ls /home/daytona/workspace/attachments/e210dfc2-9b0a-4fb2-8d78-490668104b4e/

cd /home/daytona/workspace && python3 -c " import pdfplumber, json files = [ …

python3 -c " import fitz # PyMuPDF import os base = '/home/daytona/workspace/a…

python3 -c "import pdf2image; print('pdf2image ok')" 2>&1; python3 -c "import py…

python3 << 'EOF' from pdf2image import convert_from_path import pytesseract impo…

python3 << 'EOF' import pytesseract from PIL import Image import os, glob image…

python3 << 'EOF' import os, glob ocr_dir = '/home/daytona/workspace/pdf_ocr' f…

python3 << 'PYEOF' from reportlab.lib.pagesizes import A4 from reportlab.lib imp…

~/create_spotters_pdf.py

from reportlab.lib.pagesizes import A4
from reportlab.lib import colors
from reportlab.lib.units import cm, mm
from reportlab.platypus import (
    SimpleDocTemplate, Paragraph, Spacer, PageBreak,
    Image as RLImage, Table, TableStyle, HRFlowable, KeepTogether
)
from reportlab.lib.styles import getSampleStyleSheet, ParagraphStyle
from reportlab.lib.enums import TA_LEFT, TA_CENTER, TA_JUSTIFY
from reportlab.pdfgen import canvas
from reportlab.lib.utils import ImageReader
from PIL import Image as PILImage
import io, os

OUTPUT_PATH = '/home/daytona/workspace/spotters_output/Biochemistry_Spotters_Combined_Answers.pdf'
IMAGE_DIR = '/home/daytona/workspace/pdf_images'
os.makedirs('/home/daytona/workspace/spotters_output', exist_ok=True)

# ------------------------------------------------------------------
# All spotter data
# ------------------------------------------------------------------
spotter_pages = [
    # ===== SPOTTERS 2-1.PDF =====
    {"file": "PDF 1 – Spotters 2-1", "section_header": True},
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page01.png",
        "label": "Spot – Equipment Identification",
        "question": "Identify the equipment and write its use in laboratory.",
        "answer": (
            "IDENTIFICATION: Weighing Balance (Analytical / Digital Balance)\n\n"
            "USES IN LABORATORY:\n"
            "• To accurately measure the mass/weight of chemicals before preparing solutions or reagents.\n"
            "• Used for preparing standard solutions of known concentration.\n"
            "• Essential for quantitative analysis in biochemistry labs."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page02.png",
        "label": "Spot 4 – Equipment Identification",
        "question": "Identify the above picture and write its use.",
        "answer": (
            "IDENTIFICATION: Hot Air Oven\n\n"
            "USES:\n"
            "• Used for dry heat sterilization of glassware (test tubes, Petri dishes, pipettes).\n"
            "• Temperature range: 160–180°C for 1–2 hours.\n"
            "• Also used for drying glassware after washing."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page03.png",
        "label": "Spot 1 – Triglyceride / VLDL Calculation",
        "question": "1. What is the normal range of Triglyceride level in Serum?\n2. Find out the VLDL level in serum if Triglyceride is 150 mg%.",
        "answer": (
            "1. Normal range of Serum Triglyceride: < 150 mg/dL (Desirable)\n"
            "   Borderline high: 150–199 mg/dL   |   High: 200–499 mg/dL\n\n"
            "2. VLDL Calculation (Friedewald Formula):\n"
            "   VLDL = Triglyceride / 5\n"
            "   VLDL = 150 / 5 = 30 mg%\n"
            "   ∴ VLDL level = 30 mg%"
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page04.png",
        "label": "Spot – Instrument Identification",
        "question": "Identify the Instrument and write its uses.",
        "answer": (
            "IDENTIFICATION: Colorimeter / Photoelectric Colorimeter\n\n"
            "USES:\n"
            "• Measures concentration of colored compounds using Beer-Lambert Law.\n"
            "• Used to estimate Hemoglobin, Blood Glucose, Serum Proteins, Bilirubin, Creatinine, Urea.\n"
            "• Wavelength range: 400–700 nm (visible light)."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page05.png",
        "label": "Spot – A:G Ratio Calculation",
        "question": "Concentration of Protein = 7.1 gm%\nConcentration of Albumin = 4.2 gm%\nFind out A:G ratio and write its Normal Range.",
        "answer": (
            "CALCULATION:\n"
            "  Globulin = Total Protein − Albumin = 7.1 − 4.2 = 2.9 gm%\n"
            "  A:G Ratio = Albumin / Globulin = 4.2 / 2.9 = 1.45 : 1\n"
            "  ∴ A:G Ratio ≈ 1.45\n\n"
            "NORMAL RANGE of A:G Ratio: 1.2 – 2.2\n"
            "  Decreased in: Liver disease, Nephrotic syndrome, Multiple myeloma."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page06.png",
        "label": "Spot 6 – ELISA & RIA",
        "question": "1. Full form of ELISA  2. Types of ELISA  3. Uses of ELISA  4. Full form of RIA  5. Radioactive component in RIA?",
        "answer": (
            "1. ELISA = Enzyme-Linked Immunosorbent Assay\n\n"
            "2. Types of ELISA: Direct, Indirect, Sandwich, Competitive\n\n"
            "3. Uses of ELISA: Detection of HIV, Hepatitis B/C antibodies; Hormone assays (TSH, hCG);\n"
            "   Diagnosis of infectious diseases; Detection of allergens and drugs.\n\n"
            "4. RIA = RadioImmunoAssay\n\n"
            "5. Radioactive component in RIA: ¹²⁵I (Iodine-125) – gamma emitter."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page07.png",
        "label": "Spot – Safety Measures in Laboratory",
        "question": "Write 4 safety measures in laboratory.",
        "answer": (
            "1. Wear PPE: Lab coat, gloves, safety goggles, and face mask at all times.\n\n"
            "2. Proper waste disposal: Biohazardous waste in yellow bags; sharps in puncture-proof\n"
            "   sharps containers; chemical waste in designated chemical waste bins.\n\n"
            "3. No eating, drinking, or mouth pipetting inside the laboratory.\n\n"
            "4. Emergency preparedness: Know location of fire extinguisher, eyewash station,\n"
            "   first aid kit, and emergency exit. Report spills immediately."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page08.png",
        "label": "Spot – Symbol Identification",
        "question": "Identify the symbol and write two examples covered under this category.",
        "answer": (
            "IDENTIFICATION: Biohazard Symbol (Biological Hazard Warning)\n\n"
            "Indicates presence of biological substances that pose a risk to human health.\n\n"
            "TWO EXAMPLES:\n"
            "1. Blood and blood-borne pathogens: e.g., HIV-infected blood, Hepatitis B/C samples.\n"
            "2. Microbiological cultures: e.g., bacterial cultures (Mycobacterium TB), viral stocks."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page09.png",
        "label": "Spot 6 – Machine Identification (5 Marks)",
        "question": "1. Identify the machine and write its principle (3 marks)\n2. Write its types (1 mark)\n3. Write its uses (1 mark)",
        "answer": (
            "1. IDENTIFICATION: Centrifuge\n"
            "   PRINCIPLE: Uses centrifugal force generated by rapid rotation to separate particles of\n"
            "   different densities in a liquid suspension. Denser particles sediment to the bottom\n"
            "   (pellet); lighter components remain in the supernatant. Force measured in RCF (g-force).\n\n"
            "2. TYPES: Low-speed (≤6,000 RPM), High-speed (up to 25,000 RPM),\n"
            "   Ultracentrifuge (>100,000 RPM), Microcentrifuge (for Eppendorf tubes).\n\n"
            "3. USES: Separation of serum/plasma from blood; sedimentation of cells and bacteria;\n"
            "   urinalysis (concentrating sediment); isolation of organelles."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page10.png",
        "label": "Spot – Equipment Identification",
        "question": "Identify the above picture and write its use.",
        "answer": (
            "IDENTIFICATION: pH Meter (Electronic pH Meter)\n\n"
            "USES:\n"
            "• Measures hydrogen ion concentration (acidity/alkalinity) of a solution.\n"
            "• Used to prepare buffer solutions of exact pH in the laboratory.\n"
            "• Used to check pH of reagents and biological fluids (urine, blood, CSF).\n"
            "• Principle: Glass electrode generates voltage proportional to H⁺ ion activity (pH)."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page11.png",
        "label": "Spot No. 1 – Glassware Identification",
        "question": "Identify the glassware and write its uses.",
        "answer": (
            "IDENTIFICATION: Measuring Cylinder (Graduated Cylinder)\n\n"
            "USES:\n"
            "• Used to measure approximate volumes of liquids accurately.\n"
            "• Graduated markings allow reading of volume at the bottom of the meniscus.\n"
            "• Available in various sizes: 10, 25, 50, 100, 500 mL.\n"
            "• Used in biochemistry labs to prepare solutions of required volumes."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page12.png",
        "label": "Spot No. 5 – Machine Identification",
        "question": "1. Identify the given machine.\n2. Write its use.",
        "answer": (
            "1. IDENTIFICATION: Semi-automated Biochemistry Analyzer\n\n"
            "2. USES:\n"
            "   • Performs multiple biochemical tests on serum, plasma, urine, CSF automatically.\n"
            "   • Tests include: Blood glucose, urea, creatinine, total protein, albumin, bilirubin,\n"
            "     liver enzymes (ALT, AST), lipid profile, electrolytes.\n"
            "   • Principle: Photometric/colorimetric measurement using enzyme-based reactions.\n"
            "   • Reduces manual errors and increases throughput in clinical laboratories."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page13.png",
        "label": "Spot – Instrument Identification",
        "question": "Identify the instrument and write its use.",
        "answer": (
            "IDENTIFICATION: Vortex Mixer (Vortex Shaker)\n\n"
            "USE:\n"
            "• Simple device used to rapidly mix small volumes of liquid in tubes/vials.\n"
            "• Creates a vortex (whirlpool) for thorough mixing of reagents.\n"
            "• Used to mix reagents, dissolve precipitates, and resuspend cell pellets.\n"
            "• Operates in continuous or touch mode with adjustable speed."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page14.png",
        "label": "Spot 2 – Equipment Identification",
        "question": "Identify the equipment and write its use.",
        "answer": (
            "IDENTIFICATION: Micropipette (Adjustable Automatic Micropipette)\n\n"
            "USES:\n"
            "• Accurately measures and transfers very small volumes (0.1 µL to 1000 µL).\n"
            "• Essential in molecular biology, serology, immunology, and biochemistry.\n"
            "• Available as P10 (0.5–10 µL), P200 (20–200 µL), P1000 (100–1000 µL).\n"
            "• Uses disposable sterile tips to prevent cross-contamination."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page15.png",
        "label": "Spot No. 3 – Test Identification",
        "question": "Identify the test and write its principle.",
        "answer": (
            "IDENTIFICATION: Seliwanoff's Test (Test for Ketose Sugars)\n\n"
            "PRINCIPLE:\n"
            "• Differentiates between ketose and aldose sugars based on differential dehydration by HCl.\n"
            "• Ketoses (e.g., Fructose) are rapidly dehydrated by hot HCl to form Hydroxymethylfurfural (HMF),\n"
            "  which condenses with resorcinol to produce a CHERRY RED color within 1 minute.\n"
            "• Aldoses (e.g., Glucose) react slowly and give faint pink or no color within 1 minute.\n"
            "REAGENT: Resorcinol in concentrated HCl.\n"
            "POSITIVE: Cherry red/deep red color = Ketose sugar."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page16.png",
        "label": "Spot – Vacutainer Tubes",
        "question": "Identify the above pictures and write their uses.",
        "answer": (
            "IDENTIFICATION: Vacutainer Tubes (Blood Collection Tubes)\n\n"
            "Color-coded uses:\n"
            "• Red (no anticoagulant): Biochemical & serological tests (LFT, RFT, Lipid profile, Hormones)\n"
            "• Grey (Sodium Fluoride): Glucose estimation\n"
            "• Purple/Lavender (EDTA): CBC, ESR, Blood grouping\n"
            "• Blue (Sodium Citrate 3.2%): Coagulation studies – PT, APTT, INR\n"
            "• Green (Heparin): Electrolytes, Bone marrow studies, ABG\n"
            "• Yellow (ACD/Citrate): Blood culture, Blood bank tests\n"
            "• Dark Blue (EDTA K2): Trace elements, Toxicology"
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page17.png",
        "label": "Spot – Safety Measures",
        "question": "Write 4 safety measures in laboratory.",
        "answer": (
            "1. Use PPE: Lab coat, gloves, goggles, and masks must be worn at all times to prevent\n"
            "   exposure to hazardous chemicals and biological agents.\n\n"
            "2. Proper labeling and storage: All reagents labeled clearly; flammable chemicals away\n"
            "   from heat; corrosives in acid cabinets.\n\n"
            "3. Correct waste disposal: Biohazard bags for biological waste; sharps containers\n"
            "   for needles/broken glass; chemical waste bins for chemicals.\n\n"
            "4. Emergency preparedness: Know location of fire extinguisher, eyewash station,\n"
            "   first aid kit, and emergency exits. Follow decontamination procedures for spills."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_2-1_page18.png",
        "label": "Spot – Glassware Identification",
        "question": "Identify the glassware and write its use.",
        "answer": (
            "IDENTIFICATION: Conical Flask (Erlenmeyer Flask)\n\n"
            "USE:\n"
            "• Widely used in titration experiments to hold the solution being titrated.\n"
            "• Wide base and narrow neck allows swirling without spilling.\n"
            "• Used to mix, heat, and store liquids; preparation of culture media and reagents.\n"
            "• The conical shape minimizes evaporation and splashing during mixing."
        ),
    },

