Kendali Mutu Bahan Baku dan Produk Obat Herbal Indonesia

Kendali Mutu Bahan Baku dan Produk Obat Herbal Indonesia Sebagai Bagian Standardisasi Yang Komprehensif

Webinar TropBRC IPB #7

18 Agustus 2020

• Tropical Biopharmaca Research Center • Department of Chemistry • AR-Lab

Mohamad Rafi dan Rudi Heryanto

Biosketch Dr. Mohamad Rafi

A senior lecture at the Department of Chemistry, Faculty of Mathematics and Natural sciences, IPB University Bogor, Indonesia. Now I am serving as Head od Master/Dostoral degree program in Chemistry, Deputi Manager of TropBRC Laboratory at IPB University and President of the Indonesian Chemical Society. My primary research interest is in the field of metabolomics and chemometrics especially for the development of quality control method, identifying biomarker compound and standardization of medicinal plants. Email: [email protected]

http://mra.staff.ipb.ac.id/

Education

• B.Sc - IPB University, Indonesia

• M.Sc – IPB University, Indonesia

• Ph.D – Gifu University, Japan

Standardization of Andrographis paniculata using metabolomics approach (RKI-2019, Collaboration between IPB-ITB-UGM-Unair)

mailto:[email protected]







Ak

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2020

2019

2018

Identifikasi senyawa bioaktif antikanker baru menggunakan pendekatan

metabolomik berbasis LC-MS/MS sebagai upaya mendukung

penanggulangan penyakit kanker (PDUPT)

Pendekatan metabolomik untuk standardisasi sidaguri (Sida

rhombifolia) melalui pemrofilan metabolit (Spektrum FTIR, KLT, LC-

MS/MS) serta aktivitas antioksidan dan inhibisi xantin oksidase (RKI)

Evaluasi mutu sambiloto (Andrographis paniculata) menggunakan

pendekatan metabolomik: profil fitokimia, komposisi unsur logam, dan

aktivitas biologis (RKI)

Polyethylene oxide and silica bonded stationary phase for separation of

polar compounds in medicinal plants using capillary liquid

chromatography (PKLN)

Pengembangan obat herbal antidiabetes dengan pendekatan

metabolomik berbasis kepada konsep multikomponen-multitarget dan

sinergestik (PDUPT)

Metode identifikasi dan autentikasi bahan baku obat herbal

menggunakan pendekatan metabolomik berbasis kromatografi cair

kinerja tinggi dan spektroskopi UV-Vis (PDUPT)

01

02

03

04 Multiple spectroscopic fingerprinting platforms for rapid characterization of α-glucosidase inhibitors and antioxidants from some commonly consumed Indonesian vegetables and spices (J. Food Meas. Charact. 2020)

HPLC fingerprinting coupled with linear discriminant analysis for the detection of adulteration in Orthosiphon aristatus (J. Liq. Chrom. Rel. Tech. 2019)

Quantitative Analysis of Multi-components

in Curcuma xanthorrhiza by Single Marker (Indones. J.

Pharm. 2019)

Metabolite Profiling of Andrographis

paniculata Leaves and Stem Extract Using UHPLC-Orbitrap-MS/MS (Nat. Prod. Res. 2020)

PUBLIKASI

05

06

07

08 A combination of simultaneous quantification of four triterpenes and fingerprint analysis using HPLC for rapid identification of Centella asiatica from its related plants and classification based on cultivation ages (Ind. Crops. Prod. 2018)

Discrimination of Red and White Rice Bran from Indonesia Using HPLC Fingerprint Analysis Combined with Chemometrics (Food Chem. 2017)

Metabolite profiling of tabat barito (Ficus

deltoidea) using UPLC-QTOF-MS/MS (J. Pure Appl.

