Influence of rate of inclusion of microalgae on the sensory characteristics and fatty acid composition of cheese and performance of dairy cows.

Modification of milk and cheese fat to contain long-chain n-3 fatty acids (FA) by feeding microalgae (ALG) to dairy cows has the potential to improve human health, but the subsequent effect on the sensory attributes of dairy products is unclear. The objective was to determine the effect of feeding dairy cows different amounts of ALG that was rich in docosahexaenoic acid (DHA) on milk and cheese FA profile, cheese sensory attributes, and cow performance. Twenty Holstein dairy cows were randomly allocated to 1 of 4 dietary treatments in a 4 × 4 row and column design, with 4 periods of 28 d, with cheddar cheese production and animal performance measurements undertaken during the final 7 d of each period. Cows were fed a basal diet that was supplemented with ALG (Schizochytrium limancinum) at 4 rates: 0 (control, C), 50 (LA), 100 (MA), or 150 g (HA) of ALG per cow per day. We found that both milk and cheese fat content of DHA increased linearly with ALG feed rate and was 0.29 g/100 g FA higher in milk and cheese from cows fed HA compared with C. Supplementation with ALG linearly reduced the content of saturated FA and the ratio of n-6:n-3 FA in milk and cheese. Supplementation with ALG altered 20 out of the 32 sensory attributes, with a linear increase in cheese air holes, nutty flavor, and dry mouth aftertaste with ALG inclusion. Creaminess of cheese decreased with ALG inclusion rate and was positively correlated with saturated FA content. We also observed a quadratic effect on fruity odor, which was highest in cheese from cows fed HA and lowest in LA, and firmness and crumbliness texture, being highest in MA and lowest in HA. Supplementation with ALG had no effect on the dry matter intake, milk yield, or live weight change of the cows, with mean values of 23.1, 38.5, and 0.34 kg/d respectively, but milk fat content decreased linearly, and energy-corrected milk yield tended to decrease linearly with rate of ALG inclusion (mean values of 39.6, 38.4, 37.1, and 35.9 g/kg, and 41.3, 41.3, 40.5, and 39.4 kg/d for C, LA, MA, and HA, respectively). We conclude that feeding ALG to high-yielding dairy cows improved milk and cheese content of DHA and altered cheese taste but not cow performance, although milk fat content reduced as inclusion rate increased.

Evolvability meets biogeography: evolutionary potential decreases at high and low environmental favourability.

Understanding and forecasting the effects of environmental change on wild populations requires knowledge on a critical question: do populations have the ability to evolve in response to that change? However, our knowledge on how evolution works in wild conditions under different environmental circumstances is extremely limited. We investigated how environmental variation influences the evolutionary potential of phenotypic traits. We used published data to collect or calculate 135 estimates of evolvability of morphological traits of European wild bird populations. We characterized the environmental favourability of each population throughout the species' breeding distribution. Our results suggest that the evolutionary potential of morphological traits decreases as environmental favourability becomes high or low. Strong environmental selection pressures and high intra-specific competition may reduce species' evolutionary potential in low- and high- favourability areas, respectively. This suggests that species may be least able to adapt to new climate conditions at their range margins and at the centre. Our results underscore the need to consider the evolutionary potential of populations when studying the drivers of species distributions, particularly when predicting the effects of environmental change. We discuss the utility of integrating evolutionary dynamics into a biogeographical perspective to understand how environmental variation shapes evolutionary patterns. This approach would also produce more reliable predictions about the effect of environmental change on population persistence and therefore on biodiversity.

Effects of trans-10, cis-12 conjugated linoleic acid on ovine milk fat synthesis and cheese properties.

