues. We show that the measurement of hydroxyproline provides an accurate and reliable estimate of the collagen content of such tissues and may be a useful indicator of meat tenderness.

The effect of growth path, as opposed to advancing age, on the biophysical and biochemical properties of muscle connective tissue was investigated. Nine-month old Brahman-cross steers were grown across either an uninterrupted path, or paths that incorporated weight-loss and then weight gain on two different diets: one group was realimented on pasture, whilst the other was realimented on a grain-based diet. Biophysical attributes of connective tissue toughness (Compression and Adhesion) in the semitendinosus muscle, were significantly reduced by treatment (P

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are the n-3 polyunsaturated fatty acids (n-3 PUFA) for which there is ample evidence of human health benefits, and these are also the fatty acids for which there are cut-off points for ‘source’ and ‘good source’ claims. Two consecutive experiments were conducted to determine the effect of finishing systems on the n-3 PUFA content of lamb meat in Western Australia. In experiment I, a 4-week feeding experiment was conducted using 48 Poll Dorset × Merino lambs. The lambs were divided into two lots of 24 (12 males and 12 females) and randomly assigned to either concentrate (C1) finishing on commercial lamb finishing pellets or pasture (P) finishing on kikuyu (Pennisetum clandestinum) pasture. In experiment II, 28 lambs were divided into two groups of 14 lambs (seven males and seven females) and assigned to finishing either on concentrate pellets (C2) or concentrate pellets plus linseed (C2-L). This second experiment was conducted indoors for 10 weeks. The initial liveweight (mean ± s.e.) of the lambs was 43 ± 0.6 and 32.5 ± 0.9 kg for experiments I and II, respectively. At the end of experiment II, three chops each were sampled from the leg, loin, forequarter and neck region of each carcass. The final liveweight (42 ± 0.8 v. 50 ± 1.2 kg), hot carcass weight (19 ± 0.5 v. 24 ± 0.7 kg) and GR depth (5.6 ± 0.6 v. 12.8 ± 0.6 mm) were lower (P < 0.05) for P than C1 lambs. In contrast, C2 and C2-L lambs had similar final liveweight (44 ± 0.7 v. 45 ± 0.9 kg), hot carcass weight (19 ± 0.3 v. 20 ± 0.5 kg) and GR depth (13 ± 1.3 v. 14 ± 1.2 mm). In experiment I, the total n-3 PUFA yields for C1 and P lambs in the M. longissimus lumborum were 67 ± 2.5 and 78 ± 3.2 mg per 100 g muscle, respectively. The EPA plus DHA yields were 17 and 21 mg per 100 g muscle, respectively. The sum of the long-chain (≥C20) n-3 PUFA EPA, docosapentaenoic acid and DHA for C1 and P lambs were 30 and 37 mg per 100 g, respectively. Sex had no effect on any of the n-3 fatty acids. In experiment II, the total n-3 PUFA yields for C2 lambs were 61, 54, 60 and 104 mg per 100 g for leg, loin, forequarter and neck chops, respectively. The respective values for C2-L lambs were 153, 138, 139 and 178 mg per 100 g muscle. The claimable EPA plus DHA yields for C2 lambs were 13, 10, 12 and 15 mg per 100 g of trimmed leg, loin, forequarter and neck chops, respectively. The respective values for C2-L lambs were around 2-fold higher at 27, 21, 25 and 23 mg per 100 g raw meat. All the samples from pasture-finished and linseed-supplemented groups met the 30 mg cut-off point for ‘source’ claim in Australia when the computation was based on 100 g cooked lamb serve (140 g raw). We conclude that pasture-finished lambs have more n-3 PUFA per serve than their counterparts finished indoors on commercial pellets. Further, supplementing indoor-finished lambs with linseed provided equivalent n-3 PUFA per serve to finishing lambs on pasture. Supplementation with an omega-3 source improved omega-3 per serve across the whole carcass irrespective of sex.