    # ===== SPOTTERS BIOCHEMISTRY -1.PDF =====
    {"file": "PDF 2 – Spotters Biochemistry -1", "section_header": True},
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page01.png",
        "label": "Spot No. 1 – Instrument Identification",
        "question": "Identify the Instrument and write its uses.",
        "answer": (
            "IDENTIFICATION: Colorimeter (Photoelectric Colorimeter)\n\n"
            "USES:\n"
            "• Measures concentration of colored substances using Beer-Lambert Law.\n"
            "• Used for estimation of: Hemoglobin, Blood glucose, Bilirubin, Serum proteins,\n"
            "  Urea, Creatinine, and other colored compounds.\n"
            "• Light at specific wavelength (400–700 nm) passes through sample; absorbance\n"
            "  is measured and compared to standard to calculate concentration."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page02.png",
        "label": "Spot No. 2 – Seliwanoff's Test",
        "question": "Which test is used to differentiate between ketosugar and aldosugar? Write its principle.",
        "answer": (
            "TEST: Seliwanoff's Test\n\n"
            "PRINCIPLE:\n"
            "• Ketoses (e.g., Fructose) are rapidly dehydrated by hot HCl to form Hydroxymethylfurfural (HMF),\n"
            "  which condenses with resorcinol to produce a CHERRY RED color within 1 minute.\n"
            "• Aldoses (e.g., Glucose, Galactose) react slowly and give only faint pink or no color\n"
            "  within 1 minute.\n"
            "REAGENT: Resorcinol dissolved in concentrated HCl.\n"
            "POSITIVE (Ketose): Cherry red color. NEGATIVE (Aldose): Faint pink/no color."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page03.png",
        "label": "Spot 3 – Equipment Identification",
        "question": "Identify the equipment and write its use.",
        "answer": (
            "IDENTIFICATION: Weighing Balance (Analytical/Digital Balance)\n\n"
            "USE:\n"
            "• Measures mass/weight of substances accurately (sensitivity up to 0.0001 g).\n"
            "• Used to weigh chemicals for preparing standard solutions and reagents.\n"
            "• Essential for quantitative biochemical analysis."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page04.png",
        "label": "Spot 4 – Safety Measures",
        "question": "Write 4 safety measures in laboratory.",
        "answer": (
            "1. Always wear appropriate PPE (lab coat, gloves, safety goggles) before entering the lab.\n\n"
            "2. Never pipette by mouth – always use mechanical pipette aids or micropipettes.\n\n"
            "3. Label all specimens and reagents clearly. Handle infectious samples under a biosafety cabinet.\n\n"
            "4. Know emergency procedures: location of fire extinguisher, first aid kit, eyewash station,\n"
            "   and emergency exit. Wash hands thoroughly before leaving the laboratory."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page05.png",
        "label": "Spot No. 5 – Test Identification",
        "question": "Identify the test and write its Principle.",
        "answer": (
            "IDENTIFICATION: Benedict's Test (Test for Reducing Sugars)\n\n"
            "PRINCIPLE:\n"
            "• Reducing sugars reduce Cu²⁺ (cupric ions) in alkaline Benedict's reagent\n"
            "  to Cu₂O (cuprous oxide), forming a colored precipitate when heated.\n"
            "• Color interpretation:\n"
            "  Green = trace (~0.1–0.5%)   Yellow = ~0.5–1.0%\n"
            "  Orange = ~1.0–1.5%          Brick red = >2% reducing sugar\n"
            "REAGENT: CuSO₄ + sodium citrate + Na₂CO₃ (deep blue alkaline solution).\n"
            "POSITIVE: Color change from blue → green/yellow/orange/brick red."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page06.png",
        "label": "Spot 1 – Vitamin C Deficiency",
        "question": "Write any 2 manifestations of Vitamin C deficiency.",
        "answer": (
            "VITAMIN C DEFICIENCY = SCURVY\n\n"
            "2 Manifestations:\n\n"
            "1. Bleeding gums and perifollicular hemorrhages:\n"
            "   Vitamin C is essential for collagen synthesis (hydroxylation of proline/lysine).\n"
            "   Deficiency → defective collagen → fragile capillaries → bleeding gums (scorbutic\n"
            "   gingivitis) and perifollicular hemorrhages (bleeding around hair follicles).\n\n"
            "2. Poor wound healing and corkscrew hairs:\n"
            "   Defective collagen leads to impaired wound healing. Corkscrew/swan-neck\n"
            "   deformity of hair is a classic sign of scurvy."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page07.png",
        "label": "Spot 2 – Picture Identification",
        "question": "I. Identify the picture.\nII. Mention the cause.",
        "answer": (
            "IDENTIFICATION: Pellagra (Niacin / Vitamin B3 Deficiency)\n\n"
            "I. Identification: Pellagra – characterized by the 3 D's:\n"
            "   Dermatitis (symmetric, sun-exposed rash – 'Casal's necklace'),\n"
            "   Diarrhea, and Dementia.\n\n"
            "II. CAUSE:\n"
            "   • Primary: Dietary deficiency of Niacin (Vitamin B3) and/or Tryptophan.\n"
            "   • Secondary causes: Carcinoid syndrome, Isoniazid therapy, Hartnup disease\n"
            "     (impaired tryptophan absorption).\n"
            "   • Common in populations dependent on maize/corn diet (lacks bioavailable niacin)."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page08.png",
        "label": "Spot 3 – Equipment Identification",
        "question": "Identify the equipment and write its use.",
        "answer": (
            "IDENTIFICATION: Incubator\n\n"
            "USE:\n"
            "• Provides controlled temperature, humidity, and CO₂ conditions for growing cultures.\n"
            "• Standard bacteriological incubator: 37°C for bacterial cultures.\n"
            "• CO₂ incubator: 37°C + 5% CO₂ for mammalian cell culture.\n"
            "• Used in: Microbiology (culture sensitivity tests), Biochemistry (enzyme incubation\n"
            "  assays), and Clinical labs (incubating specimens for analysis)."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page09.png",
        "label": "Spot 4 – Test Identification",
        "question": "Identify the test and write its principle.",
        "answer": (
            "IDENTIFICATION: Biuret Test (Test for Proteins)\n\n"
            "PRINCIPLE:\n"
            "• Detects the presence of peptide bonds (≥2 peptide bonds needed).\n"
            "• In alkaline conditions, Cu²⁺ from copper sulfate reacts with nitrogen atoms of\n"
            "  peptide bonds to form a violet/purple chelate complex (Biuret reaction).\n"
            "• Intensity of violet color is proportional to number of peptide bonds (protein concentration).\n"
            "REAGENT: Biuret reagent = NaOH + CuSO₄ + sodium potassium tartrate.\n"
            "POSITIVE: Violet/purple color = Proteins present."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page10.png",
        "label": "Spot No. 1 – Instrument Identification",
        "question": "Identify the instrument and write its use.",
        "answer": (
            "IDENTIFICATION: Electrophoresis Apparatus (Gel Electrophoresis Unit)\n\n"
            "USE:\n"
            "• Separates charged macromolecules (proteins, nucleic acids) based on size and charge\n"
            "  in an electric field.\n"
            "• Serum protein electrophoresis separates: Albumin, α1, α2, β, and γ-globulins.\n"
            "• Used in diagnosis of: M-band (Multiple myeloma), decreased albumin (liver disease).\n"
            "• Also used in molecular biology for DNA/RNA separation on agarose gels."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page11.png",
        "label": "Spot – Test Identification",
        "question": "I. Identify the following test.\nII. Write its principle.",
        "answer": (
            "IDENTIFICATION: Ninhydrin Test (Test for Amino Acids)\n\n"
            "I. Test: Ninhydrin Test\n\n"
            "II. PRINCIPLE:\n"
            "• Ninhydrin (triketohydrindene hydrate) reacts with free alpha-amino (NH₂) groups of amino acids.\n"
            "• Reaction: Ninhydrin + Amino acid → Oxidative decarboxylation → CO₂ + NH₃ + Aldehyde\n"
            "• Reduced ninhydrin + NH₃ + another ninhydrin → Ruhemann's Purple (violet/purple complex).\n"
            "• All alpha-amino acids → PURPLE color.\n"
            "• Proline and Hydroxyproline → YELLOW color (secondary amine, no free NH₂)."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page12.png",
        "label": "Spot – Normal Ranges",
        "question": "Write down the normal range of:\na) Total Protein\nb) A/G ratio",
        "answer": (
            "NORMAL RANGES:\n\n"
            "a) Total Protein (Serum): 6.0 – 8.0 g/dL\n"
            "   • Albumin: 3.5 – 5.0 g/dL\n"
            "   • Globulin: 2.0 – 3.5 g/dL\n\n"
            "b) A/G Ratio (Albumin to Globulin Ratio): 1.2 – 2.2\n"
            "   • Decreased in: Liver cirrhosis, Nephrotic syndrome, Multiple myeloma,\n"
            "     Chronic infections.\n"
            "   • Increased (rare): Hypogammaglobulinemia."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page13.png",
        "label": "Spot – Instrument Identification",
        "question": "Identify the instrument and write its use.",
        "answer": (
            "IDENTIFICATION: Centrifuge (Bench-top / Clinical Centrifuge)\n\n"
            "USE:\n"
            "• Uses centrifugal force to separate substances of different densities.\n"
            "• Separates serum/plasma from blood cells (2,000–3,000 RPM).\n"
            "• Concentrates urine sediment for microscopy.\n"
            "• Sediments proteins and separates cells from culture media."
        ),
    },
    {
        "image": f"{IMAGE_DIR}/Spotters_Biochemistry_-1_page14.png",
        "label": "Spot No. 5 – Test Identification",
        "question": "Identify the test and write its principle.",
        "answer": (
            "IDENTIFICATION: Fehling's Test (Test for Reducing Sugars)\n\n"
            "PRINCIPLE:\n"
            "• Fehling's Solution A (CuSO₄) + Fehling's Solution B (NaOH + sodium potassium tartrate)\n"
            "  are mixed in equal volumes → deep blue alkaline cupric tartrate complex.\n"
            "• Reducing sugars reduce Cu²⁺ to Cu₂O (cuprous oxide) → brick-red/orange precipitate on heating.\n"
            "POSITIVE: Brick red precipitate = Reducing sugar present.\n"
            "NEGATIVE: Blue color remains = Non-reducing sugar (e.g., Sucrose – must hydrolyze first).\n"
            "NOTE: Detects monosaccharides and reducing disaccharides (Maltose, Lactose)."
        ),
    },
]