Trans-10, cis-12 conjugated linoleic acid (CLA) reduces milk fat synthesis in sheep in a manner similar to that seen in dairy cows, but its effects on cheese yield and flavor are unknown. Additionally, when dietary energy supply is restricted, CLA can increase milk and milk protein yield, which may alter cheese yield and eating quality. The objectives of the study were to examine the effects of supplementing ewe diets with a rumen-protected source of CLA at a high and low dietary energy intake on milk fat and protein synthesis and on cheese yield and eating quality. Sixteen multiparous ewes were randomly allocated to 1 of 4 dietary treatments: a high (6.7 Mcal of metabolizable energy/d) or low (5.0 Mcal of metabolizable energy/d) feeding level that was either unsupplemented or supplemented with 25 g/d of a lipid-encapsulated CLA (to provide 2.4 g/d of CLA) in each of 4 periods of 21 d duration in a 4 x 4 Latin square design. There was no effect of treatment on milk yield (g/d), but milk fat percentage and milk fat yield were reduced by 23 and 20%, respectively, in ewes supplemented with CLA. Milk fatty acid concentration (g/100 g) of chain length < C16 was decreased and > C16 was increased in milk and cheese following CLA supplementation, whereas decreasing the feeding level increased fatty acids > or = C16. Milk fat contents of CLA were 0.01 and 0.12 g/100 g of fatty acids for the unsupplemented and CLA-supplemented treatments, respectively, whereas cis-9, trans-11 CLA was unaffected by CLA supplementation. There was no main effect of treatment on cheese yield, which was 0.11 +/- 0.001 kg of cheese/kg of milk, but cheese yield was highest, at 0.12 +/- 0.001 kg/kg, when made from milk of ewes fed the high feeding level + unsupplemented treatment. Cheese made from the milk of ewes supplemented with CLA, compared with the unsupplemented diet, was rated (scale 0 to 10) higher in the creaminess (2.1 vs. 1.4; SEM 0.15) and less oily (0.8 vs. 1.3; SEM 0.17) attributes, and was preferred overall (4.5 vs. 3.9; SEM 0.21). Cheese produced from sheep on the high vs. low feed level was rated less yellow (2.8 vs. 4.2; SEM 0.11), less salty (1.9 vs. 2.3; SEM 0.15), and more sour (1.5 vs. 1.1; SEM 0.13). We concluded that the effect of feeding level on animal performance and cheese characteristics was small, whereas supplementing the diets of ewes with a ruminally protected CLA source reduced milk fat yield, did not affect cheese yield, and beneficially altered the flavor characteristics of the cheese.

Nutritional evaluation of solar dried sardines as a ruminant protein supplement.

Solar dried sardines of various qualities were analyzed for nutrient content and for nutrient digestibility and nitrogen balance in sheep. Additionally, key serum enzymes and metabolites were examined to identify potential toxic effects. Dried sardine protein, ether extract and crude ash content ranged from 65 to 39%, 5.7 to 5.1%, and 22 to 51%, respectively, for high to low quality dried sardines. Visual appraisal of the dried sardines did not appear to be very reliable for determining dried sardine quality. Crude protein content was highly correlated (r(2)=0.962) with crude ash content based on the following equation (dry matter basis): crude protein%=86.0 - (0.961xcrude ash%). Therefore, crude ash could be used to estimate crude protein content and dried sardine quality. Digestibility in Omani sheep was determined on diets composed of a composite of dried sardines collected from Al-Batinah region of the Sultanate (the average crude protein content was 51%) and compared to that of a similar diet using soybean meal (also 51% crude protein) as the major protein source. The digestibility of crude protein, ether extract, total carbohydrates, digestible energy and metabolizable energy in diets utilizing dried sardines versus the soybean meal diet were 74 versus 76%, 69 versus 68%, 67 versus 69%, 64 versus 66%, and 52 versus 56%, respectively. Although, protein digestibility was similar, nitrogen balance data suggested that nitrogen in the dried sardine diet was not as efficiently utilized as nitrogen in the soybean meal diet. Key serum enzymes and metabolites did not suggest that dried sardines at 10% of the total dietary intake would cause metabolic disturbances in sheep. The study suggests that solar dried sardines may be effectively used in ruminant diets but may not have the protein by-pass value of commercial, processed fish meals.