Chem. Res.2018)

HPLC fingerprint and simultaneous quantitative analysis of phyllanthin

and hypophyllanthin for identification and authentication of Phyllanthus niruri from related

species (Braz. J. Pharmacogn. 2018)

PUBLIKASI

Isi Presentasi

Karakteristik tumbuhan obat

Standardisasi dan kendali mutu obat herbal

Contoh kendali mutu bahan baku dan produkobat herbal


Komponen

kimia

Panen dan pasca panen

Waktu panen

Lingkungan tumbuh

Kultivasi/ tumbuh liar

Komposisi yang kompleks

dengan variasi kosentrasi

dan belum diketahui total

senyawa yang terkandung

di dalamnya


mg/g

0

5

10

15

20

25

30

CUR

DMC

BDMC

M. Rafi et al. 2015. Food Analytical Methods. 8: 2185-2193.

Kadar kurkuminoid pada kunyit (CL) dan temulawak (CX)


MS AS MA AA

Bogor 7.62 7.71 2.8 2.6

Sleman 2.39 2.47 1.28 1.51

Boyolali 0.56 1.6 0.65 0.93

Kuningan 1.87 2.59 0.52 0.59

CA-1 4.22 2.99 2.1 0.69

CA-2 1.29 1.1 2.86 2.24

CA-3A1 8.09 8.23 2.09 1.52

CA-3A2 7.55 7.51 2.43 2.06

CA-3A3 6.02 6.03 1.98 1.49

CA-3A4 5.73 5.85 1.98 1.33

CA-3A5 6.18 6.25 1.8 1.32

CA-3A6 8.04 7.52 1.73 1.48

CA-3A7 5.55 5.96 2.3 2.44

CA-4A1 2.59 2.45 6.39 4.76

CA-4A2 8.44 8 5.22 3.68

CA-4A3 6.81 6.6 5.48 4.46

CA-4A4 9.92 9.77 4.14 3.1

CA-4A5 15.12 13.55 1.27 1.16

CA-4A6 7.45 7.82 4.1 3.36

CA-4A7 7.1 6.59 4.49 3.18

CA-5A1 8.22 8.21 1.25 1.11

CA-5A2 7.52 7.45 0.9 0.43

CA-5A3 7.56 7.16 0.66 0.08

CA-5A4 8.93 9.05 0.8 0.31

CA-5A5 6.78 6.85 0.61 0.04

CA-5A6 7.53 7.41 0.6 0.11

CA-5A7 7.61 7.14 0.65 0.04

Analyte content (mg/g), n = 3CA samples Kadar madekasosida, asiatikosida,

asam madekasat dan asam asiatat

pada pegagan dengan waktu panen

berbeda

M. Rafi et al. Industrial Crops & Products 122 (2018) 93–97

Karakteristik tumbuhan/produk obat herbal

Multikomponen Dapat lebih dari satu bahan --

Multikompnen

Karakteristik tumbuhan/produk obat herbal

Ma et al. Phytomedicine 45 (2018) 105–119

Problem dalam Obat Tradisional

Standardisasi obat herbal

Pemalsuan dengan bahan kimia obat atau tumbuhan

lainnya yang mirip

Menggunakan tumbuhan obat

yang salah

Dosis yang tidak tepat

Produk dengan kualitas rendah

Toksisitas

Interaksi dengan obat herbal/kimia

lainnya

Problem

Kesalahpahaman bahwa obat herbal pasti aman

Penggunaan obat herbal untuk indikasi yang

berbeda


MV Shinde, K Dhalwal, K Potdar, K Mahadik. 2009. International Journal of Phytomedicine 1: 4

Diagram sebab akibat adanya variasi khasiat obat herbal


Jadi singkatnya, standardisasi (kendali mutu) tumbuhan obat dan produknya sangat diperlukan 1. Apakah suatu produk obat herbal mengandung komponen seperti yang tertera

pada klaim label? ----- IDENTIFIKASI 2. Apakah memang obat herbal yang saya beli benar-benar yang tepat? -----

ADULTERASI 3. Apakah harga obat herbal yang dibeli sesuai khasiatnya atau tampilan luar saja?

