Skip to main content
UploadLanguage (EN)Support
BusinessMobileSocial MediaMarketingTechnologyArt & PhotosCareerDesignEducationPresentations & Public SpeakingGovernment & NonprofitHealthcareInternetLawLeadership & ManagementAutomotiveEngineeringSoftwareRecruiting & HRRetailSalesServicesScienceSmall Business & EntrepreneurshipFoodEnvironmentEconomy & FinanceData & AnalyticsInvestor RelationsSportsSpiritualNews & PoliticsTravelSelf ImprovementReal EstateEntertainment & HumorHealth & MedicineDevices & HardwareLifestyle
Uploaded byAshaGolasangimath
101 views

Unit 1. D. Leaf Constants, Lycopodium Method & Camera Lucida | Pharmacognosy & Phytochemistry-I Unit 1 | B.Pharm Semester IV

Quantitative Microscopy of Crude Drugs | Pharmacognosy and Phytochemistry-I This presentation is prepared according to the RGUHS B.Pharm Semester IV syllabus and provides a comprehensive understanding of Quantitative Microscopy of Crude Drugs, an essential topic in Pharmacognosy used for the identification, authentication, standardization, and quality control of herbal drugs. Topics Covered • Introduction to Quantitative Microscopy • Importance of Quantitative Microscopy • Leaf Constants • Stomatal Number • Stomatal Index • Vein Islet Number • Vein Termination Number • Palisade Ratio • Lycopodium Spore Method • Applications of Lycopodium Method • Camera Lucida • Procedure for Camera Lucida • Diagrams of Microscopic Objects • Pharmacognostic Evaluation of Crude Drugs • University Examination Preparation Suitable For ✔ B.Pharm Students ✔ Pharmacy Teachers ✔ Pharmacognosy & Phytochemistry Learners ✔ GPAT Aspirants ✔ Pharmacy Colleges and Universities ✔ Competitive Examination Preparation This presentation is intended for educational purposes and serves as a valuable study resource for understanding the microscopic evaluation and standardization of crude drugs in Pharmacognosy. Part 4 of 4 – Pharmacognosy Unit 1 Lecture Series Prepared by Ms. Asha Golasangimath Assistant Professor Department of Pharmacognosy Alva's College of Pharmacy, Moodubidire Affiliated to RGUHS, Karnataka

