Pregnancy refers to the fertilization and development of one or more offspring, known as a fetus or embryo, in a woman's uterus. Micronutrients are trace elements and vitamins obtained from our diet that are essential to sustain life and optimal physiological function. The aim of this study is to assess some micronutrients (total protein, calcium, albumin and glucose) in pregnant subjects attending Federal Medical Center, Asaba, Delta State. The subjects in this study comprise of pregnant women attending Federal Medical Centre, Asaba, Delta State. A total of one hundred and sixty (160) samples were recruited for this study which consist 120 pregnant women, 40 for each trimester and 40 non-pregnant women which serve as control. The results showed that total protein levels were lower (p>0.05) in the pregnant subjects (63.70±14.29 g/dl) when compared with the control (68.95±25.61 g/dl) but was not statistically significant. Albumin levels were significantly lower (p<0.05) in the pregnant subjects (35.00±10.80 g/dl) when compared with the control (40.05±14.56 g/dl). Calcium levels were also significantly higher (p<0.05) in the pregnant subjects (3.01±2.05 mmol/l) when compared with the control (1.82±0.33 mmol/l). Furthermore, Glucose levels were significantly lower (p<0.05) in the pregnant subjects (4.12±1.40 mg/dl) when compared with the control (6.16±4.25 mg/dl). Total protein levels were significantly lower (p<0.05) in 2nd trimesters (57.95±7.95 g/dl) when compared with 3rd trimesters (60.73±9.90 g/dl), control (68.95±25.61 g/dl) and 1st trimester (72.43±18.63 g/dl). Albumin levels were significantly lower (p<0.05) in 3rd trimesters (29.13±5.72 g/dl) when compared with 2nd trimesters (30.13±7.33 g/dl), control (40.05±14.56 g/dl) and 1st trimester (45.75±9.54 g/dl). Furthermore, calcium levels were significantly higher (p<0.05) in 3rd trimesters (4.54±2.52 mmol/l) when compared with 2nd trimesters (2.87±1.39 mmol/l), control (1.82±0.33 mmol/l) and 1st trimester (1.62±0.38 mmol/l). glucose levels were significantly higher (p<0.05) in control (6.16±4.25 mg/dl) when compared with 1st trimesters (4.41±2.03 mg/dl), 2nd trimesters (4.41±1.03 mg/dl) and 3rd trimester (3.77±0.77 mg/dl). The deficiencies in micronutrients that affect many women of reproductive age are now known to be associated with adverse maternal and perinatal outcomes. However, it is more beneficial to consume adequate micronutrients before conception to minimize the likely risks associated with their deficiencies in both the mother and the fetus. Also, consume it while being pregnant to meet the adequate levels of the micronutrients that the mother and fetus need.

Pregnancy refers to the fertilization and development of one or more offspring, known as a fetus or embryo, in a woman's uterus (Kalhan, 2000). The term embryo is used to describe the developing offspring during the first 8 weeks following conception, and the term fetus is used from about 2 months of development until birth (Kalhan, 2000). Adaptation to pregnancy in humans involves anatomic, physiologic and metabolic changes in the mother to support and provide her with nutritional and metabolic needs and those of growing conceptus (Kalhan, 2000). In a pregnancy, there can be multiple gestations, as in the case of twins or triplets. Childbirth usually occurs about 38 weeks after conception; in women who have a menstrual cycle length of four weeks, this is approximately 40 weeks from the last normal menstrual period. The World Health Organization (WHO) defines normal term for delivery as between 37 weeks and 42 weeks (WHO, 2006).

Pregnancy is typically divided into three periods, or trimesters, each of about three months. The first 12 weeks of pregnancy are considered to make up the first trimester. Weeks 13 to 28 of the pregnancy are called the second trimester. The third trimester of pregnancy spans from week 28 to the birth. Between the interval of 37 to 42 weeks, normal labour, rhythmic uterine contractions and birth occur and this is called Term. Pregnancy is associated with normal physiological changes that assist the nurturing and survival of the fetus. Pregnancy represents a period of major physiological and metabolicchange, in order to ensure proper fetal growth and development, as well as maternal preservation and survival (King, 2000). Adequate pregestational nutritional status, as well as proper gestational weight gain and dietary intakes are mandatory to promote these processes and to avoid potentially adverse maternal and pregnancy outcomes (Ramakrishnan et al., 2012; American College of Obstetricians and Gynecologists (ACOG), 2013; Feodor et al., 2014; Parisi et al., 2014; Shaw et al., 2014; Marangoni et al., 2016). Therefore, it is important to evaluate, monitor, and if appropriate, make changes to improve maternal nutritional status both before and during pregnancy. Moreover, inadequate and excessive dietaryintakes have been associated with long-term effects and noncommunicable diseases in the offspring (developmental model for the origins of disease, Barker Hypothesis). Particularly, fetal developmentin obesogenic intrauterine environments can permanently modify individual biological and metabolic pathways,leading to adaptive pathophysiological alterations in the offspringand to increased risks of non-communicable diseases in adulthood (Luyckx et al., 2017). This gives a critical role to preconception and pregnancy care in order to improve health of future generations and prevent transmission of obesity and non-communicable diseases in the offspring.