----- KUALITAS 4. Apakah komposisi komponen penyusun obat herbal sesuai dengan yang tertera

pada label klaim? ----- KUANTITAS 5. Apakah akan ada perubahan komposisi dengan berjalannya waktu? -----

STABILITAS


• Standardization is a system to ensure that every packet of medicine that is being sold has the correct amount and will induced its therapeutic effect (Chaudury 1992)

• Standardization refers to the body of information and controls necessary to produce material of reasonable consistency. This is achieved through minimizing the inherent variation of natural product composition through quality assurance practices applied to agricultural and manufacturing processes (American Herbal Product Association)


Main issue

kualitas

khasiat keamanan

Standardisasi

Tiga isu utama (kualitas, keamanan, dan khasiat) dalam pengembangan obat herbal dapat tercapai jika dilakukan proses standardisasi sejak menanam hingga produksinya standardisasi --- kendali mutu

Proses


The complex factors affecting quality of herbal medicine products

Liu et al. Phytomedicine 44: 247-257





Metode analitik untuk mengetahui komposisi fitokimia (kualitatif dan kuantitatif) dalam rangka menstandarkan bahan

baku/produk obat herbal

aktivitas farmakologi yang

konsisten akan dihasilkan dari

profil kimia yang konsisten pula

Obat herbal terstandar

Aktivitas biologis

konsisten

Fitokimia


Wei et al. Chin Med (2020) 15:56


Wei et al. Chin Med (2020) 15:56

Kendali mutu tumbuhan obat

Simmler, C., Graham, J.G., Chen, S.-N., Pauli, G.F. 2018. Fitoterapia, 129, pp. 401-414

Parameters affecting the determination of botanical authenticity

Kendali mutu tumbuhan obat


A three-step process for the assessment of authenticity

(step 1) documentation of traceability, which encourages supply chain transparency and helps identify risks pertaining to possible contaminations/adulterations (step 2) validation of botanical identity and integrity of the plant material or extract together with (step 3) the implementation of a variety of complementary phytochemical analyses to confirm the chemical identity and composition of the botanical, utilizing techniques that are also capable of detecting potential chemical adulterations

https://www.sciencedirect.com/topics/pharmacology-toxicology-and-pharmaceutical-science/phytochemical

Metode kendali mutu TO/OH

QC method with metabolomics

approach

Fingerprint analysis

Holistic

Profiling analysis

Targeted analysis

Commonly used

Identification, discrimination and authentication (IDA) of medicinal plants

Metode kendali mutu TO/OH

APPROACH TARGETED ANALYSIS METABOLITE PROFILING

METABOLITE FINGERPRINTING

STRENGHT • Quantitative • Low detection limit • High troughput

Global image of the samples

• Global image of the samples

• Directly applicable to pattern recognition

• Highest throughput

PITFALL • Limited number of targeted compounds

• No detection of untargeted compounds

• Need for the pure targeted compounds for the calibration

• Expensive

• Semi-quantitative • Difficulties in

informatics • Medium

throughput

No compound identification

TECHNIQUES Chromatography and spectroscopy

5 P Vernocchi et al. 2012. Frontiers in Cellular and Infection Microbiology. 2: Article 156

Different approaches and respective techniques: pitfalls and strengths5

Targeted analysis

curcumin

demethoxycurcumin

bisdemethoxycurcumin

xanthorrhizol Temulawak

E Erpina et al. Journal of Liquid Chromatography and Related Techniques 40(12): 635-639.

Targeted analysis

E Erpina et al. Journal of Liquid Chromatography and Related Techniques 40(12): 635-639.

Metabolite profiling/fingerprinting

Kunyit (C. longa) Temulawak (C.

xanthorrhiza)

Temu mangga (C. mangga)

Temu hitam (C. aeruginosa)

IDENTIFIKASI DAN

AUTENTIKASI


Spektrum UV-Vis representatif dari 4

spesies Curcuma

M Rafi et al. 2018. International Food Research Journal 25(2): 643-648.


