INTRODUCTION
Research rationale
Radishes are highly nutritious agricultural products that offer significant health benefits, serving as both food and medicine The two most commonly used varieties are white radish and red radish, each contributing unique flavors and nutritional properties to various dishes.
Vietnamese radish is valued not only for its economic benefits but also for its medicinal properties, including treatment for coughs, sore throats, and digestive issues As demand for high-quality radish increases, exporters are adopting measures such as crop enhancement and seed improvement, with plant protection drugs playing a crucial role in pest management However, while pesticides can boost crop yields, they also pose risks, including environmental pollution, human and animal poisoning, increased production costs, and harmful residues in agricultural products that jeopardize consumer health To address these concerns and ensure the safety of consumers, we propose a project focused on developing a highly sensitive method for analyzing Carbaryl residues in white radish using liquid chromatography.
Chromatography tandem Mass Spectrometry (LC-MS /MS)”.
Research’s objective
The objective for this study is evaluate the sensitive and accurate method based on: Linearity, Repeatability, Recovery, Reproducibility, Limit of detection
(LOD), Limit of quantification (LOQ)
- Why choose Liquid Chromatography tandem mass spectrometry method to analyze pesticide residues in white radish?
- How to confirm validity of the method?
Commented [A1]: 15/3/2016 món ngon vị thuốc từ củ cải
Commented [A2]: Sở tài nguyên và môi trường Vĩnh Phúc
(2008) Thuốc bảo vệ thực vật và những tác động của chúng
Due to constraints of time and sample responsiveness, the sample size is limited So, certain factors could not be studied in depth.
Limitations
Due to constraints of time and sample responsiveness, the sample size is limited So, certain factors could not be studied in depth.
LITERATURE REVIEW
Radishes
Radish (Raphanus sativus L) is an annual vegetable from the Cruciferae family, valued in various cultures Its enlarged root and hypocotyl are primarily consumed as a salted vegetable, while fresh grated radish is popular in salads and as a garnish Additionally, young leaves and seedlings, referred to as 'kaiware,' are also edible This vegetable is classified under Plantae Angiosperms, Magnoliopsida, and Brassicales.
Radish is a globally cultivated vegetable known for its spicy flavor and long tubers, which can reach up to 40 cm This versatile vegetable is available year-round and can be harvested within 45 to 55 days of planting Commonly used in various cuisines, radish can be paired with other ingredients to create delicious dishes According to the Hai Thuong book, radish offers a sweet yet slightly bitter taste and is non-toxic, providing health benefits such as relieving sputum and alleviating stasis, though it should be avoided in cases of dysentery.
2.1.2 Chemical composition and nutritional value
Modern medicine highlights the nutritional value of radishes, revealing that 100 grams of this vegetable contains 1.4 grams of protein, 3.7 grams of carbohydrates, and 1.5 grams of fiber Additionally, radishes provide essential minerals such as 40 mg of calcium and 41 mg of phosphorus, along with 1.1 mg of iron They also contain small amounts of vitamins, including 0.06 mg of vitamin B1, 0.06 mg of vitamin B2, 0.5 mg of vitamin PP, and a notable 30 mg of vitamin C.
Table 2.1: Nutritional value per 100g radish
Encyclopedia of Food Sciences and Nutrition (Second Edition), 2003
Commented [A4]: Seaman, Abby (2013-11-13) "Turnips and Radishes" Integrated crop and pest management guidelines for commercial vegetable production Retrieved 2020-05-20
Commented [A5]: 15/3/2016 món ngon vị thuốc từ củ cải
Radishes possess a spicy flavor that contributes to their antibacterial properties, promotes tendon activity, and serves as an effective analgesic During the winter months, if you experience muscle or joint pain, consider placing radishes in a cloth bag and applying it to the affected area for relief.
- Good for liver, prevent cardiovascular disease
Radishes are rich in Betaine, a bioactive compound that promotes healthy liver function A well-functioning liver effectively breaks down fat, aiding in weight loss and reducing feelings of fatigue and nausea Additionally, Betaine, also present in beets, helps lower plasma homocysteine levels, which is important for reducing the risk of cardiovascular disease.