Biochemical parameters reflect these adaptive changes in most organ system and are clearly distinct from the non-pregnant state (Tran, 2005). However, these changes become very important in the event of complications. Micronutrients are trace elements and vitamins obtained from our diet that are essential to sustain life and optimal physiological function (Bailey et al., 2015; McMillan et al., 2019). Deficiencies affect over 2 billion people and are largely associated with malnutrition or poor diet (Bailey et al., 2015). Many micronutrients are necessary to elicit an effective immune response to viral infections but are also utilized by viruses such as the hepatitis C virus (HCV) and hepatitis B virus (HBV) to propagate infections (Rashed, 2011). Essential micronutrients are involved in many metabolic pathways in the liver, such as enzymatic functions and protein synthesis, immunological competence, interferon therapy response regulations, and alterations of the virus genomes (Bhaskaram, 2002). 

Despite a small change in caloric and macronutrient requirements compared to the non-pregnant state, the need for micronutrientsupply exponentially increases during pregnancy, particularly for key elements including iron, folate, iodine, calcium and vitamin D (King, 2000). Together with a limited availability of nutrients and fortified foods in low-income countries and with an alarming decline of appropriatenutritional habits in high-income countries, this explains why micronutrient deficiencies are extremely common during pregnancy (World Health Organization, 2008). This raises the question whether dietary intake is enough to cover the increased micronutrient requirements of pregnancy. The World Health Organization (WHO) currently recommends to provide multiple micronutrient supplements to pregnant women from populations with a high prevalence of maternal nutritional deficiencies,thus reducing the risks of low birth weight (LBW) and small forgestational age (SGA) compared to iron-folic acid supplementation alone (Moos et al., 2008; Haider & Bhutta, 2017). Conversely, discordant results question the efficacy of routine multivitamin supplementation among well-nourished womenfrom high-income countries (Wolf et al., 2017). 

Of interest however is the fact that pregnant women has shown some association with various diseases (Armelle et al., 2006).  In fact, marked differences in plasma or serum concentration of some liver and intestinal enzymes (Adamo, 2010) leading to marked differences in nutritional benefit from a diet, enhances susceptibility to diseases. Pregnant women are particularly susceptible to various infections such as ascending pyelonephritis as a result of bladder infections (Epstein & Karumanchi, 2005). Therefore, the evaluation of some micronutrients in pregnant women is of major importance in monitoring the successful outcome of pregnancy.

Pregnancy is a normal physiological condition which shows many changes in the maternal environment. Many changes in renal function are associated with pregnancy and without a proper understanding of these changes, routine clinical investigations may be easily misinterpreted (Brown & Whitworth, 1992). A Norwegian study on 40,108 women showed that the nutrient contribution of dietary supplements among usersvaried from 65% for folate and vitamin D to 1% for potassium,with total intakes of vitamin D, folate, iodine and iron still lower than the national recommendations for pregnant women (Haugen et al., 2008). During pregnancy, micronutrient requirements increase more than those of macronutrients, and inadequate intakes (and, thus, a low nutritional quality of the diet) can have significant consequencesfor both the mother and the developing fetus. In particular, there is evidence to support the physiologic role played by selected minerals and vitamins. A recent meta-analysis also provided evidences that fetal gender,pregestational maternal nutritional status and adherence to supplementation represent important factors influencing the effect of multivitamin supplementation on pregnancy outcomes, showingimproved survival and outcomes in case of female newborn andundernourished/anaemic pregnant women (Smith, Shankar & Wu, 2017). Hence this study is designed to evaluate some micronutrients of pregnant women attending Federal Medical Center, Asaba, Delta State.