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    title="Biochemistry Spotters – Combined Answers",
    author="Orris AI",
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    spaceBefore=10,
    spaceAfter=8,
    textColor=colors.white,
    backColor=colors.HexColor('#1565C0'),
    fontName='Helvetica-Bold',
    borderPad=6,
    leftIndent=4,
)
style_label = ParagraphStyle(
    'SpotLabel',
    parent=styles['Normal'],
    fontSize=10,
    textColor=colors.HexColor('#0d47a1'),
    fontName='Helvetica-Bold',
    spaceBefore=4,
    spaceAfter=2,
)
style_q = ParagraphStyle(
    'Question',
    parent=styles['Normal'],
    fontSize=9.5,
    textColor=colors.HexColor('#37474f'),
    fontName='Helvetica',
    spaceBefore=2,
    spaceAfter=4,
    leftIndent=8,
)
style_ans_header = ParagraphStyle(
    'AnswerHeader',
    parent=styles['Normal'],
    fontSize=9.5,
    textColor=colors.white,
    backColor=colors.HexColor('#2e7d32'),
    fontName='Helvetica-Bold',
    spaceBefore=2,
    spaceAfter=2,
    leftIndent=4,
    borderPad=4,
)
style_ans = ParagraphStyle(
    'Answer',
    parent=styles['Normal'],
    fontSize=9.5,
    textColor=colors.HexColor('#1b5e20'),
    fontName='Helvetica',
    spaceBefore=1,
    spaceAfter=2,
    leftIndent=10,
    backColor=colors.HexColor('#f1f8e9'),
    borderPad=6,
)

story = []

# ---- COVER PAGE ----
story.append(Spacer(1, 2*cm))
story.append(Paragraph("Biochemistry Spotters", style_title))
story.append(Paragraph("Combined Questions & Model Answers", ParagraphStyle(
    'Subtitle', parent=styles['Normal'],
    fontSize=13, textColor=colors.HexColor('#455a64'),
    fontName='Helvetica', spaceAfter=4, alignment=TA_CENTER
)))
story.append(HRFlowable(width='100%', thickness=2, color=colors.HexColor('#1565C0'), spaceAfter=12))
story.append(Paragraph(
    "This document combines all three spotter PDFs into a single reference with model answers "
    "written in the space provided after each question.",
    ParagraphStyle('IntroText', parent=styles['Normal'],
                   fontSize=10, textColor=colors.HexColor('#546e7a'),
                   fontName='Helvetica', spaceAfter=6, alignment=TA_CENTER)
))
story.append(Spacer(1, 0.5*cm))

# Table of contents info
toc_data = [
    ["PDF", "Content", "Pages"],
    ["Spotters 2-1", "Lab equipment, calculations, clinical biochemistry", "18 pages"],
    ["Spotters Biochemistry -1", "Tests, instruments, vitamins, calculations", "14 pages"],
    ["Biochem Spotting 2-1", "Reference/Answer materials (integrated)", "12 pages"],
]
toc_table = Table(toc_data, colWidths=[4.5*cm, 9.5*cm, 3.5*cm])
toc_table.setStyle(TableStyle([
    ('BACKGROUND', (0,0), (-1,0), colors.HexColor('#1565C0')),
    ('TEXTCOLOR', (0,0), (-1,0), colors.white),
    ('FONTNAME', (0,0), (-1,0), 'Helvetica-Bold'),
    ('FONTSIZE', (0,0), (-1,-1), 9),
    ('ALIGN', (0,0), (-1,-1), 'CENTER'),
    ('VALIGN', (0,0), (-1,-1), 'MIDDLE'),
    ('ROWBACKGROUNDS', (0,1), (-1,-1), [colors.HexColor('#e3f2fd'), colors.white]),
    ('GRID', (0,0), (-1,-1), 0.5, colors.HexColor('#90caf9')),
    ('TOPPADDING', (0,0), (-1,-1), 6),
    ('BOTTOMPADDING', (0,0), (-1,-1), 6),
]))
story.append(toc_table)
story.append(PageBreak())

# ---- CONTENT PAGES ----
PAGE_IMG_W = 15.7 * cm  # max image width
MAX_IMG_H = 9.5 * cm    # max image height

for entry in spotter_pages:
    if entry.get('section_header'):
        story.append(Spacer(1, 0.3*cm))
        story.append(Paragraph(f"  {entry['file']}", style_section))
        story.append(Spacer(1, 0.2*cm))
        continue

    img_path = entry.get('image', '')
    label = entry.get('label', '')
    question = entry.get('question', '')
    answer = entry.get('answer', '')

    block = []

    # ---- Spot image ----
    if img_path and os.path.exists(img_path):
        try:
            pil_img = PILImage.open(img_path)
            iw, ih = pil_img.size
            aspect = ih / iw
            disp_w = PAGE_IMG_W
            disp_h = disp_w * aspect
            if disp_h > MAX_IMG_H:
                disp_h = MAX_IMG_H
                disp_w = disp_h / aspect
            rl_img = RLImage(img_path, width=disp_w, height=disp_h)
            rl_img.hAlign = 'CENTER'
            block.append(rl_img)
        except Exception as e:
            block.append(Paragraph(f"[Image: {os.path.basename(img_path)}]", styles['Normal']))
    
    block.append(Spacer(1, 3*mm))

    # ---- Label ----
    block.append(Paragraph(label, style_label))

    # ---- Question ----
    q_lines = question.replace('\n', '<br/>')
    block.append(Paragraph(f"<b>Question:</b> {q_lines}", style_q))

    block.append(Spacer(1, 2*mm))

    # ---- Answer header ----
    block.append(Paragraph(" ANSWER:", style_ans_header))

    # ---- Answer text ----
    ans_html = answer.replace('\n', '<br/>').replace('•', '&#8226;').replace('∴', '&#8756;').replace('≥', '&#8805;').replace('α', '&#945;').replace('β', '&#946;').replace('γ', '&#947;').replace('¹²⁵', '<sup>125</sup>').replace('²⁺', '<sup>2+</sup>').replace('₂', '<sub>2</sub>').replace('₃', '<sub>3</sub>')
    block.append(Paragraph(ans_html, style_ans))

    block.append(HRFlowable(width='100%', thickness=0.5, color=colors.HexColor('#b0bec5'), spaceAfter=8, spaceBefore=4))

    story.append(KeepTogether(block))

# Build
doc.build(story)
print(f"PDF created: {OUTPUT_PATH}")

cd /home/daytona/workspace && python3 create_spotters_pdf.py 2>&1

ls -lh /home/daytona/workspace/spotters_output/Biochemistry_Spotters_Combined_An…

python3 << 'EOF' # Compress images before including them - rebuild with smaller …

python3 << 'EOF' # Quick rebuild with compressed images import re with open('/h…

ls /home/daytona/workspace/pdf_images_compressed/ | head -60

pkill -f "python3 -m http.server" 2>/dev/null; cd /home/daytona/workspace/pdf_im…

python3 << 'EOF' import pytesseract from PIL import Image import os, glob # Rea…

python3 << 'EOF' import pytesseract from PIL import Image import os img_dir = '…

python3 << 'EOF' # The key insight: The Biochem_spotting_2-1 PDF = the answer ke…

python3 << 'EOF' # Better approach: crop central region (the instrument area) of…

python3 << 'EOF' # Use imagehash for perceptual matching import subprocess resul…

pip install --user imagehash -q 2>&1 | tail -3 && python3 -c "import imagehash; …

python3 << 'EOF' import imagehash from PIL import Image import os ref_dir = '/h…

python3 << 'EOF' import pytesseract from PIL import Image import os ref_dir = '…

ls /home/daytona/workspace/pdf_ocr/ | head -50 && echo "---" && cat /home/dayton…

python3 << 'EOF' from PIL import Image import numpy as np ref_dir = '/home/dayt…

python3 << 'EOF' # Read ALL existing OCR text files carefully import os, glob o…

python3 << 'EOF' from pdf2image import convert_from_path import os path = '/hom…

pkill -f "http.server 8765" 2>/dev/null; sleep 1 cd /home/daytona/workspace/new_…

python3 << 'EOF' import pytesseract from PIL import Image, ImageFilter, ImageEnh…

python3 << 'EOF' from PIL import Image import numpy as np import os out_dir = '…

python3 << 'EOF' # Let me extract just the instrument image crop from each page,…

python3 << 'EOF' # Page 11 OCR says "SPOT NO - 14 (2 mark)" and shows "90ml" and…

python3 << 'EOF' # Key findings from analysis: # Page 11: aspect=4.2, width_rati…

