Secondary elements of blood pH variation can influence the effort effectiveness based on adaptive changes within a group of elite athletes.

AIM: pH is the direct indicator of the body reaction following the activities performed. Establishing precise correlations between pH and blood biochemical parameters might support the balancing of values during periods of marked physical activity.
METHOD: We conducted a case study in a group of elite rowers. Twelve athletes were included in the study. Monitoring was carried out by collecting biological samples several times a day: in the morning, 80 minutes pre-workout, 12 hours after the last physical effort performed, at two different times, 10 days apart. Determinations were aimed at adapting the reported biochemical parameters depending on the effort performed. The following parameters were monitored: pH, HCO3, pCO2, pO2, BE, SBE, SBC, Ca++, Mg++, LDH, GPT, T-Pro, and Alb.
RESULTS: The mean value of pH found in athletes was 7.41±0.024. The value obtained was significantly correlated to biochemical parameters such as BE (2.32±1.79), SBC (1.67±1.45), SBE (2.70±1.75). However, bicarbonate (HCO3) was statistically significantly related with SBE, SBC, SBE, and pO2, but did not present a strong association with the pH value (p=0.094). However, values such as Alb, Ca++, LDH, BE, SBC are related to pH value as a result of variations in the data submitted.
CONCLUSIONS: The processed data evidence the fact that blood pH, in this case, is significantly influenced by a number of indices that correlate energy system activity, individual adaptation to effort, and the recovery process. The parameters under investigation (SBE, SBC, SBE, CPK, LDH) are associated with pH changes that could confirm the recovery efficiency of the athlete, along with a possible metabolic acidosis/alkalosis.

Introduction

pH represents the direct indicator of the body reaction following the activities performed. The acidity/alkalinity of the blood and of muscle intracellular compartments can affect athletic performance through the performed effort [1]. Over time, increased lactate concentration, both in the blood and in muscles, together with pH changes, was explained through the accumulation of lactic acid. Such an interpretation claims that due to relatively low pKa (pH=3.87) of the carboxylic acid functional group of lactic acid, there is an immediate and near total ionization of lactic acid across the range of cellular skeletal muscle pH (~6.2–7.0) [2].

Sustained intensive effort, both of average (1–8 minutes) and short durations will be directly affected by these factors [3]. Such activities result in significant metabolites concentration and hydrogen ions changes in skeletal muscle [3]. Increased hydrolysis ATP rate and glycolytic flux during maximal efforts will increase the accumulation of hydrogen ions (H+) [2], affecting both the metabolic process and muscle contraction. The hypothetical situation referred to can be confirmed by studies that present information indicating that ATP hydrolysis, which, along with glycolysis, represents the main source for hydrogen ion production, lead to a decrease in both muscle, and arterial pH [4]. Several studies demonstrate the negative role that hydrogen ions in excess have in lowering sports performance [5]. Also, within specific effort planned in sports performance, a series of systems contribute to intracellular and extracellular buffering systems in order to maintain pH homeostasis [6,7].

From a practical perspective it is claimed that protons released from ATP hydrolysis are not required for ATP resynthesis, glycolytic pathway. However, a number of protons formed from ATP glycolytic degradation are taken and transported to the mitochondria along with pyruvate. Some are used for the reduction of pyruvate to lactate, while a different part may be buffered by intracellular histidine. Therefore, non-buffered intracellular protons leave the cell, causing an alteration of the pH value and an increase of blood lactic acid associated with a decrease in muscle efficiency [8,9].

A number of compounds are provided in literature, of which NaHCO3 (sodium bicarbonate), and sodium citrate, represent buffering agents used to optimize pH [10,11].

Aim

Metabolic acidosis is the main factor of influence in muscle fatigue, reported under high intensity activity. Therefore, identification of secondary elements with a possible influence on metabolic variation and blood pH will represent an adaptation improvement during training periods, increasing muscle efficiency.

Material and method

A cross-sectional study was conducted after obtaining the approval of the ethics committee, and the informed consent of the subjects. We investigated the main relating factors which can influence pH within an elite group of rowers.

The study was conducted in November 2015 in Bucharest, Romania, in a training center for athletes. Twelve elite athletes (rowers), all males, with an average age 21.58±1.5 years, were included in the study.

