Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241128EnglishN2020April23HealthcareCan the Level of Glucose & Glycated Hemoglobin be Estimated from Same Container?
English0104Souvik KonarEnglish Amrita KarmakarEnglish Soma GuptaEnglishContext: Burden of diabetes mellitus is increasing gradually along with its complications. Nowadays along with fasting and post-prandial plasma glucose (FPG and PPPG), HbA1c is included in the diagnosis and assessing prognosis. The current practice is samples for HbA1c and FPG, PPPG are collected in different vials (EDTA and Fluoride vial respectively).
Aims: Present study was designed to find out whether any difference occurs or not in HbA1c values when collected in EDTA & fluoride vials & also to find out the stability of HbA1c in fluoride vials for five consecutive days.
Settings & Design: This case control study was conducted at service laboratory of Dept. of Biochemistry at NRS Medical College & Hospital
Methods & Material: Samples were collected from 107 subjects (44 male & 63 female) in both EDTA (E) & fluoride vials (F). Estimation of HbA1c was done from both the vials separately by Latex Turbidimetric and chromatography method (HPLC). 10 randomly chosen samples were collected and stored at 4oC in fluoride vials. HbA1c estimation was done from F vial for 3 consecutive days.
Statistical Analysis: The results were tabulated and analyzed using software.
Results: The HbA1c values from both EDTA & Fluoride vials were well correlated and no statistically significant differences (p= 0.69) existed among E & F vials. ANOVA analysis showed no significant differences (p-0.99) among HbA1c level in Fluoride vial for 3 consecutive days.
Conclusions: Estimation of blood glucose level & HbA1c level can be done from the fluoride (F) vial without compromising the quality of reports.
EnglishHbA1C estimation in Fluoride vialINTRODUCTION:
Prevalence of diabetes mellitus (DM) is increasing rapidly. In India estimated age adjusted prevalence of DM in adults is >9%. India stands second among top ten countries for number of adults with DM (1). In addition to the prognostic role of HbA1c, American Diabetes Association has recommended HbA1c for the diagnosis of Diabetes Mellitus (2)
Though fasting plasma glucose and HbA1C are advised simultaneously in almost all cases, the sample cannot be collected in same vial as HbA1c is stated to have high preanalytical stability (3) (4) when collected in EDTA vial rather than in traditional glucose vial. But no reason for it has been demonstrated. Same container is always preferred, so far time, cost and convenience is considered.
Subjects and Methods:
In this backdrop, this study was undertaken to find out whether any significant variation in HbA1c level occurs or not if the sample is estimated from the same vial which is used to estimate blood glucose level. It was also aimed to find out the stability of the level of HbA1c, when estimated from F vial.
The study was conducted in the service Laboratory of the Department of Biochemistry, Nil Ratan Sircar Medical College and Hospital, Kolkata. A total of one hundred seven subjects who were advised for blood glucose & HbA1c estimation were consecutively selected & included in this study. Individuals of both genders were included in this study. After obtaining proper consent, whole blood was drawn from the subjects and was divided separately in EDTA vial and Fluoride vial. A total of 10 samples collected in Fluoride vial were chosen randomly & were stored in 4 degree centigrade. The estimation was repeated for 3 consecutive days to find out the stability of HbA1c level.
HbA1c level was estimated from both vials separately by Latex Turbidimetric method and HPLC method. The underlying principle of latex turbidimetric method (5) is that it utilizes antigen-antibody interaction to directly determine the HbA1c in whole blood. Total hemoglobin and HbA1c have the same unspecific absorption rate to latex particles. When mouse antihuman HbA1c monoclonal antibody is added, latex-HbA1c-mouse anti human HbA1c antibody complex is formed. Agglutination is formed when goat anti-mouse IgG polyclonal antibody interacts with the monoclonal antibody. The amount of agglutination is proportional to the amount of HbA1c absorbed on to the surface of latex particles. HbA1C estimation done by Hb-Vario Analyser uses cation exchange high performance liquid chromatography (HPLC) in conjuction with gradient elution to separate hemoglobin subtypes and variants from hemolysed whole blood. The separated hemoglobin fractions are monitored by absorption of light at 415 nm. A software program analyses the chromatogram and reports are generated. (6) The results were tabulated and statistically analyzed. The significance of difference in the level of HbA1c was found out using Student’s t test (p0.05). The two methods were well correlated positively as calculated by Correlation Coefficient, which was found to be 0.9 (Fig. 1).
Table 2 shows the result of ANOVA test done to compare the level of HbA1c for 3 consecutive days. The result shows that no statistically significant difference exists among any group (p=0.905, which is >0.05).
DISCUSSION
This study was undertaken to find out whether any variation in HbA1c level occurs or not if the sample is estimated from E vial & from F vial. It was found that no significant difference (p value-0.69 in E; 0.83 in F vial) was seen among the values of HbA1c when HbA1c estimation was done from two different (E & F) vials by HPLC and Latex agglutination method. The result of this study is in accordance with the results of the study done by Mailankot M et al (4). Mailankot M et al in their study found no significant changes between the samples taken in different tubes. They excluded the absolute necessity for blood collection in EDTA tubes for HbA1c estimation. Sharma B et al (3) in their study concluded that unnecessary extra sample collection for HbA1c estimation can be avoided which can improve patient compliance & avoidance of extra vial for HbA1c can reduce the cost of HbA1c test.