Muscle respiration in rats is influenced by the type and level of dietary fat.

Experiments were conducted to elucidate the role of muscle in the enhanced thermogenic response found in rats fed diets enriched with polyunsaturated fatty acids. Isolated soleus muscle respiration and plasma cholesterol and triglyceride concentrations were determined in rats (approximately 128 g wt, 5 wk age) fed diets (minimum 3 wk) containing coconut oil, beef tallow or safflower oil at 20, 40 or 60% of the total dietary energy in a 3 x 3 factorial design (5-6 rats per treatment). Diet type did not affect plasma cholesterol concentrations but plasma triglycerides were lower (P < 0.01) in rats fed safflower oil-based diets. Greater levels of fat in the diet resulted in higher (P < 0.01) plasma cholesterol concentrations and lower (P < 0.01) plasma triglyceride concentrations. Rats fed coconut oil had lower (P < 0.05) rats of soleus muscle respiration compared with rats fed the other two diets. This lower respiration rate was not related to changes in protein synthesis (cycloheximide-sensitive respiration). However, this change may partially be related to enhanced Na+,K+ transport (ouabain-sensitive respiration). The results indicate that muscle is partially responsible for the enhanced thermogenic response found in rats fed diets enriched with polyunsaturated fatty acids and that enhanced ion transport contributes to this response in muscle.

Prepartum nutrition and solar radiation in beef cattle: II. Residual effects on postpartum milk yield, immunoglobulin, and calf growth.

Residual effects of nutrition and solar radiation during the last two-thirds of gestation on postpartum milk yield, immunoglobulin (Ig) G and M in both colostrum and calf serum, and calf growth were determined in beef cattle. Nineteen mature, multiparous crossbred cows (Bos taurus) at d 90 of pregnancy were assigned to either low (LO, 70% NRC total energy intake) or high (HI, 110% NRC total energy intake) nutritional level (sudangrass hay) and to either shade (S) or no shade (NS) treatments in a 2 x 2 factorial experiment. After parturition, all cows were moved into a large paddock and managed uniformly. Calf weights and calf serum were collected within 1 d postpartum, thereafter at 2-wk intervals for the next 12 wk, and then at 4-wk intervals until weaning. Colostrum samples were taken from the cow and milk yields were determined by the "weigh-suckle-weigh" technique. Neither prepartum nutrition nor environment influenced lactational performance of the dam. Concentrations of IgG were elevated in the colostrum of LO cows (15.3 vs 7.8 g/100 mL, LO vs HI, respectively; P < or = .05) but were not affected by shading. The patterns of IgG concentration in the calf serum were not altered by prepartum nutrition or environment; however, the pattern of IgM concentrations was greater (P < or = .01) in calves from S cows than in those from NS cows. This difference in IgM profile did not seem to be due to any residual effect from prepartum treatments. Postnatal growth of calves from birth until weaning were similar across all prepartum treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Prepartum nutrition and solar radiation in beef cattle: I. Relationships of body fluid compartments, packed cell volume, plasma urea nitrogen, and estrogens to prenatal development.