• Instrumentasi: Spektrofotometer UV-Vis

• Prapemrosesan sinyal: standar normal variate

• Metode kemometrik: a. principal component

analysis (PCA) dan b. canonical variate

analysis (CVA)

a

b

M Rafi et al. 2018. International Food Research Journal 25(2): 643-648.


Simmler, C., Graham,

J.G., Chen, S.-N., Pauli,

G.F. 2018. Fitoterapia,

129, pp. 401-414


NMR-based metabolomic model for the screening of botanical authenticity and rapid detection

of potential adulteration.

(A) The principal step towards the development of metabolomic screening model calls for inter-

laboratory collaboration in order to acquire as many (ideally: authentic) botanical reference

spectra as possible. This step requires the implementation of standard operating

procedures (SOP) for sample preparation and data acquisition.

(B) All the acquired specta are deposited in an open access repository where they can be

further processed and compared by chemometric analyses.

(C) Different laboratories can compare their own spectra from new botanical samples (prepared

and acquired according to the SOP) to the spectra in the database. The newly acquired

spectra can be tested against the statistical model of authentic spectra so as to quikly and

objectively evaluate their authenticity defined by statistical similarities when compared to a

set of referenced/digitized spectra (i.e., samples within the limits of the chemometric models

sample #1). The same process is capable of detecting major chemical differences that lead

to their classification outside the model and conclusion as being non-authentic or adulterated (sample #2).


Contoh metode kendali mutu bahan baku dan produk obat herbal

Manakah yang merupakan meniran hijau (Phyllanthus niruri)?

2

1

3

Identifikasi dan autentikasi Phyllanthus niruri

Pn9 Pd1 Ll1 Ll3 Pn1 Pn7 Pn5 Pu1 Ll2 Pu2

1. Filantin 2. Hipofilantin

1 2


Pn: Phyllanthus niruri Pd: Phyllanthus debilis Pu: Phyllanthus urinaria Ll: Leucaena leucocephala

http://biofarmaka.ipb.ac.id/news/2334-download-atlas-kromatografi-lapis-tipis-tumbuhan-obat-indonesia-volume-1-2019

WT Wahyuni, M Saharah, Z Arif, M Rafi. 2020. Journal of the Indonesian Chemical Society 3(1): 47-52.


-12

-8

-4

0

4

8

-15 -10 -5 0 5 10 15

FD2

(3

6.6

6%

)

FD1 (63.34%)

PN

PU

PD

Plot AD


1


1

2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5 25.0 27.5 30.0 32.5 35.0 37.5 min

0.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

9.0

uV(x100,000)

1

2

3 4

5

6

7 8 9 10

12

13

14 11 a b

c d

e f

g h

i

(a) PN-5 100 %

(b) PN-5:PU-5 (75:25 %)

(c) PN-5:PU-5 (50:50 %)

(d) PN-5:PU-5 (25:75 %)

(h) PU-5 100 %

(i) PD-5 100 %)

(e) PN-5:PD-5 (75:25 %)

(f) PN-5:PD-5 (50:50 %)

(g) PN-5:PD-5 (25:75 %)

3 4

a

b

c

d

e

f

g

h

i


1

Plot analisis diskriminan

PN

PU

PD


Analisis sidikjari obat herbal Glucofarmaka

Campuran simplisia

40 mL Metanol

saring

HPLC Shimadzu

volume injeksi 20 μL

Fase diam kolom ShimPack C18

Detektor PDA (Photo Diode Array)

Ekstraksi sonikasi 2×15 menit

Laju alir 1 mL/menit

Suhu kolom 35 °C

Parametes yang

divariasikan:

• jenis dan komposisi

fase gerak

• lama waktu analisis

• Panjang gelombang

deteksi H Habibie et al. Indonesian Journal of Chemistry, 2017, 17(1): 79-85.