Area of Study

This study was carried out among pregnant women visiting Federal Medical Centre, Asaba, Delta State, Nigeria. Asaba, the capital city of Delta State, Nigeria is situated within geographical co-ordinates 6°11′52.23″N6°43′42.48″E. It is situated on a terrace of the lower Niger River, overlooking the point where the Anambra River flows into it. Beyond the river banks, on the high plains which are far more extensive than the river basins, secondary forest vegetation flourishes (APU, 2016). The historic Niger River is a trans-African link beginning from West Africa and down into the Atlantic Ocean. Asaba forms a connector between western, eastern and northern Nigeria through the Niger River from the north and via the Asaba Niger Bridge, an east-west link and a Nigerian landmark. The analysis was carried out in the Chemical Pathology department of the Federal Medical Centre, Asaba, Delta State (World Gazetter, 2015).

Population of the Study

The subjects in this study comprise of pregnant women attending Federal Medical Centre, Asaba, Delta State. A total of one hundred and sixty (160) samples were recruited for this study which consist 120 pregnant women, 40 for each trimester and 40 non-pregnant women which serve as control. Subject data such as name, age, months of pregnancy were also obtained. The sample size (N) is calculated from the formula below using prevalence from previous studies (Ekejindu et al., 2002).

, where

N = the desired size

Z = 1.96 (standard score)

P = Prevalence (12%) (0.12)

q. = 1- P (0.88)

d = sample error tolerated (0.05) 

N=

Research Design

This study was designed as a prospective and cross-sectional study to assess some micronutrients (total protein, calcium, albumin and glucose) in pregnant subjects attending Federal Medical Center, Asaba, Delta State and make comparison with that of the control group. This study was carried out within three (3) months. A complete record of medical history was obtained for each subject, including name, age, and duration of pregnancy. The overall results of the study obtained was compared with the control using appropriate statistical methods.

Research Design Chat

Ethical Considerations

Ethical permission for this study was obtained from the Health Research Ethics Committee of Federal Medical Center, Asaba, Delta State, Nigeria. Also, informed consent was sought and obtained from the patients prior to the collection of samples for this study. The purpose of the study was exhaustively explained to the patients and assured of the confidentiality of the information obtained from them.

Sample Collection

Blood samples (5mls) was collected by vene-puncture into an accurately labelled lithium heparin containers for both subjects and control individuals. The blood samples were centrifuged with a laboratory centrifuge at 4000rpm for 10minutes at room temperature within two hours of collection and the serum separated into clean plain containers which are labelled corresponding to the initial blood samples containers. The serum was then stored at -20°C pending the analysis of the samples.

Sample Analysis

Serum Calcium Estimation: Calcium shall be estimated using the method described by Robertson and Mashall, (1979).

Principle: Calcium ion reacts with O-cresophthalene-complexono in an alkaline medium to form a purple coloured complex. The absorbance of this complex is proportional to the calcium concentration in the sample.

Estimation of Total Protein: The Total protein in the sample was determined using the method described by Gomall et al., (1949).

Principle: In the biuret reaction, a chelate is formed between the CU2+ and the peptide bonds of the protein in alkaline solution to form a violet coloured complex whose absorbance is measured colorimetrically. The intensity of the colour is proportional to the concentration of protein in the sample.

Estimation of Albumin: The Albumin in the sample was determined using the method described by Doumas et al., (1971).

Principle: This method is based on the specific binding of bromo- cresol green (BCG), an anion dye and the protein at acid PH with the resulting shift in absorption wavelength of the complex. The intensity of the colour formed is proportional to the concentration of albumin in the sample.

Estimation of Globulin: Plasma Globulin was estimated indirectly by subtracting the albumin concentration from total protein concentration as described by Ochie and Kolhatkar, (2000).

Plasma globulin (g/dL) = Total protein (g/dL)        Plasma Albumin (g/dL)

Determination of Glucose: The Glucose in the sample was determined using the method described by Doumas et al., (1971)

Principle: Glucose oxidase (GOD) catalyzes the oxidation of glucose to give hydrogen peroxide (H2O2) and gluconic acid. In the presence of the enzyme peroxidase (POD), the hydrogen peroxide is broken down and the oxygen released reacts with 4-aminophenazone (4-aminoantipyrine) and phenol to give a pink colour. The absorbance of the colour produced is measured in a colorimeter using a green filter 520 nm (Ilford No. 604) or in a spectrophotometer at 515 nm (Cheesebrough, 2000).

Statistical Analysis

The Mean and standard deviation of the results obtained was calculated. ANOVA (LSD) was used for the analysis using SPSS package version 21. Values with p< 0>