By biochemical tests and acid-base balancer Radiometer ABL 835 analyzer (fixed), the venous biological material was analyzed, basal, in the morning, pre-workout, 12 hours after the last physical effort performed, collected in two different sessions, over a period of 20 days. The parameters analyzed included: pH, HCO3, pCO2, pO2, BE, SBE, SBC, Ca++, Mg++, LDH, GPT, T-Pro, Alb, pursuing the statistical relationship between the average values determined in the athletes. We used the following reference values: pH (7.35–7.45), HCO3 (22–26 mEq/l), pCO2 (35–45 mmHg), pO2 (75–100 mmHg), BE (−2/+2 mEq/l) SBE (−2/+2 mEq/liter), SBC (22–26 mEq/l), Ca++ (8.5–10.2 mg/dl), Mg++ (1.9–2.2 mg/dl), LDH (120–333 IU/L), GPT (0–33 IU/L), T-Pro (6.7–8.3 g/dL), Alb (3.8–5.1 g/dL).

Statistical evaluation was performed using Graphpad Prism 6.0 software. Among the statistical indicators used we mention: standard deviation (SD), standard error (SE), and coefficient of variation (CV). Fisher exact test was conducted to determine the odds ratio (OR). In order to obtain data normalization, Pearson correlation index (r) and Student’s t-test (pairs) were used. p<0.05 was considered statistically significant.

Results

The values obtained were examined during the investigation of biochemical parameters. These included pH, HCO3 (bicarbonate), pCO2 (partial pressure of carbon dioxide), pO2 (oxygen partial pressure), BE (base excess), SBE (excess base standard), SBC (hydrogen standard), Ca++ (calcium), Mg++ (magnesium), LDH (lactate dehydrogenase), ALT (transaminase glutamine pyruvate), T-Pro (total protein), and Alb (albumin). The connection between biochemical parameters was observed in order to determine the value of the data previously mentioned. Within the studied group (12 elite athletes) the age reported was 21.58±1.5 of years, 195.1±4.86 cm height, and 96±8.28 kg body weight, alongside the median of weight (93 kg), age (22 years), and athletes’ height (196.8 cm).

Mean blood pH was 7.41±0.024. Significant interrelationships were found between pH and 7 studied biochemical parameters. Blood gas testing did not establish a certain relationship, in statistical terms, between pH and pO2. Moreover, pCO2 value representing the equivalent amount of carbon dioxide dissolved in the blood reached a maximum monitored value of 40.57±2.3, coefficient of variation (CV) 7.89%, not being statistically significant (p=0.055). HCO3, with an average value of 25.29±2.06 mEq/l, was associated with the pH value mentioned above, without presenting any statistical significance between the 2 values.

HCO3 was significantly correlated with pCO2 (40.57±2.3), pO2 (82±5.01) (Figure 1), BE (2.16±1.4 mEq/iter), SBE (2.54±1.4), SBC (26±1.14) values, along with oxygen saturation (SAT) (96±1.14%) and metabolic equivalent (R) (0.97±0.3).

Figure 1

Relationship HCO3, pCO2, pO2 (p=0.001).

Establishing a connection between blood pH value determined in the case of athletes was conducted in association with BE, SBE, SBC parameters (Figure 2). Total protein (6.47±0.38 g/dl), was associated with a CV equivalent of 21.45%, indicating good homogeneity of the obtained data. The amount of albumin in a similar position, 3.48±0.19 (mg/dL) was significantly correlated with pH value, although the reported amount was not close to that ideal value recorded in the athletes. Ca++ (8.43±0.43 mg/dL), as a basic element in acid-base changes in terms of metabolic balance and muscular efficiency, was found statistically significant with pH value (p=0.0001), associated with 16.96% CV (homogeneous distribution). Lactate dehydrogenase (276.42±37.76), essential in energy production, was significantly associated with pH value, indicating an altered acid-base balance, with an increasing total value of LDH (Figure 2).

Figure 2

Establishing significance statistical networking between pH value, LDH, BE, SBC, T-Pro.

The statistical factors that influence LDH, which is associated with a CV (13.6%) issuing improved homogeneity data, are represented by total body protein (T-Pro). Its value is changed proportionally in the presence of factors that characterize the muscle status of athletes, CPK (216±135IU/L), GPT (35.38±20.9 IU/L), LDH, Ca++, as well as Alb (3.48±0.23). Pairing analyzed parameters, such as pCO2, pO2, establishes a number of hypotheses based on which we can draw connections between metabolic efficiency and body recovery. Among the parameters associated with the two values, in the case of pO2, and pCO2 statistical relationships are established with LDH, HCO3, SBE, BE, SBC, T-Pro, Alb, as well as with the pH value. Indirectly, the presented data highlights the contact between the proposed parameters for analysis, and blood pH value (Table I). Each value taken separately emits indirect connections due to the examination manner. The base excess (BE), representing the parameter that shows the excess base status in the blood, equivalent to a median of 0.55 mEq/L, is related to pH value, pO2, pCO2, HCO3, SBE, SBC, SAT, and metabolic equivalent (R).