Education◦
Related topics:
pharmacy-notes• Crude Drugs• PCI Syllabus•
Download to read offline
Pharmacognosy and Phytochemistry-I Unit 1. D. : Quantitative Microscopy of Crude Drugs Class : IV semester B. Pharm Presented by: Ms. Asha Golasangimath Assistant Professor Department of Pharmacognosy Alva’s College of Pharmacy, Moodubidire Affiliated to RGUHS, Karnataka Academic Year: 2025–2026
Index Alva's College of Pharmacy, Moodubidire 2  Introduction to microscopy of crude drugs  Lycopodium spore method  Leaf constants  Camera lucida and diagrams of microscopic objects to scale with camera lucida
Quantitative Microscopy of Crude Drugs: Alva's College of Pharmacy, Moodubidire 3  Quantitative microscopy is the measurement and numerical evaluation of microscopic characteristics of crude drugs. It is mainly used for:  Identification and authentication of Crude and powdered drugs  Detection of adulteration  Standardization of crude drugs  Quality control of herbal drugs  Useful when chemical methods are difficult  Important in powdered drug evaluation  Example:  Senna leaf → stomatal index  Datura leaf → palisade ratio  Digitalis → vein islet number
Parameters in Quantitative Microscopy Alva's College of Pharmacy, Moodubidire 4 The following parameters are commonly studied:  Leaf constants -  Stomatal Number  Stomatal Index  Vein Islet Number  VeinTermination Number  Palisade Ratio  Lycopodium Spore Method  Camera Lucida Method
1. Stomatal Number Alva's College of Pharmacy, Moodubidire 5 Stomatal Number is theAverage number of stomata per square millimeter of leaf surface/ epidermis. Formula  Stomatal Number = Number of stomata per sq. mm  Significance  Used for identification of leaves  Helps in differentiation of species Collection of fresh leaf ↓ Preparation of epidermal peel (usually lower epidermis) ↓ Place peel on slide ↓ Add glycerin/chloral hydrate and cover with coverslip ↓ Observe under microscope ↓ Select defined area (1 sq. mm) ↓ Count number of stomata present in the area ↓ Repeat in different fields ↓ Calculate average number of stomata per sq. mm
Alva's College of Pharmacy, Moodubidire 6
2. Stomatal Index Alva's College of Pharmacy, Moodubidire 7 Stomatal index is the Percentage proportion of stomata to total epidermal cells. Formula: 𝑺𝑰 = 𝒔 𝑬+𝑺 𝑿 𝟏𝟎𝟎 Where:  SI = Stomatal Index  S = Number of stomata  E = Number of epidermal cells Significance  More constant than stomatal number  Valuable taxonomic character Collection of fresh leaf ↓ Preparation of epidermal peel ↓ Mount peel on slide with glycerin ↓ Observe under microscope ↓ Select a definite area of epidermis ↓ Count number of stomata (S) ↓ Count number of epidermal cells (E) ↓ Apply formula SI = S / (E + S) × 100 ↓ Calculate stomatal index ↓ Record the result
3. Vein Islet Number and Vein Termination Number Alva's College of Pharmacy, Moodubidire 8  AVein Islet is a small areas of green tissue enclosed by veins.  Vein Islet Number: Number of small areas of photosynthetic tissue/ vein islets enclosed by veins per square millimeter of leaf surface.  Vein Termination number: Number of veinlet terminations per square millimeter of leaf. Significance • Useful for identification of leaf drugs • Helps in quality control
Alva's College of Pharmacy, Moodubidire 9 Collection of fresh leaf ↓ Boil leaf in chloral hydrate to remove chlorophyll ↓ Wash and mount leaf on slide with glycerin ↓ Observe under microscope ↓ Select defined area (1 sq. mm) ↓ Count number of vein islets present in the area ↓ Count number of veinlet terminations in same area ↓ Repeat in different fields ↓ Calculate average values ↓ •Vein Islet Number •VeinTermination Number Procedure
Alva's College of Pharmacy, Moodubidire 10
4. Palisade Ratio Alva's College of Pharmacy, Moodubidire 11 Palisade ratio is the Average number of palisade cells beneath one epidermal cell. Formula 𝑷𝒍𝒊𝒔𝒂𝒅𝒆 𝑹𝒂𝒕𝒊𝒐 = 𝑻𝒐𝒕𝒂𝒍 𝑷𝒂𝒍𝒊𝒔𝒂𝒅𝒆 𝒄𝒆𝒍𝒍𝒔 𝑵𝒖𝒎𝒃𝒆𝒓 𝒐𝒇 𝒆𝒑𝒊𝒅𝒆𝒓𝒎𝒂𝒍 𝒄𝒆𝒍𝒍𝒔 𝒄𝒐𝒖𝒏𝒕𝒆𝒅 Significance  Characteristic for specific plants  Helps in authentication Example  Datura leaf
Alva's College of Pharmacy, Moodubidire 12 Procedure Collection of fresh leaf ↓ Prepare transverse section or epidermal peel of leaf ↓ Mount on slide with glycerin ↓ Observe under microscope ↓ Focus upper epidermis and palisade layer ↓ Count number of palisade cells beneath four epidermal cells ↓ Calculate average number of palisade cells ↓ Apply formula ↓ Record the result
Lycopodium Spore Method Alva's College of Pharmacy, Moodubidire 13 A quantitative microscopic method used to determine the purity of powdered drugs using Lycopodium spores as a reference standard. Principle  Lycopodium spores have a fixed known number of spores per mg.(94,000 spores)  Known quantity of spores is mixed with powdered drug.  By comparing counts under microscope, quantity of drug constituents can be calculated. StandardValue  1 mg Lycopodium powder contains approximately: 94,000 spores Lycopodium Clavatum (stag’s-horn clubmoss, running pine)
Alva's College of Pharmacy, Moodubidire 14 Procedure Weigh powdered crude drug ↓ Add known quantity of Lycopodium spores ↓ Mix uniformly ↓ Prepare slide mount with suspending agent (glycerine: mucilage of tragacanth: water 2:1:2) ↓ Observe under microscope ↓ Count characteristic structures of crude drug (starch grains) ↓ Count Lycopodium spores ↓ Apply formula for calculation ↓ Determine number of characteristic structures per mg of drug ↓ Record the result Formula Where: N = Number of Starch grain W =Weight of Lycopodium taken M =Weight of sample taken P =Total number of starch grains per mg of pure sample 286,000 94,000 = Number of lycopodium spores per mg % 𝑃𝑢𝑟𝑖𝑡𝑦 𝑜𝑓 𝐷𝑟𝑢𝑔 = 𝑁 𝑋 𝑊 𝑋 94,000 𝑆 𝑋 𝑀 𝑋 𝑃
Alva's College of Pharmacy, Moodubidire 15 Applications of Lycopodium Method  Quantitative evaluation of powdered drugs.  Determination of starch grains and fibers.  Detection of adulteration. Advantages  Accurate method  Useful for powdered drugs  Simple microscopic technique Limitations  Requires careful mixing  Counting errors may occur
Camera Lucida Alva's College of Pharmacy, Moodubidire 16 Camera lucida is an optical device attached to microscope used for drawing microscopic objects to scale. Principle  The device projects image of microscopic object onto paper for accurate tracing. Uses of Camera Lucida  Preparation of microscopic diagrams  Measurement of tissues  Pharmacognostic studies  Research documentation Disadvantages Time consuming. Requires skill and proper alignment. Advantages Accurate drawings. Maintains proportion. Helpful in teaching and research.
Alva's College of Pharmacy, Moodubidire 17 Procedure Attach camera lucida to microscope ↓ Place specimen on stage ↓ Focus specimen properly under microscope ↓ Adjust mirror/prism of camera lucida ↓ Place drawing sheet beside microscope ↓ Observe projected image on paper ↓ Trace outline of image carefully ↓ Label important parts ↓ Draw microscopic object to scale ↓ Record final diagram
Alva's College of Pharmacy, Moodubidire 18 Diagrams of Microscopic Objects: Stomata, Trichomes, Starch grains, Calcium oxalate crystals, Fibers, Xylem vessels, Palisade cells
Alva's College of Pharmacy, Moodubidire 19