This study was also conducted to find out the stability of HbA1c level when HbA1c estimation was done from F vial for at least three consecutive days when the samples were stored at 4 degree centigrade. It was found that no significant difference existed when HbA1c estimation was done only from the F vial for 3 consecutive days by latex agglutination method days (p value- 0.99) and HPLC method (p vaue-0.99). The comparison could not be extended beyond three days as sample in F vial was getting clotted and could not be run by HPLC method.
India is considered as the diabetic capital of the world. For diagnosis of Diabetes Mellitus, American Diabetes Association has recommended HbA1c estimation. Among Indians, proper diabetic screening is needed but for awareness of diabetes, often the adequate manpower is lacking. Moreover, HbA1c test is a costly test & for testing HbA1c using a separate E vial further increases cost. Usually, HbA1c estimation is done on the same day of blood collection but blood collection in a different E vial also requires more sample volume.
Hence, considering these facts our study is very promising when it comes to estimation of HbA1c. Our study does not encourage the need for extra E vial for estimation of HbA1c. HbA1c can be tested in the same vial which is used to estimate blood sugar (F vial).
So, estimation of HbA1c among Indians is very much needed. HbA1c should be used for diagnosis of Diabetes Mellitus in India because it can diagnose many cases which go undetected by routine blood sugar analysis. For establishing a baseline HbA1c level among Indians, a pan India study of HbA1c should be conducted based on which diagnosis & screening of diabetes could be predicted.
CONCLUSION
The study shows that HbA1c level is comparable significantly with standard practice even if it is done from vials used to estimate blood glucose level. Moreover, the concentration of HbA1c remains stable in F vials for at least 3 consecutive days, when stored in 4 degree centigrade. Usually the estimation of HbA1c is done on the same day of blood collection. Hence, the estimation of blood glucose level & HbA1c level can be done conveniently from the same vial without compromising the quality of reports.
Acknowledgement:
We are thankful to our Principal and MSVP for allowing us to conduct this research. We acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. We are also grateful to publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Source of funding: There was no source of funding
Conflict of interest: No conflict of interest
Englishhttp://ijcrr.com/abstract.php?article_id=2667http://ijcrr.com/article_html.php?did=26671. Pouya Saeedi et al.,Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045 ressults from the the International Diabetes Federation Diabetes Atlas,9th edition. Diabetes Res Clin Pract. 2019,November.volume 157
2. American Diabetes Association.Improving care and promoting health in populations: standards of Medical Care In Diabetes- 2019.Diabetes Care 2019.2019;42:S7- S12.
3. Booloo Sharma , Devajit Sarmah, Pavan Sonker .,Effect of Different Anticoagulants on HbA1C estimation and its stability. J Lab Physicians.2013; volume 5:page 143-144.
4. Maneesh Mailankot, Thusara Thomas , P Praveena , juliet Jacob , Jobin Ranjan Benjamin, DM Vasudevan. various anticoagulants and fluoride do not affect HbA1C level. Indian J Clin Biochem. 2012; volume 27, page 209.
5.Liliana A Trivelli, Helen M Ranney, Hong Tien Lai. Hemoglobin components in patients with diabetes mellitus. N Engl J Med. 1971;volume 284:page 353-357.
6. R. S Ersser, G. B Barlow, R. G. Drew, M Hjelm. Packing materials suitable for rapid, analytical, low-pressure chromatography of hemoglobins on midget columns. Biomedical Chromatography.1986;volume 1:page 183-188.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241128EnglishN2020April23HealthcareEffect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial
English0510Ilona Gracie De SouzaEnglish Pavan Kumar G.EnglishBackground: Patellofemoral pain syndrome (PFPS) is characterized by diffuse anterior knee pain. Musculoskeletal dysfunction arises from alterations in the deep fascia that pull or crowd the osseous structures out of alignment resulting in pain and dysfunction. Purpose of the study was to evaluate the effect of releasing myofascial chain in patients with patellofemoral pain syndrome.