~/build_final_pdf.py

from reportlab.lib.pagesizes import A4
from reportlab.lib import colors
from reportlab.lib.units import cm, mm
from reportlab.platypus import (
    SimpleDocTemplate, Paragraph, Spacer, PageBreak,
    Image as RLImage, Table, TableStyle, KeepTogether, HRFlowable
)
from reportlab.lib.styles import getSampleStyleSheet, ParagraphStyle
from reportlab.lib.enums import TA_LEFT, TA_CENTER
from PIL import Image as PILImage
import io, os

OUTPUT = '/home/daytona/workspace/spotters_output/Spotters_2-1_WITH_ANSWERS.pdf'
IMG_DIR = '/home/daytona/workspace/new_spotter'
os.makedirs('/home/daytona/workspace/spotters_output', exist_ok=True)

# ─── Styles ────────────────────────────────────────────────────────────────
styles = getSampleStyleSheet()

def PS(name, **kw):
    base = kw.pop('parent', styles['Normal'])
    return ParagraphStyle(name, parent=base, **kw)

S_TITLE = PS('T', parent=styles['Title'], fontSize=22, textColor=colors.HexColor('#1a237e'),
             fontName='Helvetica-Bold', spaceAfter=4)
S_SUB   = PS('Sub', fontSize=11, textColor=colors.HexColor('#455a64'),
             fontName='Helvetica', spaceAfter=6, alignment=TA_CENTER)
S_SPOTLABEL = PS('SL', fontSize=11, textColor=colors.HexColor('#0d47a1'),
                 fontName='Helvetica-Bold', spaceBefore=4, spaceAfter=2)
S_Q     = PS('Q', fontSize=10, textColor=colors.HexColor('#37474f'),
             fontName='Helvetica', spaceBefore=2, spaceAfter=4, leftIndent=8)
S_ANSH  = PS('AH', fontSize=10, textColor=colors.white,
             backColor=colors.HexColor('#1b5e20'), fontName='Helvetica-Bold',
             spaceBefore=3, spaceAfter=0, leftIndent=4, borderPad=5)
S_ANS   = PS('AN', fontSize=9.5, textColor=colors.HexColor('#1b5e20'),
             fontName='Helvetica', spaceBefore=0, spaceAfter=3,
             leftIndent=8, backColor=colors.HexColor('#f1f8e9'), borderPad=7)

PAGE_W = A4[0] - 3.6*cm   # usable width
MAX_H  = 9.0*cm            # max image height

def img_block(pg_num):
    path = f'{IMG_DIR}/page{pg_num:02d}.png'
    if not os.path.exists(path):
        return []
    pil = PILImage.open(path)
    iw, ih = pil.size
    # save compressed version
    cpath = path.replace('.png', '_c.jpg')
    # Resize to max 1400px wide
    if iw > 1400:
        ratio = 1400 / iw
        pil = pil.resize((1400, int(ih*ratio)), PILImage.LANCZOS)
        iw, ih = pil.size
    pil.convert('RGB').save(cpath, 'JPEG', quality=72)
    
    aspect = ih / iw
    disp_w = PAGE_W
    disp_h = disp_w * aspect
    if disp_h > MAX_H:
        disp_h = MAX_H
        disp_w = disp_h / aspect
    
    ri = RLImage(cpath, width=disp_w, height=disp_h)
    ri.hAlign = 'CENTER'
    return [ri, Spacer(1, 2*mm)]

def ans_block(label, question, answer):
    elems = []
    elems.append(Paragraph(label, S_SPOTLABEL))
    q_html = question.replace('\n', '<br/>')
    elems.append(Paragraph(q_html, S_Q))
    elems.append(Paragraph(' ✦ MODEL ANSWER', S_ANSH))
    a_html = (answer
              .replace('&', '&amp;')
              .replace('\n', '<br/>')
              .replace('•', '&#8226;')
              .replace('→', '&#8594;')
              .replace('≥', '&#8805;')
              .replace('≤', '&#8804;')
              .replace('²', '<sup>2</sup>')
              .replace('³', '<sup>3</sup>')
              .replace('¹²⁵', '<sup>125</sup>')
              .replace('∴', '&#8756;')
              .replace('α', '&#945;')
              .replace('β', '&#946;')
              .replace('γ', '&#947;')
             )
    elems.append(Paragraph(a_html, S_ANS))
    elems.append(HRFlowable(width='100%', thickness=0.6,
                            color=colors.HexColor('#b0bec5'),
                            spaceBefore=4, spaceAfter=6))
    return elems

# ─── SPOTTER DATA ──────────────────────────────────────────────────────────
# Each entry: (page_num, label, question_text, answer_text)
# Pages 3,5,6,7,8,15,17 have NO instrument image — just text questions
# Pages 1,2,4,9,10,11,12,13,14,16,18 have instrument images

spotters = [

  # ── PAGE 1 ─────────────────────────────────────────────────────────
  (1,
   "Spot 5  (2 marks)",
   "Identify the equipment and write its use in laboratory.",
   """IDENTIFICATION: Colorimeter (Photoelectric Colorimeter)

USES IN LABORATORY:
• Measures the concentration of colored substances in a solution using the Beer-Lambert Law: A = εcl (Absorbance ∝ concentration).
• Used to estimate: Hemoglobin, Blood glucose, Serum bilirubin, Total protein, Urea, Creatinine, Serum enzymes (ALT, AST), and other colored analytes.
• A light source emits monochromatic light through a filter; the beam passes through the colored sample and the transmitted light is detected by a photocell.
• Wavelength range: 400–700 nm (visible spectrum).
• Results are compared against a standard/blank to calculate concentration."""
  ),

  # ── PAGE 2 ─────────────────────────────────────────────────────────
  (2,
   "Spot 4  (2 marks)",
   "Identify the above picture and write its use.",
   """IDENTIFICATION: Hot Air Oven

USES:
• Used for DRY HEAT STERILIZATION of laboratory glassware (test tubes, Petri dishes, glass pipettes, flasks).
• Operating temperature: 160°C for 2 hours  OR  180°C for 1 hour.
• Also used for drying glassware after washing (at lower temperatures ~60–80°C).
• Suitable for materials that cannot be sterilized by moist heat (autoclave) — e.g., glassware, metal instruments, anhydrous oils, and powders.
• NOT suitable for rubber, plastics, or culture media."""
  ),

  # ── PAGE 3 ─────────────────────────────────────────────────────────
  (3,
   "Spot 1  (2 marks)",
   "1. What is the normal range of Triglyceride level in Serum?\n2. Find out the VLDL level in serum if Triglyceride is 150 mg%.",
   """1. NORMAL RANGE OF SERUM TRIGLYCERIDES:
   • Desirable (Normal):  < 150 mg/dL
   • Borderline high:     150–199 mg/dL
   • High:                200–499 mg/dL
   • Very high:           ≥ 500 mg/dL

2. VLDL CALCULATION:
   Formula (Friedewald):  VLDL = Triglyceride / 5
   
   VLDL = 150 / 5  =  30 mg%
   
   ∴ VLDL level = 30 mg%  (Normal VLDL: 2–30 mg/dL)"""
  ),

  # ── PAGE 4 ─────────────────────────────────────────────────────────
  (4,
   "Spot  (2 marks)",
   "Identify the Instrument and write its uses.",
   """IDENTIFICATION: Micropipette (Automatic Adjustable Micropipette / Piston Pipette)

USES:
• Accurately measures and transfers very small volumes of liquid ranging from 0.1 µL to 1000 µL.
• Available in fixed and variable volume models:
   P10 (0.5–10 µL)  |  P200 (20–200 µL)  |  P1000 (100–1000 µL)
• Uses disposable sterile tips to prevent cross-contamination between samples.
• Essential in: molecular biology (PCR setup), serology, immunology, ELISA, biochemistry, and microbiology.
• Read the volume from the digital display on the barrel; adjust using the plunger dial.
• Works by positive displacement (air displacement mechanism)."""
  ),

  # ── PAGE 5 ─────────────────────────────────────────────────────────
  (5,
   "Spot 5  (2 marks)",
   "Concentration of Protein = 7.1 gm%\nConcentration of Albumin = 4.2 gm%\nFind out A:G ratio and Write its Normal Range.",
   """CALCULATION:
   Globulin  =  Total Protein − Albumin
   Globulin  =  7.1 − 4.2  =  2.9 gm%

   A:G Ratio  =  Albumin / Globulin
   A:G Ratio  =  4.2 / 2.9  =  1.45

   ∴ A:G Ratio = 1.45 : 1

NORMAL RANGE of A:G Ratio:
   • Normal:  1.2 – 2.2  (some references: 1.0 – 2.5)
   
CLINICAL SIGNIFICANCE:
   • Decreased A:G ratio (<1.0) seen in: Liver cirrhosis, Nephrotic syndrome, Multiple myeloma, Malnutrition, Chronic infections.
   • Increased A:G ratio: Rare — seen in hypogammaglobulinemia."""
  ),

  # ── PAGE 6 ─────────────────────────────────────────────────────────
  (6,
   "Spot 6  (5 marks)",
   "1. Write the full form of ELISA  (1 mark)\n2. Write the types of ELISA  (1 mark)\n3. Uses of ELISA  (1 mark)\n4. Write the full form of RIA  (1 mark)\n5. Which Radioactive component is used in RIA?  (1 mark)",
   """1. ELISA = Enzyme-Linked Immunosorbent Assay

2. TYPES OF ELISA:
   (i)  Direct ELISA
   (ii) Indirect ELISA
   (iii) Sandwich ELISA (Double antibody sandwich)
   (iv) Competitive ELISA

3. USES OF ELISA:
   • Detection of HIV, Hepatitis B (HBsAg), Hepatitis C antibodies
   • Hormone estimation: hCG (pregnancy test), TSH, LH, FSH, Insulin
   • Diagnosis of infectious diseases (Dengue, Malaria, COVID-19)
   • Detection of drugs, allergens, and food contaminants
   • Autoimmune disease screening (ANA, anti-dsDNA)

4. RIA = RadioImmunoAssay

5. Radioactive component used in RIA:
   ¹²⁵I (Iodine-125) — a gamma (γ) ray emitter with half-life of ~60 days.
   (Also ¹³¹I, ³H-tritium, and ¹⁴C are used in some assays.)"""
  ),

  # ── PAGE 7 ─────────────────────────────────────────────────────────
  (7,
   "Spot  (2 marks)",
   "Write 4 safety measures in laboratory.",
   """4 SAFETY MEASURES IN LABORATORY:

1. Use of Personal Protective Equipment (PPE):
   Always wear a lab coat, disposable gloves, safety goggles/face shield, and mask when handling chemicals, biological specimens, or hot instruments. PPE prevents direct contact with hazardous materials.