Adaptations in body fluid pools during pregnancy were monitored in cows (n = 19) assigned to either low (LO, 70% NRC total feed intake) or high (HI, 110% NRC total feed intake) nutritional level (sudangrass hay) and to either shade (S) or no shade (NS) treatments in a 2 x 2 factorial experiment. Body water distribution (empty body water [EBW] by urea dilution; extracellular water [ECW] by thiosulfate dilution; intracellular water [ICW] by difference; plasma volume by Evans Blue dilution; interstitial water [ISW] by difference) was measured at 4-wk intervals beginning at 3 mo of pregnancy until birth and then immediately after birth. Both EBW and ICW in LO cows showed a steady decline (P < or = .05), whereas HI cows tended to maintain these body pools throughout gestation. Shading did not affect the pattern of change in EBW; however, it did produce a greater (P < or = .05) ICW in the S than in the NS cows throughout gestation. Generally, other body fluid pools (ECW, ISW, and plasma) were either not affected, or were just slightly affected, by shading or nutrition. Most of the body fluid pools (EBW, ECW, ICW, and ISW) inversely followed the seasonal changes in solar radiation. Calf birth weights were not affected by treatments but were moderately correlated to EBW (r = .49; P < or = .05) and ICW (r = .50; P < or = .05). Plasma urea nitrogen change, although not affected by nutrition, was affected (P < or = .05) by shading.(ABSTRACT TRUNCATED AT 250 WORDS)

Energy metabolism in isolated chick (Gallus domesticus) gastrocnemius and tilapia (Tilapia mossambica) epaxial muscle at various temperatures in vitro.

Muscle respiration experiments on inhibitor dosage (experiment 1), muscle preparation (tendons removed vs. unstretched vs. stretched muscles; chick muscle only; experiment 2) and media temperature (26.5, 32, 37, 42 degrees C; experiment 3) were conducted on chick (Gallus domesticus) gastrocnemius and tilapia (Tilapia mossambica) epaxial muscle in vitro. Experiment 1: The dosage of cycloheximide and ouabain required for maximum inhibition of protein synthesis and Na+,K+ ATPase, respectively, in chick and tilapia muscle was approximately 6 x 10(-5) M. Experiment 2: Removing the tendons of chick muscle decreased (% inhibition, P = 0.05) cycloheximide-sensitive respiration compared to stretched and unstretched muscles (tendons intact). However, muscle preparation had little influence on ouabain-sensitive respiration. Experiment 3: Cycloheximide-sensitive respiration tended to increase (microliter O2/mg DNA.hr, P = 0.054) with media temperature in tilapia muscle. Chick muscle was less responsive in this respect. Ouabain-sensitive respiration increased at lower temperature in chick muscle (% inhibition, cubic relationship, P = 0.001) and at higher temperature in tilapia muscle (% inhibition, quadratic relationship, P = 0.0002).

Effects of the purine biosynthesis pathway inhibitors azaserine, hadacidin, and mycophenolic acid on the developing ovine corpus luteum.

De novo synthesis precursors of the purine second messengers adenosine, guanosine and inosine are adenosine, guanosine and inosine monophosphate (AMP, GMP, IMP), respectively. Inhibitors of the de novo purinergic synthesis pathways for AMP, GMP and IMP by hadacidin, mycophenolic acid and azaserine, respectively, or adenosine, guanosine or inosine alone or in combination were given every 4 or 6 hours in vivo. Treatments were given into the ovarian vascular pedicle sheath adjacent to the luteal-bearing ovary in three separate experiments to determine whether purines were involved in development of the corpus luteum. Hadacidin lowered AMP (p < or = 0.01) and azaserine tended to lower IMP and the GMP: AMP ratio (p < or = 01) while mycophenolic acid tended to lower the GMP:AMP ratio (p < or = 0.1) in luteal tissue. Azaserine (150 mg) increased progesterone (p < or = 0.01) on some days but guanosine or inosine had no effect on profiles of progesterone in jugular blood of the developing corpus luteum (p > or = 0.1). Azaserine (500 micrograms) tended to lower progesterone in jugular blood (p < or = 0.1) while profiles of progesterone did not differ among guanosine or inosine or adenosine, guanosine and inosine plus hadacidin, mycophenolic acid and azaserine treatment groups compared to controls (p > or = 0.1). Weights of corpora lutea or composition of cell types in the corpus luteum or their viability were not affected by adenosine, guanosine, inosine, hadacidin, mycophenolic acid or azaserine (p > or = 0.1). Since profiles of jugular progesterone did not differ between treatments during development of the corpus luteum, these results suggest that progesterone production by the developing corpus luteum is a) less dependent on de novo synthesized purines or b) there may be a non-purinergic-dependent second messenger system controlling biosynthesis of steroids in the developing ovine corpus luteum.

