Name: evaluation of blood lactate and plasma insulin during high-intensity exercise by antecubital vein catheterization
Uploaded: 2018-05-18T12:30:00
Description: Watch this Scientific Journal Video about Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization at JoVE.com

The authors would like to acknowledge Dr. P. Karagiannis for reading the paper and Professor T. Hoyo for his technical support.

All methods described here have been approved by the local ethical committee (Doshisha University Ethical Committee: 15033), and were in strict accordance with the standards set by the Declaration of Helsinki. Informed consent was obtained from all participants. Participants were male recreational athletes (age, 20.1 &#177; 1.2 years; weight, 69.7 &#177; 6.2 kg; height, 176.1 &#177; 5.8 cm; body fat 13.6 &#177; 1.5% and VO2max, 52.6 &#177; 6.6 mL/min1/kg1).
1. Es...

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In this study, the antecubital vein was catheterized, and blood samples were successfully collected during a high interval exercise. As an example of the use of this methodology, blood samples were analyzed for blood lactate and (after plasma separation) for plasma insulin. The main advantage of this method is that it allows the collection of blood samples in short periods of time. At the same time, it does have some limitations. For instance, the limited exercise modalities that allow its use; the number of assistant ne...

The measurement of metabolic and endocrinal markers during physical activity is of relevance to understanding the physiological implications of different exercise modalities. During some exercise modalities (e.g., high-intensity interval exercise (HIIE)), blood metabolites and hormonal levels fluctuate in relatively short time1,2,3.
Many sports (i.e., soccer, basketball, and rugby) consist of short periods of high-intensity exercise separated by low intense exercise and/or passive periods4,5. HIIE presents similarities with the aforementioned sports and is therefore widely used as a training method for athletes6. Moreover, because of its similarity with sports, HIIE has been used as a model to study exercise physiology7. Blood sampling is critical to measure physiological variables, and it is frequently used for research purposes and the evaluation of the physical condition. However, blood sampling during HIIE can be technically difficult because of the short time between sample collections. As a solution to this problem, here we present a method to obtain blood samples by antecubital venous catheterization in short periods of time while subjects perform HIIE.

<table border="0" cellpadding="0" cellspacing="0" width="626">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Cycle ergometer</td>
			<td style="width:71px;">Monark</td>
			<td colspan="2" style="width:339px;">MONARK Ergomedic 874 E</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Catheter</td>
			<td style="width:71px;">Terumo</td>
			<td style="width:172px;">SR-FF2032</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Vacutainer</td>
			<td style="width:71px;">Terumo</td>
			<td>PZ-D03</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Tube for catheter</td>
			<td style="width:71px;">Terumo</td>
			<td style="width:172px;">SP-PTW30L02</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heparin</td>
			<td style="width:71px;">Terumo</td>
			<td>PF-10HF10UA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lactate pro2</td>
			<td style="width:71px;">Arkray</td>
			<td>&#65324;&#65332;-1730</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Insulin ELISA kit</td>
			<td>Abnova</td>
			<td>ELISA Kit KA0921</td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluation of blood lactate and plasma insulin during high-intensity exercise by antecubital vein catheterization

The measurement of metabolic and endocrinal markers during physical activity is of relevance to understanding the physiological implications of different exercise modalities. During some exercise modalities (e.g., high-intensity interval exercise), blood metabolites and hormonal levels change in short periods of time. In the present study, we describe a method to catheterize the antecubital vein, which allows the collection of several blood samples during exercise. Insulin and venous lactate concentrations were measured during high-intensity exercise by the application of the described method. The exercise consisted of three 30 s bouts of high-intensity exercise separated by 4 min of recovery. After the last recovery period, a Wingate test was performed. Blood samples from the antecubital vein were obtained before and after each 30 s bout and before and after the Wingate test. As a result, it was possible to evaluate the plasma insulin and venous blood lactate variations during the exercise.

As representative results, we used this protocol to measure blood lactate and insulin variation in six subjects during a HIIE in a cycle ergometer. Peak power and Average Power for the Wingate test were 732 &#177; 120 and 549 &#177; 84 Watts, respectively. Average Power was used to calculate the power output (494 &#177; 75 Watts) during the 30 s bouts of the HIIE.
Figure 2 shows plasma insulin and b...

Watch this Scientific Journal Video about Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization at JoVE.com

Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization

Here we present a protocol to obtain several blood samples from the antecubital vein during high-intensity interval training. This protocol may be useful for the measurement of blood metabolites and endocrinal markers during exercise.

The measurement of metabolic and endocrinal markers during physical activity is of relevance to understanding the physiological implications of different ...

The overall goal of this experiment is to obtain multiple venous blood samples from a single subject during cycle ergometer exercise. This measure can help answer key questions in porous physiology and hormonal and metabolized variation during different exercise modalities. The main advantage of this technique is that multiple blood samples can be obtained, even during exercise.Demonstrating the procedures will be Rie Oosaki who collaborate with our laboratory. To evaluate blood, hormonal, and metabolite dynamics from the antecubital vein during high intensity interval exercise protocol, have the subject first perform a five-minute warmup on the ergometer at low-intensity. After a five minute rest, have the subject perform three 30 second high-intensity exercise intervals at 90 percent intensity at the power average determined from a previously performed Wingate test, separated by four minutes of rest per exercise interval.For blood sample collection, tie a tourniquet around the arm selected for the catheterization and identify the antecubital vein. Sterilize the skin with alcohol and insert the tip of a sterilized cannula into the antecubital vein and in an approximately 15 degree angle to the skin. If the cannulation is successful, a backflash of blood should be observed.After removing the needle, secure the cannula, and place a cotton tissue under the needle. Remove heparin with an empty collection tube and connect a new container for blood collection to the cannula. Aspirate approximately 2.5 ml of blood and close the butterfly, immediately placing the sample on ice.Then, inject heparin the full length of the tube to avoid blood sample coagulation, and have the subject begin the next series of exercise intervals. After the last sample has been collected, carefully remove the cannulation tube and disinfect the skin with more alcohol. Gently press a piece of sterile cotton onto the collection site to minimize bleeding and transfer 1.5 ml of blood from each sample into individual 1.5 ml microcentrifuge tubes on ice.Use the lactate detector, measure the blood lactate of each 1 ml blood sample, and centrifuge twice the 1.5 ml blood sample aliquots. Transfer the supernatants into new mirocentrifuge tubes, and centrifuge the samples again in transfer supernatants to new tubes. Then store the purified plasma samples at minus 80 degree Celsius until plasma insulin analysis by ELISA.In this representative experiment, plasma insulin levels decreased in all six patient peripheral blood samples during the first and third 30 second high intensity interval exercise periods, but increased during the second interval in recovery periods. Plasma lactate, however, decreased during the second and third 30 second high intensity interval exercise periods and increased during the first 30 second interval and recovery periods. While attempting this procedure, it is important to remember to maintain the blood samples at no colder than 40 degrees Celsius.Following this procedure, other methods like Brodgar's Analysis can be performed to answer additional questions about pH level variations during exercise. After watching this video, you should have a good understanding of how to collect consecutive blood samples from the antecubital vein during cycle ergometer exercise.

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