2. Proper Waste Disposal:
   • Biological/infectious waste → Yellow biohazard bags
   • Sharps (needles, broken glass) → Puncture-proof sharps containers (never recap needles by two-hand technique)
   • Chemical waste → Designated chemical waste containers
   • Follow local biomedical waste management rules.

3. No Eating, Drinking, or Mouth Pipetting:
   Food and beverages are strictly prohibited inside the lab. Never pipette by mouth — use rubber bulbs, mechanical pipette aids, or micropipettes to avoid accidental ingestion of toxic/infectious material.

4. Emergency Preparedness & Fire Safety:
   Know the location and correct use of: fire extinguisher, eyewash station, first aid kit, and emergency exit. Report chemical spills immediately. Use fume hood for volatile/toxic reagents. Keep flammable solvents away from flames."""
  ),

  # ── PAGE 8 ─────────────────────────────────────────────────────────
  (8,
   "Spot  (2 marks)",
   "Identify the symbol and write two examples covered under this category.",
   """IDENTIFICATION: Biohazard Symbol (Biological Hazard Warning Symbol)

MEANING:
This internationally recognized symbol (three-lobed orange/black symbol) indicates the presence of biological substances or materials that pose a threat to the health of living organisms, primarily humans.

TWO EXAMPLES covered under this category:

1. Blood and blood-borne pathogens:
   HIV-infected blood, Hepatitis B virus (HBV), Hepatitis C virus (HCV) samples — present in specimen containers, sharps bins, and blood bags.

2. Microbiological cultures:
   Bacterial cultures (e.g., Mycobacterium tuberculosis, Staphylococcus), viral stocks, and cell cultures used in clinical and research laboratories — stored in biohazard-labeled containers."""
  ),

  # ── PAGE 9 ─────────────────────────────────────────────────────────
  (9,
   "Spot 6  (5 marks)",
   "1. Identify the given machine and write its principle  (3 marks)\n2. Write its types  (1 mark)\n3. Write its uses  (1 mark)",
   """1. IDENTIFICATION: Centrifuge

PRINCIPLE (3 marks):
A centrifuge operates on the principle of SEDIMENTATION based on centrifugal force.

When the rotor spins at high speed (RPM), centrifugal force is generated that acts outward from the center of rotation. This force is much greater than gravity (expressed as RCF — Relative Centrifugal Force, in × g).

Particles/substances of HIGHER DENSITY sediment toward the bottom of the tube (form a pellet), while particles of LOWER DENSITY remain in the supernatant.

Formula: RCF (g) = 1.118 × 10⁻⁵ × r × N²
where r = radius (cm), N = speed (RPM)

2. TYPES of Centrifuge:
   • Low-speed centrifuge (600–6,000 RPM): Routine serum/plasma separation
   • High-speed centrifuge (10,000–25,000 RPM): Bacteria, cell debris
   • Ultracentrifuge (>50,000 RPM): Viruses, organelles, macromolecules
   • Microcentrifuge (up to 14,000 RPM): Eppendorf/microtubes
   • Refrigerated centrifuge: For temperature-sensitive samples

3. USES:
   • Separation of serum or plasma from blood cells (most common)
   • Concentration of urine sediment for microscopy
   • Isolation of cells, bacteria, viruses, and subcellular organelles
   • Separation of DNA/proteins in molecular biology"""
  ),

  # ── PAGE 10 ─────────────────────────────────────────────────────────
  (10,
   "Spot  (2 marks)",
   "Identify the above picture and write its use.",
   """IDENTIFICATION: pH Meter (Electronic pH Meter)

USES:
• Measures the hydrogen ion concentration (H⁺) of a solution, expressed as pH (potential of Hydrogen).
• pH = -log[H⁺]; Range: 0 (most acidic) to 14 (most alkaline).
• Used in biochemistry lab to:
   - Prepare buffer solutions of exact pH for enzyme assays and reagent preparation
   - Measure pH of biological fluids (urine, CSF, gastric juice, blood)
   - Monitor fermentation processes
   - Quality control of reagents

PRINCIPLE:
Consists of a glass electrode (pH-sensitive) and a reference electrode dipped into the solution. The potential difference (millivolts) generated between them is proportional to the H⁺ activity; a voltmeter converts this to a pH reading."""
  ),

  # ── PAGE 11 ─────────────────────────────────────────────────────────
  (11,
   "Spot No. 1  (2 marks)",
   "Identify the glassware and write its uses.",
   """IDENTIFICATION: Measuring Cylinder (Graduated Cylinder)

USES:
• Used to measure and deliver specific volumes of liquids accurately in the laboratory (approximate, not for high-precision work).
• Graduated with volume markings (e.g., 10 mL, 25 mL, 50 mL, 100 mL).
• Volume is read at the BOTTOM of the meniscus (for transparent liquids).
• Used in:
   - Preparation of solutions and dilutions
   - Measuring reagent volumes before mixing
   - Measuring specific gravity of urine (combined with urinometer)
• NOT used for: Precise volumetric work (use volumetric flask or pipette for precision).
• Made of borosilicate glass or polypropylene."""
  ),

  # ── PAGE 12 ─────────────────────────────────────────────────────────
  (12,
   "Spot No. 5  (2 marks)",
   "1. Identify the given machine.\n2. Write its use.",
   """1. IDENTIFICATION: Semi-automated Biochemistry Analyzer
   (Semi-automated Clinical Chemistry Analyzer)

2. USES:
   • Performs multiple biochemical/clinical chemistry tests on serum, plasma, urine, CSF, and whole blood semi-automatically.
   • Tests performed include:
     - Liver function tests: Total bilirubin, ALT, AST, Alkaline phosphatase, Total protein, Albumin
     - Kidney function tests: Urea (BUN), Creatinine, Uric acid
     - Metabolic tests: Blood glucose, Cholesterol, Triglycerides, HDL, LDL
     - Cardiac enzymes: CK, LDH
   • The operator manually loads the sample/reagent; the analyzer performs mixing, incubation, photometric reading, and calculation automatically.
   • PRINCIPLE: Photometric/colorimetric measurement (absorbance at specific wavelengths) using enzymatic or chemical reactions."""
  ),

  # ── PAGE 13 ─────────────────────────────────────────────────────────
  (13,
   "Spot  (2 marks)",
   "Identify the instrument and write its use.",
   """IDENTIFICATION: Vortex Mixer (Vortex Shaker)

USE:
• A simple benchtop device used to rapidly mix small volumes of liquid in test tubes, microtubes, or vials using an oscillating eccentric motion.
• Creates a vortex (whirlpool/cyclone) inside the tube for thorough and even mixing.
• Operating modes: Touch mode (momentary mixing) or Continuous mode (constant mixing).
• Speed: Adjustable from ~100 to 3,200 RPM.

APPLICATIONS:
   - Mixing reagents and samples before analysis
   - Dissolving precipitates or powders in solution
   - Resuspending cell pellets after centrifugation
   - Mixing blood samples with anticoagulants
   - Preparing serial dilutions in microbiology/serology"""
  ),

  # ── PAGE 14 ─────────────────────────────────────────────────────────
  (14,
   "Spot 2  (2 marks)",
   "Identify the equipment and write its use.",
   """IDENTIFICATION: Weighing Balance (Digital Analytical Balance / Electronic Balance)

USES:
• Measures the mass/weight of chemical substances, specimens, and reagents accurately.
• Sensitivity: Analytical balance → 0.0001 g (0.1 mg); Top-loading balance → 0.01 g.
• Essential for:
   - Preparing standard solutions of known concentration (accurate weighing of solutes)
   - Weighing specimens and tissue samples
   - Calibrating and quality control purposes
   - Pharmaceutical compounding
• Precautions:
   - Level the balance before use (check bubble level)
   - Tare (zero) the balance with the weighing container
   - Never exceed maximum capacity
   - Protect from air currents (use draught shield for analytical balances)"""
  ),

  # ── PAGE 15 ─────────────────────────────────────────────────────────
  (15,
   "Spot No. 3  (2 marks)",
   "Identify the test and write its principle.",
   """IDENTIFICATION: Seliwanoff's Test (Test to differentiate Ketoses from Aldoses)

PRINCIPLE:
Seliwanoff's test is based on the differential rate of DEHYDRATION of sugars by hot concentrated HCl.

Step 1 — Dehydration:
   Ketoses (e.g., Fructose) contain a ketone group at C-2 position, making them more reactive. They are rapidly dehydrated by hot HCl to form Hydroxymethylfurfural (HMF).
   Aldoses (e.g., Glucose, Galactose) dehydrate much more slowly under the same conditions.

Step 2 — Condensation:
   HMF condenses with Resorcinol (the reagent) to produce a deep CHERRY RED colored complex.

RESULT:
   • Ketose (Fructose): CHERRY RED color within 1 minute → POSITIVE
   • Aldose (Glucose): Faint pink or no color within 1 minute → NEGATIVE (may turn faint red after prolonged heating)

REAGENT: Resorcinol dissolved in concentrated HCl (Seliwanoff's reagent)."""
  ),

  # ── PAGE 16 ─────────────────────────────────────────────────────────
  (16,
   "Spot  (2 marks)",
   "Identify the above pictures and write their uses.",
   """IDENTIFICATION: Burette (50 mL Burette)

USES:
• A long, graduated cylindrical tube with a stopcock at the bottom, used to deliver precise variable volumes of liquid (titrant) during titration.
• Graduated from 0 at the top to 50 mL at the bottom (reads downward).
• Reading: At the bottom of the meniscus for transparent solutions; top of meniscus for dark solutions (e.g., KMnO₄).
• Smallest graduation: 0.1 mL; can be read to 0.05 mL precision.

APPLICATIONS IN BIOCHEMISTRY LAB:
   - Acid-base titrations (HCl vs NaOH)
   - Redox titrations (potassium permanganate)
   - Complexometric titrations (EDTA)
   - Estimation of Vitamin C (by iodometric titration)
   - Total protein estimation (by Kjeldahl method titration)

Types: Glass burette (with glass stopcock) or Automatic/Digital burette."""
  ),

  # ── PAGE 17 ─────────────────────────────────────────────────────────
  (17,
   "Spot  (2 marks)",
   "Write 4 safety measures in laboratory.",
   """4 SAFETY MEASURES IN LABORATORY:

1. Personal Protective Equipment (PPE):
   Always wear a lab coat, gloves (nitrile/latex), safety goggles, and closed-toe shoes before entering the laboratory. Never remove PPE while handling specimens, chemicals, or infectious material.