Methods: 40 patients were randomly allocated into 2 groups. Experimental group (n=20) mean age 36.60±7.36 was treated with Myofascial chain release and exercise therapy. Control group (n=20) mean age 32.25±9.11 was treated with exercise therapy, three sessions in a week for a period of 4 weeks. Kujala, Visual analogue scale (VAS) and Patient specific functional scale (PSFS) were used as outcome measures. Paired & unpaired t test was used for statistical analysis
Results: Both groups showed statistically significant change in all the three outcome measures. The experimental group showed (pEnglishExercise therapy, Fascia, Myofascial release, Patellofemoral pain syndrome (PFPS), Physical therapyINTRODUCTION
Patellofemoral pain syndrome (PFPS) is the most prevalent musculoskeletal condition seen in general practice and sports medicine clinics which is characterized as anterior knee pain.[1] The patellofemoral joint is the most heavily loaded in the human frame and its articular cartilage is the thickest.[2] Incidence ranges from 21 to 40% in clinical setting and females are affected more than males in the ratio 2:1. [3] The symptoms include diffuse pain over anterior aspect of knee that is aggravated by activities such as ascending or descending stairs, prolonged sitting and squatting.[4] Physical impairments include destruction of articular cartilage, pain, diminished muscular control, patella mal-alignment, inflexibility of the hamstrings, gastrocnemius, iliotibial band, tensor fasciae latae, and biomechanical alterations in the lower extremity, thus indicating multifactorial causes. The causes and mechanisms involved in patellofemoral pain are extrinsic and intrinsic.[5]
Clinical experience and emerging data proved that weakness of hip musculature and limited lower extremity flexibility are the key factors for the conservative management of patellofemoral pain syndrome.[6] Tight lateral retinaculum causes lateral patellar compression that puts excessive stress on lateral patellofemoral joint. Tight quadriceps becomes less efficient to absorb energy eccentrically which transmits the load to adjacent structures that is patella and quadriceps tendon subjecting them to stress. A tight iliotibial band (ITB) causes increase tension on lateral retinaculum and lateral patellofemoral joint stress. In addition with a tight quadriceps, hamstrings has to overcome this resistance during knee flexion thus generating an increase in patellofemoral joint reaction force.[7]
In accordance with Fascial Manipulation theory, uncoordinated quadriceps contraction produces anomalous fascial tension in the thigh causing pain in the patella. Therefore the focus of treatment was on the muscular fascia of the thigh.[8] Musculoskeletal dysfunction arise from alterations in the deep fascia.[9] Fascia is tough connective tissue from head to toe spreading throughout the body in a three-dimensional web pattern.[10]
As a response to injury, disease, inactivity or inflammation fascial tissue loses its elasticity and become dehydrated. Fascia binds around traumatized areas causing fibrous adhesions that prevents normal muscle mechanics and decreases soft tissue extensibility. Fascial restrictions crowd or pull the osseous structures out of proper alignment creating abnormal strain patterns, resulting in joint compression producing pain and/or dysfunction.[10]
‘Myofascial chain’ is the bundled together inseparable nature of muscle tissue (myo) and its accompanying web of connective tissue (fascia).[11] 'Myofascial continuity' describes the connection between two longitudinally adjacent and aligned structures within the structural webbing. Quadriceps runs from ASIS, shaft of femur, then as subpatellar tendon, patella and then continues as Tibialis Anterior towards the inside of the ankle. This Quadriceps, Tibialis anterior chain affects knee tracking. The tensor fascia latae blends with the iliotibial band on the outer aspect of the knee on the lateral condyle of tibia and then onto the tibialis anterior crossing the lower shin bone.[12] Thus Quadriceps, Tibialis Anterior and Iliotibial band chains influence the patellofemoral joint the most.
The current rehabilitation studies for patellofemoral pain syndrome are Kinesio taping, custom-fitted foot orthosis, lumbopelvic manipulation and exercise therapy.[13-16] Exercise therapy has proved effective in management of patellofemoral pain syndrome but has limitations as it doesn’t correct the fascial restrictions and a recent systematic review has stated its poor adherence in patients suffering with knee pain due to lack of time, lack of knowledge about the exercise, apathetic condition, poor behavior to exercise and impaired general health status.[17] Despite the success of conservative treatment, patients continue to experience pain and dysfunction making patellofemoral pain syndrome a challenging condition to treat.[18]
Myofascial therapy is defined as “the facilitation of mechanical, neural and psycho physiological adaptive potential as interfaced by the myofascial system”. Myofascial release (MFR) is a hands on technique which stretches the fascia and releases, bonds between muscles, integuments and fascia in order to eliminate pain, improve motion and to maintain myofascial balance within the body. [19]
Fascia is manipulated by direct technique that involves the use of knuckles, elbows, ulnar border of the hands and fist, indirect or self-myofascial technique, indirect myofascial release uses hands and the self myofascial release uses a soft roll or ball (tennis ball). The purpose of myofascial release is to focus on the deeper layers of fascia by elongating the muscular elastic component, cross-links and altering the viscosity of ground tissue.[19]
Studies have shown the effectiveness of myofascial release in reducing chronic pain, neck pain, shoulder pain, muscular spasm and muscle tightness.[19] Studies proved the effectiveness of myofascial release on Iliotibial band flexibility and patellar alignment in patients with knee osteoarthritis.[20] Recent study has shown effectiveness of static stretching versus myofascial release in Iliotibial band tightness in long distance runners.[21] Also a recent study have shown the effectiveness of chiropractic mechanical assisted adjusted techniques (MAT), soft tissue therapy involving deep effleurage and myofascial release and specific strengthening and stretching rehabilitation program in a professional basketball player with chronic patellar tendinopathy complicated with patellofemoral pain syndrome.[22]
As inflexibility of soft tissues have been an important factor in causing patellofemoral pain syndrome due to the interconnected fascia, many treatment techniques have been successful in treating patients with patellofemoral pain syndrome but no study has focused on the treatment of muscular fascia involvement. Exercise therapy has been the most effective treatment in patellofemoral pain syndrome. However, myofascial therapy can be an added benefit in order to overcome the poor adherence to exercise therapy. Research on myofascial release has shown promising results in the area of soft tissue inflexibility and could potentially be a form of treatment. As per the existing evidence of soft tissue inflexibility causing patellofemoral pain syndrome. Though the efficacy of myofascial release has been effective in releasing inflexibility of soft tissues, no studies have been retrieved of its effectiveness in patellofemoral pain syndrome patients. Thus the purpose of this study is to know the effect of myofascial chain release compared to exercise therapy on patellofemoral pain syndrome patients.