Table I

Correlations between the analyzed parameters in relation to blood pH value.

Biochemical parameters determined		Paired Differences				Sig. (2-taled)	Shapiro-Wilk normality test	Passed normality test
Analyzer 1	Analyzer 2	CV%	N	95% Confidence Interval of the Difference
				Lower	Upper
pH	BE	166	24	0.1952	0.7831	0.004	0.936	YES
	SBE	150	24	0.2742	0.8134	0.001	0.943	YES
	SBC	5.23	24	0.1277	0.7548	0.011	0.954	YES
	T-Pro	10.1	24	0.4181	0.8620	0.000	0.940	YES
	Alb	6.62	24	0.1216	0.7521	0.012	0.961	YES
	Ca++	7.56	24	−0.8426	−0.3580	0.000	0.887	NO
	LDH	26.1	24	−0.8778	−0.4702	0.000	0.886	NO
HCO3	pCO2	7.89	24	0.3841	0.8512	0.000	0.931	YES
	pO2	8.88	24	−0.7570	−0.1328	0.011	0.925	YES
	R		24	−0.8910	−0.5160	0.000	0.912	YES
	BE	166	24	0.8347	0.9680	0.000	0.9366	YES
pCO2 pO2	BE	166	24	0.1637	0.7702	0.007	0.9366	YES
			24	−0.7959	0.2278	0.003
	SBE	150	24	0.08885	0.7374	0.018	0.9431	YES
			24	−0.7848	−0.1995	0.004
	SAT	0.94	24	−0.7677	−0.1577	0.008	0.9020	NO
			24	0.7989	0.9604	0.000
	SBC	5.23	24	0.2048	0.7869	0.004	0.9542	YES
			24	−0.8043	0.2498	0.002
	R	3.45	24	−0.9157	−0.6083	0.000	0.9124	NO
			24	−0.01077	0.6884	0.15*
T-Pro	Hb	9.02	24	0.2405	0.8008	0.002	0.9602	YES
	LDH	26.1	24	−0.7041	−0.01985	0.000	0.9404	YES
	CPK	154	24	−0.7474	−0.1109	0.014	0.4672	NO
	GPT	59	24	−0.8275	0.3136	0.000	0.7476	NO

Values obtained by applying the Shapiro-Wilk test determines the normal significance for α=0.05 data. Fisher’s exact test determines a difference in the analyzed data by the alternative method described in the materials and methods section. Therefore, the odds ratio was determined from two different biochemical tests at intervals of 10 days viewing the biochemical value variation during a training program, established in order to develop the aerobic capacity.

pH value, primary among the determinations, is influenced as a risk factor (<1), or protective factor (>1) in the interpretation of the biochemical determined values. Parameters such as Alb, Ca++, SEB, SBC, LDH, BE, and T-Pro are statistically significant after applying the Pearson test, with established relationships relating to pH variation (Table II). Thus, the statistical relationship with serum albumin did not represent significant correlations (Odds Ratio) between the reported amount of biochemical parameters, and pH variation in the 7.35–7.45 range. A relationship which has not been identified even in the case of serum calcium, of which negative value does not influence the blood pH variation in the studied group. Statistical significance was found in the case of lactate dehydrogenase whose range does not fit into the normal biochemistry values is supported by blood pH ranged between 7.41–7.45. However, T-Pro whose biochemical value was located outside the acceptance range, as a risk factor, positively influences the obtaining of a pH within 7.41–7.45 range. Neither SBE nor SBC data present a significant association with pH value. But in the case of excess base, we can highlight that negative values of the biochemical determined parameter were obtained at a pH value ranging between 7.41–7.45 (Table II).

Table II

Connections established after applying Fisher’s exact test.

Fischer Test (Odds ratio)
Analyzer 1	Analyzer 2	Odds ratio	95% Confidence Interval of the Difference between proportions		Statistical significance (alpha<0.05)	P value
			Inferior	Superior
pH	BE	21.67	1.061	442.3	Yes*	0.014
	SBE	8.478	0.4135	173.9	No	0.130
	SBC	8.478	0.4135	173.9	No	0.130
	T-Pro	0.02118	0.001003	0.4470	Yes*	0.001
	Alb	0.1005	0.004875	2.071	No	0.11
	Ca++	5.000	0.8060	31.02	No	0.09
	LDH	0.0400	0.003350	0.4776	Yes*	0.006

Discussion

Establishing connections following biochemical determinations will positively influence the work performed during general periods of training, as well as the entire recovery process of the athletes. In the studied group, the pH value variations were taken into account along with the secondary parameters that were determined. As a result, significant correlations were found between the main parameters represented by pO2, pCO2, pH, HCO3, T-Pro, LDH.