2. Proper Labeling and Storage of Chemicals:
   All reagent bottles must be clearly labeled (name, concentration, date, hazard symbols). Store flammable chemicals away from heat and open flames; store acids and alkalis separately; refrigerate temperature-sensitive reagents.

3. Safe Handling of Sharps and Biological Waste:
   Dispose of used needles, lancets, and broken glassware immediately into puncture-resistant sharps containers. Never recap needles by two hands. Discard biological waste in properly sealed biohazard bags following biomedical waste management guidelines.

4. Know Emergency Procedures:
   Be familiar with the location of: fire extinguisher, eyewash station, safety shower, first aid kit, and emergency exits. Report all accidents, spills, and near-misses to the supervisor. For chemical eye/skin exposure — flush with water for 15 minutes immediately."""
  ),

  # ── PAGE 18 ─────────────────────────────────────────────────────────
  (18,
   "Spot  (2 marks)",
   "Identify the glassware and write its use.",
   """IDENTIFICATION: Conical Flask (Erlenmeyer Flask)

USE:
• A conical (triangular) shaped flask with a flat base and narrow neck, typically available in sizes of 50 mL, 100 mL, 250 mL, and 500 mL.
• WIDE BASE allows stable placement; NARROW NECK allows swirling without spillage.

USES IN BIOCHEMISTRY LABORATORY:
   • Primary use: Holding the analyte solution during TITRATION — the wide base and narrow neck allow easy swirling to mix without spilling.
   • Preparation and storage of chemical solutions, buffers, and culture media.
   • Heating of liquids (with bunsen burner or hot plate, using a wire gauze).
   • Carrying out chemical reactions (mixing reagents for tests like Benedict's, Biuret).
   • Preparing microbiological culture media (autoclaved in conical flask with cotton plug).

Note: For very precise volume measurements, use a volumetric flask instead."""
  ),
]

# ─── BUILD PDF ────────────────────────────────────────────────────────────
doc = SimpleDocTemplate(
    OUTPUT,
    pagesize=A4,
    topMargin=1.5*cm, bottomMargin=1.5*cm,
    leftMargin=1.8*cm, rightMargin=1.8*cm,
    title="Spotters 2-1 – Questions with Model Answers",
    author="Orris AI",
)

story = []

# Cover
story.append(Spacer(1, 1.5*cm))
story.append(Paragraph("Spotters 2-1", S_TITLE))
story.append(Paragraph("Biochemistry Lab Spotters — Questions with Model Answers", S_SUB))
story.append(HRFlowable(width='100%', thickness=2,
                         color=colors.HexColor('#1565C0'), spaceAfter=10))
story.append(Spacer(1, 0.3*cm))

# Quick summary table
info = [
    ['Total Spotters', '18', 'Total Marks', '5+5+2×16 = 42'],
    ['5-mark questions', '2 (Spot 6 ×2)', '2-mark questions', '16'],
    ['Theory questions', '4 (No image)', 'Image-based questions', '12'],
]
it = Table(info, colWidths=[4.5*cm, 3*cm, 4.5*cm, 5.5*cm])
it.setStyle(TableStyle([
    ('BACKGROUND', (0,0), (-1,-1), colors.HexColor('#e3f2fd')),
    ('FONTNAME', (0,0), (-1,-1), 'Helvetica'),
    ('FONTSIZE', (0,0), (-1,-1), 9),
    ('GRID', (0,0), (-1,-1), 0.5, colors.HexColor('#90caf9')),
    ('TOPPADDING', (0,0), (-1,-1), 5),
    ('BOTTOMPADDING', (0,0), (-1,-1), 5),
    ('ALIGN', (1,0), (1,-1), 'CENTER'),
    ('ALIGN', (3,0), (3,-1), 'CENTER'),
    ('FONTNAME', (0,0), (0,-1), 'Helvetica-Bold'),
    ('FONTNAME', (2,0), (2,-1), 'Helvetica-Bold'),
]))
story.append(it)
story.append(PageBreak())

# Add each spotter
for (pg, label, question, answer) in spotters:
    block = []
    # Original page image
    block.extend(img_block(pg))
    # Q + A
    block.extend(ans_block(label, question, answer))
    story.append(KeepTogether(block))

doc.build(story)
print(f"PDF saved: {OUTPUT}")
import os
print(f"Size: {os.path.getsize(OUTPUT)/1024/1024:.2f} MB")

cd /home/daytona/workspace && python3 build_final_pdf.py 2>&1

python3 << 'EOF' from pdf2image import convert_from_path import pytesseract from…

python3 << 'EOF' from PIL import Image import numpy as np import pytesseract, os…

python3 << 'EOF' # Key observations from shape analysis: # P7: horiz=832, vert=…

python3 << 'EOF' # Now let me cross-reference this PDF with the "Biochem_spottin…

~/build_biochem_pdf.py

from reportlab.lib.pagesizes import A4
from reportlab.lib import colors
from reportlab.lib.units import cm, mm
from reportlab.platypus import (
    SimpleDocTemplate, Paragraph, Spacer, PageBreak,
    Image as RLImage, Table, TableStyle, KeepTogether, HRFlowable
)
from reportlab.lib.styles import getSampleStyleSheet, ParagraphStyle
from reportlab.lib.enums import TA_CENTER
from PIL import Image as PILImage
import os

OUTPUT = '/home/daytona/workspace/spotters_output/Spotters_Biochemistry_-1_WITH_ANSWERS.pdf'
IMG_DIR = '/home/daytona/workspace/biochem_spotter'
os.makedirs('/home/daytona/workspace/spotters_output', exist_ok=True)

styles = getSampleStyleSheet()

def PS(name, **kw):
    base = kw.pop('parent', styles['Normal'])
    return ParagraphStyle(name, parent=base, **kw)

S_TITLE  = PS('T', parent=styles['Title'], fontSize=22,
              textColor=colors.HexColor('#1a237e'), fontName='Helvetica-Bold', spaceAfter=4)
S_SUB    = PS('Sub', fontSize=11, textColor=colors.HexColor('#455a64'),
              fontName='Helvetica', spaceAfter=6, alignment=TA_CENTER)
S_LABEL  = PS('SL', fontSize=11, textColor=colors.HexColor('#0d47a1'),
              fontName='Helvetica-Bold', spaceBefore=4, spaceAfter=2)
S_Q      = PS('Q', fontSize=10, textColor=colors.HexColor('#37474f'),
              fontName='Helvetica', spaceBefore=2, spaceAfter=4, leftIndent=8)
S_ANSH   = PS('AH', fontSize=10, textColor=colors.white,
              backColor=colors.HexColor('#1b5e20'), fontName='Helvetica-Bold',
              spaceBefore=3, spaceAfter=0, leftIndent=4, borderPad=5)
S_ANS    = PS('AN', fontSize=9.5, textColor=colors.HexColor('#1b5e20'),
              fontName='Helvetica', spaceBefore=0, spaceAfter=3,
              leftIndent=8, backColor=colors.HexColor('#f1f8e9'), borderPad=7)

PAGE_W = A4[0] - 3.6*cm
MAX_H  = 9.0*cm

def img_block(pg_num):
    path = f'{IMG_DIR}/page{pg_num:02d}.png'
    if not os.path.exists(path): return []
    pil = PILImage.open(path)
    iw, ih = pil.size
    cpath = path.replace('.png', '_c.jpg')
    if iw > 1400:
        r = 1400/iw; pil = pil.resize((1400, int(ih*r)), PILImage.LANCZOS); iw,ih = pil.size
    pil.convert('RGB').save(cpath, 'JPEG', quality=72)
    aspect = ih/iw
    dw = PAGE_W; dh = dw*aspect
    if dh > MAX_H: dh = MAX_H; dw = dh/aspect
    ri = RLImage(cpath, width=dw, height=dh); ri.hAlign = 'CENTER'
    return [ri, Spacer(1, 2*mm)]

def fmt(text):
    return (text.replace('&','&amp;').replace('\n','<br/>')
            .replace('•','&#8226;').replace('→','&#8594;')
            .replace('≥','&#8805;').replace('≤','&#8804;')
            .replace('∴','&#8756;').replace('∝','&#8733;')
            .replace('α','&#945;').replace('β','&#946;').replace('γ','&#947;')
            .replace('¹²⁵','<sup>125</sup>').replace('²⁺','<sup>2+</sup>'))

def ans_block(label, question, answer):
    e = []
    e.append(Paragraph(label, S_LABEL))
    e.append(Paragraph(fmt(question).replace('\n','<br/>'), S_Q))
    e.append(Paragraph(' ✦  MODEL ANSWER', S_ANSH))
    e.append(Paragraph(fmt(answer), S_ANS))
    e.append(HRFlowable(width='100%', thickness=0.6,
                        color=colors.HexColor('#b0bec5'), spaceBefore=4, spaceAfter=6))
    return e

# ─── All 14 spotters ──────────────────────────────────────────────────────
spotters = [

  # PAGE 1 ─────────────────────────────────────────────────────────────────
  (1,
   "Spot No. 1  (2 marks)",
   "Identify the Instrument and write its uses.",
   """IDENTIFICATION: Colorimeter (Photoelectric Colorimeter)

USES:
• Measures the concentration of colored substances in solution using Beer-Lambert Law:
  A = ε × c × l   (Absorbance is directly proportional to concentration)
• Used to estimate: Haemoglobin, Blood glucose, Serum bilirubin, Total protein,
  Urea, Creatinine, ALT, AST, Alkaline phosphatase, and other colored analytes.
• Works by passing a specific wavelength of light (400–700 nm) through a filter,
  then through the colored sample; the transmitted light is measured by a photocell.
• Concentration is calculated by comparing absorbance of sample to a known standard.
• Two types: Filter photometer (colorimeter) and Spectrophotometer (broader wavelength range)."""
  ),

  # PAGE 2 ─────────────────────────────────────────────────────────────────
  (2,
   "Spot No. 2  (2 marks)",
   "Which test is used to differentiate between ketosugar and aldosugar?\nWrite its principle.",
   """TEST: Seliwanoff's Test

PRINCIPLE:
This test is based on the differential DEHYDRATION rate of ketoses and aldoses
by hot concentrated Hydrochloric acid (HCl).

Step 1 – Dehydration:
• Ketoses (e.g., Fructose) have a ketone group at C-2 and are RAPIDLY dehydrated
  by hot HCl to form Hydroxymethylfurfural (HMF).
• Aldoses (e.g., Glucose, Galactose) dehydrate SLOWLY under the same conditions.

Step 2 – Condensation:
• HMF condenses with Resorcinol (present in the reagent) to produce a
  CHERRY RED / deep red colored complex within 1 minute.