METHOD
Forty subjects were recruited from Srinivas college of physiotherapy and research center OPD and Srinivas Hospital, Mangalore from April 2016 to March 2017. Inclusion criteria 1) Age between 18-50 years 2) both gender 3) visual analogue scale (VAS) rating of atleast 30mm on a 100mm scale over the previous week. 4) anterior knee or retropatellar pain during atleast three of the following activities like stair ascent or descent, squatting, kneeling, hopping, jumping and prolonged sitting. 5) Gradual onset of symptoms 6) Subject experiences pain during palpation of patellar facets or while performing a 25-cm step down test or double legged squat.[3,6] Exclusion criteria included meniscal or intra-articular pathological conditions, cruciate or collateral ligaments involvement, tenderness over the patellar tendon, iliotibial band or pes anserinus tendons, sign of patellar apprehension, patellar tendinopathy, hip or lumbar referred pain, history of patellar dislocation, evidence of knee joint effusion, history of previous surgery, pre-patellar tendinitis, fat pad bursitis, quadriceps tendinopathy, over sensitive patients, systemic or local infections, healing fractures, anti-coagulant therapy and under corticosteroids and or anti-inflammatory drugs.
PROCEDURE
The approval was obtained from the scientific committee and college ethical committee. Subjects with patellofemoral pain syndrome were screened for inclusion and exclusion criterion. A total of 53 subjects were screened. 40 subjects met the inclusion criteria. The purpose of the study was explained to all the subjects and was asked to sign the written consent form stating the voluntary acceptance to participate in this study. Demographic information was collected. Purposive sampling was done. 40 subjects were assigned randomly into experimental or control groups by block randomization. Pre -treatment score was evaluated by using Kujala Questionnaire [23], Patient specific functional scale [24] and visual analogue scale[25]. Experimental group (n=20) was treated with myofascial chain release for the Quadriceps, Iliotibial band and Tibialis anterior muscle along with exercise therapy. Control group (n=20) was treated with exercise therapy that included stretching and strengthening of hip and knee musculature. Treatment was given for 3 days in a week for a period of 4 weeks. Post treatment score were evaluated after the last treatment session
DESCRIPTION OF TREATMENT
Brief assessment was conducted.
Evaluation of fascial restriction was assessed using the therapists elbow. Any restrictions felt were assessed and given treatment.
Experimental group: Myofascial chain release and exercise therapy
Myofascial chain release- [26]
Position of the patient- Supine lying, prone lying and side lying
Position of the therapist- Side of the patient limb to be treated
Technique-
Using the therapist elbow a vertical release proximal to the attachment of the muscle belly or fascia was applied.
Once an end-feel was reached a slow stroke down the length of the target tissue was performed along with monitoring the indirect feedback and tissue tension to identify any additional restrictions.
The long stroke was repeated in a line parallel to the first stroke. It was continued until an end-feel is reached throughout the entire muscle belly or fascia on the most restricted/painful points.
Myofascial chain release was given to Quadriceps, Iliotibial band and Tibialis anterior for three days in a week for a period of four weeks of 5 to 10 min duration.[16]
Figures 1: Myofascial chain release for Quadriceps.
Maximum resistance were given that enables 10 repetitions
Load was 70% of the 1-repetition maximum.
Treatment was given for 3 days in a week for a period of 4 weeks, total 12 sessions.
STATISTICAL ANALYSIS[27, 28]
Descriptive analysis was done by finding mean and standard deviation. The data was then subjected to test of normality. Data was analysed using Paired t-test to compare the outcomes within the group and Unpaired t- test for between the groups. The confidence interval was kept at 95%.
RESULT:
Both groups showed a statistically highly significant change (P Englishhttp://ijcrr.com/abstract.php?article_id=2668http://ijcrr.com/article_html.php?did=26681. Richard B, Christopher M. Differences in hip kinematics, muscle strength and muscle activation between subjects with and withour patellofemoral pain. J Orthop Sports Phys Ther 2009;39:12-9.
2. S.T.Green. Patellofemoral syndrome. J Bodyword Move Ther 2005;9:16-26.
3. Nakagawa TH, Muniz TB, Baldon RM, Maciel CD, Reiff RBM, Serrao FV. The effect of additional strengthening of hip abductor and lateral rotator muscles in patellofemoral pain syndrome: a randomized controlled pilot study. Clin Rehabil 2008;22:1051-60.
4. Wilson T, Carter N, Thomas G. A multicenter, single-masked study of medial, neutral and lateral patellar taping in individuals with patellofemoral pain syndrome. J Orthop Sports Phys Ther 2003;33(8):437-48.