A study conducted at Wake Forest University reveals that the high nitrate content in radishes can boost blood flow to the brain, leading to improved mental function and enhanced cognitive abilities, while also providing long-term protection for brain cells.
Commented [A6]: 6/7/2016 tac dung cua cu cai
The natural Vitamin B12 in Radish helps promote absorption of iron, involved in the synthesis of Hemoglobin, so the increased amount of Oxygen
Hemoglobin that good for the body, prevent anemia
Radish leaves are rich in vitamins A and C, with particularly high levels of vitamin C compared to other vegetables, which aids in preventing skin aging and the formation of dark spots, promoting a brighter and softer complexion Additionally, using radish paste can be an effective natural remedy for facial cleansing.
Radishes are a good source for Anthocyanins that keep our hearts functioning properly, reducing the risk of cardiovascular diseases Addition, Radish are high on
Vitamin C, Folic acid, and Flavonoids also.
Plant protection products
Plant protection drugs, also known as pesticides or plant protection products, are toxic substances derived from natural sources or synthesized chemicals They play a crucial role in agriculture by safeguarding plants and crops from pests in fields and gardens.
Plant protection drugs can be divided into 4 main groups:
- Chlorine group: Are Chlorine derivatives of some organic compounds such as
Diphenyletane, Cyclodiene, Benzene and Hexane This group includes Organic compounds that very stable in the natural environment and have a long half-life
Representatives of this group are Aldrin, Dieldrin, Heptachlo, Lindan, Methoxychlor
The organophosphorus group primarily consists of esters, which are organic derivatives of phosphoric acid These compounds have a shorter half-life compared to organic chlorine and are more widely used They impact insect nerves by inhibiting the formation of the Cholinesterase enzyme, leading to disrupted nerve function.
The Department of Natural Resources and Environment of Vinh Phuc Province (2008) highlights the harmful effects of pesticides, which can lead to muscle weakness, dizziness, and even death Notable compounds in this category include Parathion, Malathion, Diclovos, and Clopyrifos.
Carbamates are organic derivatives of carbamic acid that include unstable chemicals harmful to humans and animals They function by directly inhibiting the enzyme cholinesterase in the nervous system, exhibiting a toxic mechanism similar to that of organophosphates.
Representing for this group as Carbofuran, Carbaryl, Carbosulfan, Isoprocarb,
- Pyrethroids: These are natural-derived insecticides, mixtures of different esters with complex structures separated from flowers of certain daisy varieties
Representatives of this group include Cypermethrin, Permethrin, Fenvalarate,
Deltamethrin, etc In addition, there are some other groups such as: Inorganic insecticides, biological insecticides derived from Bacteria, Fungi, Viruses
(Fungicides, Bacteria ), Inorganic compounds (compounds of Copper, Mercury)
2.2.3 List of pesticides allowed to be used and banned in Vietnam.[4]
According to Circular No 10/2019/TT Ministry of Agriculture and Rural
Development about promulgating the list of pesticides allowed to be used and banned in Vietnam
Commented [A8]: Theo thông tư số 10/2019/TT
BNNPTNT ban hành danh mục thuốc BVTV được phép sử dụng, cấm sử dụng tại Việt Nam
Table 2.2: List number of common name and trade name for pesticides allowed to be used and banned in Vietnam
Type of pesticides Number of Common name Number of Trade name
List of pesticides banned in Vietnam
Pesticides and forest product preservative
- List of pesticides allowed to be used in Vietnam
2.2.4 Effect of pesticide to health [4]
Most pesticides are toxic to humans and animals at difference levels
- Acute toxins: Below the lethal dose, they are not capable of causing death, gradually resolved and excreted
Chronic toxins, such as plant protection drugs, can persist in the body for extended periods due to their durability and resistance to breakdown and excretion These pesticides can enter humans and animals through three primary routes: respiratory, digestive, and direct contact Exposure to these substances may lead to either acute or chronic poisoning, depending on the severity of the pesticide's impact.
Acute intoxication can lead to severe symptoms such as vomiting, headache, dizziness, and sweating If the toxicity is significant, it may affect liver metabolism, resulting in neurological disorders, insomnia, memory issues, blurred vision, and hearing loss Women may face increased risks of miscarriage, premature birth, and congenital malformations in their children Other symptoms include fatigue, skin itching, hoarseness, increased salivation, and watery eyes, with severe cases potentially resulting in death.