metanol

asetonitril

Pemodifikasi organik untuk fase gerak

280 nm

254 nm

210 nm

Panjang gelombang deteksi


5-95% ACN-air

5-70% ACN-air

10-80% ACN-air

Komposisi fase gerak


50 menit

60 menit

80 menit

90 menit

Waktu analisis

Profil kromatogram KCKT daun sembung (a), batang brotowali (b), rimpang

jahe (c), daun pare (d), ekstrak jamu (e), dan campuran simplisia(f)



Evaluasi Kinerja Analitik Metode Analisis Sidik Jari KCKT

Parameter Presisi Kedapatulangan Stabilitas

SDR WRR <0.3% <0.3% <0.3%

SDR LAR <3.9% <6.0% <4.7%

Framework for quality control of Ayurvedic drugs by the Ministry of AYUSH (India)

Katoch et al. Journal of Ethnopharmacology 197 (2017) 25–31

Q-marker in TCM

• The Q-marker of TCM refers to the intrinsic or processing/ preparation-resultant

chemical substances closely associated with the functional properties that exist

in the raw materials and products of TCM (involving the decoction pieces,

decoctions, extractives, and Chinese patent medicines), which can be used as

the indicators for quality control of TCM to embody the safety and effectiveness.

• Q-marker was proposed by considering the complex factors from the basic

theory of TCM, the formulation, preparation technology, dosage form, and usage

of formula preparations, to standardize TCM quality research and quality

standards elaboration, to enhance the quality consistency, controllability, and

traceability.

• The establishment of Q-marker integrates multi- disciplinary technologies like

natural products chemistry, analytical chemistry, bionics, chemometrics,

pharmacology, systems biology, and pharmacodynamics, etc

Yang et al. Acta Pharmaceutica Sinica B (2017) 7(4):439–446

Q-marker in TCM

Yang et al. Acta Pharmaceutica Sinica B (2017) 7(4):439–446

The basic properties of Q-marker can be outlined in four aspects:

1) They are the intrinsic chemical components in TCM materials and products, or

processing/ preparation-resultant;

2) They are functional properties-associated, with definite chemical structures;

3) They can be qualitatively characterized and quantitatively determined; and

4) For the formulae, there presentative substances of the Monarchare firstly

considered, and those from the Minister, Assistant, and Guide, should be

considered as well, following the compatibility theory. The factors, involving 1)cell

and tissue specificity, 2) organ specificity, 3) developmental specificity for

biosynthesis, 4) the extrinsic factors of growing process (drugmaterials), and 5)

the preparation factors (formula preparations), can affect Q-markers.

In establishment of Q-markers, special attention should be paid to the components

that ensure the authenticity (identity marker), differentiate the quality difference

(superiority/inferiority marker), and identify the geo-authenticity (geo-authentic

marker).

Q-marker in TCM

Xie et al. RSC Adv., 2018, 8, 812

Q-marker in TCM

Strategy of Q-biomarkers screening and validation

Xin et al. Phytomedicine 44: 103-108

Q-marker in TCM

Identification of the quality markers for quality and process control of herbal medicinal products


Q-marker in TCM

The network

database for

the traceability

platform of

herbal

medicinal products


Q-marker in TCM

Identify quality

markers based on

integration

methodology to

establish the

quality markers

for production

process system

from raw

materials to products


Summary

• Standardisasi menjadi bagian penting dalam menghasilkan obat herbal yang konsisten khasiat, kualitas, dan keamanannya.

• Konsep dalam standardisasi obat herbal Indonesia perlu dikembangkan menyesuaikan dengan karakteristik yang ada di Indonesia

Ucapan Terima Kasih

Terima kasih untuk

• Hanifullah Habibie, Raudatul Jannah, Meri Saharah, Epi Erpina, dan Roni Nasrulloh

• Hibah Penelitian Unggulan Perguruan Tinggi tahun 2016, 2018, dan 2019 dari Kemenristekdikti RI

• TropBRC, Departemen Kimia dan Advance Lab IPB

TERIMA KASIH THANK YOU

Tropical Biopharmaca Research Center Department of Chemistry Advance Research Laboratory

Documents

Kendali Mutu Bahan Baku dan Produk Obat Herbal Indonesia