5. Fairbank JCT, Pynsent PB, Poortvliet JAV, Phillips H. Mechanical factors in the incidence of knee pain in adolescents and young adults. J Bone Joint Surg 1984;66(5):685-93.
6. Piva SR, Goodnite EA, Childs JD. Strength around the hip and flexibility of soft tissues in individuals with and without patellofemoral pain syndrome. J Orthop Sports Phys Ther 2005;35(12):793-801.
7. LaBella C. Patellofemoral pain syndrome: evaluation and treatment. Prim Care Clin Office Practl 2004;31:977-1003.
8. Pedrelli A, Stecco C, Day JA. Treating patellar tendinopathy with fascial manipulation. J Bodyw Mov Ther 2009;13:73-80.
9. John W. the effectiveness of myofascial release techniques in the treatment of myofascial pain: a literature review. Prim Care Clin Office Practl 2004; 2:12-9
10. Barnes, MF. The basic science of myofascial release: morphologic changes in connective tissue. J Bodyword Move Ther 1997;1:231-8.
11. Vlacilova I. Cervical-thoracic spine and change of its shape due to posterior muscle chain and tilt of the pelvis. Ceska kinantropologie 2015;19(1):39-45.
12. Myers TW. Myofascial meridians for manual and movement therapist. 2nd ed. Sydney Toronto: Churchill Livingstone Elsevier; 2009. p. 136-144, 98-101.
13. Chen PL, Hong WH, Lin CH, Chen WC. Biomechanics effects of Kinesio taping for persons with patellofemoral pain syndrome during stair climbing. Biomed 2008;21:395-7.
14. Johnston LB, Michael T. Gross. Effects of foot orthoses on quality of life for individuals with patellofemoral pain syndrome. J Orthop Sports Phys Ther 2004;34(8):440-8.
15. Iverson CA, Sutlive TG, Crowell MS, Morrell RL, Perkins MW, Garber MB et al. Lumbopelvic Manipulation for the treatment of patients with patellofemoral pain syndrome: development of a clinical prediction rule. J Orthop Sports Phys Ther 2008;6:297-312.
16. Fukuda TY, Rossetto FM, Magalhaes E, Bryk FF, Lucareli PRG, Carvalho NAA. Short-term effects of hip abductors and lateral rotators strengthening in females with patellofemoral pain syndrome: a randomized controlled clinical trial. J Orthop Sports Phys Ther 2010;40(11):736-42.
17. Marks R. Knee osteoarthritis and exercise adherence: a review. Curr Aging Sci 2012;5(1):72-83.
18. Peters JS, Tyson NL. Proximal exercises are effective in treating patellofemoral pain syndrome: A systematic review. Int J sports Phys Ther 2013;8(5):689-700.
19. Shah S, Bhalara A. Myofascial release. Int J Health Sci Res 2012;2(2):69-77.
20. Gomaa EF, Zaky LA. Effect of iliotibial band myofascial release on flexibility and patellar alignment in patients with knee osteoarthritis. Int J Adv Res 2015;3(4):399-410.
21. Muragod A, Patil VR, Nitsure P. Immediate effects of static stretching versus myofascial release in iliotibial band tightness in long distance runners- a randomised clinical trial. Eur J Sport Med 2014;2(1):31-8.
22. Jarosz, Brett S. Tretatment of chronic patellar tendinopathy in a professional basketball player: a case report. Chiropr J Aust 2010;40(1):3-8.
23. Urho M.K, Seppo K.K, Simo T et al. Scoring of patellofemoral disorders. Arthroscopy. 1993;9(2):2-6.
24. Chatman AB, Hyam CP, Neel JM, Binklq JM, Stratfwd PW, Schornberg A. The patient-specific functional scale: measurement properties in patients with knee dysfunction. Phys Ther 1997;77(8):820-9.
25. Bijur PE, Silver W, Gallagher JE. Reliability of the visual analog scale for measurement of acute pain. Acad Emerg Med 2001;8(12):1153-6.
26. Manheim CJ. The myofascial release manual. 3rd ed. Thorofare New Jersey: Slack; 2000. p. 194.
27. Portney LG, Watkins MP. Foundation for clinical research. 2nd ed. New Jersey: Prentice Hall Health; 2005. p. 475-8.
28. Daniel WW. Biostatistics a foundation for analysis in the health sciences. 8th ed. New York: Wiley; 2005. p. 680-762.
29. Hruska R. Pelvic stability: influences of lower extremity kinematics. Biomechs1998;5:23-9.
30. Paloni, John. Review of myofascial release as an effective massage therapy technique. Athletic Ther Today. 2009;14:30-4.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241128EnglishN2020April23HealthcareQuantitative Analysis of Nutrients in the Gum Exudates of Acacia nilotica
English1116Rukshana IraniEnglish Kazi Layla KhaledEnglishAcacia nilotica is an evergreen tree found throughout Indiaand it belongs to the leguminosae family. The gum of this tree has been used in some parts of the country for its salutary therapeutic benefits. The gum was obtained from Acacia nilotica trees of West Bengal, India and was investigated for macronutrients and micro nutrients. The results of the present study showed that some essential minerals like calcium, iron, sodium, potassium, phosphorus were present in significant quantities which were 366.37, 25.41, 11.91, 124.87, 2.96 mg per 100 gram of pulverised gum respectively. The quantity of minerals like chromium, zinc, selenium, copper, magnesium, manganese was 0.67, 10.28, 0.54, 0.66, 22.61, 5.73 mg/kg gum respectively. It was found to be an energy dense gum containing 87.05 per cent of carbohydrate with substantially low fat, protein and vitamin content. The outcome of proximate analysis and high mineral content may account for the wide spectrum of medicinal and pharmaceutical properties of gum of Acacia nilotica which has been claimed in the pertinent literature.