- Chronic poisoning: Stimulates cancer cells to grow, cause birth defects, deformities, impaired memory and concentration, serious weakness, affecting the nervous system and brain
The Poison Control Center at Bach Mai Hospital highlights key symptoms of pesticide poisoning, which can affect the digestive system, leading to nausea and severe diarrhea Additionally, neurological symptoms may include drowsiness, coma, convulsions, and muscle paralysis These conditions can result in hypotension and respiratory failure, ultimately contributing to fatalities caused by plant protection drug poisoning.
Cholinergic syndrome, often associated with poisoning from organic phosphorus pesticides and carbamates, can lead to significant urinary issues Symptoms may include vomiting, shortness of breath, chest pain, excessive sweating, body tremors, and seizures, affecting both adults and children.
2.2.5 Current situation of pesticide residues in vegetables
- Determined 790 samples of 52 units of safe vegetables business in the city,
In the past 11 months, the Ho Chi Minh City Plant Protection Department identified 26 samples of pesticide residue, representing 3.29% of their inspections Notably, straw mushrooms from Tra Vinh, along with vegetables such as celery, lettuce, broccoli, cauliflower, and amaranth, exhibited high levels of pesticide residue.
(3.94%) Including 5 green onion samples of Da Lat, 2 samples of morning glory and choysum of Ho Chi Minh City, which over maximum residue level [5]
- 31/67 people in Hanoi were tested with blood levels of pesticides The results were published by the Institute of Occupational Health and Environment
(Ministry of Health) when conducting random tests at a class of the Center for
Community learning initiatives in the suburban districts of Hanoi, including Soc Son, Dong Anh, Me Linh, and Hoai Duc, have highlighted significant health risks, with 31 individuals identified as having pesticides stored in their blood Additionally, one individual is classified as being at a more severe risk level.
Over 221 pesticide types are detected in vegetables, with nearly half exceeding permissible levels In the last decade, countries like Australia, the US, Japan, and the EU have rejected 483 Vietnamese vegetable products, totaling over $1 billion, primarily due to excessive use of fertilizers and pesticides by farmers Additionally, many enterprises fail to adhere to standards such as VIETGAP and Global GAP, impacting the export potential of Vietnamese agricultural products.
- In recent years, the authorities took 7593 samples of vegetables and found
A total of 393 samples were found to be contaminated with banned substances, with pesticide residues exceeding permitted limits in 5.17% of cases This high level of violations raises concerns about food safety and indicates that public trust is not being met The lack of compliance by manufacturers with plant protection product regulations poses a significant risk of unsafe food products.
Commented [A9]: Danchi (2006) rau xạch thừa thuốc trừ sâu
Commented [A10]: Danchi (31/07/2018) Quá nửa số người xét nghiệm bị nhiễm thuốc trừ sâu trong máu
Commented [A11]: Trung kiên 30/4/2017 Nông sản xuất khẩu "gặp khó" vì tồn dư thuốc bảo vệ thực vật
Commented [A12]: Diệu thu (09/06/2016) Hơn 5% lượng rau bán trên thị trường nhiễm chất cấm
- In Ho Chi Minh City, in the first six months of 2009, more than 2,200 samples of vegetables and fruits were tested in three wholesale markets (Binh Dien,
In the first eight months of 2009, Hoc Mon and Thu Duc reported 50 positive samples of plant protection drug residues, reflecting a 2.4% prevalence, which is 1.3% higher compared to the same period in 2008 Additionally, Binh Duong analyzed nearly 310 vegetable samples from markets, production areas, and collective kitchens, revealing that almost 80 samples contained residues of these chemicals.
2.2.6 Food poisoning situation due to pesticides
- According to a survey of the Department of Preventive Medicine and
Vietnam faces significant environmental challenges, with over 5,000 annual hospitalizations and more than 300 deaths linked to toxic chemical pesticides The detrimental effects of these plant protection drugs on human health and the environment arise primarily from their improper use, which often deviates from established safety standards.