Chronic heat stress and prenatal development in sheep: II. Placental cellularity and metabolism.

Aspects of placental protein and energy metabolism were examined in pregnant ewes subjected to either thermoneutral (TN, 18 to 20 degrees C, 30% humidity, n = 7) or hot (H, 30 to 40 degrees C, 40% humidity, n = 5) temperatures through mid and late gestation. Fetal and placental weights and total content of protein, RNA, and DNA were reduced (P less than .001) in H ewes. Placental protein and RNA concentrations (mg/g) were not different, and DNA concentrations were slightly greater (P less than .1), in H vs TN ewes. Thus, heat seemed to greatly reduce total cell number and placentome size and only slightly decrease cell size. Ratios of RNA to DNA indicated a reduced capacity for protein synthesis in H placenta. However, in vitro fractional rates of protein synthesis in tissue slices from the fetal and maternal placenta and from the myoendometrium were not different between TN and H ewes. The H ewes had greater placental protein concentrations of hydroxyproline and glycine, perhaps suggesting a greater collagen content. In vitro oxygen consumption of fetal placenta, but not of maternal placenta or myoendometrium, was lower in H than in TN ewes. This lower oxygen consumption was partially due to a lower Na+,K+ ATPase-dependent oxygen consumption.

Growth and metabolism in somatotropin-treated steers: I. Growth, serum chemistry and carcass weights.

Growth, feed conversion, serum chemistry and gross slaughter characteristics were determined in 20 steers (initially 9 mo of age, 231 +/- 18 kg) receiving daily injections of either saline (S) or recombinantly derived bovine somatotropin (rBST, 20.6 mg/d) for 112 d. Live weight gains were 15% greater for steers treated with rBST than for those treated with S. Feed intake was not different between S- and rBST-treated steers; thus, feed conversion was 12% more efficient in rBST steers. Scanogram backfat measurements were not affected by treatments. Serum electrolytes, protein, glucose and most enzyme activities were similar in S and rBST steers. Serum urea, creatinine and cholesterol (toward the end of treatment) concentrations, however, were lower (P less than .05) in rBST steers, suggesting that nitrogen retention was increased and lipid turnover was decreased by rBST. Total (P less than .1) and conjugated (P less than .05) bilirubin concentrations and glutamate-pyruvate transaminase activity (P less than .05) were lower in rBST steers. Carcass weights were not altered, but dressing percentages were lower (P less than .05) in rBST steers. This indicated that weight gain response to rBST was primarily in noncarcass components; further examination showed that this gain was predominantly in gut fill (approximately 2/3 of the greater live weight gain in rBST steers). Alternative protocols, such as administering the hormone to younger animals and (or) for a longer duration, may be necessary in order to achieve desirable responses in carcass growth.

Growth and metabolism in somatotropin-treated steers: II. Carcass and noncarcass tissue components and chemical composition.

Carcass and noncarcass tissue compositional characteristics were determined in growing Hereford steers treated with daily subcutaneous injections (20.6 mg/d) of recombinantly derived bovine somatotropin (rBST) for 112 d. For carcass primal cuts, weights and rates of gain of bone, lean and total fat and site of fat deposition were not significantly affected by rBST treatments with the exception of a few tissues (loin total weight, flank total and lean weight and shank total weight). Lean to fat ratios, however, were greater (P less than .1) for the loin, flank, chuck and brisket. Weights and growth of individual muscles from the hindquarter were not affected by rBST administration. Weights and(or) average daily gains of the liver, kidneys, lungs and trachea and head were greater (P less than .05) in rBST-treated animals. Weights and (or) average daily gains were greater (P less than .1) in rBST-treated steers for water in the total body and carcass, for CP in the total body and noncarcass, and for ash in the total body, carcass and noncarcass. Ratios of CP to ether-extractable fat in the total body were greater (P less than .1) in rBST steers. These data indicate that rBST modified carcass lean and fat composition in cattle, but responses were modest compared to effects of somatotropin treatment of swine.