EnglishAcacia nilotica, Minerals, Nutrients, Vitamins, Medicinal, PropertiesINTRODUCTION
Plants have been a major source of medicines in all cultures since the beginning of human civilisation. In the traditional system, various indigenous plants are used in the diagnosis, prevention and treatment of many ailments. In this modern era, there is a growing demand of plant based medicines, health products, pharmaceuticals, food supplements, cosmetics etc. Acacia nilotica is a pioneer species, relatively high in bioactive secondary compounds and can exert a variety of functions [1,2].
Plant Details
The plant parts of Acacia nilotica has been reported to have therapeutic uses arising from its wide spread folkloric and customary uses(3). Ethnobotanical studies suggested the usage of Acacia nilotica gum for the treatment of skin irritation and smoothening of the inflamed membranes of the pharynx, alimentary canal and genito-urinary tracts [4,5]. The bark or gum of the plant Acacia nilotica is used in West Africa to treat cancers and tumours of ear, eye or testicles[5]. The gum of the babul (Acacia nilotica) has also been used by the Bhils of Rajasthan to fill the dental cavities and to maintain oral health[6].The beneficial role Acacia nilotica Gum for promoting health of women after parturition has been revealed in a project titled Biomedical Studies and IPR (Intellectual Property Rights) where medicinal plants which were used in the treatment of Women Diseases in Sindh were also documented. In some regions of the Indian subcontinent, it furnishes the prime important ingredient of the nourishing food used for lactating mothers especially during the first three months of postnatal period[7].
Acacia nilotica (Babul tree) is one of the major gum-yielding acacia species found in India [8]. The composition of a gum varies with the plant species as well as geographical conditions and hence investigation of the indigenous Acacia nilotica gum can play an important role in exploring its potential for a wide range of applications [9].
In literature a very few reports are available on the gum exudate of A. Nilotica of West Bengal region in India. The present study aimed to quantify the various nutrients in this underutilised plant commodity that can facilitate us to comprehend its nutritive, curative and restorative properties.
MATERIALS AND METHODS
Sample collection
The gum of Acacia nilotica was collected from village Matchpota in Nadia district of West Bengal, India. They were authenticated for their unambiguous identity by Prof. N.D.Paria, Department of Botany, University of Calcutta and Botanical Society of India, Central National Herbarium, Howrah, West Bengal.
Preparation of Sample
The handpicked lumps of gum were cleaned to remove if any dirt present by wiping the surface of the nodule with water. It was then air dried and pulverized in a mechanical grinder. The powdered material was then stored in a clean and sterile bottle made up of glass. It was then kept in a dry and dark place at room temperature.
Analysis of Nutrients:
1. Moisture Content [10,11]
The porcelain crucibles were first washed and cleaned properly. It was then dried in a hot air oven at a temperature of 110 ºC for 10 minutes. The crucibles were then carefully transferred into a desiccator and then were kept in it for 30 minutes for cooling; it was then weighed again. This process was repeated until a constant weight was attained. The weight so obtained was labelled as W1.
2 grams of powdered sample was accurately weighed and put into previously weighed crucibles and then reweighed; and the weight thus obtained was labelled as W2.
The crucibles containing the samples were placed in an oven maintained at 103ºC for 14 hours. They were removed and transferred to desiccators to cool and again weighed. The process was repeated for several times until the constant weight was reached to get final weight (W3). The percentage of moisture content was calculated.
% Moisture= Initial Weight (W2)- Final Weight (W3) × 100
Weight of the sample
2. Ash Content [10,11]
Porcelain crucibles were washed and dried in an oven to a constant weight at 100ºC for 10 minutes. They were allowed to cool in a desiccator and then weighed (W1). 2.0 grams of each sample were weighed into each of the previously weighed porcelain crucibles and reweighed (W 2). The crucibles containing the samples were transferred into a Muffle furnace, which was set at 550ºC for 8 hours to make it into ash. They were then removed and allowed to cool in the desiccators then finally weighed (W 3). The percentage of ash content was calculated.