- On 14/11, Xuan Non Kindergarten's collective kitchen organized cooking for 796 lunches and snacks at 14:00 pm on the same day Lunch menu includes:
Enjoy a delicious meal featuring sausage, beef in wine sauce, Duong Chau fried rice, mixed boiled vegetables, and refreshing orange juice For the afternoon, indulge in yogurt and cakes Following lunch, three teachers and nearly 200 students were welcomed at Dong.
Anh General Hospital and North Thang Long Hospital with the symptoms of abdominal pain, vomiting, diarrhea, headache and fever [11].
Carbaryl
Carbaryl, known chemically as 1-naphthyl methylcarbamate, is a pesticide belonging to the carbamate group This white crystalline solid is widely recognized by its brand name, Sevin, which is a trademark of Bayer Originally discovered by Union Carbide, carbaryl was introduced to the market in 1958, and Bayer later acquired Aventis, further solidifying its presence in the agricultural sector.
CropScience in 2002, a company that includes the union carbide pesticide activities
The insecticide ranks as the third most popular choice in the United States for home gardens, commercial agriculture, and forest and rangeland protection, with approximately 11 million kilograms utilized in U.S agricultural crops in 1976.
Commented [A13]: Công An Ninh Thuận ( 2009), Hơn hai triệu người ăn rau bẩn
Commented [A14]: Nguyễn Thị Thanh Huyền 01/11/2010
Tác hại của thuốc hóa chất bảo vệ thực vật đối với sức khỏe con người và môi trường
Carbaryl have chemical formula is C12H11NO2, molar mass is 201.225 gãmol−1, melting point at 142 °C (288 °F; 415 K), the solubility in water very low
Figure 2.1 structure formula of Carbaryl
Carbaryl, the active ingredient in Carylderm shampoos, effectively combats head lice until the infestation is eliminated While it is approved for use on more than 100 crops in the United States, Carbaryl is banned in several countries, including the UK, Austria, Denmark, Sweden, and Iran.
The Acceptable Daily Intake (ADI) for carbaryl is set at 0.008 mg/kg of body weight In studies involving mice, the average lethal dose (LD50) ranges from 250 to 850 mg/kg, while the average lethal concentration (LC50) is between 0.005 and 0.023 mg/kg Additionally, the Maximum Residue Level (MRL) of carbaryl in beets is established at 0.1 mg/kg.
• Carbaryl is used to control a wide range of pests, including beetles, cockroaches, ants, ticks and mosquitoes
• Carbaryl products can be made in the form of dust, absorbent powders, liquid concentrates, granules
• Carbaryl products are used on fruits, vegetables, rangeland, lawns, ornamental plants, trees, and building foundations
- Effect of carbaryl to health
Early symptoms of Carbaryl exposure can manifest as headaches, muscle weakness, nausea, stomach cramps, sweating, and restlessness The impact of Carbaryl on human health and the environment is influenced by the level of exposure.
Commented [A15]: Robert L Metcalf “côn trùng Control” trong Bách khoa toàn thư Ullmann của Hóa học công nghiệp”Wiley-VCH, Weinheim, 2002 doi : 10,1002 / 14356007.a14_263
Commented [A16]: Dữ liệu nguy hiểm về thuốc trừ sâu
Carbaryl được lưu trữ ngày 11 tháng 5 năm 2010, tại Wayback Machine
Commented [A17]: 2003 NPIC national pesticide inf center present and the length and frequency of exposure The reaction also depends on the health of each person or certain environmental factors
Greater exposure to Carbaryl can lead to mydriasis, tearing, excessive salivation, runny nose, vomiting, diarrhea, muscle twitching, and coordination problems Severe poisoning can lead to convulsions, coma and death
• Carbaryl is practically non-toxic to birds such as ducks, quails, geese and pheasants [20]
• Carbaryl ranges from toxic to extremely toxic to some fish [20]
Carbaryl poses significant risks to beneficial insects, particularly bees, which may be adversely affected in areas where it is applied Although there have been reports of bee fatalities linked to its use, a specific application, apart from one instance involving asparagus, has not been directly associated with these incidents Concerns have been voiced about carbaryl's role in thinning fruit in orchards; however, a field study demonstrated that its application did not significantly affect bee mortality or behavior.