The physical capacity of the body can be limited by reducing the possibility of generating muscular strength [6]. Repetitive skeletal muscle activity is part of the mechanism that balances the pH, representing a direct influence of sports activity and muscle efficiency [6]. Specialized data indicate that venous pO2 will drop due to increased muscle activity during endurance exercise [12]. Thus, O2 must be accessible at the start of the effort, while pCO2 value normally will slightly fall during early effort. The action is carried out as a result of compensation towards the development of lactic acidosis [13]. Increasing pCO2 seems to be directly proportional to pH decrease, suggesting blood acidity during physical activity [14]. The pH value and its variations are related to the actual concentration of hydrogen ions (H+) [15]. Body fluids require a stable pH for proper functioning of enzymes, maintaining the protein structure, and ionic distribution. As a result, for a pH value of 7.44 we can find a H+ concentration of 36 mmol/l, while a pH of 7.36 reports a concentration of 44 mmol/l [16].

Blood buffer systems of the body are represented by parameters such as carbonic acid, bicarbonate, hemoglobin, plasma proteins and phosphates [17]. From this point of view, the state of acidosis or body alkalosis are identified [18]. Respiratory acidosis is a drop in pH (<7.35), due to an increased pCO2 value, because of hypoventilation, identifying a normal HCO3 value. Respiratory alkalosis is associated with an increase in pH (>7.45) following a pCO2 drop due to hyperventilation, and a system that removes a large amount of CO2, associated with normal HCO3 [19]. Decreases in pCO2 within 30–35 mmHg can cause a reduction in H+, thus resulting in an increased pH≥7.45 [15]. Metabolic acidosis represents the lowering process of pH to a pCO2 value located within normal limits, and a decrease in HCO3 value with the possible factors of influence increasing the production of acids, failure in the removal of H+, or loss of an elevated amount of bicarbonate [20]. Metabolic alkalosis represents an increased pH value to pCO2, which is within the normal range, indicating a decrease of acidity, or bicarbonate increase [21]. These data suggest a direct or indirect relationship between the studied parameters, to normalize or imbalance pH values, aspect identified within the paper.

In physiological terms, pH variation can be interpreted through pCO2, HCO3, BE values. Thus, a pCO2 value exceeding 45 mmHg is associated with respiratory inefficiencies and pH decrease in acid values [22]. pCO2 value which falls below 35 mmHg is associated with hyperventilation, and an increase in pH, in alkalosis stage [23]. Base excess (BE) or HCO3 at a value of −2 mEq/l and <22 mEq/l respectively is associated with acid pH. An increase in bicarbonate and base excess more than 2 mEq/l, and 22 mEq/liter, is linked to an alkaline pH [19]. At the same time blood pH alters the binding of Ca++ to serum proteins. In alkalosis, an increased amount of calcium is bound to plasma proteins, resulting in a lower percentage of ionized calcium. Thus, an increased pH is associated with a decreased calcium value [24], confirmed by our the results. However, low albumin levels are associated with pH changes, depending on the transport mechanism, osmosis, filtration, diffusion, active transport, and plasma proteins, which are associated with blood as buffers [25].

As a result, a number of changes in acid-base balance in our study group were highlighted at a pH range of 7.42–7.45. From a practical standpoint, balancing pH along with normal respiratory values, and inducing a state of metabolic cost reduction will increase the adaptation of the athlete during effort. Ongoing studies may reveal that metabolic recovery of the body will not necessarily represent respiratory muscle recovery, a state that will determine a decreasing intensity of the performed effort and decrease in muscular efficiency. Thus, biochemical, cardio respiratory, and metabolic analysis concerning energy sources will support the body’s recovery stage, given that existing data between the analyzed systems will often be inversely proportional, based on the activity performed and the body’s response.

Conclusions

We confirm the connection established between pH and BE, HCO3, PCO2, PO2, Alb, Ca++, T-Pro. The obtained values that dictate acidity or alkalinity are directly proportional to factors that influence the body’s state of recovery after training. Thus, the combination of bicarbonate, excess base, and partial pressure with pH values will provide concrete information on how the body reacts, directly related to metabolic recovery and respiratory recovery of the athlete.