% of Ash= Weight of Ash (W3-W1) × 100
Weight of the Sample (W2-W1)
3. Estimation of Carbohydrates [12]
100 mg of the sample was weighed into a boiling tube. It was then hydrolysed in a boiling water bath for 3 hours with 5 ml of 2.5N HCl and cooled to room temperature. It was then neutralised with sodium carbonate until the effervescence ceases and the volume was made up to 100 ml. It was centrifuged, supernatant was collected and 1ml aliquot was taken for analysis. Standard glucose Stock was prepared by dissolving 100mg in 100 ml water. Working standard was prepared by mixing 10 ml of stock solution with 100 ml of distilled water. So, the concentration of working standard was 10mg/ 100ml. A series of standards were prepared by taking 0, 0.2, 0.4, 0.6, 0.8 and 1 mL of the working standard solution containing 0, 0.02, 0.04, 0.06, 0.08 and 0.1 mg of glucose respectively. 0.1 ml of sample or test solution (aliquot) was also taken in test tube. The volumes of all the test tubes were made up to 1 ml by adding distilled water. Then 4ml of anthrone reagent (200mg anthrone was dissolved in 100 mL of ice-cold 95% H2SO4) was added to all the test tubes which were followed by heating for 8 minutes in a boiling water bath. It was then cooled rapidly and the green to dark green colour was read at 630 nm.
4. Fat Estimation[10,11]
Fat was estimated as crude ether extract of the dry material. 10 grams of dried sample wasweighed accurately into a thimble and plugged with cotton. The thimble was then placed in a Soxhlet apparatus and extracted with anhydrous ether for about 16 hours. The ether extract was filtered into a weighed conical flask. The flask containing the ether extract was washed 4 to 5 times with small quantities of ether and the washings were also transferred. The ether was then removed by evaporation and the flask with the residue dried in an oven at 80-100 ? C, cooled in a desiccator and weighed.
Fat content (g/100g sample) = ( we×100)/ ws
Where weisthe weight of ether extract and wsis the weight of the sample.
5. Protein Content[10, 13]
2 grams of sample was weighed into a dry Kjeldahl flask. About 5g of digestionmixture (4.9 grams K2SO4+ 0.1 gram of CuSO4) and 20ml of pure concentrated H2SO4 were added to the same sample and themixture digested by heating for 4-5 hours. Glass beads were added to preventbumping. After the contents of the flask became clear, the process of digestion in the Kjeldahl flask was continued for atleast 1 hour more. The contents of the Kjeldahl flask were cooled, diluted withdistilled water and the mixture made alkaline by adding an excess of 40% NaOH(about 75ml). A small quantity of pumice powder was added to prevent bumpingduring distillation. The ammonia liberated was distilled into a receiver containing25ml of N/10 H2SO4. The excess of acid in the receiver was back titrated againstN/10 NaOH using 3 drops of methyl red indicator (0.1 gram of Methyl Red Indicator was dissolved in 60 ml alcohol and water was added to make the volume to 100 ml) . A reagent blank was similarlydigested and distilled. This titre value of Blank was subtracted from the value obtained for thesample to get the true titre value ‘b’.
Calculation:
If ‘a’ g of the sample was taken and if ‘b’ and ‘c’ ml of alkali of normality ‘d’ wererequired for back titration and to neutralise 25ml of N/10 H2SO4 respectively then
Protein content (g/100g of sample)= [(c-b) * (14 * d) * 6.25 * 100] / a * 1000
6. Crude Fibre[10,11]
About 2g of the moisture free sample was weighed into a 500ml beaker and 200 ml of boiling 0.255 N (1.25% W/V) sulphuric acid was added. The mixture was boiled for 30 minutes with bumping chips keeping the volume constant by addition of water at frequent intervals. A glass rod was placed in the beaker to ensure smooth boiling. At the end of this period, the mixture was filtered through a muslin cloth and the residue washed with hot water till free from acid. It was then boiled with 200ml of 1.25 % sodium hydroxide solution for 30 minutes. It was filtered through muslin cloth again and washed with 25 ml of boiling 1.25% H2SO4, three 50 ml portions of water and 25ml of alcohol. The residue was removed and transferred to ash dish (pre weighed dish W1). The residue was dried for 2 hours at 130±2 ? C. The dish was cooled in a desiccator and weighed (W2). It was then ignited for 30 minutes at 600±15 ? C. Cooled in a desiccator and reweighed (W3).
% crude fibre in ground sample= [Loss in weight in ignition(W2-W1)-(W3-W1)/ Weight of the sample] × 100
7. Energy Value [14]
The energy value of the 100g of food sample was calculated by using the followingformula.
Energy content = 4×wc+ 4×wp+ 9×wf
Where wc is the carbohydrate content (g/100g),
wp is the protein content (g/100g) and
wf is the fat content (g/100g).
8. Mineral Estimation
The working solution was prepared for measuring the minerals like Ca, Fe, Na, K, Cr, Se, Cu, Co, Mg, Mn, Pb, Zn, P, Mo using AOAC (Association of Official Analytical Chemists)method. The instrument used for mineral estimation was ICP (OES) or Inductively Coupled Plasma / Optical Emission Spectrometry, model ICAP6800, serial number ICP 20073108, calibrated with NIST (National Institute of Standards and Technology) certified multi standards[15].