- Maximum residue limit of carbaryl [15]
- Maximum Residue Level - MRL is the maximum content of a pesticide residue in a food (unit: mg/kg of food)
- Acceptable Daily Intake – ADI is the daily intake of a chemical throughout life without harm to human health (unit: mg/kg body weight)
Table 2.3: MRL and ADI of Carbaryl in some vegetables
Pesticides ADI (mg/kg) Products MRL (mg/kg)
Commented [A18]: A World Compendium: The Pesticide
Manual, 12th ed.; Tomlin, C D S., Ed.; British Crop Protection Council: Farnham, Surrey, UK, 2000; pp 67-68
The Environmental Fate and Effects Division of the Agency's Office of Pesticide Programs has conducted a revised risk assessment of Carbaryl to support its Reregistration Eligibility Decision (RED) This assessment is crucial for evaluating the environmental impact and safety of Carbaryl, a widely used pesticide For more detailed information, refer to the full document available at the provided link.
Commented [A20]: R David Jones, Ph.D., Senior
Agronomist Thomas Steeger, Ph.D., Senior Biologist March 18, 2003 Revised EFED Risk Assessment of Carbaryl in Support of the Reregistration
Commented [A21]: Theo thông tư số 50-2016 TT/BYT về Quy định giới hạn tối đa dư lượng thuốc bảo vệ thực vật trong thực phẩm.
Current methods for determination of Carbaryl in vegetable
Some traditional methods are used to determine pesticide residues in fruits and vegetables such as: Liquid–Liquid Extraction, Microwave Assisted Extraction,
Solid Phase Extraction, Supercritical Extraction Most of which take a long time
Liquid-liquid extraction, also known as solvent extraction or partitioning, is a technique used to separate compounds or metal complexes based on their solubility in two immiscible liquids This process involves transferring the analyte from one solvent to another while recovering the original solvent to obtain the analyte Selecting the appropriate solvent is crucial to minimize analyte loss and enhance extraction efficiency Key factors influencing the extraction process include the solubility of the analyte, temperature, and the presence of other solutes Additionally, the right solvent choice can aid in eliminating impurities from the sample Auxiliary physical methods such as mechanical shaking, high-speed agitation, and ultrasonic waves can further improve extraction, especially for samples with high impurity levels, which may require coordination with other purification processes.
Solid phase extraction (SPE) is a technique used to separate compounds from liquid mixtures based on their physical and chemical properties, making it essential in analytical laboratories for concentrating and purifying samples This method effectively isolates analytes from various matrices, including blood, water, beverages, soil, and animal tissue While SPE is cost-effective and offers high accuracy, it may sometimes result in incomplete removal of interferences and variability in results.
The QUECHERS method it means quick, easy, cheap, effective, rugged and safe It was expanded by Michelangelo Anastassiades in the years 2001 and 2002
The first time, the methodology was developed for the analysis of veterinary drugs
Commented [A22]: Berk, Zeki (2013) "Food Process
Engineering and Technology (Second Edition)" Advances in Molecular Toxicology, 2014 ISBN 978-0-12-415923-5
The QUECHERS technique, initially recognized for its effectiveness in extracting polar and basic compounds from animal tissues, has proven valuable in pesticide residue analysis of plant materials, including fruits and vegetables Today, it serves as the foundation for widely accepted testing methods globally, such as AOAC 2010 and EN standards.
15662 and is being applied in the large and modern laboratories in our country [25]
A straightforward, cost-effective method for detecting pesticide residues in fruits and vegetables is presented This procedure begins with the extraction of a 10 g sample using 10 ml of acetonitrile, followed by a liquid-liquid partition by adding 4 g of anhydrous MgSO4 and 1 g of NaCl The simultaneous removal of residual water and cleaning is achieved through a rapid technique known as dispersed solid phase extraction (dispersion-SPE), utilizing 150 mg of anhydrous MgSO4.
The process involves mixing 25 mg of MgSO4 and a secondary amine absorbent (PSA) with 1 ml of acetonitrile extract This method of Dispersion-Solid Phase Extraction (SPE) using PSA effectively removes various polar matrix components, including organic acids, certain polar pigments, and sugars, from food extracts Subsequently, Gas Chromatography Mass Spectrometry (GC/MS) is employed for the quantitative analysis and confirmation of GC-amenable pesticides.
A recovery of 85 to 101% (mainly> 95%) and repeatability