RESULTS:
• Positive (Ketose – Fructose): Cherry red/deep red color within 1 minute
• Negative (Aldose – Glucose): Faint pink or no color within 1 minute

REAGENT: Resorcinol dissolved in concentrated HCl (Seliwanoff's reagent)."""
  ),

  # PAGE 3 ─────────────────────────────────────────────────────────────────
  (3,
   "Spot 3  (2 marks)",
   "Identify the equipment and write its use.",
   """IDENTIFICATION: Weighing Balance (Digital Analytical Balance / Electronic Balance)

USES:
• Accurately measures the mass/weight of chemical substances, specimens, and reagents.
• Sensitivity: Analytical balance → 0.0001 g (0.1 mg); Top-loading → 0.01 g.
• Used for:
  - Weighing chemicals to prepare standard solutions of known concentration
  - Weighing biological specimens, tissue samples, and powders
  - Pharmaceutical compounding and quality control
  - Calibration of volumetric glassware

PRECAUTIONS:
  - Level the balance (check bubble spirit level) before use
  - Tare (zero) with empty container before weighing
  - Close draught shield doors during weighing (analytical balance)
  - Never exceed the maximum capacity
  - Wipe the pan clean after use to prevent corrosion"""
  ),

  # PAGE 4 ─────────────────────────────────────────────────────────────────
  (4,
   "Spot 4  (2 marks)",
   "Write 4 safety measures in laboratory.",
   """4 SAFETY MEASURES IN LABORATORY:

1. Use of Personal Protective Equipment (PPE):
   Always wear a lab coat, disposable gloves (nitrile/latex), safety goggles/face shield,
   and closed-toe shoes when handling chemicals, biological specimens, or hot instruments.
   PPE is the first line of defense against chemical burns, infections, and splashes.

2. Proper Disposal of Biohazardous and Chemical Waste:
   • Infectious/biological waste → Yellow biohazard bags (autoclave before disposal)
   • Sharps (needles, lancets, broken glass) → Puncture-proof sharps containers
   • Chemical waste → Designated labeled chemical waste containers
   Never mix different categories of waste.

3. No Eating, Drinking, or Mouth Pipetting in the Laboratory:
   Food and beverages are strictly prohibited in the lab to prevent accidental ingestion
   of toxic, infectious, or chemical substances. Always use mechanical pipette aids,
   rubber bulbs, or micropipettes — never pipette by mouth.

4. Emergency Preparedness and Fire Safety:
   Know the location and proper use of: fire extinguisher, eyewash station,
   safety shower, first aid kit, and emergency exits. Report all spills, accidents,
   and near-misses immediately. Use fume hoods for volatile/toxic reagents.
   Keep flammable solvents away from open flames."""
  ),

  # PAGE 5 ─────────────────────────────────────────────────────────────────
  (5,
   "Spot No. 5  (2 marks)",
   "Identify the test and write its Principle.",
   """IDENTIFICATION: Benedict's Test (Test for Reducing Sugars)

PRINCIPLE:
Benedict's test detects the presence of reducing sugars (free aldehyde or
alpha-hydroxy ketone groups) in a solution.

The reducing sugar REDUCES cupric ions (Cu²⁺) in the alkaline Benedict's reagent
to cuprous oxide (Cu₂O), which precipitates out as a colored deposit.

BENEDICT'S REAGENT composition:
  Copper sulphate (CuSO₄) + Sodium citrate + Sodium carbonate (Na₂CO₃)
  → Forms a clear deep-blue alkaline copper citrate complex.

REACTION:
  Reducing sugar + Cu²⁺ (blue)  →  Cu₂O (cuprous oxide) + Oxidized sugar

COLOUR INTERPRETATION (semi-quantitative):
  • Blue (no change)    = No reducing sugar (negative)
  • Green precipitate   = Trace sugar (~0.1–0.5 g/dL)
  • Yellow precipitate  = 0.5–1.0 g/dL
  • Orange precipitate  = 1.0–1.5 g/dL
  • Brick red/red       = >2.0 g/dL reducing sugar (strongly positive)

REDUCING SUGARS detected: Glucose, Fructose, Galactose, Lactose, Maltose
NON-REDUCING (negative): Sucrose (must hydrolyze first with HCl)"""
  ),

  # PAGE 6 ─────────────────────────────────────────────────────────────────
  (6,
   "Spot 1  (2 marks)",
   "Write any 2 manifestations of Vitamin C deficiency.",
   """VITAMIN C (ASCORBIC ACID) DEFICIENCY = SCURVY

Vitamin C is essential for COLLAGEN SYNTHESIS (hydroxylation of proline and lysine
residues in pro-collagen → forms cross-links → stable triple helix).
Deficiency causes defective collagen → fragile capillaries and connective tissue.

2 MANIFESTATIONS OF VITAMIN C DEFICIENCY:

1. Perifollicular Haemorrhages and Bleeding Gums (Scorbutic Gingivitis):
   • Defective collagen in blood vessel walls causes capillary fragility.
   • Results in: bleeding and swollen gums (Scorbutic gingivitis), small hemorrhages
     around hair follicles (perifollicular hemorrhages), and petechiae.
   • 'Corkscrew' or 'swan-neck' deformity of hair is a pathognomonic sign.
   • Gums become spongy, inflamed, and bleed easily on touch.

2. Impaired Wound Healing and Subperiosteal Haemorrhages:
   • Defective collagen synthesis → wounds fail to heal properly (reopening of old scars).
   • In children: Subperiosteal haemorrhages cause severe bone and joint pain
     (Barlow's disease in infants); the child lies in 'frog-leg' position.
   • Generalized weakness, fatigue, and anaemia (due to bleeding + impaired iron absorption).
   • Haemarthrosis (bleeding into joints) causing painful swelling."""
  ),

  # PAGE 7 ─────────────────────────────────────────────────────────────────
  (7,
   "Spot 2  (2 marks)",
   "I. Identify the picture.\nII. Mention the cause.",
   """IDENTIFICATION: Pellagra (Niacin / Vitamin B3 Deficiency)

I. IDENTIFICATION: Pellagra
   • The image shows characteristic symmetric hyperpigmented, rough, scaly dermatitis
     on sun-exposed areas of skin — especially the neck ('Casal's Necklace'),
     hands, forearms, and feet.
   • Classic presentation: The 4 D's of Pellagra:
     - Dermatitis (photosensitive, symmetric, hyperpigmented, scaly rash)
     - Diarrhea (watery, sometimes bloody)
     - Dementia (confusion, memory loss, psychosis, peripheral neuropathy)
     - Death (if untreated — the 4th D)

II. CAUSE:
   PRIMARY CAUSE:
   • Dietary deficiency of Niacin (Nicotinic acid / Vitamin B3) and/or its
     precursor amino acid Tryptophan.
   • Common in populations subsisting on maize (corn) as the staple diet — maize
     contains niacin in bound form (niacytin) which is biologically unavailable.

   SECONDARY CAUSES:
   • Carcinoid syndrome: Tryptophan diverted to serotonin synthesis, leaving
     less for niacin production.
   • Isoniazid (INH) therapy: Inhibits pyridoxal phosphate (needed for
     tryptophan → niacin conversion).
   • Hartnup disease: Impaired intestinal absorption of tryptophan.
   • Alcoholism (malabsorption and poor diet).

   NOTE: 60 mg dietary tryptophan = 1 mg niacin (niacin equivalent)."""
  ),

  # PAGE 8 ─────────────────────────────────────────────────────────────────
  (8,
   "Spot 3  (2 marks)",
   "Identify the equipment and write its use.",
   """IDENTIFICATION: Incubator (Laboratory Incubator / Bacteriological Incubator)

USE:
• An enclosed device that provides a controlled environment (temperature, humidity,
  CO₂ level) for growing and maintaining microbiological cultures or cell cultures.

KEY SETTINGS:
  • Standard bacteriological incubator: 37°C (human body temperature) for
    bacterial cultures (E. coli, Staphylococcus, Streptococcus, etc.)
  • CO₂ incubator: 37°C + 5% CO₂ + 95% humidity → for mammalian cell culture
  • Mycobacterial culture: 37°C for up to 6–8 weeks (Lowenstein-Jensen medium)

APPLICATIONS:
  • Microbiology: Culture of bacteria, fungi, and mycobacteria on solid/liquid media
  • Culture sensitivity testing (Kirby-Bauer disk diffusion method at 37°C)
  • Biochemistry: Enzyme incubation assays (maintain optimal temperature for reactions)
  • Serology: Incubation of antigen-antibody reactions (ELISA, agglutination tests)
  • Cell biology: Maintaining mammalian cell lines in tissue culture

Temperature range: 5°C above ambient to 70°C (standard); CO₂ incubator: 37°C."""
  ),

  # PAGE 9 ─────────────────────────────────────────────────────────────────
  (9,
   "Spot 4  (2 marks)",
   "Identify the test and write its principle.",
   """IDENTIFICATION: Biuret Test (Test for Proteins)

PRINCIPLE:
The Biuret test detects the presence of PEPTIDE BONDS (–CO–NH–) in proteins
and polypeptides (requires at least 2 peptide bonds = 3 amino acids minimum).

In ALKALINE conditions:
Cupric ions (Cu²⁺) from copper sulphate react with the nitrogen atoms of
peptide bonds to form a stable VIOLET/PURPLE coordinate complex (chelate).

REACTION:
  Cu²⁺ + Peptide bonds (alkaline) → Violet/Purple Cu²⁺-peptide chelate complex

REAGENT: Biuret reagent = NaOH (or KOH) + CuSO₄ + Sodium potassium tartrate
  (Tartrate prevents precipitation of Cu(OH)₂ in alkaline conditions)

RESULTS:
  • Violet/Purple color = POSITIVE → Proteins present (≥3 amino acids / ≥2 peptide bonds)
  • Pink/Mauve = Dipeptides or small polypeptides
  • Blue = NEGATIVE → No peptide bonds (free amino acids, monosaccharides)

USES:
  • Total protein estimation (semi-quantitative and quantitative)
  • Confirms presence of protein in biological samples (urine, serum, CSF)
  • Biuret method is the basis for automated protein assays in clinical chemistry analyzers

NOTE: The name 'Biuret' comes from the compound Biuret (H₂N–CO–NH–CO–NH₂)
which gives the same reaction; proteins mimic this structure via peptide bonds."""
  ),

  # PAGE 10 ──────────────────────────────────────────────────────────────────
  (10,
   "Spot No. 1  (2 marks)",
   "Identify the instrument and write its use.",
   """IDENTIFICATION: Gel Electrophoresis Apparatus (Agarose / Polyacrylamide Gel Electrophoresis)

USE:
• Separates charged macromolecules (proteins, DNA, RNA) based on their SIZE
  and CHARGE in an electric field.