9. β-Carotene Estimation [16]
β-Carotene content of sample was determined by using the colorimetric method of Srivastava and Kumar (2002). Five grams of the sample were crushed in 10–15 ml acetone by adding a few crystals of anhydrous Na2SO4. The supernatant was decanted into a beaker and the process was repeated twice. The combined supernatant was transferred to a separating funnel and 10–15 ml of petroleum ether were added and mixed thoroughly. After discarding the lower layer, the upper layer was collected in a 100 ml volumetric flask and the volume was made up with petroleum ether. The OD (Optical Density) was recorded at 452 nm and β-carotene was expressed as mg/100 g by the following formula.
β-Carotene (mg/100g) = Optical Density ×13.9×104×100
Weight of the Sample× 560×1000
10. Vitamin-C Content[17]
The blue colour produced by the reduction of 2,6-dichlorophenol indophenol by ascorbic acid was estimated colorimetrically.2 gram of sample was weighed and blended with convenient volume of 6% HPO3 to make 50 ml. The mixture was filtered and 5 ml of filtrate is placed in a 50 ml separating funnel. The same amount of extractant (6% HPO3) was taken in 2 more separating funnels, B and C. Funnel B served as the Dye Blank and to funnel C which served as Standard was added 0.1 ml (equivalent to 0.1 mg ascorbic acid) of the ascorbic acid standard solution. An amount of acetate buffer (pH 4) equal to the volume of the extract taken was then added to all the three funnels, followed by 2ml of the dye solution (25 mg of the sodium salt of 2,6-dichlorophenol was dissolved in distilled water and made up to 200 ml). 10 ml of Xylene was then added quickly and the contents shaken for 6-10 sec. After the layer separation, the lower water layer was removed and the colour in the Xylene extract was measured in a colorimeter at 500 nm.
11. Estimation of B-Vitamins
Vitamin B1 and Vitamin B2 were estimated by Fluorometric method [18,19] and Vitamin B3 was estimated by Colorimetric Method [20].
RESULTS
DISCUSSION
In a developing country like India, food scarcity is a colossal problem and has been plaguing the growth and development of precious human resource adversely. The rich heritage of flora and fauna which nature has bestowed us need to be explored and properly utilised as a source of nutrients. Plants and plant products have been widely used since time immemorial. Variety of gum exudates are used for their nutritional, culinary and therapeutic values throughout the world.
Natural gums are promising biodegradable polymeric materials which can serve as attractive alternatives to synthetic products because of their biocompatibility, low toxicity, environmental friendliness and low price[21].
The study undertaken here suggests that the Gum exudate obtained from Acacia nilotica is a very useful commodity which is not only energy dense but also an excellent reservoir of many minerals. Protein Energy Malnutrition is a very common phenomenon in our nation affecting children of rural and slum areas and thus weakening the backbone of the society. This gum can be utilised for producing edible products and can also be incorporated with other food ingredients to enhance its nutritive value. Literature studies indicated its utility and benefits in physiological conditions like pregnancy and lactation. Inadequate nutrient intake is a major cause of maternal mortality and Intrauterine Growth Retardation in foetuses. Thus, these traditional food products need to be advertised more and people should be encouraged to use this underutilised gum. The high carbohydrate content can exert hepatoprotective activity and stimulating property thus can be used in medicinal and pharmaceutical formulations.
Diarrhoea is a leading cause of childhood mortality in India and literature studies suggested it to play a protective role in its treatment. Acacia nilotica gum due to its high carbohydrate and mineral content can be beneficial in this condition by enhancing availability of glucose and absorption of solutes.
Its low protein and low fat content makes it useful in conditions associated with renal and cardiac functionalities. Its high mineral content makes it a magical ingredient. Minerals are indispensable in our diet because they serve as cofactors for many physiological and metabolic functions. It plays pivotal role in human life provides healthy growth and developments. High calcium and iron content of this gum exudate is can combat deficiency diseases associated with it. Presence of high amount of minerals like magnesium, manganese, chromium, Zinc etc. can help in the prevention of many degenerative diseases.
It contains appreciable amount of β-carotene, Vitamin C but negligible amount of thiamine, riboflavin and niacin.
Its low moisture content influences its perishability and thus can be stored for a long time period. It’s lower water activity hinder microbial activity and enhances shelf life.
CONCLUSION
A majority of the plant products remain virtually unexplored and thus new insight has to be developed into their potential use as therapeutic agents. It has high calorific value and the substantial mineral content can make it a viable supplement and immense source of dietary mineral in human food to fight various diseases. This natural gum can be used for encapsulation and production of several synthetic drugs with decreased side-effects. Its nutrient composition shows that it can have several physiological and nutritional benefits which need to be investigated. The ethno medicinal and traditional claims need reconnaissance and if properly employed can impart positive affect on nation’s economy.
Declaration:
Competing interests: The authors declare no potential conflicts of interest.
Funding:Department of Home Science, University of Calcutta, West Bengal, India & UGC (University Grant Commission), New Delhi, India.
Acknowledgements: I would like to thank Department of Home Science, University of Calcutta for providing me the necessary infrastructural facilities. I would also like to thank all my teachers and my parents for always guiding and motivating me. I would like to thank UGC, New Delhi, India for providing the financial assistance. I am extremely thankful to the authors, editors and publishers of articles, journals and books from where the literature is reviewed which in turn directed us to conduct this analytical study.
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