Growth and metabolism in somatotropin-treated steers: III. Protein synthesis and tissue energy expenditures.

Rates of in vivo protein synthesis in intercostal, sartorius and semitendinosus muscle and in the heart, liver, kidneys, rumen and jejunum were determined in 20 growing Hereford steers treated for 112 d with daily subcutaneous injections of either saline (S) or recombinantly derived bovine somatotropin (rBST; 20.6 mg/d). In vitro rates of protein synthesis and energy expenditures associated with Na+, K+ transport also were determined in external intercostal muscle, liver, kidneys and jejunum. Neither in vivo nor in vitro tissue protein fractional synthetic rates (mg/[g protein.d]), using either the plasma (P) or intracellular fluid (ICF) phenylalanine specific radioactivity for the precursor pool, were affected by rBST treatments. Energy expended on Na+, K+ transport was greater (P less than .1) in the livers of rBST-treated steers, which would increase the maintenance energy expenditures of these steers. Protein accretion rates in the liver, kidneys, stomach, hide, and head, feet and tail of rBST-treated steers were greater (P less than .05) than in S steers. Tissue amino acid profiles were not affected by rBST treatments except in the rumen, where profiles suggested that less collagen was present in rumen wall tissue protein of rBST steers. Plasma phenylalanine entry rates also were not affected by rBST treatment; muscle protein synthesis accounted for a minimum of 20% of this entry rate.

Chronic heat stress and prenatal development in sheep: I. Conceptus growth and maternal plasma hormones and metabolites.

Pregnant ewes were chronically exposed to thermoneutral (TN; 20 degrees C, 30% relative humidity) or hot (H; 40 degrees C 9 h/d, 30 degrees C 15 h/d, 40% relative humidity) environments between d 64 and 136 to 141 of pregnancy. They were sampled for blood at 14-d intervals during this period for measurement of plasma metabolites and hormones, then slaughtered and dissected to measure conceptus weights, dimensions and fetal organ weights. Rectal temperatures of H ewes were elevated .3 to 1.0 C degrees above those of TN ewes throughout the experiment. Voluntary feed intakes were not altered by heat exposure except after 120 d of pregnancy, when feed intake was about 25% lower (P less than .10) by H than by TN ewes. Blood 3-hydroxybutyrate concentrations were not affected by heat, but plasma glucose concentrations were greater in H than in TN animals after 120 d (P less than .05). Placental weight, reduced by 54% (P less than .001) by heat exposure of ewes, was correlated positively with fetal weight and correlated negatively with fetal/placental weight ratio, fetal brain/liver weight ratio and fetal relative heart weight. Late in pregnancy, plasma concentrations of progesterone, cortisol and placental lactogen were reduced (P less than .01) in H ewes, whereas triiodothyronine levels were markedly lower (P less than .03) at all stages of pregnancy. Plasma concentrations of prolactin were elevated dramatically (P less than .01) and a modest increase (P less than .03) in somatotropin levels was recorded in H ewes. These results are consistent with our hypothesis that heat-induced fetal growth retardation is secondary to a primary reduction in placental growth; this could be mediated partly by reduced peripheral activity of thyroid hormones. Heat-induced reductions in secretion of progesterone and ovine placental lactogen more likely were a consequence than a cause of placental stunting.

Energy expenditure associated with sodium/potassium transport and protein synthesis in skeletal muscle and isolated hepatocytes from hyperthyroid sheep.