PRINCIPLE:
  When an electric current is applied, negatively charged molecules (DNA/RNA at
  neutral pH; proteins denatured by SDS) migrate toward the positive electrode (anode).
  Smaller molecules migrate FASTER through the gel matrix; larger ones migrate SLOWER.
  Molecules separate into distinct bands which are visualized by staining.

APPLICATIONS IN BIOCHEMISTRY:
  • SERUM PROTEIN ELECTROPHORESIS: Separates serum proteins into bands:
    Albumin, α₁-globulin, α₂-globulin, β-globulin, γ-globulin
    → Diagnoses: M-band (Multiple myeloma), decreased albumin (liver disease/nephrotic syndrome)
  • DNA/RNA separation for molecular biology (PCR products, restriction fragments)
  • Western blot (protein identification after electrophoresis)
  • Isoenzyme analysis (e.g., LDH isoenzymes, CK-MB)
  • Haemoglobin electrophoresis (Hb S, Hb A, Hb F)

Types: Agarose gel electrophoresis (DNA), SDS-PAGE (proteins), Cellulose acetate electrophoresis (serum proteins)."""
  ),

  # PAGE 11 ──────────────────────────────────────────────────────────────────
  (11,
   "Spot No. 2  (2 marks)",
   "I. Identify the following test.\nII. Write its principle.",
   """IDENTIFICATION: Ninhydrin Test (Test for Amino Acids)

I. TEST NAME: Ninhydrin Test

II. PRINCIPLE:
Ninhydrin (Triketohydrindene hydrate) reacts with FREE ALPHA-AMINO GROUPS (–NH₂)
of amino acids to produce a characteristic purple/violet color called
RUHEMANN'S PURPLE.

MECHANISM (3 steps):
Step 1 – Oxidative decarboxylation:
  Ninhydrin oxidizes the alpha-amino acid → CO₂ + NH₃ + an aldehyde
  (The amino acid loses its amino group and carboxyl group)
  Ninhydrin itself is reduced to Hydrindantin.

Step 2 – Reaction:
  The released NH₃ reacts with another molecule of ninhydrin + hydrindantin
  → Forms a colored Schiff's base complex.

Step 3 – Color:
  This complex = Ruhemann's Purple (deep purple/violet)

COLOUR RESULTS:
  • PURPLE/VIOLET = Alpha-amino acids (all standard amino acids EXCEPT below)
  • YELLOW color  = Proline and Hydroxyproline (secondary amines — no free NH₂ group)
  • BLUE color    = Ammonia and primary amines
  • No color      = Non-amino acid compounds

USES:
  • Detection and identification of amino acids in biological fluids
  • Quantitative amino acid analysis (spectrophotometric measurement at 570 nm)
  • Paper chromatography — spray reagent to locate amino acid spots
  • Urine amino acid screening (aminoaciduria)"""
  ),

  # PAGE 12 ──────────────────────────────────────────────────────────────────
  (12,
   "Spot No. 5  (2 marks)",
   "Write down the normal range of:\na) Total Protein\nb) A/G ratio",
   """NORMAL RANGES (Serum):

a) TOTAL PROTEIN:
   • Normal range: 6.0 – 8.0 g/dL  (some references: 6.3–8.3 g/dL)
   
   Components:
   • Albumin:  3.5 – 5.0 g/dL
   • Globulin: 2.3 – 3.5 g/dL

b) A/G RATIO (Albumin to Globulin Ratio):
   • Normal range: 1.2 – 2.2

CLINICAL SIGNIFICANCE OF CHANGES:

DECREASED A/G ratio (<1.2) — seen in:
   • Liver disease (cirrhosis) → decreased albumin synthesis
   • Nephrotic syndrome → albumin lost in urine
   • Multiple myeloma → markedly raised immunoglobulins (γ-globulins)
   • Chronic infections / Inflammatory states → raised globulins
   • Malnutrition → decreased albumin

INCREASED A/G ratio (>2.5) — rare, seen in:
   • Hypogammaglobulinemia (decreased immunoglobulin production)
   • Agammaglobulinemia"""
  ),

  # PAGE 13 ──────────────────────────────────────────────────────────────────
  (13,
   "Spot  (2 marks)",
   "Identify the instrument and write its use.",
   """IDENTIFICATION: Centrifuge (Bench-top / Clinical Centrifuge)

USE:
• Uses CENTRIFUGAL FORCE generated by rapid rotation to separate substances
  of different densities in a liquid suspension.

PRINCIPLE:
  Denser particles sediment to the BOTTOM of the tube (pellet) while lighter
  components remain in the SUPERNATANT.
  Force generated = RCF (Relative Centrifugal Force) measured in × g.

COMMON USES IN BIOCHEMISTRY LAB:
  • Separation of serum or plasma from blood cells (2,000–3,000 RPM, 10 min)
  • Concentration of urinary sediment for microscopy
  • Separation of cells and cellular debris from culture supernatant
  • Preparation of platelet-poor plasma (PRP vs PPP)
  • Sedimentation of precipitated proteins

TYPES:
  • Low-speed (≤6,000 RPM): Routine clinical use
  • High-speed (10,000–25,000 RPM): Bacteria, cell organelles
  • Ultracentrifuge (>50,000 RPM): Viruses, ribosomes, lipoproteins
  • Microcentrifuge (up to 14,000 RPM): Eppendorf microtubes (1.5/2.0 mL)"""
  ),

  # PAGE 14 ──────────────────────────────────────────────────────────────────
  (14,
   "Spot No. 5  (2 marks)",
   "Identify the test and write its principle.",
   """IDENTIFICATION: Fehling's Test (Test for Reducing Sugars)

PRINCIPLE:
Fehling's test detects REDUCING SUGARS (those with a free aldehyde or
alpha-hydroxy ketone group capable of reducing Cu²⁺ to Cu⁺).

REAGENT PREPARATION:
  Fehling's Solution A: 7% aqueous Copper sulphate (CuSO₄) — deep blue
  Fehling's Solution B: Alkaline sodium potassium tartrate (Rochelle salt in NaOH)
  Mix A + B in equal volumes just before use → forms alkaline cupric tartrate complex (deep blue)

(Sodium potassium tartrate acts as a CHELATING AGENT to keep Cu²⁺ in solution
 and prevent Cu(OH)₂ precipitation in the alkaline medium.)

REACTION:
  Reducing sugar + Cu²⁺ (blue)  →  Cu₂O (cuprous oxide) ↓ + Oxidized sugar
  (Brick-red/orange precipitate forms on heating)

RESULT:
  • POSITIVE: Brick red/orange precipitate on heating = Reducing sugar present
    (Glucose, Fructose, Galactose, Maltose, Lactose)
  • NEGATIVE: Solution remains deep blue = Non-reducing sugar
    (Sucrose — must be hydrolyzed by HCl first to give positive result)

DIFFERENCE from Benedict's test:
  Fehling's uses TWO separate solutions mixed before use;
  Benedict's is a single stable reagent (preferred in clinical labs — more sensitive)."""
  ),

]

# ─── BUILD PDF ──────────────────────────────────────────────────────────────
doc = SimpleDocTemplate(
    OUTPUT, pagesize=A4,
    topMargin=1.5*cm, bottomMargin=1.5*cm,
    leftMargin=1.8*cm, rightMargin=1.8*cm,
    title="Spotters Biochemistry -1 — Questions with Model Answers",
)
story = []

# Cover
story.append(Spacer(1, 1.5*cm))
story.append(Paragraph("Spotters Biochemistry -1", S_TITLE))
story.append(Paragraph("Biochemistry Spotters — All 14 Questions with Model Answers", S_SUB))
story.append(HRFlowable(width='100%', thickness=2,
                         color=colors.HexColor('#1565C0'), spaceAfter=10))
story.append(Spacer(1, 0.3*cm))

# Quick ref table
rows = [
    ['Page', 'Spot', 'Topic', 'Marks'],
    ['1',  'Spot 1',  'Colorimeter (Identify instrument)',         '2'],
    ['2',  'Spot 2',  'Seliwanoff\'s Test (ketosugar vs aldosugar)','2'],
    ['3',  'Spot 3',  'Weighing Balance (Identify equipment)',     '2'],
    ['4',  'Spot 4',  '4 Safety Measures in Laboratory',           '2'],
    ['5',  'Spot 5',  'Benedict\'s Test (Identify test)',          '2'],
    ['6',  'Spot 1',  'Vitamin C Deficiency — 2 Manifestations',   '2'],
    ['7',  'Spot 2',  'Pellagra — Identify picture & cause',       '2'],
    ['8',  'Spot 3',  'Incubator (Identify equipment)',            '2'],
    ['9',  'Spot 4',  'Biuret Test (Identify test)',               '2'],
    ['10', 'Spot 1',  'Gel Electrophoresis (Identify instrument)', '2'],
    ['11', 'Spot 2',  'Ninhydrin Test (Identify test)',            '2'],
    ['12', 'Spot 5',  'Normal ranges: Total Protein & A/G ratio', '2'],
    ['13', 'Spot',    'Centrifuge (Identify instrument)',          '2'],
    ['14', 'Spot 5',  'Fehling\'s Test (Identify test)',           '2'],
]
t = Table(rows, colWidths=[1.4*cm, 2.2*cm, 10.5*cm, 1.8*cm])
t.setStyle(TableStyle([
    ('BACKGROUND', (0,0), (-1,0), colors.HexColor('#1565C0')),
    ('TEXTCOLOR',  (0,0), (-1,0), colors.white),
    ('FONTNAME',   (0,0), (-1,0), 'Helvetica-Bold'),
    ('FONTNAME',   (0,1), (-1,-1), 'Helvetica'),
    ('FONTSIZE',   (0,0), (-1,-1), 8.5),
    ('ALIGN',      (0,0), (-1,-1), 'CENTER'),
    ('ALIGN',      (2,1), (2,-1), 'LEFT'),
    ('ROWBACKGROUNDS', (0,1), (-1,-1), [colors.HexColor('#e3f2fd'), colors.white]),
    ('GRID',       (0,0), (-1,-1), 0.4, colors.HexColor('#90caf9')),
    ('TOPPADDING', (0,0), (-1,-1), 4),
    ('BOTTOMPADDING', (0,0), (-1,-1), 4),
]))
story.append(t)
story.append(PageBreak())

# Content
for (pg, label, question, answer) in spotters:
    block = []
    block.extend(img_block(pg))
    block.extend(ans_block(label, question, answer))
    story.append(KeepTogether(block))

doc.build(story)
print(f"PDF saved: {OUTPUT}")
print(f"Size: {os.path.getsize(OUTPUT)/1024/1024:.2f} MB")

cd /home/daytona/workspace && python3 build_biochem_pdf.py 2>&1