The object of the present study was to determine the effect of thyroxine (T4) treatment of sheep on protein synthesis and associated energy costs in skeletal muscle and hepatocytes. Protein synthesis, and ouabain-sensitive and cycloheximide-sensitive respiration in isolated intercostal muscle and hepatocytes were determined in sheep after 5 weeks of daily injections of either saline or T4. Plasma T4 and total triiodothyronine (T3) concentrations were doubled and free T3 concentrations were quadrupled by T4 injections. The fractional rates of protein synthesis increased in isolated external intercostal muscle and hepatocytes from hyperthyroid sheep. Fractional rates of protein synthesis in isolated external intercostal muscle and hepatocytes were linearly correlated with plasma free T3 concentrations. Total oxygen consumption of muscle and hepatocytes was unaffected by T4 injections. Ouabain-sensitive respiration increased in hepatocytes and muscle of T4-treated animals. Cycloheximide-sensitive respiration was elevated in hepatocytes from hyperthyroid sheep. Cycloheximide-sensitive respiration in muscle was unaffected by T4 treatment. The present experiment demonstrates that T4 increases protein synthesis in ruminants. The energy expenditure in support of Na+, K(+)-ATPase and protein synthesis in skeletal muscle and hepatocytes may account for 34-60% of total cellular energy expenditure.

Breathing pattern, lung inflation reflex and airway tone in acrylamide neuropathy.

Slowly adapting pulmonary stretch receptors (SARs) participate in numerous respiratory reflexes, including the Hering-Breuer lung inflation reflex (HBR), and reflex control of respiratory rate and depth. In addition, SAR discharge may modulate airway tone. We studied the effect of acrylamide neuropathy, which causes reversible dysfunction of SARs and their myelinated vagal afferents, on the breathing pattern, HBR and airway tone in conscious dogs. As neuropathy evolved, breathing became slow and deep, and both the apnoeic and bronchodilatory responses to lung inflation were markedly reduced, findings consistent with SAR dysfunction. Recovery of clinical neuropathy and respiratory reflexes followed acrylamide withdrawal. Despite the obvious abnormalities of SAR-mediated reflexes in acrylamide-affected dogs, airway tone remained normal and sinus arrhythmia prominent in the presence of neuropathy, suggesting that small diameter vagomotor efferents are relatively resistant to acrylamide. Acrylamide neuropathy provides a useful preparation for the study of vagomotor function in animals in which feedback from myelinated pulmonary afferents is attenuated.

Capillary gas chromatographic analysis of amino acids in blood and protein hydrolysates as tert.-butyldimethylsilyl derivatives.

A method is given for a one-step derivatization and gas chromatography of amino acids in blood and protein hydrolysates. Blood samples are partially purified by solvent extraction. Protein hydrolysates are neutralized with a triethylamine solution. Then tert.-butyldimethylsilyl derivatives of the amino acids are prepared in a one-step procedure and separated on a 30-m fused-silica SE-30 capillary column. Except for tryptophan and cystine, amino acids are eluted within 30 min. Amino acids are derivatized more rapidly than their corresponding trimethylsilyl derivatives and do not degrade on the long fused-silica columns.

Branched-chain alpha-keto acid analysis in biological fluids: preparative clean-up by anion-exchange and analysis by capillary gas chromatography.

A method is given for the quantitative analysis of the alpha-keto derivatives of the branched-chain amino acids in physiological fluids. A sample containing alpha-ketovalerate and alpha-ketocaproate as internal standards is passed through a weak anion-exchange resin at neutral pH. After washing the resin with distilled water, the alpha-keto acids are eluted with 4 M hydrochloric acid--ethanol (50:50). Quinoxalinol derivatives are prepared directly in the eluent, extracted with methylene chloride, and trimethylsilylated. Separation of the derivatives is by capillary gas chromatography on a 30 m fused-silica SE-30 column. Chromatographic separation is superior to that reported for packed column methods, thereby permitting the use of alpha-ketovalerate and alpha-ketocaproate as internal standards.